EP1089769A1 - Albumine reticulee de carbodiimide pour bioadhesifs chirurgicaux de scellement et dispositifs a implanter - Google Patents
Albumine reticulee de carbodiimide pour bioadhesifs chirurgicaux de scellement et dispositifs a implanterInfo
- Publication number
- EP1089769A1 EP1089769A1 EP99930613A EP99930613A EP1089769A1 EP 1089769 A1 EP1089769 A1 EP 1089769A1 EP 99930613 A EP99930613 A EP 99930613A EP 99930613 A EP99930613 A EP 99930613A EP 1089769 A1 EP1089769 A1 EP 1089769A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- albumin
- carbodiimide
- preparation
- cross
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/108—Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/043—Mixtures of macromolecular materials
Definitions
- the present invention is directed to the development and use of carbodiimide cross- linked proteins for use in vitro or in vivo during experimental or surgical procedures to bond a tissue to another tissue and/or to a prosthetic device; to seal incisions, perforations, and/or fluid or gaseous leaks in tissues; or to form implantable devices for drug delivery or prostheses, or to form coatings on inert devices for biocompatibility and other functions.
- Biomedical researchers have attempted to develop various methods and products to bind tissues to tissues, and/or tissues to prosthetic devices, for a wide array of in vitro or in vivo experimental or surgical procedures, including the implantation of devices such as prostheses for drug delivery systems.
- researchers have attempted to develop safe and efficient means to bind or seal tissues cut, torn or perforated as a result of trauma and/or surgical procedures.
- suturing and stapling have been the most common methods for binding or sealing tissues.
- suturing and stapling may require significant skill and/or extensive surgical time. More important, the results are often less than satisfactory due to gaps between sutures or staples, tearing of delicate tissue around sutures or staples, and the possibility of progressive weakening of the bindings that may result in leaks of biological fluids or bacterial infections.
- safer and more efficient methods have been sought to bind or seal tissues.
- bioadhesives or surgical sealants have been developed to adhere to tissue surfaces and to form bonds between tissues until healing is complete.
- fibrin-based adhesives have commanded considerable attention in this area (see, e.g. , Epstein et al. (1986), Ann. Otol. Rhinol. Laryngol. 95:40-45; Siedentop et al. (1983), Laryngoscope 93:1310-1313). Fibrin adhesives, however, have low mechanical (bond) strength, as well as a lengthy set-up time, which limits their utility.
- Non-biological adhesives such as cyanoacrylates (e.g., isobutyl-2-cyanoacrylate) have also been examined for their potential adhesive properties. However, these materials are difficult to apply in vivo because cyanoacrylate adhesives require application in a dry field.
- gelatin must be applied at a temperature significantly above the human body temperature, and requires significant attention during mixing and application to achieve successful results.
- Carbodiimides have been utilized in a variety of contexts as cross-linking reagents.
- carbodiimides have been used as cross-linking reagents with the polysaccharides hyaluronic acid and pectin (see, e.g. , Tomihata et al. (1997), J. Biomed. Mater. Res. 37:243- 251).
- carbodiimides were used to cross-link aqueous mixtures of gelatin and poly(L-glutamic acid) (PLGA) as adhesive agents (see, e.g., Otani et al. (1996), J. Biomed. Mater. Res. 31:157-166).
- the invention provides methods and compositions that are useful for adhering biological tissues and/or prosthetic devices, sealing fluid and/or gaseous leaks in biological tissues, and preparing implants for drug delivery, including bio-erodable implants.
- the invention comprises protein and cross-linking preparations that are useful for cross-linking biological tissues and/or prosthetic devices.
- Compositions of the invention comprise protein solutions that are suitable for tissue cross-linking.
- protein preparations of the invention comprise albumin preparations.
- the albumin is bovine serum albumin (BSA).
- BSA bovine serum albumin
- the albumin is a human albumin.
- the albumin preparations are provided at a pH of between 5.0 and 8.0, more preferably between 5.5 and 7.5.
- the albumin preparation comprises a protein that is a naturally occurring albumin protein, a recombinant albumin protein, a major fragment of an albumin protein, or a chemically modified albumin.
- the albumin is a mammalian albumin protein or a major fragment of a mammalian albumin protein.
- the albumin protein comprises an amino sequence of at least 100 amino acid residues having at least 60% homology to an amino acid sequence of human albumin.
- the albumin comprises an amino-acid sequence which has been recombinantly modified relative to a naturally occurring albumin sequence to enhance one or more physical properties such as solubility, reactivity with a carbodiimide cross-linker, stability, viscosity in an aqueous solution, and immunocompatibility.
- the albumin preparation comprises additives to increase or decrease the crosslinking reaction rate or to promote interaction between the protein solution and the tissue or material at the site of application.
- the albumin may provided along with one or more surfactant, lipid, and/or fatty acid.
- the albumin may be covalently bound to a molecule selected from the group consisting of polysaccharides (e.g., glycosaminoglycans, dextrans, hyaluronic acid, chondroitin sulfate, heparan sulfates), polyethers (e.g., polyethylene glycol, polypropylene glycol, polybutylene glycol), polyesters (e.g., polylactic acid, polyglycolic acid, polysalicylic acid), and aliphatic, alicyclic, aromatic, perfluorinated or non-perfluorinated, and acylating or sulfonating agents.
- the albumin preparation may contain a chlorinated, fluorinated, brominated or iodinated albumin protein.
- the albumin protein is provided in an aqueous solution at a concentration of about 10-50 % by weight, more preferably about 20-40% by weight, most preferably 35-40% by weight.
- the albumin protein is provided in solution of secondary or a tertiary alcohol.
- the alcohol solution comprises isopropyl alcohol (IP A) or isobutyl alcohol (IBA).
- IP A isopropyl alcohol
- IBA isobutyl alcohol
- the albumin is dissolved in a 20% IPA solution or an 8% IBA solution.
- the protein may be derivatized to alter its properties in the cross- linking reaction or to promote its interaction with tissue or substrate at the site of application, as explained and illustrated in the following description and examples.
- the carbodiimide is selected from the group consisting of ethyl dimethylaminopropyl carbodiimide (EDC ⁇ C1); l-(3- dimethylaminopropyl)-3-ethylcarbodiimide; 1 ,3-di-p-tolylcarbodiimide; 1 ,3- diisopropylcarbodiimide; 1 ,3-dicyclohexylcarbodiimide; 1 -cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluenesulfonate; polycarbodiimide; l-tert-butyl-3-ethylcarbodiimide; 1 ,3-dicyclohexylcarbodiimide; 1 ,3-bis(trimethylsilyl)carbodiimide; 1 ,3-di-tert- butylcarbodiimide; l-
- the invention provides a method for producing a cross-linked albumin composition for use in a bioadhesive, surgical sealant or implantable device, comprising the steps of providing an albumin preparation; providing a carbodiimide preparation; and mixing the albumin preparation and the carbodiimide preparation under conditions which permit cross- linking of the albumin.
- the albumin and carbodiimide preparations are mixed in one of the following ratios : between approximately 1 : 1 and 1 : 100, between approximately 1:5 and 1:50, between approximately 1:10 and 1:20, between approximately 100:1 and 1:1, between approximately 36:1 and 4:1, and approximately 18:1.
- Preferred methods of the invention are useful for sealing incisions, perforations, and/or fluid or gaseous leaks in biological tissues during a surgical procedure, and comprise contacting the tissue with an effective amount of an albumin preparation and a carbodiimide preparation under conditions that promote cross-linking of the albumin preparation to the tissue thereby sealing the incision, perforation, or fluid or gaseous leak.
- Such methods are particularly useful for surgical procedures such as cardiovascular, pulmonary, renal, and hepatic surgeries.
- a fluid or gaseous leak can be sealed by cross- linking the tissues surrounding the leak.
- a cross-linked gel of the invention can seal a leak by physically occluding it without cross-linking the surrounding tissues.
- the albumin and carbodiimide preparations are mixed before applying them to a tissue locus.
- the mixing step is performed at a tissue locus.
- an accessory molecule is provided and mixed with the albumin and carbodiimide preparations.
- the accessory molecule is preferably selected from the group consisting of viscosity-enhancing agents, cross-linkers, buffers, hormones, growth factors, antibiotics, surfactants, lipids, fatty acids, and anti-inflammatory agents. - o -
- a primer solution is applied to the tissue or prosthetic device before adding the mixture of albumin and cross-linker.
- the primer solution may be a saline solution.
- the primer solution is an albumin solution. More preferably, the primer solution is identical to the albumin preparation used in the cross-linking reaction, and most preferably the primer solution is a dilute solution of the albumin preparation.
- the albumin and carbodiimide preparations are preferably provided at a pH between 5.0 and 8.0, more preferably between 5.5 and 7.5, most preferably between 6.0 and 7.0.
- the pH of the preparations is preferably adapted to the desired rate of cross-linking. Rapid cross-linking is useful in cardiovascular applications to rapidly seal blood leaks. Rapid cross-linking involvin EDC ⁇ Cl is obtained at pH 5.0 to 5.5. Alternatively, for pulmonary applications, rapid cross-linking is not as important and the preparations can be provided at a higher pH.
- the cross-linking reaction rates can also be modified using additives, derivatized albumin, or by altering the ratio or concentrations of albumin and carbodiimide, as described in the following description and as illustrated by the following examples.
- the preparations may be provided in hydrophobic solutions.
- the solutions do not interfere with the cross-linking reaction.
- Preferred solutions include secondary and tertiary alcohols, including IPA and IBA.
- IPA and IBA secondary and tertiary alcohols
- a 30% solution of BSA is made using 20% IPA or 8% IBA.
- the protein and cross-linker may be provided as dry powders. The protein and cross-linker powders are preferably mixed prior to applying them to a tissue site. In a most preferred embodiment, the protein and cross-linker absorb body fluids from the site of application, and this fluid absorption starts the cross-linking reaction.
- additional fluid may be provided as necessary to the site of application.
- the site of application is primed with a solution of saline or dilute protein before the dry protein and cross-linker mixture is applied. The primer solution is then absorbed by the dry mixture, thereby starting the cross-linking reaction.
- the invention provides methods and compositions that bind or adhere to synthetic material such as artificial blood vessels (for example PTFE material) or biological implants (for example polyethylene material).
- synthetic material such as artificial blood vessels (for example PTFE material) or biological implants (for example polyethylene material).
- the invention provides a method for adhering a first biological tissue to a second tissue and/or prosthetic device, comprising contacting the first tissue and the second tissue or prosthetic device with a mixture of an albumin preparation and a carbodiimide preparation under conditions that promote cross-linking of said albumin preparation to the first tissue and second tissue or prosthetic device.
- methods are provided for forming an implantable device comprising.
- the methods comprise providing an albumin preparation, providing a carbodiimide preparation, providing a mold, and mixing the albumin preparation and the carbodiimide preparation under conditions which permit cross-linking of the albumin in the mold.
- Products of the invention also include a bioadhesive, surgical sealant or implantable device produced according to any of the foregoing methods.
- the invention provides a kit for producing a bioadhesive, surgical sealant or implantable device comprising, in separate containers, an albumin preparation, and a carbodiimide preparation.
- the kit further comprises an accessory molecule, preferably a molecule selected from the group consisting of viscosity-enhancing agents, cross-linkers, buffers, hormones, growth factors, antibiotics, anti-inflammatory agents, hydrophobicity increasing agents, and surfactants.
- the albumin preparation and the carbodiimide preparation are provided in a binary delivery device that delivers the preparations in a ratio which is either predetermined or regulatable.
- the invention provides methods and products relating to protein-based bioadhesives and surgical sealants, and implantable devices for drug delivery and prostheses.
- the invention relates to albumin-based methods and products.
- albumin-based methods and products it should be understood that other proteins can be substituted in the following description of albumin-based methods and products.
- Compositions of the invention are formed by cross- linking albumins by reacting them with carbodiimides under specified conditions. The general reaction proceeds in two steps (see, e.g., Damink et al., 1996).
- a free carboxyl group of an albumin molecule e.g., the carboxy terminus, or a side chain of a glutamic or aspartic acid residue
- a free amine group of the same or another albumin molecule e.g., the amino terminus, or a side chain of a lysine or arginine residue
- a free amine group of the same or another albumin molecule attacks one of these intermediates to yield the cross linked albumin and a substituted urea by-product as follows:
- the reaction may be performed in vitro in a laboratory or manufacturing facility to produce products including cross-linked albumin as a bioadhesive or surgical sealant material, or as part of an implantable drug delivery device or prosthesis, or it may be performed in vivo during an experimental or surgical procedure to bond a tissue to another tissue and/or to a prosthetic device, or to seal incisions, perforations, and/or fluid or gaseous leaks in tissues.
- the albumin starting material may be a chemically modified form of albumin
- the carbodiimide cross-linker may include other functional or reactive groups, including additional carbodiimide groups such that the cross-linker is a poly(carbodiimide), additional non-carbodiimide cross-linkers may be employed, and/or various accessory molecules may be added to modify the course of the cross-linking reaction or the characteristics of the final product.
- the methods of the invention comprise the steps of providing an appropriate albumin preparation and an appropriate carbodiimide, and optionally accessory molecules, and mixing these components under conditions which permit the carbodiimide to promote the formation of intermolecular (as well as intramolecular) cross-links amongst the albumin molecules.
- albumin refers to any mammalian albumin protein, including allelic variants, and to modified albumins and albumin fragments which comprise a sequence of at least 100 amino acid residues having at least 60% homology, preferably at least 70 or 80% homology, to the human albumin sequence as disclosed in Minghetti et al. (1986) J. Biol. Chem. 261 :6747-6757.
- homology means "homology” or "similarity” as calculated by the BLAST amino acid sequence comparison programs (including XBLAST, Altschul et al. (1990), J. Mol. Biol. 215:403-10) using default parameters for the appropriate programs (see http://www.ncbi.nlm.nih.gov). Alternatively, homology is calculated using the algorithm of Myers and Miller, CABIOS (1989), which is incorporated into the ALIGN program (version 2.0). Exemplary parameters for use when comparing amino acid sequences are a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.
- the bioadhesives, surgical sealants, and implantable devices of the present invention comprise a cross-linked form of the protein albumin.
- the albumin is of mammalian origin, but other sources of albumin also may be employed. It is believed that most albumins are readily cross-linked according to the methods of the invention. However, an albumin with low immunogenicity is preferred for in vivo applications. Accordingly, for uses in humans, it is preferred that the albumin is human albumin. Bovine serum albumin (BSA) may also be used in humans, and is more readily available. Alternatively, the albumin may be recombinant albumin, isolated from cells expressing a recombinant albumin gene, using methods known in the art.
- BSA bovine serum albumin
- the albumin gene is preferably a human or bovine gene.
- Major fragments of albumin comprising at least 100 residues of an albumin sequence, whether produced by partial proteolysis or be recombinant means, may also be used instead of intact albumin.
- useful fragments may contain at least 50 residues, and more preferably at least 75 residues of an albumin sequence.
- mixtures of different forms of albumin e.g., human, bovine, recombinant, fragmented
- albumin e.g., human, bovine, recombinant, fragmented
- Albumin may be purified directly from tissues or cells, using methods well known in the art (see, e.g., Cohn et al. (1946) J. Amer. Chem. Soc. 68:459; Cohn et al. (1947) J. Amer. Chem. Soc. 69:1753; Chen (1967) J. Biol. Chem. 242:173).
- albumin may be purchased from a commercial supplier.
- albumin preparations from various mammalian and avian species may be purchased from Sigma Chemical Company (St. Louis, MO) in the form of solutions or lyophilized powders.
- a preferred commercially available albumin preparation is a 30%) human albumin solution (e.g., Sigma catalog no. A 9080) or a 30%> BSA solution (e.g., Sigma catalog number A 8327).
- albumin is provided as an aqueous solution of 10-50%), preferably 20-40%, and most preferably about 35%-40% albumin by weight.
- concentrations of albumin may be employed when viscosity-enhancing agents are added.
- the solution is substantially purified to remove contaminants such as immunogens or proteases which would disrupt or interfere with the bioadhesive or sealant properties of the cross-linked albumin.
- proteins such as collagen, elastin, laminin, fibrin, and thrombin, can be tolerated.
- albumin may be provided as a dry powder.
- the dry albumin is solubilized at the site of administration.
- body fluids such as blood
- additional fluids may be provided along with the dry albumin.
- the cross-linker may also be provided as a dry powder that is solubilized at the site of administration.
- the dry protein and cross-linker are mixed prior to administration.
- a wetting reagent is added to the protein and cross-linker mixture in order to increase fluid absorbance.
- the wetting reagent absorbs water from the available body fluids and speeds up solubilization of the protein and cross-linker.
- Albumin may be modified or derivatized to increase viscosity.
- albumin viscosity may be increased by covalently attaching relatively large (10-100 kD), substantially linear molecules such as polysaccharides (e.g., glycosaminoglycans, dextrans, hyaluronic acid, chondroitin sulfate, heparan sulfates), polyethers (e.g., polyethylene glycol, polypropylene glycol, polybutylene glycol), polyesters (e.g., polylactic acid, polyglycolic acid, polysalicylic acid), and aliphatic, alicyclic or aromatic acylating or sulfonating agents.
- polysaccharides e.g., glycosaminoglycans, dextrans, hyaluronic acid, chondroitin sulfate, heparan sulfates
- polyethers e.g., polyethylene glycol, polypropylene
- Preferred acylating agents including aliphatic, alicyclic and aromatic anhydrides or acid halides, particularly acid anhydrides of dicarboxylic acids.
- Non-limiting examples of these include glutaric anhydride, succinic anhydride, lauric anhydride, diglycolic anhydride, methacrylic anhydride, phthalic anhydride, succinyl chloride, glutaryl chloride, and lauroyl chloride.
- the acylating agents may also include various substituents and secondary functionalities such as aliphatic, alicyclic, aromatic and halogen substituents, as well as amino, carboxy, keto, ester, epoxy, and cyano functionalities, and combinations thereof.
- preferred sulfonating agents useful in the invention include aliphatic, alicyclic and aromatic sulfonic acids and sulfonyl halides, which may also include various substituents and secondary functionalities as described above.
- Albumin also may be modified or derivatized to increase its hydrophobicity in order to promote interactions with hydrophobic tissues or prosthetic materials.
- albumin may be derivatized with branches or straight chain alkyl, alkenyl, or aromatic reagents, including long chain alkyl or alkenyl and alkyl aldehydes or carboxylic acids such as octyl or dodecyl aldehyde or carboxylic acid.
- albumin or modified albumin may be halogenated, preferably fluorinated, by standard methods well known in the art.
- albumin may be derivatized with polyfluoro dicarboxylic acid anhydrides (e.g., hexafluoro glutaric anhydride), polyfluoro aklyl ethers (e.g., perfluoroalkyl glycidyl ethers), or other halogen containing reagents.
- a recombinant albumin may be produced by standard techniques of site- directed mutagenesis in which one or more amino acid residues are inserted, deleted or substituted to increase the viscosity of the albumin, to alter the hydrophobicity of the protein, to provide more side chains for derivatization, or to provide more free carboxyl or amine groups for the cross-linking reaction.
- albumin contains an adequate (and roughly equal) number of free carboxyl and amine groups for cross-linking.
- modifications of the albumin sequence will be most useful for increasing the viscosity of the protein by replacing small or hydrophilic residues (e.g., glycine, alanine) with larger and/or more hydrophobic and/or charged residues which can more participate in non-covalent intermolecular bonds through charge-charge or hydrophobic interactions.
- small or hydrophilic residues e.g., glycine, alanine
- Carbodiimides are cross-linking reagents having the general formula:
- R, and R 2 may be essentially any chemical group, provided it does not contain a strong nucleophile.
- Carbodiimides are very reactive, and the presence of a nucleophilic group in either R, or R 2 would be destabilizing due to intermolecular (or intramolecular) reactions amongst carbodiimide molecules.
- R, and R 2 are independently selected from the group consisting of any straight chain or branched, saturated or unsaturated, alkyl, alkenyl, aryl, aralkyl, or aralkenyl group, or variants thereof with halogen, tertiary amino, ester, keto, or other substituents.
- one or both of R, and R 2 may include an additional carbodiimide group, such that the cross-linker is a polycarbodiimide.
- carbodiimides are employed which are water soluble, and reactive with albumin under physiological conditions.
- a suspension of water insoluble carbodiimide such as ethyl dimethylaminopropyl carbodiimide (EDC)
- EDC ethyl dimethylaminopropyl carbodiimide
- R groups the solubility and reactivity of the carbodiimide may be varied.
- the choice of R groups will affect the immunogenicity and toxicity of the cross-linker, as well as its ability to interact with albumin molecules.
- the R groups may be chosen to have additional cross-linking groups, such as photoactivatable cross-linking groups.
- the carbodiimides are provided as a solution or suspension.
- the carbodiimides may be provided in dry form, such as a powder.
- the dry carbodiimide is solubilized or suspended either before it is administered to the tissue, or by body fluids present at the site of administration.
- One preferred cross-linking agent is ethyl dimethylaminopropyl carbodiimide hydrochloride (EDC ⁇ Cl).
- EDC ⁇ Cl ethyl dimethylaminopropyl carbodiimide hydrochloride
- carbodiimides include l-(3-dimethylaminopropyl)-3- ethylcarbodiimide (Aldrich catalog no. 42433-1); 1,3-di-p-tolylcarbodiimide (Aldrich catalog no. D21980-0); 1,3-diisopropylcarbodiimide (Aldrich catalog no. D12540-7); 1,3- dicyclohexylcarbodiimide (Aldrich catalog no. D8000-2); l-cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluenesulfonate (Aldrich catalog no.
- the cross-linking agent is a polyethylene glycol (PEG) based water soluble carbodiimide of the general formula:
- R, through R 4 are independently selected as described above.
- R 2 and R 3 may also include hydrolytically or enzymatically cleavable groups so that a cross- linked product is biodegradable via cleavage of the crosslinks.
- a PEG based carbodiimide is synthesized by reacting alkoxy PEG isocyanates in the presence of an appropriate catalyst.
- the cross-linker e.g., EDC ⁇ Cl
- the cross-linker solution and albumin solution have similar viscosities and volumes in order to promote efficient mixing and delivery.
- inert polymers include poly(vinyl alcohol), PEG, non-ionic surfactants, including PEG-based surfactants such as pluronic polymers, poly(saccharides) and other inert polymers.
- PEG-based surfactants such as pluronic polymers, poly(saccharides) and other inert polymers.
- the carbodiimide cross-linkers of the invention can react spontaneously with albumin (and other proteins) to form cross-links between amino acid side chains of the proteins.
- initiators of the cross-linking reaction are not required.
- the reaction can occur under conditions of pH and temperature which are compatible with in vivo applications.
- the carbodiimide cross-linking reaction of the invention is very sensitive to pH, and is preferably conducted at a pH between 5 and 7, more preferably between pH 5 and 6. At pH values below 5, unmodified albumin may precipitate, and at pH values above 8, the speed of the cross-linking reaction is greatly reduced.
- Physiological pH is approximately 7.0-7.4 and, therefore, the reaction mixture may be allowed to occur at physiological pH, or the reaction mixture may be slightly acidified by the addition of acidic accessory molecules.
- the reaction may be conducted at room temperature for in vitro uses, and may be conducted at body temperature for in vivo applications.
- the molar ratio of the carbodiimide to albumin will significantly affect both the rate of the cross-linking reaction and the characteristics of the final cross-linked product. That is, the higher the ratio carbodiimide to albumin, the faster the reaction will occur and the more highly cross-linked the product will be.
- the bioadhesives, surgical sealants and implantable devices of the invention are preferably prepared by mixing a carbodiimide with an albumin at a molar ratio of between 1 and 100 moles of carbodiimide per mole of albumin monomer.
- carbodiimide and albumin are mixed at a molar ratio of approximately 36:1.
- the ratio is approximately 18:1.
- sufficient cross-linking can be achieved with a carbodiimide:albumin ratio of approximately 4:1.
- a ratio of approximately 0.4:1 does not provide sufficient cross-linking for most applications of the invention.
- EDC-HCl has a molecular weight of approximately 191.7
- an albumin monomer has a molecular weight of, on average, about 69,293. Therefore, a bioadhesive prepared by mixing EDC ⁇ Cl with BSA at a molar ratio of approximately 36:1 corresponds to a mixture with approximately (36 moles x 191.7 grams/mole) 6901.2 g of EDC-HCl per (1 mole x 69,293 grams/mole) 69,293 g of BSA, or approximately 1 : 10 by weight EDC ⁇ Ckalbumin. Similar calculations may, of course, be used for other carbodiimide reagents to determine the appropriate weight:weight ratios based on the desired molar ratios.
- EDC-HCl cross-linked albumin preparations may be produced with weigh weight ratios of EDC-HCl:albumin varying from about 1 :1 to 1 :100, preferably about 1:5 to 1 :50, and most preferably about 1 : 10 to 1 :20.
- the reaction proceeds extremely rapidly and the product may cure before application has been completed.
- the resulting product is extremely tough and relatively non-plastic or inflexible.
- the reaction is much slower and the product may be shaped and molded while the reaction proceeds.
- the resulting product is weaker, but is more plastic or flexible.
- a ratio of 1 : 100 is believed to be useful for some applications, but ratios as low as 1 : 1000 are expected to be inoperative due to insufficient cross-linking.
- a weightweight ratio of EDC-HCl:albumin of 1 :5 is currently preferred.
- a lower EDC-HCl ratio perhaps 1 :10 to 1 :20 may be appropriate, although this will produce a weaker and more gel-like sealant.
- a 1 :20 ratio is expected to be too weak for cardiovascular applications, but a 1 : 1 ratio is expected to be too inelastic.
- both the albumin and carbodiimide are provided in aqueous preparations.
- an aqueous preparation of albumin may be mixed with a suspension of an insoluble carbodiimide.
- albumin and carbodiimide will begin to cross-link upon mixing, and because the reaction may be quite rapid for some formulations, it may be important to mix them immediately before application, or to mix them at the site where the bioadhesive or surgical seal is desired to be formed.
- carbodiimides are not stable for extended periods in aqueous solution, they may be prepared by dissolving carbodiimide powder or lyophilized carbodiimide in an aqueous solution immediately prior to mixing with albumin.
- carbodiimides may be provided in an inert, water miscible, biocompatible organic solvent. Therefore, a binary delivery device, having separate compartments holding the albumin preparation and the carbodiimide cross-linker prior to dispensing and mixing, may be particularly useful.
- a double-barreled syringe which simultaneously dispenses and mixes the components is used. Such a double-barreled syringe may be quite convenient for in vivo applications where the bioadhesive or surgical sealant is applied to the site of tissue injury or incision.
- a double-barreled syringe comprises a first barrel containing an aqueous solution of albumin, and a second barrel containing a carbodiimide powder separated from an aqueous solution by a breakable membrane.
- the membrane is first broken, causing the carbodiimide to dissolve in the aqueous solution of the second barrel, and then the syringe is used as described above.
- a double barrel comprises a first barrel containing an aqueous albumin solution at a pH at which the cross- linking reaction may occur (e.g., pH 5.0-6.0), and a second barrel containing a carbodiimide solution which has been adjusted to an alkaline pH to reduce the conversion of the carbodiimide to disubstituted urea compounds by water in the solution.
- a pH at which the cross- linking reaction may occur e.g., pH 5.0-6.0
- the pH and or buffer systems in the two barrels must be selected such that, upon mixing, the pH of the resultant solution is sufficiently acidic to permit the cross-linking reaction to proceed efficiently.
- a single barreled syringe contains an albumin solution separated from a carbodiimide powder by a breakable membrane.
- the cross-linking reaction is started by breaking the membrane, and the resulting mixture is applied as described above.
- the two components and syringe may be provided in a sterile and disposable kit.
- the two components may be applied as a spray from a device with separate reservoirs for the two components.
- the two components may be applied sequentially. This method suffers from the disadvantage that the components will not be as thoroughly mixed, and only a thin coat of cross-linked albumin may form at their interface.
- accessory molecules are added to the reaction.
- Such accessory molecules may include viscosity-enhancing agents, solubility-enhancing agents, non- carbodiimide cross-linking agents, anti-inflammatory agents, hormones, growth factors, antibiotics, buffers, and the like.
- viscosity-enhancing agents are added to the mixture and, therefore, the concentration of albumin which is employed may be decreased.
- concentration of albumin is preferably at least 10%, and more preferably at least 20%>.
- the viscosity-enhancing agent is itself cross-linked in the reaction.
- Viscosity-enhancing agents may include substituted or unsubstituted polysaccharides (e.g., glycosaminoglycans or heparin sulfates), fibrous proteins (e.g., collagen, elastin, fibrin, fibrinogen, thrombin, laminin), or other compounds which polymerize under physiological conditions or in the presence of the carbodiimides of the invention (e.g., polyacids and polyamines).
- Preferred viscosity-enhancing agents include glycosaminoglycans, dextran, hyaluronic acid, collagen, chondroitin sulfate, and elastin.
- accessory molecules are added in order to alter the rate and or degree of cross-linking.
- a carboxylic acid may reduce the rate or degree of cross-linking by competing with a protein carboxylic group in the first step of the carbodiimide cross-linking reaction.
- an amine may reduce the rate or degree of cross-linking by competing with a protein amine group in the second step of the carbodiimide cross-linking reaction.
- polycarboxylic acids, polyamines, poly(carboxy/amino)compounds may increase the rate of gel formation by reacting with carbodiimides to form cross-links with two or more protein molecules, thereby participating in the gel formation
- polycarboxylic acids, polyamines, and/or poly(carboxyl/amino)compounds should have a relatively high density of carboxy and/or amino groups.
- accessory molecules have molecular weights that are preferably less than 1,000, more preferably less than 500, and most preferably less than 250 Daltons per carboxy and/or amino group.
- an accessory molecule having a molecular weight of 2,000 Daltons and having four carboxy and/or amino groups would have a molecular weight of 500 Daltons per carboxy or amino group.
- Polycarboxylic acids include citric acid and poly(acrylic acid).
- Polyamines include poly(lysine) and chitosan.
- other acids may be included to accelerate the EDC mediated protein cross-linking reaction by lowering the pH
- other bases may be used to slow the cross- linking reaction by raising the pH.
- the optimal pH for a carbodiimide cross-linking reaction is about 5.0 to 6.0.
- the reaction rate may also be increased at lower pH due to denaturation of the protein at the low pH.
- the hydrophobicity of the albumin solution is increased by solubilizing the albumin in a solution that is more hydrophobic than water.
- the albumin is solubilized in a solution comprising a secondary or tertiary alcohol.
- albumin is provided in a solution of isopropyl alcohol (IPA) or isobutyl alcohol (IBA).
- IPA isopropyl alcohol
- IBA isobutyl alcohol
- a 30% solution of BSA is prepared with 20% IPA or 8% IBA.
- buffers such as phosphate buffers and Tris buffers are not preferred.
- Currently preferred buffers are inorganic acids such as hydrochloric acid.
- the cross-linked albumin preparations of the invention have a large number of applications.
- they may be employed as surgical sealants, bioadhesives, hemostats, coating materials and drug delivery media. When formed ex vivo into films and then implanted, they may also serve to prevent surgical adhesions by forming a resorbable barrier between healing tissues.
- the cross-linked albumins are expected to be particularly useful in sealing air and blood leaks in the lungs, blood leaks in the liver, spleen and kidneys, cerebrospinal fluid leaks in dura, and blood leaks in the heart and vascular system.
- the carbodiimide cross-linked albumins must be designed to have sufficient flexibility for pulsatile stretching, but must also have sufficient strength to withstand cardiovascular pressures.
- the reaction mixture rapidly forms a seal when it is applied to a vascular leak.
- cross-linking occurs before the reaction mixture can slide off or wash away from the site of vascular application, for example a blood vessel.
- cross-linking preferably occurs before the reaction mixture is diluted or washed away by blood or other body fluids.
- a cross-linked gel forms within 5 minutes from the time at which the components are mixed and applied to the vascular site. In a most preferred embodiment, the cross-linked gel forms within 30-45 seconds.
- Rapid gel formation is also important to prevent excessive blood loss. This is especially important when a patient, such as a trauma patient in an emergency situation, has multiple sites of vascular injury.
- the reaction mixture may be applied to stop bleeding when the precise source of blood loss has not been identified.
- the gel formation time should be slow enough for the reaction mixture to be applied over a relatively large area of bleeding, but fast enough to seal the leak or leaks before the reaction mixture is diluted or washed away.
- the gel reaction mixture is used to seal an air or blood leak in a lung.
- the reaction mixture is designed to take longer to form a gel than a reaction mixture that is used for a vascular application.
- the reaction mixture may be spread across the surface of the lung before it cross-links to form a gel.
- the reaction mixture may contain additives such as lipids or surfactants that help spread the mixture over the lung surface.
- the mixture is less likely to slide off or wash away from the application site, because the lung provides a relatively large and flat surface area when compared to the surface area of a blood vessel.
- a primer solution is applied to the tissue before the gel mixture.
- the primer solution is a dilute solution of BSA. More preferably, the primer solution is a dilute solution of the cross-linking reaction mixture without the cross-linker.
- the primer mixture is hydrophobic.
- a hydrophobic derivation of albumin is used.
- albumin is mixed with a fatty acid.
- albumin may be complexed with palmityl anhydride.
- the molar ratio of fatty acid to albumin is greater than 1:1, and preferably 3:1.
- albumin may be purchased in association with a fatty acid.
- BSA may be purchased in association with octanoic acid (Sigma catalog number A3174, 10-15mg octanoic acid per g of protein).
- the primer solution is applied to the tissue to replace body fluids and provide a good environment for the cross-linking reaction.
- a brush is a useful applicator to spread the primer at the tissue location where the cross-linking reaction mixture will be administered.
- the albumin solution being cross-linked comprises additional reagents to promote interaction with the tissues at the site of application.
- an albumin solution comprises a surfactant and/or a lipid when it is used in a pulmonary context.
- the surfactant and lipid component is similar to the natural surfactant and lipid composition of the lung.
- synthetic surfactants and lipids may be used.
- the cross-linked albumin compositions of the invention may also be used to produce implantable drug deliver)' devices.
- the cross-linked albumin compositions may be used to produce bioerodable implants which contain drugs interspersed throughout the cross- linked matrix of albumin.
- the degree of cross-linking will determine both the ability of drugs to diffuse into and out of the matrix, and the rate at which the device erodes.
- the degree of cross-linking will determine the rigidity or flexibility of the device. By controlling the degree of cross-linking, therefore, devices may be made which deliver drugs at different rates, and which have different degrees of flexibility.
- the devices are produced by introducing an albumin preparation and carbodiimide of the invention, optionally with accessory molecules such as pharmaceuticals, into a mold under conditions which promote cross-linking of the albumin to form the implantable device.
- a solution of BSA at pH 6.02 was prepared by adding drops of 0.5 N HC1 to 1 ml of 37.5 % BSA dissolved in water. As described above, 0.25 ml of 8%> EDC-HCl was added while the solution was stirring. Stirring stopped 1 minute after complete addition of EDC-HCl, and a very cohesive, rubbery, non-tacky gel was produced.
- pH 5.5 pH 5.5
- a solution of BSA at pH 5.5 was prepared by adding drops of 0.5 N HC1 to 1 ml of 37.5 % BSA dissolved in water. As described above, 0.25 ml of 8% EDC-HCl was added while the solution was stirring. Stirring stopped after 31 seconds, and again a very cohesive gel was produced.
- the total volume of the gel was 1.25 ml.
- the gel contained 0.375 g of BSA, 0.020 g of EDC-HCl, and 0.855 g of water. Solids therefore represented 31.6 %> of the gel weight, with EDC-HCl representing only 1.6 %> of the gel weight.
- the cross-linked protein contained approximately 5.3 %> EDC-HCl by weight.
- a 30% solution of BSA was purchased from Sigma (catalog number A8327).
- the pH of 1.5 ml of this BSA solution was adjusted to between 5.38 and 5.40 with 3 drops of 0.5 N HC1.
- a 1 ml aliquot of the pH adjusted BSA was stirred with a magnetic bar using a stirrer at a constant setting.
- a 0.25 ml volume of an 8% EDC-HCl solution 80 mg EDC-HCl dissolved in 1 ml water was added. Stirring became sluggish after 1 minute, and stopped after 1 A minutes. The resulting gel was less rigid than the gel obtained using a 37.5%) BSA solution.
- Example 3 A Two Syringe Mixing System for Delivering BSA/EDC-HCl Bioadhesive
- a two syringe mixing system delivering a 9.3:1 ratio of BSA:EDC-HC1 was tested.
- a 3 ml volume of BSA at pH 5.5 taken up in a first syringe (a 10 cc syringe).
- a 0.5 ml volume of 18.12% EDC-HCl (90 mg EDC-HCl dissolved in 0.500 ml of distilled water) was taken up in a second syringe (a 1 cc syringe).
- the syringes were connected to a modified Micromedix (Eagan, MN) applicator mixing nozzle.
- Example 5 The Effect of Additives on the Rate of Gel Formation
- a 1 ml volume of a 35%> solution of BSA pH 5.5
- polyacids such as dicarboxylic acids, citric acid, polyacrylic acid, glutaric anhydride derivatized BSA, and other polyacids
- polyacids are able to accelerate the gelation either by changing the local pH, by reacting with EDC-HCl at a faster rate than unmodified BSA, or by denaturing the BSA and thereby increasing the rate of BSA cross-linking.
- a monocarboxylic acid in high proportions may result in a longer gel time by interfering with the protein cross-linking reaction.
- 1 ml of 35%> BSA was mixed with 0.1 ml of an aqueous 20%> solution of EDC-HCl, resulting in a rigid gel in 45-50 seconds.
- ethylene diamine dihydrochloride 0.6% of ethylene diamine dihydrochloride was added along with the cross-linker, and the gelation was slowed to 70-85 seconds.
- a diamine can reduce the rate of gelation.
- a polyamine e.g., polyethylene amine, chitosan, poly(l-lysine)
- Other additives such as N-hydroxy succinimide (NHS), a water soluble analog (sulfo NHS, the sulfonic acid salt of NHS), or hydroxy benzotriazole (HOBT) were shown to reduce the rate of gelation. These effects were pH sensitive.
- the crosslinker can be provided as a powder that can be stored at room temperature for over 6 months.
- an aqueous solution of cross-linker should be refrigerated, and is fully active for less than one month.
- Example 7 Adhering an End-to-Side Arterial Anastomosis of ePTFE Graft to Artery
- a sealant mixture of BSA and EDC-HCl was delivered in vitro onto an end-to-side anastomosis of an ePTFE (expanded polytetrafluoroethylene) graft onto a porcine aorta.
- the mixture was delivered through a static mixing nozzle, and contained a 9.3:1 ratio of 35% BSA (pH 5.55) : 40% EDC-HCl.
- the glue mixture was slowly extruded onto all sides of the anastomosis. The mixture cross-linked fairly rapidly. Indeed, the gel could be neatly cut with scissors - 30 seconds after its application.
- the artery was pressurized by introducing saline via a large syringe, and the glue treated graft provided a good seal.
- a 40%> solution was prepared by using a 25/10 ratio of BSA and glutaric anhydride derivatized BSA. This solution (pH 6) was used with a 16.67% aqueous solution of EDC-HCl at a ratio of 8 : 1 (vol/vol) on a porcine lung (ex vivo) to seal a planar wedge resection.
- the solution on curing adhered to the lung tissue and withstood a static air pressure in excess of 60 mm of Hg (average lung pressure during surgery is in the range of 20-25 mm of Hg).
- Example 8 Effect of Derivatization of BSA BSA was derivatized to increase its hydrophobicity with reactive molecules with hydrophobic tails.
- the pH of the resulting solution was adjusted to 8.5, and 6 g of tetra fluoro phthalic anhydride was added as a solution in acetone. The pH was maintained at 8-9 using dilute NaOH. After several hours of stirring, the reaction mixture was diafiltered, pH adjusted to 6.0, and dried. The dry derivatized BSA exhibited a more hydrophobic nature as evidenced from contact angle studies. In one experiment, 20 g of BSA was dissolved in 500ml of a 65/35 mixture of de-ionized water and methanol.
- the pH of the pale yellow solution was adjusted to 9.0, and 8 ml of (2, 2, 3, 3, 4, 4, 5, 5, 6, 6, 7, 7, 8, 8, 9, 9, 9-heptadecafluorononyl)-oxirane was added all at once in a 50% acetone solution.
- the reaction was allowed to stir for 2 days, maintaining the pH at 9.
- the slightly turbid solution was centrifuged, dialyzed, pH adjusted to 6.0, and allowed to dry.
- the dry modified BSA solution exhibited higher viscosity and an improved wettability towards ePTFE graft, and on cross-linking with the appropriate amount of EDC-HCl, adhered very well to the graft and natural tissue.
- Gelatin 300 Bloom, Sigma
- EDC-HCl a ratio of 10: 1.
- This powder was applied on a canine lung in a planar wedge resection model. The material absorbed the neighboring fluids, rapidly gelled into a rubbery mass, and proceeded to stop the leak.
- BSA solution was mixed with Tyloxapol and dipalmitoyl, phosphatidyl choline (DPPC) such that they were 1 mg/ml and 14 mg/ml, respectively.
- the dispersion was mixed (10: 1) with an aqueous solution of EDC HC1 (20%) and applied to a porcine lung in a planar wedge resection model.
- the site was previously primed with a 30% solution of BSA (pH 5.5).
- the material was allowed to attain maximum strength (about 4-5 minutes), and then tested.
- the material withstood a dynamic pressure of about 100mm of Hg before lung tissue rupture occurred.
- Gelatin 300 Bloom was mixed with DPPC and tyloxapol in a similar ratio.
- the material was a gel at room temperature.
- the material was warmed to about 45 ° C and mixed appropriately with an aqueous EDC-HCl solution.
- the material gelled rapidly and adhered satisfactorily to the wound site.
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- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
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Abstract
Cette invention a trait à des techniques ainsi qu'aux produits correspondants en rapport avec des bioadhésifs et des substances de scellement à usage chirurgical à base de protéine ainsi qu'à des dispositifs à implanter aux fins d'une libération de médicament et à des prothèses. Elle concerne, plus particulièrement, des produits à base d'albumine et les techniques afférentes. Ces produits sont utilisés pour faire adhérer des tissus biologiques les uns aux autres ou bien pour les faire adhérer à des prothèses, pour empêcher des fuites de liquides ou de gaz dans des tissus biologiques ou encore pour élaborer des dispositifs bio-érodables. Ces techniques ainsi que les produits de l'invention se révèlent des plus utiles s'agissant d'applications cardio-vasculaires et pulmonaires.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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US9060998P | 1998-06-23 | 1998-06-23 | |
US90609P | 1998-06-23 | ||
PCT/US1999/014232 WO1999066964A1 (fr) | 1998-06-23 | 1999-06-23 | Albumine reticulee de carbodiimide pour bioadhesifs chirurgicaux de scellement et dispositifs a implanter |
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EP1089769A1 true EP1089769A1 (fr) | 2001-04-11 |
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Application Number | Title | Priority Date | Filing Date |
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EP99930613A Withdrawn EP1089769A1 (fr) | 1998-06-23 | 1999-06-23 | Albumine reticulee de carbodiimide pour bioadhesifs chirurgicaux de scellement et dispositifs a implanter |
Country Status (5)
Country | Link |
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EP (1) | EP1089769A1 (fr) |
JP (1) | JP2002518135A (fr) |
AU (1) | AU4711599A (fr) |
CA (1) | CA2335952A1 (fr) |
WO (1) | WO1999066964A1 (fr) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN109054000A (zh) * | 2018-07-24 | 2018-12-21 | 中山大学 | 一种基于聚水杨酸的纳米载药体系及其制备方法和应用 |
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US7615373B2 (en) | 1999-02-25 | 2009-11-10 | Virginia Commonwealth University Intellectual Property Foundation | Electroprocessed collagen and tissue engineering |
US20020081732A1 (en) | 2000-10-18 | 2002-06-27 | Bowlin Gary L. | Electroprocessing in drug delivery and cell encapsulation |
AU2596401A (en) * | 1999-12-22 | 2001-07-03 | Surgical Sealants, Inc. | Methods and compositions for sealing tissue leaks |
CN1211076C (zh) * | 2000-02-17 | 2005-07-20 | 3M创新有限公司 | 使用预制的生物可降解聚合物组合物的输递装置和方法 |
WO2001062312A1 (fr) * | 2000-02-25 | 2001-08-30 | Monterey Biomedical, Inc. | Pansement a formation de mousse |
US7192766B2 (en) | 2001-10-23 | 2007-03-20 | Medtronic Minimed, Inc. | Sensor containing molded solidified protein |
DE60328899D1 (de) * | 2003-12-18 | 2009-10-01 | Antoine Turzi | Bioresorbierbares vernetztes Hydrogel bestehend aus Albumin |
GB0403406D0 (en) * | 2004-02-17 | 2004-03-24 | Biotransys Ltd | Preparation of carriers for drug delivery and other therapeutic applications |
US20050276863A1 (en) * | 2004-05-27 | 2005-12-15 | Medtronic Vascular, Inc. | Methods for treatment of aneurysmal tissue |
US20110171167A1 (en) * | 2004-06-30 | 2011-07-14 | Tyco Healthcare Group Lp | Isocyanate-Based Compositions and Their Use |
JP2006051121A (ja) * | 2004-08-10 | 2006-02-23 | National Institute For Materials Science | アルブミンと生体低分子誘導体から構成される生体内分解吸収性粘着性医用材料 |
US7837986B2 (en) | 2004-12-01 | 2010-11-23 | E. I. Du Pont De Nemours And Company | Protein-based polymer tissue adhesives for medical use |
GB0615377D0 (en) * | 2006-08-03 | 2006-09-13 | Advanced Protein Systems Ltd | Gels for use in wound management |
AU2007294808A1 (en) | 2006-09-13 | 2008-03-20 | Southeastern Medical Technologies, Llc | Methods and compositions for sealing and adhering biological tissues and medical uses thereof |
CA2818728C (fr) * | 2010-11-23 | 2020-09-22 | Elastagen Pty Ltd | Preparations et/ou formulations a base de proteines reticulees avec des polysaccharides |
US8846849B2 (en) | 2011-05-13 | 2014-09-30 | Ethicon, Inc. | Tissue sealants from plasma derived proteins |
US9446166B2 (en) | 2013-01-24 | 2016-09-20 | Ethicon, Inc. | Fibrin sealant compositions with chemical crosslinking |
KR101794401B1 (ko) | 2015-03-26 | 2017-11-06 | 윤원준 | 접착성 펩타이드 및 그 용도 |
CN116328050A (zh) | 2016-05-09 | 2023-06-27 | 生命连结有限公司 | 基于网状补片的原位可交联组合物 |
CA3127191A1 (fr) | 2019-01-21 | 2020-07-30 | Eclipse Medcorp, Llc | Procede, systeme et appareil pour separer des composants d'un echantillon biologique |
EP4052034A4 (fr) | 2019-10-31 | 2023-12-20 | Crown Laboratories, Inc. | Systèmes, procédés et appareil de séparation de constituants d'un échantillon |
CN112933251B (zh) * | 2021-02-05 | 2023-03-17 | 广东粤港澳大湾区国家纳米科技创新研究院 | 四氧化三锰纳米颗粒制备方法及用途 |
CN113440644B (zh) * | 2021-06-10 | 2023-01-17 | 广东省科学院健康医学研究所 | 一种弹性白蛋白胶粘剂及其制备方法 |
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US4046871A (en) * | 1975-04-08 | 1977-09-06 | Ortho Diagnostics Inc. | Compositions and serological methods including bovine serum albumin polymers |
US4526714A (en) * | 1982-12-13 | 1985-07-02 | Cordis Europa N.V. | Conjugates of anticoagulant and protein |
JP2953702B2 (ja) * | 1989-01-20 | 1999-09-27 | 生化学工業株式会社 | 合成コンドロイチン硫酸プロテオグリカンを含有する組織癒着防止剤 |
US5632776A (en) * | 1990-11-22 | 1997-05-27 | Toray Industries, Inc. | Implantation materials |
IT1245217B (it) * | 1991-03-12 | 1994-09-13 | Baldacci Lab Spa | Coniugato di azidotimidina ed albumina umana procedimento per la sua preparazione e composizioni farmaceutiche che lo contengono |
US5376692A (en) * | 1992-05-15 | 1994-12-27 | Purdue Research Foundation | Method of binding using irradiation and product with albumin bound to biomaterials |
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1999
- 1999-06-23 WO PCT/US1999/014232 patent/WO1999066964A1/fr not_active Application Discontinuation
- 1999-06-23 JP JP2000555650A patent/JP2002518135A/ja active Pending
- 1999-06-23 EP EP99930613A patent/EP1089769A1/fr not_active Withdrawn
- 1999-06-23 CA CA002335952A patent/CA2335952A1/fr not_active Abandoned
- 1999-06-23 AU AU47115/99A patent/AU4711599A/en not_active Abandoned
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN109054000A (zh) * | 2018-07-24 | 2018-12-21 | 中山大学 | 一种基于聚水杨酸的纳米载药体系及其制备方法和应用 |
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CA2335952A1 (fr) | 1999-12-29 |
WO1999066964A1 (fr) | 1999-12-29 |
AU4711599A (en) | 2000-01-10 |
JP2002518135A (ja) | 2002-06-25 |
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LINE | Tissue adhesives have many potential medical applications, including wound closure, supplementing or replacing sutures or staples in surgical procedures, adhesion of synthetic onlays or inlays to the cornea, drug delivery devices, and as anti-adhesion barriers to prevent post-surgical adhesions. Conventional tissue adhesives include fibrin sealants, cyanoacrylate based |
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