CN105233343A - Manufacturing method for acellular dermal matrix tympanic membrane and external ear tissue repair material - Google Patents
Manufacturing method for acellular dermal matrix tympanic membrane and external ear tissue repair material Download PDFInfo
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- CN105233343A CN105233343A CN201510825102.1A CN201510825102A CN105233343A CN 105233343 A CN105233343 A CN 105233343A CN 201510825102 A CN201510825102 A CN 201510825102A CN 105233343 A CN105233343 A CN 105233343A
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Abstract
The invention discloses a manufacturing method for an acellular dermal matrix tympanic membrane and an external ear tissue repair material. The method comprises the following steps of A, selecting mammal body or human body dermis tissue; B, determining three adjacent layers of dermis tissue alpha layer structures similar to the structure of tympanic membrane tissue in the dermis tissue structure by comparing the dermis tissue with the tympanic membrane tissue structure; C, removing the other layers except for the dermis tissue alpha layer structures in the dermis tissue structure to obtain a dermis tissue alpha layer preliminary material. According to the specificity of the dermis characteristics and the tympanic membrane tissue structure for development and improvement, and the tympanic membrane repair material similar to the human body tympanic membrane structure and the tissue characteristics is obtained. On the basis that the tissue integrity repair is completed, the tympanic membrane function is basically repaired, and the clinic effect is greatly improved; the raw material sources of the material for manufacturing the acellular dermal matrix tympanic membrane and the external ear tissue repair material are broadened, and the cost for manufacturing the repair material is lowered.
Description
Technical field
The present invention relates to the preparation method of acellular dermal matrix tympanum and external ear tissue renovation material.
Background technology
At present, mainly adopt autologous Temporal fascia to repair clinically for tympanum reparation, also have and adopt animal derived biomaterial to repair, owing to there is the problem of secondary damage and ectopic tissue scarring, repairing effect is difficult to satisfied.Fascia tissue and tympanum organizational structure differ greatly, and are therefore repairing iris action result only completing external ear and middle ear, and repair as scar repairing, very large to tympanum original sound aerial conduction function effect.
As can be seen here, tympanum repair materials repairing effect of the prior art is poor, only can play the effect of obstruct, very large to tympanum original sound aerial conduction function effect, urgently research and develop a kind of new tympanum preparation method of restoration materials, to overcome the defect of the method manufacturing tympanum repair materials at present.
Summary of the invention
The technical problem to be solved in the present invention is for providing the preparation method of a kind of acellular dermal matrix tympanum and external ear tissue renovation material, this preparation method has widened the raw material sources of acellular dermal matrix tympanum and external ear tissue renovation material, reduce the cost of acellular dermal matrix tympanum and external ear tissue renovation material, acellular dermal matrix tympanum and the external ear tissue renovation material of preparation have the good compatibility, can complete the reparation organizing complete correction and tympanum function.
For solving the problems of the technologies described above, the preparation method of acellular dermal matrix tympanum of the present invention and external ear tissue renovation material, it comprises the following steps:
Steps A: the dermal tissue choosing mammalian body or human body;
Step B: by contrasting the structure of described dermal tissue and tympanum tissue, determines the close and dermal tissue α Rotating fields of adjacent 3 layers of the structure organized with described tympanum in described dermal tissue structure;
Step C: remove other layers except dermal tissue α Rotating fields in described dermal tissue, obtains dermal tissue α layer primary raw materials;
Step D: carry out de-cell process to described dermal tissue α layer primary raw materials, obtains acellular dermal and organizes α layer primary raw materials;
Step e: organized by the acellular dermal through step D process α layer primary raw materials under aseptic, constant temperature, mixedly in balance stock solution shakes certain hour, restores the biological property that described acellular dermal organizes collagen protein, active factors in α layer primary raw materials;
Step F: by the α layer primary raw materials of the acellular dermal tissue after process in step e, process in balance liquid, by changing the permeability of albuminous membranae and the mode of supplementary equilibrium ion, close the potential point of the protein of dermal tissue α layer primary raw materials, obtain acellular dermal matrix tympanum and external ear tissue renovation material.
In one embodiment of the present of invention, in described step F, carried out the potential difference of the dermal tissue α layer primary raw materials after de-cell described in leveling by combined ionic or single ionic, reach the object of the potential point of the protein of the dermal tissue α layer primary raw materials after closed described de-cell.
In one embodiment of the present of invention, in described step D, in the process of de-cell process, I, V, VI collagen type, basic fibroblast growth factor, transforming growth factor-β, fibronectin are fixed.
In one embodiment of the present of invention, in described step C, fixative is utilized to be fixed dermal tissue α layer primary raw materials.
In one embodiment of the present of invention, in described step D, remove dermal tissue α layer primary raw materials immunity, retain collagen protein and the elastic fibers material of natural tissues space framework, and maintain the frame structure of its natural material.
In one embodiment of the present of invention, in described step e, in described constant temperature, thermal creep stress scope is 16 ~ 37 degrees Celsius.
In one embodiment of the present of invention, in described step e, in balance liquid, the mixed time of shaking is 4 ~ 24 hours.
In one embodiment of the present of invention, prepared acellular dermal matrix tympanum and external ear tissue renovation material are directly used in the transplanting of human or animal.
The beneficial effect of the preparation method of acellular dermal matrix tympanum of the present invention and external ear tissue renovation material is:
The preparation method of acellular dermal matrix tympanum of the present invention and external ear tissue renovation material, particularity according to corium feature and tympanum organizational structure improves, prepare the tympanum repair materials very close with human body tympanum structure, tissue characteristic, on the basis completing tissue integrity reparation, tympanum function obtains basic reparation, clinical effectiveness is greatly enhanced, widen the raw material sources preparing acellular dermal matrix tympanum and external ear tissue renovation material, reduce the cost of this repair materials of preparation.
Accompanying drawing explanation
Above-mentioned is only the general introduction of technical solution of the present invention, and in order to better understand the present invention, below in conjunction with accompanying drawing and detailed description of the invention, the present invention is described in further detail.
Fig. 1 is that acellular dermal matrix tympanum of the present invention subsidizes the photo figure rebuilding after tympanum 2 weeks;
Fig. 2 and Fig. 3 is that acellular dermal matrix tympanum Leap frog of the present invention rebuilds the tympanum photo figure of postoperative 6 weeks;
Fig. 4 is after the perforation of ear drum, and the acellular dermal matrix tympanum utilizing the present invention to prepare and external ear tissue renovation material take interplantation to carry out the photo figure of tympanum reconstruction;
Fig. 5 is the photo figure that the acellular dermal matrix tympanum interplantation prepared of the present invention is rebuild after tympanum 2 weeks;
Fig. 6 is the narrow screen photo figure that the acellular dermal matrix tympanum interplantation prepared of the present invention is rebuild after tympanum 6 weeks;
Fig. 7 is that acellular dermal matrix tympanum interplantation prepared by the present invention rebuilds the tympanum photo figure of postoperative 6 weeks;
Fig. 8 is the photo figure before the acellular dermal matrix prepared of the present invention is transplanted;
Fig. 9 is that acellular dermal matrix prepared by the present invention implants the Cavia porcellus ear reconstruction tympanum photo figure of postoperative 2 weeks;
Figure 10 implants Cavia porcellus ear after the acellular dermal matrix process prepared of the present invention to rebuild tympanum postoperative 6 weeks photo figure;
Figure 11 implants Cavia porcellus ear to rebuild the tympanum photo figure of postoperative 8 weeks after the acellular dermal matrix process of preparation of the present invention;
Figure 12 is the photo figure before the external ear tissue renovation material prepared of the present invention is transplanted;
Figure 13 is the photo figure of the front local surfaces of external ear tissue renovation material transplanting prepared by the present invention;
Figure 14 is the photo figure in normal human tympanic membrane external auditory meatus face;
Figure 15 is the photo figure in the rear 8 weeks external auditory meatus faces of external ear tissue renovation material transplanting prepared by the present invention;
Figure 16 is the photo figure of normal human tympanic membrane facies tympanica;
Figure 17 is the photo figure of the rear 8 weeks facies tympanicas of external ear tissue renovation material transplanting prepared by the present invention;
Figure 18 is that external ear tissue renovation material prepared by the present invention transplants rear 8 weeks facies tympanica photo figure.
Detailed description of the invention
Below by drawings and Examples, technical scheme of the present invention is described in further detail.
The preparation method of acellular dermal matrix tympanum of the present invention and external ear tissue renovation material, it comprises the following steps:
Steps A: the corium choosing mammalian body or human body.
Mammalian body is pig, cattle, sheep etc., preferably, chooses the corium on human body, and the structure composition of the corium of mammal different parts and function there are differences, and the type of stromatin and the various factor contained by different parts corium, quantity are different.According to the composition structure of different parts corium, the difference of function and various factor content, selects the corium of corresponding site targetedly.
Step B: by contrasting the structure of described dermal tissue and tympanum tissue, determines the close and dermal tissue α Rotating fields of adjacent 3 layers of the structure organized with described tympanum in described dermal tissue structure;
Concretely, tympanum organizational structure can be divided into 3 layers substantially, respectively: epithelium layer, consolidated fibers, endothelium mucomembranous cell layer.Wherein, strata externum membranae tympani is skin epithelial cell, and main component is collagen protein and Skin Cell, and I, V collagen type layering arranges in an orderly manner, wherein, the NTx albumen that diameter is unified and regularly arranged maintains the key factor that tympanum epithelial cell growth arranges; Intermediate layer is I, V fine and close collagen type, arranges elastic fibers and glycoprotein according to certain rules therebetween; VI collagen type is formed the hypothallus of mucous layer in tympanum with network structure, plays an important role to the ordered arrangement of tympanum substrate, mucomembranous cell growth.
The present invention by ultramicroscope to tympanum organizational structure and a large amount of skin corium organizational structure comparisons, the qualitative comparison of collagen types, the comparison of collagen protein orientation order, the 3-tier architecture very close and adjacent with tympanum organizational structure is determined, the 3-tier architecture called after dermal tissue α Rotating fields adjacent by this in 250 layers of dermal tissue structure.
Step C: remove other layers except dermal tissue α Rotating fields in described dermal tissue, obtains dermal tissue α layer primary raw materials;
Physics is utilized to be separated with the cell membrane of other cells with biochemical method the epidermis cell in dermal tissue, to destroy, and remove the material that cytoclasis produces, reach and remove other layers in corium, only retain the object of the α Rotating fields of dermal tissue.
In removal hypodermal cell substance process, biochemistry is adopted to the specific factor etc. of the collagen protein and attachment that form frame structure in material and tissue engineering method is protected, fixing and reduction (patent No. is the technology in CN1116794C, material being fixed to process).
When mammalian tissues sustains damage (comprise pathologic damage and artificially initiatively destroy damage), body starts self-regeneration function, changes, chemotactic factor, basic fibroblast growth factor (FibroblastgrowthfactorFGF), NTx protein promoter etc. have inducing cell to move to damage location, break up and value-added function at the cell of damage local.The present invention protects emphatically and creates above-mentioned environment in the preparation process of acellular dermal matrix tympanum and external ear tissue renovation material, and after acellular dermal matrix tympanum and external ear tissue renovation material are implanted, inducing cell enters material frame.
Step D: carry out de-cell process to described dermal tissue α layer primary raw materials, obtains acellular dermal and organizes α layer primary raw materials;
After prepared by dermal tissue α layer primary raw materials, patent is utilized to take off cell technology, prepare on this basis in liquid (patent No.: disclose this basis in CN1116794C and prepare liquid) and add certain enzyme and chemical reagent is prepared into de-cell and extracellular matrix fixes preparation, fixing formulation soln and dermal tissue α layer primary raw materials mix postpone to carry out constant temperature and mixes and shake operation, carries out de-cell process to dermal tissue α layer primary raw materials.Simultaneously; prepare in liquid at fixing preparation and add specific enzyme and chemical reagent by I, V, VI collagen type, basic fibroblast growth factor, transforming growth factor-β (Transforminggrowthfactor-β TGF-β), fibronectin (FibronectinFN) to being fixed, protect it to avoid destroying in de-cell processes.
Then, the dermal tissue α layer primary raw materials after de-cell process aseptically, in balance reducing solution, under constant temperature, carries out mixed shaking and operates the some time.Through above-mentioned de-cell processing procedure, eliminate the immunity of dermal tissue, remain the material such as collagen protein and elastic fibers of natural tissues space framework, and maintain the frame structure of its natural material.
Step e: organized by the acellular dermal through step D process α layer primary raw materials under aseptic, constant temperature, concrete temperature range is 16 ~ 37 degrees Celsius, mix in balance stock solution and shake 4 ~ 24 hours, restore the biological property that described acellular dermal organizes collagen protein, active factors in α layer primary raw materials;
In rapid E, preferably, be 25 degrees Celsius in temperature, mix in balance liquid and shake operation 20 hours.
As disposable embodiment, in this step, under 32 degrees Celsius of constant temperatures, in balance liquid, mix and shake operation 24 hours.
As disposable embodiment, in this step, under 16 degrees celsius, carry out mixing in balance liquid and shake operation 4 hours.
Dermal tissue α layer primary raw materials processes further in balance reducing solution, wherein, corresponding reductase and neutralization reagent is added in balance reducing solution, reduction I, III, VI collagen type, basic fibroblast growth factor, transforming growth factor-β, fibronectin, make it can be activated under certain condition after implanting.
Step F: by the α layer primary raw materials of the acellular dermal tissue after process in step e, process in balance liquid further, by changing the permeability of albuminous membranae and the mode of supplementary equilibrium ion, close the potential point of the protein of dermal tissue α layer primary raw materials, obtain acellular dermal matrix tympanum or external ear tissue renovation material.
Close the potential point in dermal tissue, the difference of the collagen component contained in mammalian tissues foundation position, place and function of organization, has the electricity physiological signal of diversity.The generation of the signal of telecommunication is the potential difference because the difference of albuminous coat two ends ionic species concentration and composition is formed, varied in the frequency of the same area signal of telecommunication, pressure reduction intensity, its synthesis result is exactly that the precursor making to enter this region transforms to required adult cell.Dermal tissue α layer primary raw materials itself not derives from tympanum, and therefore the collagen protein signal of telecommunication can enter the precursor of framework to tympanum cell transformation by severe jamming.By changing the permeability of albuminous membranae and the mode of supplementary equilibrium ion, in dermal tissue α layer primary raw materials preparation process, balance liquid, all potential points of the α layer primary raw materials albumen of dermal tissue are made to be 0, reach the object of the α layer primary raw materials potential point of closed dermal tissue, enter the host cell divisions direction of this repair materials frame gap after making acellular dermal matrix tympanum or external ear tissue renovation material implant into body not by the interference of material itself.
Concrete, the present invention carrys out the potential difference of the protein of the dermal tissue α layer primary raw materials after de-cell described in leveling by combined ionic or single ionic, reach the object of the dermal tissue α layer primary raw materials potential point after closing de-cell.
After acellular dermal matrix tympanum and external ear tissue renovation material implant host's tympanum position, in host tissue liquid and blood, self stem cell and precursor enter in acellular dermal matrix tympanum and external ear tissue renovation material frame gap to stick and are detained, stimulate and under correlation factor combined effect at tympanic membrane surrounding tissue physiological signal, stem cell and precursor multiple fission to the cell transformation forming tympanum, secrete new extracellular matrix, strengthen repairing tympanum tissue, epithelial cell is creeped along framework top layer covering, carries out tympanum Regeneration and Repair.Meanwhile, setting up vasoganglion is tissue with nutrient, and peripheral nervous fiber penetrates newly-built tympanum tissue of growing into, and completes the Regeneration and Repair of tympanum tissue.
Embodiment 1
Utilize acellular dermal matrix tympanum of the present invention and external ear tissue renovation material Leap frog, subsidize in Cavia porcellus 28 ears of tympanic membrane perforation lateral surface, have 25 ears 2 weeks after surgery all closed, wherein tympanum congested, thicken.3 ears are had to occur infecting.After 2 weeks, tympanum hyperemia is faded, to part ear drum membrane when 6 weeks without congested, transparent, smooth surface, surperficial visible vessels hypertrophy.
Please coordinate shown in Fig. 1, acellular dermal matrix tympanum prepared by the present invention subsidizes to be rebuild after tympanum 2 weeks, tympanum hyperemia, thickens (↓), × 40.Shown in Fig. 2, acellular dermal matrix tympanum prepared by the present invention subsidizes to be rebuild after tympanum 6 weeks, and tympanum color is greyish white, smooth surface (←), similar to normal human tympanic membrane.The visible blood capillary (▲) in surface, × 40.Shown in Fig. 3, acellular dermal matrix tympanum Leap frog prepared by the present invention rebuilds tympanum postoperative 6 weeks, visible surface squamous epithelial cancer (↓), the fibrous layer (▲) of lower floor, the visible a small amount of cuboid cell (↑) in bottom, H-E, × 400.Shown in Fig. 4, after the perforation of ear drum, the acellular dermal matrix tympanum (↓) utilizing the present invention to prepare takes interplantation to carry out tympanum reconstruction, × 40.Shown in Fig. 5, acellular dermal matrix tympanum interplantation prepared by the present invention to be rebuild after tympanum 2 weeks, and tympanum thickens (↗), merge with remaining tympanum, × 40.Shown in Fig. 6, acellular dermal matrix tympanum interplantation prepared by the present invention to be rebuild after tympanum 6 weeks, tympanum (↗) color is greyish white, smooth surface, similar to normal human tympanic membrane, and × 40.Shown in Fig. 7, acellular dermal matrix tympanum interplantation prepared by the present invention rebuilds tympanum postoperative 6 weeks, visible surface squamous epithelial cancer (↓), the fibrous layer (▲) of lower floor, H-E, × 400.The ironed device of acellular dermal matrix tympanum operation consent application tympanum is ironed, observes visible acellular dermal matrix tympanum repair materials smooth surface under environmental scanning electronic microscope, and local has a small amount of hole to occur, without any cell attachment.Implant Cavia porcellus ear reconstruction tympanum after the process of acellular dermal matrix tympanum repair materials to draw materials for postoperative 2 weeks, observing visible acellular dermal matrix tympanum repair materials surface under environmental scanning electronic microscope has a small amount of cell attachment.Implant Cavia porcellus ear reconstruction tympanum after the process of acellular dermal matrix tympanum repair materials to draw materials for postoperative 6 weeks, observing visible acellular dermal matrix tympanum repair materials surface under environmental scanning electronic microscope has a large amount of cell attachment, forms cell monolayer.Implant Cavia porcellus ear reconstruction tympanum after the process of acellular dermal matrix tympanum to draw materials for postoperative 8 weeks, observing visible acellular dermal matrix tympanum repair materials surface attachment under environmental scanning electronic microscope has a large amount of cell attachment, forms multilamellar.
Shown in Fig. 8, it is smooth that acellular dermal matrix of the present invention transplants front surface, and local has a small amount of hole to occur, without any cell attachment, scale: 50 μm, running voltage (HV): 15.0kv, bundle speckle size controling parameters (Spot): 5.0, amplification (Mag): 755 times, operating distance (WD): 11.3mm, operating pressure (Pressure): 515.7Pa, operating temperature (Tem): 0 DEG C, ADM the 1st day.Shown in Fig. 9, implant Cavia porcellus ear after acellular dermal matrix process of the present invention and rebuild tympanum postoperative 2 weeks, visible surface has a small amount of fibroblast to adhere to (→), scale: 20 μm, running voltage (HV): 15.0kv, bundle speckle size controling parameters (Spot): 5.0, amplification (Mag): 1906 times, operating distance (WD): 12.5mm, operating pressure (Pressure): 475.1Pa, operating temperature (Tem): 0 DEG C, ADM the 2nd week.Shown in Figure 10, implant Cavia porcellus ear after acellular dermal matrix process of the present invention and rebuild tympanum postoperative 6 weeks, visible surface has a large amount of cell attachment, forms monolayer (→), scale: 50 μm, running voltage (HV): 15.0kv, bundle speckle size controling parameters (Spot): 5.0, amplification (Mag): 973 times, operating distance (WD): 12.7mm, operating pressure (Pressure): 515.7Pa, operating temperature (Tem): 0 DEG C, ADM the 6th week.Shown in Figure 11, implant Cavia porcellus ear after acellular dermal matrix process of the present invention and rebuild tympanum postoperative 8 weeks, visible surface is attached with a large amount of cell, forms multilamellar (→), scale: 20 μm, running voltage: 15.0kv, bundle speckle size controling parameters: 5.0, amplification: 2200 times, operating distance: 11.5mm, operating pressure: 556.1Pa, operating temperature 0 DEG C, ADM the 8th week.Scanning electron microscopic observation visible surface is smooth, and there is nipple-like elevation local, there are the collagen fiber of a large amount of interleaved arrangement, has no cell spline structure.Normal guinea pig tympanum scanning electron microscopic observation can by the flat epithelial cell of the squamous cell in external auditory meatus face, middle fibrous layer and facies tympanica.Implant acellular dermal matrix tympanum repair materials after the Cavia porcellus perforation of ear drum to carry out after tympanum Reconstruction 8 weeks, visible collagen fiber, there is a large amount of squamous cells in external auditory meatus face, facies tympanica has a large amount of flat epithelial cell to exist, visible a large amount of immature flat epithelial cell in the middle part of acellular dermal matrix tympanum repair materials.
Embodiment 2
Shown in Figure 12, it is smooth that external ear tissue renovation material of the present invention transplants front surface, the collagen fiber (→) of visible a large amount of interleaved arrangement, without any cell attachment, and scale: 30 μm.Shown in Figure 13, external ear tissue renovation material of the present invention transplants front local surfaces a small amount of nipple spline structure (→), the collagen fiber of visible a large amount of interleaved arrangement, without any cell attachment, and scale: 30 μm.Shown in Figure 14, normal human tympanic membrane external auditory meatus face, the squamous cell (▲) of visible squamous epithelial cancer desquamation (→) and demyelination, scale: 30 μm.Shown in Figure 15, external ear tissue renovation material of the present invention transplants rear 8 weeks external auditory meatus faces, and visible collagen fiber, there is a large amount of squamous cell (→) on surface, scale: 30 μm.Shown in Figure 16, normal human tympanic membrane facies tympanica, visible radial fiber, there is a large amount of flat epithelial cell (→) on surface, scale: 30 μm.Shown in Figure 17, external ear tissue renovation material prepared by the present invention transplants rear 8 weeks facies tympanicas, visible collagen fiber, and there is ripe flat epithelial cell (→) on surface, scale: 30 μm.Shown in Figure 18, external ear tissue renovation material of the present invention transplants rear 8 weeks facies tympanicas, and visible collagen fiber, there is a large amount of immature flat epithelial cell (→) on surface, scale: 30 μm.
The acellular dermal matrix tympanum utilizing preparation method of the present invention to obtain and external ear tissue renovation material, prove can complete tissue integrity reparation completely by clinical trial, and the functional of tympanum can obtain basic recovery, clinical effectiveness is greatly improved, solve the problem of the scarcity of ear drum membrane repair materials clinically, widen the source of ear drum membrane and external ear tissue renovation material.
The above, be preferred embodiment of the present invention, not impose any restrictions the present invention, those skilled in the art utilize the content of above-mentioned announcement to make a little simple modification, equivalent variations or modification, all drop in protection scope of the present invention.
Claims (8)
1. a preparation method for acellular dermal matrix tympanum and external ear tissue renovation material, is characterized in that comprising the following steps:
Steps A: the dermal tissue choosing mammalian body or human body;
Step B: by contrasting the structure of described dermal tissue and tympanum tissue, determines the close and dermal tissue α Rotating fields of adjacent 3 layers of the structure organized with described tympanum in described dermal tissue structure;
Step C: remove other layers except dermal tissue α Rotating fields in described dermal tissue, obtains dermal tissue α layer primary raw materials;
Step D: carry out de-cell process to described dermal tissue α layer primary raw materials, obtains acellular dermal and organizes α layer primary raw materials;
Step e: organized by the acellular dermal through step D process α layer primary raw materials under aseptic, constant temperature, mixedly in balance stock solution shakes certain hour, restores the biological property that described acellular dermal organizes collagen protein, active factors in α layer primary raw materials;
Step F: by the α layer primary raw materials of the acellular dermal tissue after process in step e, process in balance liquid, by changing the permeability of albuminous membranae and the mode of supplementary equilibrium ion, close the potential point of the protein of dermal tissue α layer primary raw materials, obtain acellular dermal matrix tympanum and external ear tissue renovation material.
2. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step F, carried out the potential difference of the dermal tissue α layer primary raw materials after de-cell described in leveling by combined ionic or single ionic, reach the object of the potential point of the protein of the dermal tissue α layer primary raw materials after closed described de-cell.
3. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step D, in the process of de-cell process, I, V, VI collagen type, basic fibroblast growth factor, transforming growth factor-β, fibronectin are fixed.
4. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step C, fixative is utilized to be fixed dermal tissue α layer primary raw materials.
5. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step D, remove dermal tissue α layer primary raw materials immunity, retain collagen protein and the elastic fibers material of natural tissues space framework, and maintain the frame structure of its natural material.
6. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step e, in described constant temperature, thermal creep stress scope is 16 ~ 37 degrees Celsius.
7. the preparation method of acellular dermal matrix tympanum according to claim 1 and external ear tissue renovation material, is characterized in that,
In described step e, in balance liquid, the mixed time of shaking is 4 ~ 24 hours.
8. the acellular dermal matrix tympanum according to any one of claim 1-7 and the preparation method of external ear tissue renovation material, is characterized in that,
Prepared acellular dermal matrix tympanum and external ear tissue renovation material are directly used in the transplanting of human or animal.
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| CN106983909A (en) * | 2017-03-03 | 2017-07-28 | 北京博辉瑞进生物科技有限公司 | A kind of otology repair materials, preparation method and application |
| EP3572103A4 (en) * | 2017-03-03 | 2020-03-04 | Beijing Biosis Healing Biological Technology Co., Ltd. | Biological tissue matrix material, preparation method therefor and use thereof in otological repair material |
| CN115245601A (en) * | 2022-01-18 | 2022-10-28 | 郑州大学第一附属医院 | Tympanic membrane repair material and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106983909A (en) * | 2017-03-03 | 2017-07-28 | 北京博辉瑞进生物科技有限公司 | A kind of otology repair materials, preparation method and application |
| EP3572103A4 (en) * | 2017-03-03 | 2020-03-04 | Beijing Biosis Healing Biological Technology Co., Ltd. | Biological tissue matrix material, preparation method therefor and use thereof in otological repair material |
| US11529437B2 (en) | 2017-03-03 | 2022-12-20 | Beijing Biosis Healing Biological Technology Co., Ltd. | Biological tissue matrix material, preparation method therefor and use thereof in otological repair material |
| CN115245601A (en) * | 2022-01-18 | 2022-10-28 | 郑州大学第一附属医院 | Tympanic membrane repair material and preparation method thereof |
| CN115245601B (en) * | 2022-01-18 | 2023-08-22 | 郑州大学第一附属医院 | Tympanic membrane repair material and preparation method thereof |
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