MXPA98007928A - Promotion of regeneration of organized tissues - Google Patents

Abstract

A system, method and device for promoting the regeneration of tissue in an injured area in an organized tissue structure, in a human or animal living body, from a lesioned surface of the injured area, in a predetermined direction is described. An enclosing structure (5) encloses the injured area to inhibit the entry of injured granulation tissue and mechanical guiding means (11) for the regeneration of the growing tissue, are arranged in the enclosed lesion area, to expand in the predetermined direction. In one aspect, an inhibiting agent of fibrin network formation is administered concurrently to the injured surface of the enclosed lesioned area. In another aspect, the mechanical guiding means takes the form of a gel structure provided with one or more guiding channels for regenerating the growing tissue, which channels extend in the pre-derminate direction

Description

PROMOTION OF REGENERATION OF ORGANIZED TISSUES Description of the invention The present invention relates to the promotion of the regeneration of organized tissue in an injured area, in an organized tissue structure, of a human or animal living body, such as for example nerves ( spinal and cranial), tendons, ligaments, skeletal muscles, bones, joint capsules, cartilages and aponeurosis. It will be noted that the term "regeneration" and derivatives thereof mentioned herein, do not necessarily mean the repair of an injured area in an organized tissue structure by the formation of organized replacement tissue in the injured area, which is identical to the original organic tissue, but simply repair the injured area by forming the organized tissue replacement in the injured area per se. Repair of injured areas in organized tissue structures after trauma resulting from surgical procedures or injuries or injuries, such as transection, bruising or otherwise, is impeded due to incomplete restoration of structure and function. In the case of nerve repair and regeneration, they have been proposed in the art REP. : 28350 previous several implant structures to treat this problem. For example, the use of guidewires for the repair of divided or split nerves has been suggested so far (Alexander et al: Proc. Soc. Exp. Biol. Med. 68: 380-383 (1948); Stopford: Lancet, 10 1296-1297, (1920)). This normally consists of sewing a suture through the space between the proximal and distant ends of the split or split nerve. The coating of the guide filaments with materials such as laminin, collagen and fibronectin has also been disclosed. However, the use of guide filaments has had limited success. The use of an open-ended tube to join the proximal and distant ends of a split or split nerve to promote regeneration of nerve tissue through the space between the proximal and distant ends of the nerve has also been documented (Glück: Arch. Klin., Chir 25: 606-616, (1880), Forssman, Ziegler 's Beitragge zur Pathol, Anato ie 27: 407 (1900)). The use of such a nerve guiding tube results in an increase in the number and / or size of regenerated axons and a decrease in the time required for the regenerative nerve tissue to extend into the damaged region. The provision of a collagen gel matrix in the lumen or lumen of a nerve guiding tube has been shown to further improve the rate of regeneration of the nerve tissue in the space between the proximal and distant ends of the split or split nerve (T. Satou et al: Acta pathologica Japónica 36, 199-208, 1986 and Acta pathologica Japónica 38 (12), 1489-1502, 1988). In addition, it has been proposed in the Canadian patent No. 1328710 (Aebischer et al) form a nerve guiding tube with a non-porous external surface and a porous internal surface, such that an active factor that induces nerve growth can be incorporated in it, for the slow release of it in the injured area. Similarly, International Patent Application Publication No. W092 / 13579 (Fidia SPA) describes a biodegradable and bioabsorbable guide tube for the repair and regeneration of nerve tissue, in which a growth factor is held at the boundary wall of the nerve. lumen to improve nerve regeneration, growth and repair. The incorporation of mechanical guiding structures into the lumen of a nerve guiding tube has also been suggested. For example, International Patent Application Publication No. WO88 / 06871 (Astra Meditec) discloses the provision of a plurality of guide channels extending axially in the lumen of an open ended tube for the tubing of the tubes. proximal and distant ends of a divided nerve. The channels are defined between and / or through fibers which extend axially in the lumen of the tube. Also contemplated is a nerve guide tube having a plurality of isolated lumens formed by laser drilling a plurality of holes or perforations through a cylinder body. The detrimental factors in the use of tubing for the repair of damaged nerves, however, such as inflammation and / or extensive compression of the nerve, have led some experts in the field to conclude that the only damaged nerve regions, which are successfully joined by tubing techniques, are those which can be closed more appropriately (Sunderland: Peripheral Nerve Injuries and Their Repair, pp. 605 (1978) and Nerve Injuries and Their Repair, Churchill Livingstone, pp. 431 ff (1991)). The use of guide tubes has also been suggested for the regeneration of other body structures. For example, International Patent Application Publication No. O88 / 06872 (Astra Meditec) discloses an implant structure for promoting regeneration of cruciform tendons, ligaments and ligaments, comprising an open ended tube, to the lumen of which the free end of a twisted tendon, ligament or cruciate ligament is inserted and through which a plurality of tissue guiding channels extend axially. The guide channels in the lumen of the tube are defined by the spacing between filaments or elements that extend axially through the lumen. However, none of the hitherto proposed structures takes into account the fibrin network and cells in which platelets are included (hereinafter referred to as the "fibrin network") that inevitably forms on the surface of a structure of damaged or injured (injured) tissue. On the other hand, the applicant has appreciated the significance of the fibrin network in the process of repair and regeneration of organized tissues. Growing cells, such as regenerated tissue structures, demand a physical support for their adhesion and migration. In the case of regenerated tissue in a traumatized area, this mechanical support structure is provided or determined, in the stages of growth of formation, in any case, by the fibrin network established in the area of trauma as the base of a clot . The effect of this is that the structure of the fibrin network exerts a crucial influence on the direction of the cells that invade the injured area, with respect to the invading granulation tissue cells and the specific cells that characterize the healing structure ( or cure), that is, the fibrin network constitutes a template or model for the direction and distribution of the cells that characterize the healing structure in a traumatized area. For example, in the case of a damaged or injured nerve, the track or trajectory for the regenerating axons and supporting Schwann cells, through a space or a contused area of a nerve is extensively related to the distribution and organization of the complex fibrin network. The tendons, ligaments, aponeurosis, skeletal muscle, cartilage, bone and other organized tissue structures all show a similar dependence on the configuration created by the fibrin network in the clot-filling substance after the wound. With this in mind, the fibrin network formed in a traumatized area of an organized tissue structure has a highly complex, irregular, three-dimensional (3D) structure of branched fibrin strands or filaments. Thus, in the case of repair of growing nerve tissue, Schwann cells and growing axons progress along the fibrin filaments and branch when the fibrin strands cross or cross each other. The same configuration is true for connective tissue cells that accompany nerve regeneration, that is, its course extensively follows the configuration of the fibrin network that fills the space between the ends of the divided nerve or the area of the bruised nerve. This dependence is equally evident in divided or severed structures, such as after injuries to contusions or surgical procedures. The result of this dependence, in the case of the regeneration of nerve tissues, is that the high capacity of impressive regeneration is compromised extensively, because the vast majority of the new growing axons take a highly aberrant course, branch widely and fail to take their trajectory appropriate, to result in the new growing axons being unable to reach their proposed objectives and to re-establish functional connections. The consequence of the axons randomly directed and misaligned, highly branched, in the apparently scarred nerve, which is unable to reach the innervation of the opposite objectives, is that neuromas are formed (tumor developed in a nerve). The growth is thus prematurely terminated with a fraction of the nerve cells that are inevitably lost. The presence, distribution and organization of the fibrin network is thus a key factor in determining the subsequent organization of tissue regeneration formed during the repair process and thus the ability for the damaged or damaged structure to function properly again. Accordingly, the present invention proposes to improve wound healing in an organized tissue structure in a human or animal living body, by providing means to control the direction of tissue regeneration growth. According to a first aspect of the invention, there is provided a system for promoting the growth of tissue regeneration in a lesioned area in an organized tissue structure, in a human or animal living body, of an injured surface of the injured area in a predetermined direction, comprising an enveloping structure adapted, during use, to be implanted in the human or animal living body, to enclose the injured area, mechanical guidance means for tissue regeneration, adapted, during use, to be arranged in the enclosed lesioned area, to extend in the predetermined direction and an inhibiting agent of the formation of the fibrin network, administrable to the injured surface of the enclosed lesioned area. Normally, the inhibitory agent of fibrin network formation will be administered to the injured area systemically or locally enclosed.

It will be understood that the term "inhibitor" in the characteristics "agent inhibiting the formation of the fibrin network", covers the case of the inhibition of the formation of a fibrin network in the injured area and also the degradation of a network of pre-existing fibrin in the injured area. According to a second aspect of the invention, there is provided a method for promoting the growth of tissue regeneration in an injured area of an organized tissue structure in a human or animal living body, from a lesioned surface of the injured area in a direction predetermined, comprising the steps of enclosing the injured area with a wraparound structure, providing mechanical guiding means for regenerating the tissue in the enclosed injured area, such that the mechanical guiding means extend in the predetermined direction and administer a inhibitor agent of the formation of the fibrin network to the enclosed injured area. In one embodiment of the invention, the inhibiting agent of fibrin network formation comprises a thrombin inhibitor. The thrombin inhibitor can be a thrombin inhibitor based on a low molecular weight peptide. The term "low molecular weight peptide-based thrombin inhibitor" is understood by those skilled in the art, in which thrombin inhibitors are included with one to four peptide bonds and / or with a molecular weight less than 1000 and include those described in the document reviewed by Claesson in "Blood Coagul, Fibrin." (1994) 5,411 also as those described in U.S. Patent No. 4346078, International Patent Application Publication Nos. W093 / 11152, WO95 / 23609, O95 / 35309, W096 / 25426, W094 / 29336, WO93 / 18060 and WO95 / 01168 and European patent application publications Nos. 648780, 468231, 559046, 641779, 185390, 526877, 542525, 195212, 362002, 364344, 530167, 293881, 686642, 669317 and 601459. Peptide-based thrombin inhibitors of low molecular weight preferred include those known collectively as "gatrans", examples are melagatran (H00C-CH2- (R) Cgl-Aze-Pab-H: see International Patent Application Publication No. 094/29336 and the list of abbreviations therein) and inogatran (HOOC-CH2- (R) Cha-Pic-Nag-H: see International Patent Application Publication No. W093 / 11152 and the list of abbreviations therein). The thrombin inhibitor can also be a bisulphated or oligosaccharide polysaccharide, such as a chondroitin sulfate, a dermatan sulfate, a dextron sulfate, a cheratan sulfate, a heparan sulfate or heparin. Alternatively, the thrombin inhibitor can be a hirudin, a biosynthetic analog of hirudin, a fragment of hirudin such as a fragment consisting of at least the last 8 C-terminal amino acids of the known sequence in hirudin or the NAPc2 protein. In another embodiment of the invention, the inhibiting agent of fibrin network formation comprises a fibrinolytic agent. The fibrinolytic agent can be a plasminogen activator (tPA), for example, a recombinant human plasminogen activator (hrtPA) such as Actilyse®, streptokinase or urokinase. In further embodiments of the invention, the fibrin network formation inhibiting agent comprises an inhibitor of Factor X, a trypsin inhibitor or a protease inhibitor, ie, other compounds that affect the activity of the thrombin-thrombin system that they instigate the formation of the fibrin network. In one embodiment of the invention, the inhibiting agent of fibrin network formation is immobilized to the inner surface of the envelope structure, which during use faces the injured area. In one embodiment of the invention, the inhibiting agent of fibrin network formation is a solution and a pump is provided for administering the inhibiting agent of fibrin network formation to the enclosed injured area. The pump may be an osmotic minipump which may be further adapted to be implanted subcutaneously in the living human or animal body. In an embodiment of the invention, the inhibiting agent of fibrin network formation is incorporated into a matrix material for its disposal or administration to the enclosed injured area. As an example, the matrix material can be formed of a material comprising a polysaccharide, such as a chitosan or a hyaluronan such as hyaluronic acid, an agar gel, a hydrogel, such as methyl cellulose gel, Matrigel®, Biomatrix I® , water, saline solution, pH regulated phosphate solution, a lipid or a protein such as collagen. According to a third aspect of the invention, an implantable device is provided for promoting the growth of tissue regeneration to a lesioned area in an organized tissue structure, in a human or animal living body, of a lesioned surface of the injured area in a predetermined direction, comprising an outer envelope structure which, when the device is implanted in the human or animal living body encloses the injured area and an internal gel structure provided with one or more guide channels for tissue regeneration, which , when the device is implanted, it is disposed in the enclosed injured area, such that the guide channels extend in the predetermined direction. The implantable device can be used in conjunction with an inhibiting agent of fibrin network formation according to the invention, although this is not strictly necessary. In one embodiment of the invention, the wraparound structure is a patch for a bruised lesioned area or the like of the organized tissue structure. In an alternative embodiment according to the first and second aspects of the invention, the enclosing structure is a tube having an open end adapted to receive the injured surface and the mechanical guiding means are adapted, during use, to extend into the predetermined direction in the lumen of the tube. If the inhibiting agent of fibrin network formation is administered locally to the lumen of the tube by infusion of a pump, then a uniform distribution of the agent can be promoted, by use of a tube comprising an external continuous tube element, to inhibit the entrance of granulation tissue to the injured area, through which passes a tube connected to the pump and an inner tube element formed of a plurality of longitudinally spaced tube sections, to axially distribute the agent discharged from the pump to the lumen In an alternative embodiment according to the third aspect of the invention, the enclosing structure is a tube having an open end adapted to receive the injured surface with the, or, each guide channel extending in the predetermined direction in the lumen of the tube. In one embodiment of the invention according to the first and second aspects, the injured surface of the injured area is a first injured surface, the open end of the tube is a first open end, the tube has a second open end adapted to receive a second The injured surface of the injured area and the mechanical guiding means are adapted, during use, to extend into the tube lumen between the first and second open ends in the predetermined direction. In one embodiment of the invention, according to the third aspect, the injured surface of the injured area is a first injured surface, the open end of the tube is a first open end, the tube has a second open end adapted to receive a second injured surface of the injured area and the, or each, guide channel extends into the lumen of the tube between the first and second open ends in the predetermined direction.

In these cases, the invention may be to promote the growth of tissue regeneration through a space between the free ends divided or cut transversely of an organized tissue structure, such as a nerve, tendon, skeletal muscle or ligament, the ends open ends of the tube are each adapted to receive one of the free ends divided or cut transversely. The enclosing structure is preferably a biocompatible material and can be biodegradable or non-biodegradable. However, biodegradable material is preferred. The shell structure can be constructed from a material comprising a polysaccharide, for example, a chitosan, heparin, a heparanoid or a hyaluronan such as hyaluronic acid. The envelope structure can also be constructed of a material comprising collagen or other protein complexes. Alternatively, the shell may be constructed from a material comprising a polymer or copolymer, for example, polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, polyglucuronic acid, or poly-N-acetylglucosamine or copolymers thereof as a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid. Where the shell structure is constructed of a non-biodegradable material, the possible materials are silicone and ethylene-vinyl acetate. In one embodiment of the invention, according to first and second aspects, the mechanical guiding means are held or presented by the internal surface of the wrapping structure, which, in use, faces the injured area. In this instance, the mechanical guiding means and the envelope structure could be integrally formed as an implantable body. In one embodiment of the invention, according to the first and second aspects, the mechanical guiding means take the form of guiding channels in the enclosed injured area. One way to obtain this would be to have a wraparound structure which, when implanted, is a tube-like structure having a transverse, spiral cross-section formed for example by lamination to a flat membrane. Then, the guide channels are defined by the longitudinally extending spaces presented by the spiral cross section.

Another way is to have the mechanical guiding means which take the form of a gel structure, which are provided with one or more guiding channels therethrough, the gel structure is adapted, during use to be disposed in the injured area enclosed, such that the guide channels extend in the predetermined direction. Wherein the invention is to promote the regeneration of the nerve tissue, the or each guide channel has a cross-sectional dimension in the range of 50 μm - 1 mm and preferably a cross-sectional dimension in the range of 150 - 500 μm to allow the growth of the functional units of the nerve through the channels, that is, fascicles of the nerve. For other organized tissue structures, the cross-sectional dimension of the, or each, guide channel would be chosen in such a manner as to allow the growth of corresponding functional units therethrough. In one embodiment of the invention, the gel structure is formed from agar, a hydrogel such as methyl cellulose gel, albumin or other proteins, which can be gel-formed, a polysaccharide such as chitosan or a hyaluronan such as acid hyaluronic, a lipid which can be formed in gel, Matrigel® or Biomatrix I®.

In another embodiment of the invention according to the first and second aspects, the mechanical guidance means comprise one or more guide filaments or fibers adapted during use to extend through the aforesaid area enclosed in the predetermined direction. The mechanical guiding means can be for example, monofilaments, multifilaments or woven / non-woven fibers. Preferably, the, or each, guide filament or fiber is of a biocompatible material, which is also a biodegradable material. The, or each, filament or guide fiber can nevertheless be formed from a non-biodegradable material. In one embodiment of the invention, according to the first and second aspects, the or each guide filament or fiber is formed of a material comprising a polysaccharide, such as a chitosan, heparin, a heparanoid or a hyaluronan, such as acid hyaluronic In an alternative embodiment of the invention, according to the first and second aspects, the or each guide filament or fiber is formed from a material comprising a polymer or copolymer. As examples, the or each guide filament or fiber can be formed of a polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, poly-N-acetylglucosamine or copolymers thereof such as for example a copolymer of polyhydrobutyric acid and hydroxyvaleric acid . In another embodiment of the invention according to its first and second aspects, the or each guide filament or fiber is formed of collagen or other protein complexes. In one embodiment of the invention according to the first and second aspects, the mechanical guiding means comprise one or more suture filaments formed for example from vicryl, catgut, polyamide, chitin or nylon. Where non-biodegradable guide filaments are to be used, silicone is appropriate. In one embodiment of the invention, a growth factor or mixtures of growth factors can be administered to the enclosed lesioned area. The growth factor can, for example, be immobilized to the inner surface of the envelope structure. The growth factor may comprise insulin-like growth factors I, insulin-like growth factors II, platelet-derived growth factors, fibroblast growth factors, transforming growth factors, transforming growth factors, neurotrophins, factors ciliary neurotrophs, EGF or glial growth factors. The growth factor can also comprise Schwann cells, endothelial cells, fibroblasts, macrophages or inflammatory cells or genetically altered cells, which can express a growth factor. According to a fourth aspect of the invention, there is provided the use of a system, according to the first aspect of the invention, to promote the growth of tissue regeneration in an injured area of an organized tissue structure, in a living body. human or animal, from a damaged surface of the injured area in a predetermined direction. According to a fifth aspect of the invention, there is provided the use of an implantable device according to the third aspect of the invention to promote the growth of tissue regeneration in an injured area of an organized tissue structure in a human living body or animal from a damaged surface of the injured area in a predetermined direction. Examples of organized tissue structures in which the invention can be used to promote the growth of tissue regeneration in an injured area are the nerves, tendons, ligaments, joint capsules, cartilages, bones, aponeurosis and skeletal muscles.

The invention thus provides inhibition or control of the formation of the fibrin network in the injured tissue, coupled with the provision of mechanical guiding means to allow emerging cells in the injured area of the injured surface to follow the track or path offered. by mechanical guidance means. The advantage of this is that the cells do not show any branching or irregular trajectory since a coherent trajectory for growth is provided. Therefore, it is possible to obtain the regeneration of specific organized cells, such as Schwann cells from peripheral nerves or tenocytes from tendons, to join the defect induced by a lesion with minimal deviation and irregularity in the organization of the newly tissue. formed. To illustrate the invention, experiments were carried out on adult rats under the permits O 293/93, 0 69/95 and 0 70/95 granted by the "Animal Experiments Ethical Committee of the University of Gothenburg", which will now be described with reference to the accompanying drawings of the drawings, in which: Figure 1 is a cross-sectional side view of a guide tube having a lumen at opposite ends of which the proximal and distal ends of a sciatic nerve cut out transversely of an adult rat are sutured and through which the guide filaments extend schematically, illustrating the fibrin network formed in the lumen between the proximal and distant ends in the absence of administration of an inhibitory agent of the network formation. fibrin. Figure 2 is a cross-sectional side view of a guidewire tube and guidewire assembly corresponding to that of Figure 1, to opposite ends of the tube of which the proximal and distal ends of a transversally cut sciatic nerve of An adult rat is sutured, schematically illustrating the fibrin network formed with the administration of an inhibiting agent of fibrin network formation. Figure 3 is a cross-sectional side view of a guide tube corresponding to that of Figure 1, to the lumen of which only the proximal end of a transversely cut sciatic nerve of an adult rat is sutured, schematically illustrating the network of fibrin formed in the absence of administration of an inhibiting agent of fibrin network formation. Figure 4 is a cross-sectional side view of a guide tube corresponding to that of Figure 1, to the lumen from which only the proximal end of a transversely cut sciatic nerve of an adult rat is sutured, schematically illustrating the network of fibrin formed with administration of an agent inhibiting the formation of the fibrin network. Figure 5 is a cross-sectional side view of a guiding filament guide and mounting tube corresponding to that of Figure 1 to the lumen of the tube from which only the proximal end of a transversely cut sciatic nerve of an adult rat is sutured , which schematically illustrates the fibrin network formed in the absence of administration of an inhibitor of fibrin network formation. Figure 6 is a cross-sectional side view of a guiding filament guide and mounting tube corresponding to that of Figure 1, to the lumen from which only the proximal end of a transversely cut sciatic nerve of an adult rat is sutured, which schematically illustrates the fibrin network formed with administration of an inhibitor of fibrin network formation. Fig. 7 is a cross-sectional side view of a guide tube comprising an external continuous tube element and an inner tube element formed from longitudinally spaced sections having a lumen at one end of which the proximal end of a transversally cut sciatic nerve of an adult rat sutured, and through which a guide wire extends, schematically illustrating the fibrin network formed with local administration of an inhibitor of fibrin network formation by infusion of an implanted osmotic minipump. Figure 8 is a schematic cross-sectional side view of a guide tube having a lumen to which the proximal and distal ends of a transversely cut sciatic nerve of an adult rat are sutured at opposite open ends and filled with a gel provided with longitudinally extending guide channels. With reference to Figures 1 and 2, a silicone guide tube 5 is shown, at the opposite open ends of which the proximal end 1 and the distal end 3 of a transversely cut sciatic nerve of an adult rat have been inserted and fixed in place by sutures 7. The guide tube 5 prevents or inhibits access of the granulation tissue to the injured area or surfaces by isolating the damaged nerve structure from the surrounding tissue, usually tissue connecting blood vessels. This makes it easier to control the repair and modeling of tissue regeneration formed between the split ends of the damaged sciatic nerve. The guide tube 5 additionally acts as a delivery system for compounds that interfere with the formation of a fibrin network in the injured area, as will become apparent and can also act as a slow delivery system for growth promoting agents. While Figures 1 and 2 show the damaged rib extending only a short distance to the guide tube 5, both proximal and distal ends 1, 3 can be covered by longer lengths of the guide tube 5. This distance is it can extend to the point where the division of a fascicle of the nerve at the distant end prevents further wrapping. A guide wire 11 formed from an ophthalmic suture material extends between the proximal and distant ends 1, 3 of the damaged sciatic nerve through the lumen of the guide tube 5. Of course, instead of this, a plurality of guide filaments. When the guidewire 11 is surgically inserted, a segment of filament projects outwardly of the guide tube 5. Then, this segment is cut off when the surgical repair procedure is complete. In Figure 1, the damaged sciatic nerve is left to regenerate in the presence of the saline introduced into the guide tube 5 in surgery. The result after a few days is a large complex fibrin network 13 of branched filaments formed along guide filament 11 in the space between the proximal and distant ends 1, 3 of the sciatic nerve, so that subsequent regeneration of the sciatic nerve continues. nerve tissue. As in the assembly of Figure 1, the guide tube 5 in the assembly of Figure 2 is also filled with saline in surgery. However, in addition, melagatran is administered systemically to the injured area by subcutaneous infusion with an implanted minipump (not shown). As shown schematically, a narrower outlet network 14 contours the center guide filament 11 in the space between the transversely cut ends of the rib. Importantly, the fibrin network 14 exhibits a coherent structure of fibrin platelets and fragments aligned in the direction of the filament 11 so that subsequent regeneration of the nerve tissue proceeds. In Figures 3 and 4 there is shown a silicone guide tube 105 corresponding to the guide tube 5 of Figures 1 and 2. In these assemblies, however, only the proximal end 101 of the transversely cut sciatic nerve of an adult rat is it retains in place in tube 105 with a suture 107 and no guidewire extends through the lumen. Thus, one of the open ends 102 of the tube is left open. Like before, saline is introduced into the lumen of tube 105 in surgery. In the assembly of figure 3, no additional treatment is provided. A small clot 115 consisting of fibrin, platelets and other blood cells is formed, which only covers the proximal end but the fibrin network is not formed in the rest of the tube. Thus, none of the substantial fibrin network is formed to support the growth of tissue regeneration in the subsequent nerve. In contrast, in the assembly of Figure 4, melagatran is administered systemically to the injured area by subcutaneous infusion with an implanted minipump. A small clot 116 of fibrin and cells is formed, which only covers the proximal end of the nerve. There is no fibrin network in the rest of the tube. Thus, no substantial fibrin network is formed to support the regeneration growth of the subsequent nerve tissue. With reference to Figures 5 and 6, there is shown a silicone guide tube 205 corresponding to the guide tube 5 of Figures 1 and 2. As in Figures 3 and 4, the proximal end 201 of a transversally cut sciatic nerve of an adult rat is retained in tube 205 with a suture 207 and the lumen of the tube filled with saline in surgery. In contrast to Figures 3 and 4, however, a guide wire 211 in the form of an ophthalmic suture extends from the proximal end 201 to the lumen. In the case of Figure 5, no additional treatment is administered and a narrow complex clot 217 consisting of irregularly organized fibrin filaments, platelets and other blood cells are formed along guide filament 211 throughout tube 205 for continue the subsequent regeneration of the nerve tissue. On the other hand, in the case of Figure 6, melagatran is administered systemically to the area administered by subcutaneous infusion with an implanted minipump. A small clot 218 of coherent fibrin platelets, fragments of fibrin and cells is formed along and aligned with the guide filament 211 in the center of the tube, so that subsequent regeneration of the nerve tissue proceeds. In Figure 7, an open-ended guide tube 305 is shown, comprising an outer continuous tube element 306 and an inner tube element 308 formed from a plurality of longitudinally spaced sections 310. As shown, the proximal end 301 of a transversely cut sciatic nerve of an adult rat is inserted into the tube and held in place by a suture 307. A guide filament 311 in the form of an ophthalmic suture extends through the lumen. of the tube from the near end. The tube is filled with saline in the surgery and then local infusion of melagatran is done to the enclosed lesioned area, with an implanted 312 osmotic minipump. The longitudinally spaced sections 310 help distribute the melagatran throughout the space in the lumen. A narrow coherent clot 319 of coherent fibrin platelets, fibrin fragments and cells is formed along and aligned with the length of the guide filament 311 in the center of the tube for subsequent regeneration of the nerve tissue. The construction of the guide tube shown in Figure 7 can be varied in such a way that each isolated longitudinal section is provided with one or more guide filaments. Thus, the guide tube and filament assembly consists essentially of smaller individual assemblies of similar construction. This facilitates the separation of the regenerative nerve fascicles since the longitudinal sections act as a separate guide for each fascicle. Turning now to FIG. 8, there is shown an open ended guide tube 405, at opposite ends of which the proximal and distant ends 401, 403 of a transversely cut or divided sciatic nerve of an adult rat have been inserted and fixed. with sutures 407. The tube lumen 405 in this case, it is filled with agar gel 402 through which a plurality of axial guide channels 404 extend. The gel structure 402 is shown spaced from the end surfaces of the cross-cut nerve structure for the purpose of clarity, although needless to say that it is not a strict requirement for the gel structure to be attached to the end surfaces of the rib. The result of the assembly is the formation of a coherent fibrin network in the guide channels having long fibrin filaments or cables axially aligned with the guide channels for further regeneration of the nerve tissue to proceed. The same systems as those described above with reference to the attached figures of the drawings, modified with respect to shape and dimensions, have also been tested on tendons, ligaments, abdominal aponeuroses and skeletal muscles of adult rats, with corresponding results. The use of silicone tubes and ophthalmic sutures for the guide filaments are selected in the assemblies described hereinabove with reference to the attached figures of the drawings only due to the fact that they are suitable for use in experimental animals.

The guide tube can be formed from biocompatible bioabsorbable or non-bioabsorbable materials, which can be permeable or impermeable to materials soluble in aqueous solutions. However, the use of a bioabsorbable or biodegradable material is preferred. The materials from which the guide tube can be constructed include, but are not limited to, collagen complex, heparin and heparonoids, chitosan and related polysaccharides, polylactic acid, polyglycolic acid, polyhydroxybutyric acid, permselective polytetraethylene, poly-N -acetylglucosamine or polymers to which growth factors can be directly incorporated (eg, ethylene-vinyl acetate). With respect to the guidewire filaments, these can be formed from biocompatible material, similar or identical to the materials used for the guide tube. Other materials that may be used include suture materials available in the present, such as vicryl, catgut, nylon, chitin, as well as other materials, which may act as a compatible substrate for the formation of a regenerating tissue cable such as nerve axons. The invention will now be illustrated, but by no means limited, by the following examples carried out on adult rats under the ethical permits identified and referenced hereinafter where appropriate with the assemblies described hereinabove with reference to the figures of the attached drawings.

Example 1 The sciatic nerve of adult rats is unilaterally exposed in the middle muscle and cut transversely. Then, the proximal and distant ends are inserted into a silicone guide tube of the type shown in Figures 1 to 6 of the accompanying drawings, to leave a space of 10 mm between them filled with phosphate buffered saline ( PBS). The silicone tube is sutured to the perineurium of the nerve ends inserted with 9-0"atraumatic" ophthalmic sutures (Ethicon). The regenerated formed in the space was after 2 and 4 weeks, examined with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were quite few and showed extensive aberration, often arranged as rings. The Schwann cells were identified in the central parts of the regenerated arranged in an apparently random configuration. The fibroblasts formed a structure similar to wrapped perineurium. Numerous macrophages were distributed throughout the region of space, such as scattered erythrocytes. Thus the regeneration of the nerve was blocked in the absence of a mechanical guide structure that connects the space between the ends of the nerve, such as a guide wire covered by a fibrin network. Thus, a poor repair of the nerve is obtained.

Example 2 The sciatic nerve of adult rats was exposed unilaterally in the middle thigh and cut transversely. Then, the proximal and distant ends are inserted into a silicone guide tube, to leave a gap of 10 mm between them. The silicone tube is sutured to the perineurium of the inserted ends of the nerve with 9-0"atraumatic" ophthalmic sutures (Ethicon). As in the assembly described hereinabove with reference to Figure 1, the space is filled with PBS and a single central guide filament (monofilament nylon 10-0) positioned in the lumen of the tube to join the proximal and distal ends of the tube. nerve. The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were slightly more numerous than in Example 1, but they still showed extensive aberration, again arranged frequently as rings or rings. The Schwann cells were identified in the central parts of the regenerated, arranged in a seemingly random configuration. The fibroblasts formed a structure similar to an enclosed perineurium. Numerous macrophages were distributed throughout the region of space, as scattered erythrocytes. Thus, a poor repair of the nerve is obtained, because the regeneration of the nerve tissue follows the path laid by a complex fibrin network formed in the early stages of the repair process.

Example 3 The sciatic nerve of adult rats was exposed unilaterally in the middle thigh and cut transversely. Then, the proximal and distant ends were inserted by 2 mm into a silicone guide tube, having a diameter of 1.5 mm, to leave a space of 10 mm between them. The silicone tube is sutured to the perineurium of the inserted ends of the nerve with 9-0"atraumatic" ophthalmic sutures (Ethicon).

As in the experiment described hereinabove with reference to Figure 2, a single central guide filament (monofilament nylon 10-0) is positioned in the lumen of the tube to join the proximal and distant ends of the nerve, the space fills up with PBS and systemic infusion of melagatran via the peritoneum with the help of an osmotic minipump (Alza 2001, volume = 200 μl, pumping speed = 1 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Melagatran, Astra Hássle AB, Molndal, Sweden) implanted into the perifonea cavity for a week and a free positioned pump outlet in the perifonea cavity. The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were at least as numerous as in Examples 1 and 2, showed little aberration and were arranged parallel to the central guidewire, that is, exhibited a very high degree of coherence. Schwann cells exhibited a high degree of coherence as they were arranged mainly parallel to the central guidewire and few cells diverged from that direction. There was a hardly random configuration in the organization of the Schwann cells. The fibroblasts formed a structure similar to an enclosed perineurium. An outstanding feature was the absence of macrophages and blood cells outside the regenerated. An excellent regeneration is thus obtained through a space of 10 mm, because the regeneration of the nerve tissue follows the path laid by a coherent fibrin network, formed in the early stages of the repair process.

Example 4 The sciatic nerve of adult rats was exposed unilaterally in the middle thigh and cut transversely. Then, the proximal and distant ends are inserted by 2 mm into a silicone guide tube, to leave a gap of 10 mm between them. The silicone tube is sutured to the perineurium of the inserted ends of the nerve with 9-0"atraumatic" ophthalmic sutures (Ethicon). As in example 3, a single central guide filament (monofilament nylon 10-0) is positioned in the lumen of the tube to join the proximal and distant ends of the nerve, the space is filled with PBS and melagatran is administered. However, in this case, a tube of an osmotic minipump (Alza 2001, volume = 200 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor. Melagatran, Astra Hássle AB, Molndal, Sweden) implanted subcutaneously on the back of the animal, provides localized melagatran solution to the space, for at least 8 days, in the manner shown in figure 7. The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were at least as numerous as in Examples 1 and 3, showed little aberration and were arranged parallel to the central guidewire, that is, exhibited a very high degree of coherence. The Schwann cells were arranged mainly parallel to the central guide filament and few cells diverged from that direction. There was no disordered or random configuration in the organization of the Schwann cells. The fibroblasts formed a structure similar to an enclosed perineurium. There were no macrophages and blood cells outside the regenerated. Thus, an excellent regeneration of coherent axons is obtained again through a space of 10 mm.

Example 5 As in the assembly described above with reference to Figure 3, the sciatic nerve of adult rats is cut transversely and the proximal end was inserted into a silicone guide tube filled with PBS. No regeneration is formed from the proximal end of the nerve after 2 or 4 weeks. Macrophages and blood cells could be recognized near the proximal end of the nerve. The regeneration of the nerve was thus blocked in the absence of a mechanical guide structure extending from the proximal end of the nerve, such as a guide wire covered by a fibrin network.

Example 6 As in the experiment described hereinabove with reference to Figure 4, the sciatic nerve of adult rats is cut transversely and the proximal end was inserted into a silicone guide tube filled with PBS and to which an infusion is made Systemic melagran, with the help of an implanted osmotic minipump. As in example 5, however, no regeneration is formed from the proximal end of the nerve after 2 or 4 weeks. Macrophages and blood cells could be recognized, near the proximal end of the nerve.

Thus, nerve regeneration was blocked in the absence of a mechanical guide structure extending from the proximal end of the nerve, such as a guidewire covered by a fibrin network.

Example 7 The sciatic nerve was exposed unilaterally in the middle thigh of adult rats and cut transversely. Immediately after this, the proximal and distal ends were inserted by 2 mm into a silicone guide tube, of the type shown in Figures 1 to 6, pre-filled with pH-regulated saline. The space between the ends of the nerve was 10 mm. The silicone tube is sutured to the perineurium of the inserted ends of the nerve with 9-0"atraumatic" ophthalmic sutures (Ethicon). An osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Melagatran, Astra Hássle AB, Molndal, Sweden) It was implanted subcutaneously on the back of the animal and the outlet of the pump was connected via a tube, to the middle portion of the silicone tube enclosing the transversely cut nerve for the local supply of melagran to the space, as shown in the figure 7 The nerves and the tube with their space are examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the axons and the Schwann, as visualized by immunological histochemistry with the help of neurofilament antibodies (N 0142 &N 5389, Sigma) and the S-100 neurological proteins (S 2644, Sigma; Z 0311, Dakopatts). There was no regeneration that united the space. No axons or Schwann cells filled the space. Strains and cells of scattered amorphous proteins could be recognized in the fluid that filled the space. The regeneration of the nerve was thus blocked in the absence of a mechanical guiding structure that joined the space between the ends of the nerve, such as a guide wire covered by a fibrin network.

Example 8 The sciatic nerve was exposed unilaterally in the middle thigh of adult rats and cut transversely. Immediately after this, the proximal and distal ends were inserted by 2 mm into a silicone guide tube, of the type shown in Figures 1 to 6, pre-filled with pH-regulated saline. However, there was no guidewire, for example a sutured thread, in the center of the silicone tube. The space between the ends of the nerve was 10 mm. The silicone tube is sutured to the perineurium of the nerves inserted with 9-0"atraumatic" ophthalmic sutures (Ethicon). An osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Melagatran, Astra Hássle AB, Mólndal, Suecia) was implanted in the peritoneal cavity and the pump outlet was left open for the systemic supply of melagatran via the peritoneal cavity and the blood system. The nerve and the tube with its space are examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the axons and Schwann cells, as visualized by immunological histochemistry with the help of antibodies to neurofilaments ( N 0142 & N 5389, Sigma) and the neurogical proteins S-100 (S 2644, Sigma; Z 0311, Dakopatts). There was no regeneration that united the space. No axons or Schwann cells filled the space. Strains and cells of scattered amorphous proteins could be recognized in the fluid that filled the space. The regeneration of the nerve was thus blocked in the absence of a mechanical guide structure that joined the space between the ends of the nerve, such as a filament covered with a fibrin network.

Example 9 The sciatic nerve of adult rats is cut transversely and the proximal end is inserted 2 mm into a silicone guide tube. As in the experiment described above with reference to Figure 4, a single central guidewire filament (monofilament nylon 10-0) was sewn through the proximal end of the sciatic nerve to extend into the lumen and the lumen was filled with PBS, with the help of an implanted osmotic minipump. The distant end of the transversally cut sciatic nerve was positioned between the muscles of the thigh, in such a way that it could not interfere with the growth in the silicone tube. The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons adhered to the central guidewire and many appeared parallel to it. However, axons showed extensive aberration and rings or rings were common. The Schwann cells were, to a large extent, arranged parallel to the central guidewire, but many cells diverged from that direction. The fibroblasts formed a structure similar to an enclosed perineurium. There were numerous macrophages and blood cells outside the regenerated.

Thus, a poor repair of the nerve is obtained, because the regeneration of the nerve tissue follows the path laid by the complex fibrin network formed in the early stages of the repair process.

Example 10 The sciatic nerve of adult rats is cut transversely and the proximal end is inserted 2 mm into a silicone guide tube. As in the experiment described above with reference to Figure 6, a single central guidewire filament (monofilament nylon 10-0) is sewn through the proximal end of the sciatic nerve to extend into the lumen, the lumen is filled with PBS and made infusion of melagatran systemically, with the help of an implanted osmotic minipump. The distant end of the transversally cut sciatic nerve was positioned between the muscles of the thigh, in such a way that it could not interfere with the growth in the silicone tube. The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were arranged parallel to the central guidewire and very few diverged. There was no axonal aberration and there was hardly any ring or ring. The Schwann cells were, to a large extent, arranged parallel to the central guidewire. The fibroblasts formed a structure similar to an enclosed perineurium. There were few macrophages and blood cells outside the regenerated. Thus, an excellent regeneration of coherent axons is obtained, because the regeneration of the nerve tissue follows the trajectory laid down by a coherent fibrin network formed in the early stages of the repair process.

Example 11 The sciatic nerve of adult rats is cut unilaterally and the proximal end is inserted 2 mm into a silicone guide tube that takes the shape of the tube shown in Figure 7. As in the experiment described hereinabove with reference to Figure 7, a single central guidewire filament (monofilament nylon 10-0) is sewn through the proximal end of the sciatic nerve to extend into the lumen, the lumen is filled with PBS and systemic infusion of melagatran is made, with the help of an implanted osmotic minipump. The distal end of the transversally cut sciatic nerve is positioned between the thigh muscles at a distance from the silicone tube and thus can not interfere with the regenerated one.

The regenerated formed in space is examined after 2 and 4 weeks with respect to the distribution and direction of axons and Schwann cells, as visualized by immunological histochemistry. The axons were arranged parallel to the central guidewire and very few diverged, except in the immediate vicinity of the proximal end of the nerve. There was no aberration of the axons and there was hardly any ring or ring, except for a few at the proximal end of the nerve. The Schwann cells were mainly arranged parallel to the central guidewire. The fibroblasts formed a structure similar to an enclosed perineurium. There were few macrophages and blood cells outside the regenerated. Thus, an excellent regeneration of coherent axons is obtained, because the regeneration of the nerve tissue follows the trajectory laid down by a coherent fibrin network formed in the early stages of the repair process.

Example 12 The assembly for this example was the same as that of example 11, unlike the local infusion of streptokinase (Hoescht) with the help of an osmotic minipump.

Similar results were obtained to those described in example 11. However, the osmotic minipump was changed every second day to obtain sufficient streptokinase activity for a period of 8 days.

Example 13 The assembly for this example was the same as that of example 11, in contrast to the local infusion of the recombinant human plasminogen activator (hrtPA) Actilyse® (Boehringer Ingelheim) with the help of an osmotic minipump. Results similar to those described above in Example 11 were also obtained.

Example 14 The assembly for this example was the same as that of example 11, unlike the local infusion of urokinase locally, with the help of an osmotic minipump. Results similar to those described above in Example 11 were also obtained.

Example 15 The sciatic nerve was exposed unilaterally in the middle thigh of adult rats and cut transversely. As in example 3, the proximal and distant ends are inserted 2 mm into a silicone tube, pre-filled with saline of pH regulated and having through its center a guide filament, for example, a suture thread. The space between the ends of the nerve is 10 mm. The silicone tube is sutured to the perineurium of the nerves inserted with 9-0"atraumatic" ophthalmic sutures (Ethicon). However, in this case, an osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Hirudin, (Sigma)) is implanted subcutaneously on the back of the animal and the outlet of the pump is connected, via a tube, to the middle portion of the silicone tube that encloses the transversely cut nerve for local administration of Hirudin to space, in the manner shown in Figure 7. The regenerated formed in the space is examined after 2 and 4 weeks with respect to the distribution, direction and coherence of axons and Schwann cells, as visualized by immunological histochemistry. The axons were numerous and highly coherent, showing only minor aberrations and were poorly arranged as rings. The Schwann cells were identified in the central parts of the regenerated and showed a high degree of coherence as well. The fibroblasts formed a structure similar to an enclosed perineurium. Scarce macrophages were noted in the space region. This results in an excellent regeneration of coherent axons through a space of 10 mm.

Example 16 The sciatic nerve was exposed unilaterally in the middle thigh of adult rats and cut transversely. As in example 7, the proximal and distal ends are inserted 2 mm into a silicone tube, pre-filled with pH-regulated saline, but having no guide filament, for example a suture, in the center of the tube of sylicon. The space between the ends of the nerve is 10 mm. The silicone tube is sutured to the perineurium of the nerves inserted with 9-0"atraumatic" ophthalmic sutures (Ethicon). In contrast to example 7, an osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Hirudin, (Sigma)) is implanted subcutaneously on the back of the animal and the outlet of the pump is connected, via a tube, to the middle portion of the silicone tube that encloses the transversally cut nerve for local administration to space, as shown in Figure 7. The nerve and the tube with its space is examined after 2 and 4 weeks with respect to the distribution, direction and coherence of axons and Schwann cells, as visualized by immunological histochemistry with the help of antibodies to neurofilaments (N 0142 &N 5389, Sigma) and the neurogical proteins S-100 (S 2644, Sigma; Z 0311, Dakopatts). There was no regeneration that united the space. No axon or Schwann cells fills space. Strains and cells of scattered amorphous proteins could be recognized in the fluid that filled the space. The regeneration of the nerve was thus blocked in the absence of a guide structure, such as a filament covered with a fibrin network.

Example 17 The sciatic nerve is unilaterally exposed in the middle thigh of adult rats and cut transversely. As in example 16, the proximal and distal ends of the rib are inserted 2 mm into a silicone tube, pre-filled with pH-regulated saline, but having no guide filament, for example a suture, in the center of the silicone tube. The space between the ends of the nerve is 10 mm. The silicone tube is sutured to the perineurium of the nerves inserted with 9-0"atraumatic" ophthalmic sutures (Ethicon). In contrast to example 16, an osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Hirudin, (Sigma)) is implanted in the peritoneal cavity and The output of the pump is left open for the systemic delivery of Hirudin, via the peritoneal cavity and the blood system. The nerve and the tube with its space are examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the axons and Schwann cells, as visualized by immunological histochemistry with the help of neurofilament antibodies.

(N 0142 &N 5389, Sigma) and the S-100 neurological proteins (S 2644, Sigma; Z 0311, Dakopatts). There was no regeneration that united the space. No axons or Schwann cells filled the space. Strains and cells of scattered amorphous proteins could be recognized in the fluid that filled the space. The regeneration of the nerve was thus blocked in the absence of a mechanical guide structure, such as a filament covered with a fibrin network.

EXAMPLE 18 In this example, parts of the Achilles tendon of adult rats are cut longitudinally and the proximal and distant ends are introduced 1 mm into a silicone tube that takes the shape of that shown in Figures 1 to 6. A single tube is provided. central guide filament (monofilament nylon 10-0) to extend through the lumen of the tube, to join the proximal and distant stump of the tendon. Then, a systemic infusion of PBS is made or melagatran, with the help of an osmotic minipump. After 3 weeks more parallel fibroblasts and a higher degree of ordered coherence of the collagen fibers could be recognized in the tubes treated with melagatran, as compared to the tubes treated with PBS.

Example 19 The Achilles tendon is exposed unilaterally in adult rats and a portion thereof is cut transversely. As in example 3, the proximal and distal ends of the tendon, cut transversely and in isolation, are inserted 2 mm into a silicone tube, pre-filled with pH-regulated saline solution and having a guide filament through its center. example a suture thread. The space between the ends of the tendon is 4 - 6 mm. The silicone tube is sutured to the paratenon with 9-0"atraumatic" ophthalmic sutures (Ethicon). An osmotic minipump (Alza 2002, volume = 220 μl; pumping speed = 0.5 μl / hour; Alza Corp .; Palo Alto, CA, USA; pre-filled with a solution of the thrombin inhibitor Melagatran, Astra Hássle) is implanted subcutaneously in the back of the animal and the outlet of the pump is connected, via a tube, to the middle portion of the silicone tube that encloses the transversely cut tendon, to the local supply of Melagatran to the space, as shown in Figure 7. The regenerated formed in the silicone tube is examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the fibers and collagen cells. The space is filled with collagen fibers that show high degrees of coherence. Collagen structures randomly organized are scarce. Fibroblasts and vascular wall cells were common, whereas macrophages and inflammatory cells were rare. A structure similar to paratenon could be recognized. A good regeneration of the tendon is obtained.

Example 20 The Achilles tendon is exposed unilaterally in adult rats and a portion thereof is cut transversely. As in example 7, the proximal and distal ends of the tendon, cut transversely and insulated, are inserted 2 mm into a silicone tube, pre-filled with saline of pH regulated, but lacking a guide filament, such as a nylon thread. suture inserted in the center of the silicone tube. The space between the ends of the tendon is 4 - 6 mm. The silicone tube is sutured to the paratenon with 9-0"atraumatic" ophthalmic sutures (Ethicon). An osmotic minipump (Alza 2002, volume = 220 μl, pumping speed = 0.5 μl / hour, Alza Corp, Palo Alto, CA, USA, pre-filled with a solution of the thrombin inhibitor Melagatran, Astra Hássle) is implanted subcutaneously in the The animal's back and the outlet of the pump is connected, via a tube, to the middle portion of the silicone tube that encloses the transversely cut tendon, for the local supply of Melagatran to space. The regenerated formed in the silicone tube is examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the fibers and collagen cells. The space is incompletely filled with randomly arranged collagen filaments, highly variable in size and direction. There was a lack of good coherence. Thus, randomly organized collagen structures predominated. Fibroblasts and vascular wall cells were common, whereas macrophages and inflammatory cells were rare. Thus, a poor repair of the tendon is obtained.

Example 21 The sciatic nerve is unilaterally exposed in the middle thigh of adult rats and cut transversely. Then, the proximal and distal ends of the rib were inserted 2 mm into a 10 mm silicone guide tube (inner diameter 1.8 mm) of the type shown in Figures 1 to 6, to leave a space in the lumen between the ends of the mandrel. 6 mm nerve, filled with a 1% homogenous agar gel. The silicone tube is sutured to the epineurium of the inserted ends of the nerves with 10-0"atraumatic" Ethilone® sutures (Ethicon). The nerve and the tube with its space were examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the axons and Schwann cells, as visualized by immunological histochemistry with the help of antibodies to neurofilaments ( N 0142 &N 5389, Sigma) and the S-100 neurogylic proteins (S 2644, Sigma; Z 0311, Dakopatts).

The tissue that fills the space between the ends of the nerve and the gel, which lacks channels, is composed of highly irregular connective tissue with rings or rings of cells and filaments. The Schwann cells were randomly aligned. There were no minifascicles or different neurite bundles. The axons exhibited intense aberration and were part of structures similar to neuromas. Axons organized randomly between the inner surface of the silicone tube and the agar gel were observed. A positive prick test occurs after 4 weeks, at a distance of 15 mm in one of four animals, the other three are negative. Thus, the regeneration of the nerve was blocked in the absence of a mechanical guide structure.

Example 22 The sciatic nerve is unilaterally exposed in the middle thigh of adult rats and cut transversely. Then, the proximal and distant ends of the nerve were inserted 2 mm into a 10 mm silicone guide tube (inner diameter 1.8 mm) to leave a space in the lumen, between the ends of the 6 mm nerve, pre-filled with a gel of homogeneous agar at 1% having 3 or 5 longitudinal channels of nominal diameter of 0.4 mm formed by temporary insertion of filaments during the gelation process, for example as shown in Figure 8. The silicone tube is sutured to the epineurium of the inserted ends of the nerves with 10-0"atraumatic" Ethilone® sutures (Ethicon). The nerve and the tube with its space are examined after 2 and 4 weeks with respect to the distribution, direction and coherence of the axons and Schwann cells, as visualized by immunological histochemistry with the help of antibodies to neurofilaments ( N 0142 & N 5389, Sigma) and the neurogylic proteins S-100 (S 2644, Sigma; Z 0311, Dakopatts). The interfaces between the ends of the nerve and the gel and between the gel and the surrounding silicone tube are filled with tissue, reintegrated and restructured in a regular manner with the passage of time. The macroscopic inspection after 1 week reveals that a fibrin network is convergent towards the opening of one of the channels, both close and distant. Scanning electron microscopy (scanning) verifies the presence of a network of axially aligned, coherent fibrin cell lines, rich in platelets, converging towards the openings of the channels and extending therethrough. After 2 weeks and more obviously after 4 weeks, the axons and Schwann cells are identified at the interface between the ends of the nerve and the gel and in the channels delimited by the granulation tissue and its blood vessels. The cells that enter the area of space follow the trajectory of the fibrin and platelet cables. Only a minor inflammatory cell reaction is noted in the gel channels and at the interface between the agar and the surrounding silicone tube. The nerve tissue that joins the duct space from the proximal end to the distant end of the nerve through the intermediate channels is arranged in mini-fascicles, composed mainly of axons and Schwann cells enclosed by a thin perineurium. The axons and Schwann cells were arranged in bundles of different fascicles, which are joined between the ends of the nerve through the channels. In addition, randomly oriented Schwann cells could be recognized between fairly coherent bundles of cells in the region of space. The number of axons per channel is increased by a factor of the order of at least 3 from 2 weeks to 4 weeks. The perineural connective tissue enclosing the minifascicles was less abundant than that observed in the regenerated tubes filled with PBS, that is, example 1. The axons were also observed along the interface between the gel and the tube. The vast majority of these axons were randomly oriented, lacking the high degree of demonstrable axonal coherence in the gel channels. The axons in the 5-channel system exhibited good coherence even when they enter the distant nerve. Positive puncture tests were produced after 4 weeks at a greater distance than when homogeneous agar gel filled the space in the lumen of the tube between the ends of the tube, the 5-channel system produces positive puncture tests at a distance greater than the 3-channel system. In addition, the number of coherent axons advancing at approximately twice the speed is obtained almost twice as compared to the homogeneous gel assembly of Example 21 and also the assembly of Example 1. Thus, excellent regeneration is obtained through the space. Similar results have also been obtained to those obtained in the previous examples, which use melagatran and streptokinase, in experiments of regeneration of tubular abdominal aponeurosis and skeletal muscles of the thigh, that is, the treatment with melagatran and streptokinase results in a better order structure in the regenerated tissue, compared to that after infusion of PBS subcutaneously. In addition, corresponding experiments carried out with inogatran, heparin, the recombinant human plasminogen activator Actilyse® and urokinase have produced similar improvements in the repair and regeneration of a damaged area in an organized tissue structure. Accordingly, it can be seen that the use of a wrapping structure, such as the guide tubes of the examples, with mechanical guiding means for tissue regeneration, such as the central guiding filaments of the examples, in combination with a agent inhibitor of fibrin network formation, such as fibrinolytic agents and / or a thrombin inhibitor, causes an improvement in the regeneration of the wound structure, when compared with the systems proposed hitherto. There is also an improvement when a gel, which has guide channels, is provided with a damaged area enclosed by a wraparound structure. In accordance with the present invention, the materials that can be used as a carrier matrix for the inhibiting agent of fibrin network formation include any material or system in which the agent can be inserted into or transported to the enclosed injured area, such as any biocompatible material from which the agent can be suspended, dissolved or released. Such carrier materials include, but are not limited to: collagen, methylcellulose gel, chitosan and other polysaccharides, fibrin or other proteins, extracellular TM matrix materials, such as Matrigel (Collaborative Research, Inc., Waltham, MA, USA), Biomatrix ™ (Biomedical Technologies, Inc., Stoughton, MA) or other related materials. The carrier can also comprise saline solution, water or, as in the examples described above, pH regulated solutions which can be delivered to the enclosed lesioned area by using a continuous delivery system, such as an osmotic minipump or externally accessible catheter connected to the device for continuous supply. The inhibiting agent of fibrin network formation could also be immobilized in the envelope structure. The present invention also achieves that growth promoting agents are placed or delivered to the enclosed injured area. Such agents include trophic, chemotactic, mitogenic or similar substances or combinations and mixtures thereof, which are capable of stimulating growth, directly or indirectly. More preferably, these agents include insulin-like growth factors I, insulin-like growth factors II, platelet-derived growth factors, interleukins, cytokines, fibroblast growth factors, transforming growth factors, transforming growth factors. a, epidermal growth factors, neurotrophic growth factors derived from the brain, neurotrophins, ciliary growth factors, EGF and glial growth factors. The therapeutic agents can also include whole cells or their parts, which can be delivered to the enclosed lesioned area. These include Schwann cells, endothelial cells, fibroblasts, monocytes, macrophages, inflammatory cells or genetically altered cells or mixtures thereof. As an example, the growth promoting agent can be incorporated into the carrier matrix for the agent that inhibits the formation of fibrin network or on its own carrier, for example altered or non-genetically altered cells capable of delivering growth promoting agents can be incorporated as a supply system for agents that stimulate growth to the damaged structure. The agent that stimulates growth can also be immobilized in the envelope structure for the slow release of it. It is noted that, in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.Having described the invention as above, it is claimed as property, what is contained in the following

Claims (142)

1. Claims 1. A system for promoting the growth of tissue regeneration to an injured area, in a tissue structure organized in a human or animal living body, of a lesioned surface of the injured area, in a predetermined direction, characterized in that it comprises: envelope structure adapted during use to be implanted in the human or animal living body, to enclose the injured area, mechanical guide means for tissue regeneration, adapted during use to be disposed in the enclosed lesioned area, to extend into the predetermined direction and an inhibiting agent of fibrin network formation, administrable to the injured surface of the enclosed injured area.
2. 2. A system in accordance with the claim 1, characterized in that the inhibiting agent of fibrin network formation comprises a thrombin inhibitor.
3. 3. A system in accordance with the claim 2, characterized in that the thrombin inhibitor is a thrombin inhibitor based on low molecular weight peptide.
4. 4. A system according to claim 3, characterized in that the thrombin inhibitor is a gatran.
5. 5. A system according to claim 4, characterized in that the thrombin inhibitor is melagatran or inogatran.
6. 6. A system according to claim 2, characterized in that the thrombin inhibitor is a bisulphated polysaccharide or oligosaccharide, such as a chondroitin sulfate, a dextron sulfate, a cheratan sulfate, a dermatan sulfate, a herapan sulfate or heparin.
7. A system according to claim 2, characterized in that the thrombin inhibitor is a hirudin, a biosynthetic analog of hirudin, a fragment of hirudin, such as a fragment consisting of at least the last 8 C-terminal amino acids of the known sequence in hirudin or the NAPc2 protein.
8. 8. A system according to claim 1, characterized in that the inhibiting agent of fibrin network formation comprises a fibrinolytic agent.
9. 9. A system according to claim 8, characterized in that the fibrinolytic agent is a plasminogen activator (tPA), streptokinase or urokinase.
10. 10. A system according to claim 8, characterized in that the fibrinolytic agent is a recombinant human plasminogen activator (hrtPA) such as Actilyse®.
11. 11. A system according to claim 1, characterized in that the inhibiting agent of fibrin network formation comprises a factor X inhibitor.
12. 12. A system according to claim 1, characterized in that the inhibiting agent of the formation of The fibrin network comprises a trypsin inhibitor.
13. 13. A system according to claim 1, characterized in that the inhibitor of fibrin network formation comprises a protease inhibitor.
14. 14. A system according to any of claims 1 to 13, characterized in that the inhibiting agent of the fibrin network formation is immobilized to the internal surface of the enclosing structure, which during use faces the injured area.
15. 15. A system according to any of claims 1 to 13, characterized in that the agent inhibiting the formation of the fibrin network is in solution and because the system also comprises a pump for administering the agent that inhibits the formation of the fibrin network. fibrin network to the injured area enclosed.
16. 16. A system according to claim 15, characterized in that the pump is an osmotic minipump.
17. 17. A system according to claim 15 or 16, characterized in that the pump is adapted to be implanted subcutaneously in the human or animal living body.
18. 18. A system according to any of claims 1 to 13, characterized in that the inhibiting agent of fibrin network formation is incorporated in a matrix material for its disposition or supply to the enclosed injured area.
19. 19. A system according to claim 18, characterized in that the matrix material is formed of a material comprising a polysaccharide, such as a chitosan or a hyaluronan, such as hyaluronic acid, an agar gel, a hydrogel such as methyl cellulose gel, Matrigel®, Biomatrix I®, water, saline solution, phosphate buffered saline, a lipid or a protein such as collagen.
20. 20. A system according to claim 1, characterized in that the inhibiting agent of fibrin network formation is adapted to be administered to the enclosed, systemically or locally injured area.
21. 21. A system according to any of the preceding claims, characterized in that the enclosing structure is a patch for a bruised lesioned area or the like of the organized tissue structure.
22. 22. A system according to any of claims 1 to 20, characterized in that the enclosing structure is a tube having an open end adapted to receive the injured surface and because the mechanical guiding means are adapted, during use, to extend in the predetermined direction in the lumen of the tube.
23. 23. A system according to claim 22, characterized in that the tube comprises an external continuous tube element and an inner tube element that is formed of a plurality of spaced apart longitudinally spaced tube sections.
24. A system according to claim 22 or 23, characterized in that the injured surface of the injured area is a first injured surface, the open end of the tube is a first open end, the tube has a second open end adapted to receive a second The injured surface of the injured area and the mechanical guiding means are adapted, during use, to extend into the lumen of the tube, between the first and second open ends in the predetermined direction.
25. A system according to claim 24, characterized in that the system is for promoting the regeneration of tissue through a space between the free ends divided or cut transversely of an organized tissue structure, such as a nerve, tendon , skeletal muscle or ligament and because the open ends of the tube are each adapted to receive one of the free ends divided or cut transversely.
26. 26. A system according to any of the preceding claims, characterized in that the enclosing structure is of a biocompatible material.
27. 27. A system according to any of the preceding claims, characterized in that the enclosing structure is made of a biodegradable material.
28. 28. A system according to any of claims 1 to 26, characterized in that the enclosing structure is a non-biodegradable material.
29. 29. A system according to any of claims 1 to 25, characterized in that the shell structure is constructed of a polysaccharide.
30. 30. A system according to claim 29, characterized in that the shell structure is constructed of a material comprising a chitosan, heparin, a heparoinoid or a hyaluronan such as hyaluronic acid.
31. 31. A system according to any of claims 1 to 25, characterized in that the shell structure is constructed of a material comprising collagen or other protein complexes.
32. 32. A system according to any of claims 1 to 25, characterized in that the enclosing structure is constructed of a material comprising a polymer or copolymer.
33. 33. A system according to claim 32, characterized in that the enclosing structure is constructed of a material comprising polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, polyglucoronic acid or poly-N-acetylglucosamine or copolymers thereof.
34. 34. A system according to claim 32, characterized in that the enclosing structure is constructed of a material comprising a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid.
35. 35. A system according to claim 28, characterized in that the shell structure is constructed from silicone or ethylene vinyl acetate.
36. 36. A system according to any of the preceding claims, characterized in that the mechanical guiding means are held or presented by the internal surface of the enclosing structure which, in use, faces the injured area.
37. 37. A system according to claim 36, characterized in that the mechanical guiding means and the enclosing structure are integrally formed as an implantable body.
38. 38. A system according to any of the preceding claims, characterized in that the mechanical guiding means take the form of guiding channels in the enclosed injured area.
39. 39. A system according to claim 38, as it depends on any of claims 1 to 20, characterized in that the shell structure, when implanted, is a tube-like structure, having a spiral cross-section formed for example by rolling to a flat membrane and in that the guide channels are defined by the longitudinally extending spaces presented by the spiral cross-section.
40. 40. A system according to claim 38, characterized in that the mechanical guide means take the form of a gel structure that is provided with one or more guide channels therethrough, the gel structure is adapted to be disposed, during use, in the enclosed injured area, such that the guide channels extend in the predetermined direction.
41. 41. A system according to claim 38, 39 or 40, characterized in that the system is for promoting regeneration growth of the nerve tissue and because the, or each, guide channel has a cross-sectional dimension in the range of 50 μm - 1 mm.
42. 42. A system according to claim 41, characterized in that the, or each, guide channel has a cross-sectional dimension in the range of 150-500 μm.
43. 43. A system according to claim 40, characterized in that the gel structure is formed of agar, a hydrogel such as methylcellulose gel, albumin or other proteins which can be formed into a gel, a polysaccharide such as chitosan or a hyaluronan such as hyaluronic acid, a lipid which can be formed in a gel, Matrigel® or Biomatrix I®.
44. 44. A system according to any of claims 1 to 37, characterized in that the mechanical guide means comprise one or more guide filaments or fibers adapted, during use, to extend through the injured area enclosed in the predetermined direction.
45. 45. A system according to claim 44, characterized in that the mechanical guide means comprise fibers of one or more monofilaments, multifilaments or woven / non-woven fibers.
46. 46. A system according to claim 44 or 45, characterized in that the, or each, guide filament or fiber is of a biocompatible material.
47. 47. A system according to claim 44, 45 or 46, characterized in that the, or each, guide filament or fiber is formed from a biodegradable material.
48. 48. A system according to claim 44, 45 or 46, characterized in that the, or each, guide filament or fiber is formed of a non-biodegradable material.
49. 49. A system according to claim 44 or 45, characterized in that the, or each, guide filament or fiber is formed from a material comprising a polysaccharide.
50. 50. A system according to claim 49, characterized in that the, or each, guide filament or fiber is formed from a material comprising a chitosan, a heparin, a heparanoid or a hyaluronan, such as hyaluronic acid.
51. 51. A system according to claim 44 or 45, characterized in that the, or each, guide filament or fiber is formed from a material comprising a polymer or copolymer.
52. 52. A system according to claim 51, characterized in that the, or each, guide filament or fiber is formed from polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, poly-N-acetylglucosamine or copolymers thereof , such as, for example, a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid.
53. 53. A system according to claim 44 or 45, characterized in that the, or each guide filament or fiber is formed from collagen or other protein complexes.
54. 54. A system according to claim 44, characterized in that the mechanical guide means comprise one or more suture filaments.
55. 55. A system according to claim 54, characterized in that the, or each, suture filament is formed from vicril, catgut, polyamide, chitin or nylon.
56. 56. A system according to claim 48, characterized in that the, or each, filament or guide fiber is formed from silicone.
57. 57. A system according to any of the preceding claims, characterized in that the system further comprises a growth factor or mixture of growth factors for administration to the enclosed injured area.
58. 58. A system according to claim 57, characterized in that the growth factor is immobilized to the internal surface of the enclosing structure.
59. 59. A system according to any of claims 57 or 58, characterized in that the growth factor comprises insulin-like growth factors I, insulin-like growth factors II, platelet-derived growth factors, fibroblast growth factors. , transforming growth factors β, transforming growth factors a, neurotrophins, ciliary neurotrophic factors, EGF or glial growth factors.
60. 60. A system according to claim 59, characterized in that the growth factor comprises Schwann cells, endothelial cells, fibroblasts, macrophages or inflammatory cells or genetically altered cells which can express a growth factor.
61. 61. A system according to claim 1, characterized in that the system is for promoting the regeneration of tissue in an injured area in a nerve, tendon, ligament, joint capsule, cartilage, bone, aponeurosis or skeletal muscle.
62. 62. An implantable device for promoting the regeneration of tissue in an injured area in an organized tissue structure in a human or animal living body, from an injured surface of the injured area in a predetermined direction, characterized in that it comprises: an enclosing structure external which, when the device is implanted in the human or animal living body, encloses the injured area and an internal gel structure provided with one or more guide channels for tissue regeneration which, when the device is implanted, is disposed in the enclosed injured area, such that the guide channels extend in the predetermined direction.
63. 63. A device according to claim 62, characterized in that the enclosing structure is a patch for a bruised injured area or the like of the organized tissue structure.
64. 64. A device according to claim 62, characterized in that the envelope structure is a tube having an open end adapted to receive the injured surface, the or each guide channel extends into the lumen of the tube in the predetermined direction.
65. 65. A device according to claim 64, characterized in that the injured surface of the injured area is a first injured surface, the open end of the tube is a first open end, the tube has a second open end adapted to receive a second injured surface. of the injured area and the mechanical guiding means are adapted, during use, to extend into the lumen of the tube, between the first and second open ends in the predetermined direction.
66. 66. A device according to claim 65, characterized in that the device is for promoting the growth of tissue regeneration through a space between the free ends divided or cut transversely of an organized tissue structure, such as a nerve, tendon, muscle Skeletal or ligament and because the open ends of the tube are each adapted to receive one of the free ends divided or cut transversely.
67. 67. A device according to any of claims 62 to 66, characterized in that the enclosing structure is of a biocompatible material.
68. 68. A device according to any of claims 62 to 67, characterized in that the enclosing structure is made of a biodegradable material.
69. 69. A device according to any of claims 62 to 67, characterized in that the enclosing structure is made of a non-biodegradable material.
70. 70. A device according to any of claims 62 to 66, characterized in that the enclosing structure is constructed of a polysaccharide.
71. 71. A device according to claim 70, characterized in that the enclosing structure is constructed of a material comprising a chitosan, heparin, a heparoinoid or a hyaluronan such as hyaluronic acid.
72. 72. A device according to any of claims 62 to 66, characterized in that the shell structure is constructed of a material comprising collagen or other protein complexes.
73. 73. A device according to any of claims 62 to 66, characterized in that the shell structure is constructed of a material comprising a polymer or copolymer.
74. 74. A device according to claim 73, characterized in that the enclosing structure is constructed of a material comprising polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, polyglucoronic acid or poly-N-acetylglucosamine or copolymers thereof.
75. 75. A device according to claim 73, characterized in that the shell structure is constructed of a material comprising a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid.
76. 76. A device according to claim 69, characterized in that the shell structure is constructed from silicone or ethylene vinyl acetate.
77. 77. A device according to any of claims 62 to 76, characterized in that the device is for promoting the regeneration of the nerve tissue and that the, or each, guide channel has a cross-sectional dimension in the range of 50 μm - 1 mm
78. 78. A device according to claim 77, characterized in that the, or each, guide channel has a cross-sectional dimension in the range of 150-500 μm.
79. 79. A device according to any of claims 62 to 78, characterized in that the gel structure is formed of agar, a hydrogel such as methyl cellulose gel, albumin or other proteins which can be formed in gel, a polysaccharide such as chitosan or a hyaluronan such as hyaluronic acid, a lipid which can be formed in a gel, Matrigel® or Biomatrix I®.
80. 80. The use of a system according to any of claims 1 to 61, characterized in that it is used to promote the regeneration of tissue in an injured area of an organized tissue structure in a human or animal living body of an area injured from the injured area in a predetermined direction.
81. 81. The use of an implantable device according to any of claims 62 to 79, characterized in that it is used to promote the growth of tissue regeneration in an injured area of an organized tissue structure in a living human or animal body of a surface injured from the injured area in a predetermined direction.
82. 82. A method for promoting the regeneration of tissue in an injured area of an organized tissue structure in a human or animal living body of a lesioned surface of the injured area in a predetermined direction, characterized in that it comprises the steps of: enclosing the injured area with a wraparound structure, provide mechanical guiding means for regenerating the tissue in the enclosed injured area, such that the mechanical guiding means extend in the predetermined direction and administer an inhibiting agent of fibrin network formation to the injured area enclosed.
83. 83. A method according to claim 82, characterized in that the fibrin network-forming inhibiting agent comprises a thrombin inhibitor.
84. 84. A method in accordance with the claim 83, characterized in that the thrombin inhibitor is a thrombin inhibitor based on a low molecular weight peptide.
85. 85. A method in accordance with the claim 84, characterized in that the thrombin inhibitor is a gatran.
86. 86. A method according to the claim 85, characterized in that the thrombin inhibitor is melagatran or inogatran.
87. 87. A method according to claim 83, characterized in that the thrombin inhibitor is a bisulphated polysaccharide or oligosaccharide, such as a chondroitin sulfate, a dextron sulfate, a cheratan sulfate, a dermatan sulfate, a herapan sulfate or heparin.
88. 88. A method according to claim 83, characterized in that the thrombin inhibitor is a hirudin, a biosynthetic analog of hirudin, a fragment of hirudin, such as a fragment consisting of at least the last 8 C-terminal amino acids of the known sequence in hirudin or the NAPc2 protein.
89. 89. A method according to claim 82, characterized in that the inhibiting agent of fibrin network formation comprises a fibrinolytic agent.
90. 90. A method according to claim 89, characterized in that the fibrinolytic agent is a plasminogen activator (tPA), streptokinase or urokinase.
91. 91. A method in accordance with the claim 89, characterized in that the fibrinolytic agent is a recombinant human plasminogen activator (hrtPA) such as Actilyse®.
92. 92. A method according to claim 82, characterized in that the inhibiting agent of fibrin network formation comprises a factor X inhibitor.
93. 93. A method according to claim 82, characterized in that the inhibitor of the formation of The fibrin network comprises a trypsin inhibitor.
94. 94. A method in accordance with the claim 82, characterized in that the inhibiting agent of fibrin network formation comprises a protease inhibitor.
95. 95. A method according to any of claims 82 to 94, characterized in that the inhibiting agent of fibrin network formation is immobilized to the internal surface of the enclosing structure, which during use faces the injured area.
96. 96. A method according to any of claims 82 to 94, characterized in that the inhibiting agent of fibrin network formation is in solution and because the method further comprises the step of providing a pump for administering the inhibiting agent. the formation of the fibrin network to the injured area enclosed.
97. 97. A method according to claim 96, characterized in that the pump is an osmotic minipump.
98. 98. A method according to claim 96 or 97, characterized in that the pump is implanted subcutaneously in the human or animal living body.
99. 99. A method according to any of claims 82 to 94, characterized in that the inhibiting agent of fibrin network formation is incorporated in a matrix material for its disposition or supply to the enclosed injured area.
100. 100. A method according to claim 99, characterized in that the matrix material is formed of a material comprising a polysaccharide, such as a chitosan or a hyaluronan, such as hyaluronic acid, an agar gel, a hydrogel such as gel of methylcellulose, Matrigel®, Biomatrix I®, water, saline, phosphate buffered saline, a lipid or a protein such as collagen.
101. 101. A method according to claim 82, characterized in that the inhibiting agent of fibrin network formation is adapted to be administered to the enclosed, systemically or locally injured area.
102. 102. A method according to any of claims 82 to 101, characterized in that the enclosing structure is a patch for a bruised lesioned area or the like of the organized tissue structure.
103. 103. A method according to any of claims 82 to 101, characterized in that the enclosing structure is a tube having an open end adapted to receive the injured surface and because the mechanical guiding means are adapted, during use, to extend. in the predetermined direction in the lumen of the tube.
104. 104. A method according to claim 103, characterized in that the tube comprises an external continuous tube element and an inner tube element which is formed of a plurality of spaced apart longitudinally spaced tube sections.
105. 105. A method according to claim 103 or 104, characterized in that the injured surface of the injured area is a first injured surface, the open end of the tube is a first open end, the tube has a second open end in which it is received a second injured surface of the injured area and the mechanical guiding means extend into the lumen of the tube between the first and second open ends in the predetermined direction.
106. 106. A method according to claim 105, characterized in that the method is for promoting the growth of tissue regeneration through a space between the free ends divided or cut transversely of an organized tissue structure, such as a nerve, tendon , skeletal muscle or ligament and the open ends of the tube receive one of the free ends divided or cut transversely.
107. 107. A method according to any of claims 82 to 106, characterized in that the shell structure is of a biocompatible material.
108. 108. A method according to any of claims 82 to 107, characterized in that the enclosing structure is made of a biodegradable material.
109. 109. A method according to any of claims 82 to 107, characterized in that the enclosing structure is made of a non-biodegradable material.
110. 110. A method according to any of claims 82 to 106, characterized in that the shell structure is constructed of a polysaccharide.
111. 111. A method according to claim 110, characterized in that the shell structure is constructed of a material comprising a chitosan, heparin, a heparoinoid or a hyaluronan such as hyaluronic acid.
112. 112. A method according to any of claims 82 to 106, characterized in that the shell structure is constructed of a material comprising collagen or other protein complexes.
113. 113. A method according to any of claims 82 to 106, characterized in that the shell structure is constructed of a material comprising a polymer or copolymer.
114. 114. A method according to claim 113, characterized in that the shell structure is constructed of a material comprising polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, polyglucoronic acid or poly-N-acetylglucosamine or copolymers thereof.
115. 115. A method according to claim 113, characterized in that the shell structure is constructed of a material comprising a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid.
116. 116. A method according to claim 109, characterized in that the shell structure is constructed from silicone or ethylene vinyl acetate.
117. 117. A method according to any of claims 82 to 116, characterized in that the mechanical guide means are supported or presented by the internal surface of the enclosing structure which, in use, faces the injured area.
118. 118. A method according to claim 117, characterized in that the mechanical guiding means and the enclosing structure are integrally formed.
119. 119. A method according to any of claims 82 to 118, characterized in that the mechanical guiding means take the form of guiding channels in the enclosed injured area.
120. 120. A method according to claim 119, as it depends on any of claims 82 to 101, characterized in that the shell structure is a tube-like structure, having a spiral cross-section formed, for example, by laminate a flat membrane and because the guide channels are defined by the longitudinally extending spaces presented by the spiral cross-section.
121. 121. A method according to claim 119, characterized in that the mechanical guide means take the form of a gel structure that is provided with one or more guide channels and arranged in the enclosed injured area, such that the guide channels they extend in the predetermined direction.
122. 122. A method according to claim 119, 120 or 121, characterized in that the method is for promoting regeneration growth of nerve tissue and that the, or each, guide channel has a cross-sectional dimension in the range of 50 μm - 1 mm.
123. 123. A method according to claim 122, characterized in that the, or each, guide channel has a cross-sectional dimension in the range of 150-500 μm.
124. 124. A method according to claim 121, characterized in that the gel structure is formed of agar, a hydrogel such as methylcellulose gel, albumin or other proteins which can be formed into a gel, a polysaccharide such as chitosan or a hyaluronan such as hyaluronic acid, a lipid which can be formed in a gel, Matrigel® or Biomatrix I®.
125. 125. A method according to any of claims 82 to 118, characterized in that the mechanical guiding means comprise one or more guide filaments or fibers that extend through the injured area enclosed in the predetermined direction.
126. 126. A method according to claim 125, characterized in that the mechanical guide means comprise fibers of one or more monofilaments, multifilaments or woven / non-woven fibers.
127. 127. A method according to claim 125 or 126, characterized in that the, or each, filament or guide fiber is of a biocompatible material.
128. 128. A method according to claim 125, 126 or 127, characterized in that the, or each, guide filament or fiber is formed from a biodegradable material.
129. 129. A method according to claim 125, 126 or 127, characterized in that the, or each, guide filament or fiber is formed of a non-biodegradable material.
130. 130. A method according to claim 125 or 126, characterized in that the, or each, guide filament or fiber is formed from a material comprising a polysaccharide.
131. 131. A method according to claim 130, characterized in that the, or each, guide filament or fiber is formed from a material comprising a chitosan, a heparin, a heparanoid or a hyaluronan, such as hyaluronic acid.
132. 132. A method according to claim 125 or 126, characterized in that the, or each, guide filament or fiber is formed from a material comprising a polymer or copolymer.
133. 133. A method according to claim 132, characterized in that the, or each, guide filament or fiber is formed from polylactic acid, polyhydroxybutyric acid, polyglycolic acid, permselective polytetraethylene, poly-N-acetylglucosamine or copolymers thereof , such as, for example, a copolymer of polyhydroxybutyric acid and hydroxyvaleric acid.
134. 134. A method according to claim 125 or 126, characterized in that the, or each guide filament or fiber is formed from collagen or other protein complexes.
135. 135. A method according to claim 125 or 126 ,. characterized in that the mechanical guiding means comprise one or more suture filaments.
136. 136. A method according to claim 135, characterized in that the, or each, suture filament is formed from vicril, catgut, polyamide, chitin or nylon.
137. 137. A method according to claim 129, characterized in that the, or each, filament or guide fiber is formed from silicone.
138. 138. A method according to any of claims 82 to 137, characterized in that the method further comprises the step of administering a growth factor to the enclosed lesioned area.
139. 139. A method according to claim 138, characterized in that the growth factor is immobilized to the inner surface of the envelope structure.
140. 140. A method according to any of claims 138 or 139, characterized in that the growth factor comprises insulin-like growth factors I, insulin-like growth factors II, platelet-derived growth factors, fibroblast growth factors , transforming growth factors β, transforming growth factors a, neurotrophins, ciliary neurotrophic factors, EGF or glial growth factors.
141. 141. A method according to claim 138 or 139, characterized in that the growth factor comprises Schwann cells, endothelial cells, fibroblasts, macrophages or inflammatory cells or genetically altered cells which can express a growth factor.
142. 142. A method according to claim 82, characterized in that the method is for promoting the regeneration of tissue in an injured area in a nerve, tendon, ligament, joint capsule, cartilage, bone, aponeurosis or skeletal muscle.