CN105233343B - The preparation method of acellular dermal matrix eardrum and external ear tissue renovation material - Google Patents
The preparation method of acellular dermal matrix eardrum and external ear tissue renovation material Download PDFInfo
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- CN105233343B CN105233343B CN201510825102.1A CN201510825102A CN105233343B CN 105233343 B CN105233343 B CN 105233343B CN 201510825102 A CN201510825102 A CN 201510825102A CN 105233343 B CN105233343 B CN 105233343B
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Abstract
The invention discloses the preparation method of a kind of acellular dermal matrix eardrum and external ear tissue renovation material, it comprises the following steps：Step A：Choose the dermal tissue of dynamic lactation object or person body；Step B：By contrasting the structure of dermal tissue and the eardrum tissue, close with the structure of the eardrum tissue in the dermal tissue structure and adjacent 3 layers dermal tissue α Rotating fields are determined；Step C:Other layers in addition to dermal tissue α Rotating fields in the dermal tissue are removed, obtain dermal tissue α layer primary raw materials.The present invention is according to corium feature and the particularity of eardrum institutional framework, carry out development improvement, prepare and the extremely similar eardrum repair materials of human body eardrum structure, tissue characteristic, on the basis of completing tissue integrity and repairing, eardrum function has obtained basic reparation, is greatly improved clinical effectiveness；The raw material sources for preparing acellular dermal matrix eardrum and external ear tissue renovation material have been widened, have reduced the cost for preparing the repair materials.
Description
Technical field
The present invention relates to the preparation method of acellular dermal matrix eardrum and external ear tissue renovation material.
Background technology
At present, clinically mainly repaired, also had using animal derived using autologous Temporal fascia for eardrum reparation
Biomaterial is repaired, and due to the problem of secondary damage and ectopic tissue scarring being present, repairing effect is difficult to be satisfied with.Manadesma
Tissue and eardrum institutional framework are differed greatly, therefore the iris action of external ear and middle ear is only completed on repairing multiple junction fruit, and
Repair as scar repairing, it is very big to the original sound air transmitted function effect of eardrum.
As can be seen here, eardrum repair materials repairing effect of the prior art is poor, is only capable of playing a part of barrier, to drum
The original sound air transmitted function effect of film is very big, it would be highly desirable to a kind of new eardrum preparation method of restoration materials is researched and developed, to overcome
The defects of method of currently manufactured eardrum repair materials.
The content of the invention
The technical problem to be solved in the present invention is a kind of acellular dermal matrix eardrum of offer and external ear tissue renovation material
Preparation method, the preparation method widened the raw material sources of acellular dermal matrix eardrum and external ear tissue renovation material, drops
The low cost of acellular dermal matrix eardrum and external ear tissue renovation material, the acellular dermal matrix eardrum and external ear of preparation
Tissue renovation material has good compatibility, can complete the reparation of tissue complete correction and eardrum function.
In order to solve the above technical problems, the preparation of the acellular dermal matrix eardrum and external ear tissue renovation material of the present invention
Method, it comprises the following steps：
Step A：Choose the dermal tissue of mammalian body or human body；
Step B：By contrasting the structure of dermal tissue and the eardrum tissue, determine in the dermal tissue structure with institute
State eardrum tissue structure is close and adjacent 3 layers of dermal tissue α Rotating fields；
Step C:Other layers in addition to dermal tissue α Rotating fields in the dermal tissue are removed, obtain dermal tissue α layers
Primary raw materials；
Step D:De- cell processing is carried out to the dermal tissue α layers primary raw materials, at the beginning of obtaining acellular dermal tissue α layers
Level raw material；
Step E：By the acellular dermal tissue α layers primary raw materials handled through step D under sterile, constant temperature, Yu Ping
Mixed in weighing apparatus stoste and shake certain time, restore the life of collagen, active factors in the acellular dermal tissue α layer primary raw materials
Physical performance；
Step F：By the α layer primary raw materials of the acellular dermal tissue after being handled in step E, locate in stoste is balanced
Reason, by way of the permeability and supplement ion balance that change albuminous membranae, close the albumen of dermal tissue α layer primary raw materials
The potential point of matter, obtain acellular dermal matrix eardrum and external ear tissue renovation material；
Wherein, in the step F, the dermal tissue after cell is taken off by combined ionic or single ionic described in leveling
The potential difference of the protein of α layer primary raw materials, reach the mesh of the dermal tissue α layer primary raw materials potential points after the de- cell of closing
So that all potential points of the α layer primary raw materials albumen of dermal tissue are 0.
In one embodiment of the present of invention, in the step F, taken off by combined ionic or single ionic described in leveling
The potential difference of dermal tissue α layer primary raw materials after cell, reach the dermal tissue α layer primary raw materials after closing the de- cell
Protein potential point purpose.
In one embodiment of the present of invention, in the step D, during the processing of de- cell, by I, V, VI Collagen Type VI
Albumen, basic fibroblast growth factor, transforming growth factor-β, fibronectin are fixed.
In one embodiment of the present of invention, in the step C, dermal tissue α layers primary raw materials is carried out using fixer
It is fixed.
In one embodiment of the present of invention, in the step D, dermal tissue α layer primary raw materials immunitys are removed, retain day
So collagen and elastic fibers material of tissue space framework, and maintain the frame structure of its natural material.
In one embodiment of the present of invention, in the step E, in the constant temperature, temperature range of choice is 16~37
Degree Celsius.
It it is 4~24 hours in balancing the mixed time shaken in stoste in the step E in one embodiment of the present of invention.
The preparation method of acellular dermal matrix eardrum and external ear tissue renovation material of the present invention has the beneficial effect that：
The preparation method of acellular dermal matrix eardrum and external ear tissue renovation material of the present invention, according to corium feature and drum
The particularity of membrane tissue structure is improved, and is prepared and the extremely similar eardrum reparation material of human body eardrum structure, tissue characteristic
Material, on the basis of completing tissue integrity and repairing, eardrum function has obtained basic reparation, clinical effectiveness is carried significantly
Height, the raw material sources for preparing acellular dermal matrix eardrum and external ear tissue renovation material are widened, has reduced and prepares the reparation
The cost of material.
Brief description of the drawings
Above-mentioned is only the general introduction of technical solution of the present invention, in order to better understand the present invention, below in conjunction with accompanying drawing with
The present invention is described in further detail for embodiment.
Fig. 1 is the photo figure that the acellular dermal matrix eardrum of the present invention subsidizes 2 weeks after reconstruction eardrum；
The acellular dermal matrix eardrum Leap frog that Fig. 2 and Fig. 3 is the present invention rebuilds the eardrum photo figure of postoperative 6 weeks；
Fig. 4 is the acellular dermal matrix eardrum and external ear tissue renovation material that are prepared using the present invention after the tympanic membrane perforation
Interplantation is taken to carry out the photo figure of eardrum reconstruction；
Fig. 5 is the photo figure that acellular dermal matrix eardrum interplantation prepared by the present invention is rebuild 2 weeks after eardrum；
Fig. 6 is the narrow screen photo figure that acellular dermal matrix eardrum interplantation prepared by the present invention is rebuild 6 weeks after eardrum；
Fig. 7 is that acellular dermal matrix eardrum interplantation prepared by the present invention rebuilds the eardrum photo figure of postoperative 6 weeks；
Fig. 8 is the photo figure before acellular dermal matrix transplanting prepared by the present invention；
Fig. 9 is that acellular dermal matrix prepared by the present invention is implanted into the cavy ear reconstruction eardrum photo figure of postoperative 2 weeks；
Figure 10 is the implantation cavy ear reconstruction postoperative 6 weeks photos of eardrum after acellular dermal matrix processing prepared by the present invention
Figure；
Figure 11 is to be implanted into cavy ear after the acellular dermal matrix of the preparation of the present invention is handled to rebuild the eardrum photograph of postoperative 8 weeks
Piece figure；
Figure 12 is the photo figure before external ear tissue renovation material transplanting prepared by the present invention；
Figure 13 is the photo figure of local surfaces before 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 8 weeks external auditory meatus faces after 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 8 weeks facies tympanicas after external ear tissue renovation material transplanting prepared by the present invention；
Figure 18 is 8 weeks facies tympanica photo figures after external ear tissue renovation material transplanting prepared by the present invention.
Embodiment
Below by drawings and examples, technical scheme is described in further detail.
The acellular dermal matrix eardrum of the present invention and the preparation method of external ear tissue renovation material, it includes following step
Suddenly：
Step A：Choose the corium of mammalian body or human body.
Mammalian body is pig, ox, sheep etc., preferably, choosing the corium on human body, the corium of mammal different parts
Structure composition and function have differences, and the type of stromatin and the various factors, quantity contained by different parts corium are not
Together.According to the composition structure of different parts corium, the difference of function and various factor contents, corresponding portion is targetedly selected
The corium of position.
Step B：By contrasting the structure of dermal tissue and the eardrum tissue, determine in the dermal tissue structure with institute
State eardrum tissue structure is close and adjacent 3 layers of dermal tissue α Rotating fields；
Concretely, eardrum institutional framework can be divided into 3 layers substantially, be respectively：It is epithelium layer, consolidated fibers, interior
Mucocutaneous membrane cellular layer.Wherein, strata externum membranae tympani is skin epithelial cell, and main component is collagen and Skin Cell, I, V type
Collagen layering arranges in an orderly manner, wherein, the unified and regularly arranged NTx albumen of diameter is to maintain eardrum epithelial cell
An important factor for growth arrangement；Intermediate layer is I, V fine and close collagen type, arrange according to certain rules therebetween elastic fibers and
Glycoprotein；VI collagen type is formed the hypothallus of mucous layer in eardrum with network structure, ordered arrangement to eardrum matrix, viscous
Theca cell growth plays an important role.
The present invention is compared by electron microscope to eardrum institutional framework and substantial amounts of skin corium institutional framework, collagen
The qualitative comparison of type, collagen orientation order compare, and are determined in 250 layers of dermal tissue structure and eardrum knot of tissue
The extremely close and adjacent 3-tier architecture of structure, the adjacent 3-tier architecture is named as dermal tissue α Rotating fields.
Step C:Other layers in addition to dermal tissue α Rotating fields in the dermal tissue are removed, obtain dermal tissue α layers
Primary raw materials；
The epidermal cell in dermal tissue and the cell membrane of other cells are divided using physics and biochemical method
From, destroy, and remove material caused by cytoclasis, reach and remove other layers in corium, only retain the α of dermal tissue
The purpose of Rotating fields.
In dermal cell substance process is removed, in material form frame structure collagen and attachment it is special because
Son etc. is protected, fixed and reduced (to material in Patent No. CN1116794C using biochemistry and organizational engineering method
The technology of processing is fixed in material).
Mammalian tissues sustain damage (including pathologic damage and artificial actively destroy damage) when, body starts self
Repair function, change in the local cell of damage, chemotactic factor (CF), basic fibroblast growth factor (Fibroblast
Growth factor FGF), NTx protein promoter etc. have inducing cell to damage location is mobile, differentiation and value-added
Function.The present invention is protected and created emphatically in the preparation process of acellular dermal matrix eardrum and external ear tissue renovation material
Above-mentioned environment so that inducing cell enters material frame after acellular dermal matrix eardrum and the implantation of external ear tissue renovation material.
Step D:De- cell processing is carried out to the dermal tissue α layers primary raw materials, at the beginning of obtaining acellular dermal tissue α layers
Level raw material；
After the completion of the preparation of dermal tissue α layers primary raw materials, cell technology is taken off using patent, in the basic preparation solution (specially
Profit number：The basic preparation solution is disclosed in CN1116794C) in add certain enzyme and chemical reagent and be prepared into de- cell and cell
Epimatrix fixes preparation, and fixed formulation soln and dermal tissue α layers primary raw materials be mixed to be postponed and carry out that constant temperature is mixed to shake operation, to corium
Tissue α layers primary raw materials carries out de- cell processing.Meanwhile add specific enzyme and chemical reagent in fixation preparation preparation solution and incite somebody to action
Ith, V, VI collagen type, basic fibroblast growth factor, transforming growth factor-β (Transforming growth
Factor- β TGF-βs), fibronectin (Fibronectin FN) to being fixed, protect its in de- cell processes from
Destroy.
Then, the dermal tissue α layers primary raw materials after cell processing is taken off aseptically, in balance in reducing solution, perseverance
Under the conditions of temperature, carry out mixed shake and operate some time.By above-mentioned de- cell processing procedure, the immunity of dermal tissue is eliminated,
The materials such as collagen and the elastic fibers of natural tissues space framework are remained, and maintain the framework knot of its natural material
Structure.
Step E：By the acellular dermal tissue α layers primary raw materials handled through step D under sterile, constant temperature, specifically
Temperature range be 16~37 degrees Celsius, in balance in stoste it is mixed shake 4~24 hours, restore the acellular dermal tissue α layers
The biological property of collagen, active factors in primary raw materials；
In rapid E, preferably, being 25 degrees Celsius in temperature, mixed in stoste is balanced and shake operation 20 hours.
It is mixed to shake in stoste is balanced under 32 degrees Celsius of constant temperatures in this step as disposable embodiment
Operation 24 hours.
As disposable embodiment, in this step, under 16 degrees celsius, carry out mixed shaking behaviour in stoste is balanced
Make 4 hours.
Dermal tissue α layers primary raw materials is further handled in reducing solution is balanced, wherein, add phase in reducing solution is balanced
The reductase and neutralization reagent answered, reduction I, III, VI collagen type, basic fibroblast growth factor, conversion growth because
Son-β, fibronectin, it is set to be activated under certain condition after implanting.
Step F：By the α layer primary raw materials of the acellular dermal tissue after being handled in step E, further stoste is being balanced
Middle processing, by way of the permeability and supplement ion balance that change albuminous membranae, closing dermal tissue α layer primary raw materials
The potential point of protein, obtain acellular dermal matrix eardrum or external ear tissue renovation material.
Close the potential point in dermal tissue, the collagen component contained in mammalian tissues according to place position and
The difference of function of organization, variant electricity physiological signal be present.The generation of electric signal is due to protein film both ends ionic species
The potential difference that concentration and the difference of composition are formed, the frequency, pressure difference intensity in the same area electric signal are varied, and it is integrated
As a result it is exactly the precursor into the region is converted to required adult cell.Dermal tissue α layer primary raw materials
Eardrum itself is not originating from, therefore the precursor of collagen electric signal meeting severe jamming into framework turns to eardrum cell
Change.By way of the permeability and supplement ion balance that change albuminous membranae, in dermal tissue α layer primary raw materials preparation process
In, use balance stoste so that all potential points of the α layer primary raw materials albumen of dermal tissue are 0, reach closing corium group
The purpose for the α layer primary raw materials potential points knitted, after making acellular dermal matrix eardrum or external ear tissue renovation material implantation human body
Into the not interference by material of the host cell divisions direction of the repair materials frame gap in itself.
Specifically, the present invention takes off the dermal tissue α layers after cell by combined ionic or single ionic described in leveling
The potential difference of the protein of primary raw materials, reach the purpose of the dermal tissue α layer primary raw materials potential points after the de- cell of closing.
After acellular dermal matrix eardrum and external ear tissue renovation material implantation host's eardrum position, host tissue liquid and blood
The stem cell of itself and precursor enter in acellular dermal matrix eardrum and external ear tissue renovation material frame gap in liquid
Stick and be detained, under the stimulation of tympanic membrane surrounding tissue physiological signal and correlation factor collective effect, stem cell and precursor propagation
Divide and to the cell transformation for forming eardrum, secrete neoblast epimatrix, strengthen repairing eardrum tissue, epithelial cell is along frame table
Layer is creeped covering, progress eardrum Regeneration and Repair.Meanwhile it is tissue with nutrient to establish rete vasculosum, peripheral nervous fiber penetrates length
Enter newly-built eardrum tissue, complete the Regeneration and Repair of eardrum tissue.
Embodiment 1
Using the acellular dermal matrix eardrum and external ear tissue renovation material Leap frog of the present invention, subsidize in tympanic membrane perforation
In 28 ears of cavy of lateral surface, there are 25 ears to close for 2 weeks after surgery, wherein eardrum is congested, thickens.There are 3 ears to infect.2
Eardrum hyperemia is faded after week, and at 6 weeks, part ear drum membrane is without hyperemia, and transparent, surface is smooth, and surface visible vessels increase
It is raw.
It please coordinate shown in Fig. 1, acellular dermal matrix eardrum prepared by the present invention is subsidized after reconstruction eardrum 2 weeks, and eardrum fills
Blood, (↓) is thickened, × 40.Please refer to shown in Fig. 2, acellular dermal matrix eardrum prepared by the present invention subsidizes reconstruction eardrum
6 weeks afterwards, eardrum color was greyish white, and surface is smooth (←), similar to normal human tympanic membrane.The visible capillary (▲) in surface, × 40.It please coordinate
As shown in fig.3, acellular dermal matrix eardrum Leap frog prepared by the present invention rebuilds eardrum postoperative 6 weeks, it is seen that surface squamous
Epithelium (↓), the fibrous layer (▲) of lower floor, the visible a small amount of cuboid cell (↑) in bottom, H-E, × 400.Please refer to shown in Fig. 4,
After the tympanic membrane perforation, interplantation is taken to carry out eardrum reconstruction using the acellular dermal matrix eardrum (↓) for preparing of the present invention, × 40.
Rebuild please refer to the acellular dermal matrix eardrum interplantation that shown in Fig. 5, prepared by the present invention after eardrum 2 weeks, eardrum thickens
(↗), merged with remaining eardrum, × 40.Please refer to the acellular dermal matrix eardrum interplantation that shown in Fig. 6, prepared by the present invention
Method is rebuild after eardrum 6 weeks, eardrum (↗) color is greyish white, and surface is smooth, similar to normal human tympanic membrane, and × 40.Please refer to Fig. 7 institutes
Show, acellular dermal matrix eardrum interplantation prepared by the present invention rebuilds eardrum postoperative 6 weeks, it is seen that surface scaly epithelium (↓), under
The fibrous layer (▲) of layer, H-E, × 400.The ironed device of acellular dermal matrix eardrum operation consent application eardrum is ironed, environmental scanning
The visible acellular dermal matrix eardrum repair materials surface of electric Microscopic observation is smooth, locally has a small amount of hole to occur, without any thin
Born of the same parents adhere to.Cavy ear is implanted into after the processing of acellular dermal matrix eardrum repair materials and rebuilds postoperative 2 weeks materials of eardrum, environmental scanning
The visible acellular dermal matrix eardrum repair materials surface of electric Microscopic observation has a small amount of cell to adhere to.Acellular dermal matrix eardrum
Cavy ear is implanted into after repair materials processing and rebuilds postoperative 6 weeks materials of eardrum, visible acellular dermal is observed under environmental scanning electronic microscope
Matrix eardrum repair materials surface has a large amount of cells to adhere to, and forms cell monolayer.It is implanted into after the processing of acellular dermal matrix eardrum
Cavy ear is rebuild eardrum and drawn materials for postoperative 8 weeks, and visible acellular dermal matrix eardrum repair materials table is observed under environmental scanning electronic microscope
Face is attached with a large amount of cell attachments, forms multilayer.
Please refer to shown in Fig. 8, the preceding surface of acellular dermal matrix of the invention transplanting is smooth, locally there is a small amount of hole
Occur, adhere to without any cell, scale：50 μm, operating voltage (HV)：15.0kv, beam spot size control parameter (Spot)：5.0
Multiplication factor (Mag)：755 times, operating distance (WD)：11.3mm, operating pressure (Pressure)：515.7Pa, operating temperature
(Tem)：0 DEG C, ADM the 1st day.Please refer to shown in Fig. 9, cavy ear weight is implanted into after acellular dermal matrix of the invention processing
Postoperative 2 weeks of drum film, it is seen that there are a small amount of fibroblast attachment (→), scale in surface：20 μm, operating voltage (HV)：15.0kv,
Beam spot size control parameter (Spot)：5.0, multiplication factor (Mag)：1906 times, operating distance (WD)：12.5mm operating pressure
(Pressure)：475.1Pa, operating temperature (Tem)：0 DEG C, ADM the 2nd week.Please refer to shown in Figure 10, of the invention is de- thin
Cavy ear is implanted into after the processing of born of the same parents' dermal matrix and rebuilds eardrum postoperative 6 weeks, it is seen that surface there are a large amount of cells to adhere to, and forms individual layer
(→), scale：50 μm, operating voltage (HV)：15.0kv, beam spot size control parameter (Spot)：5.0, multiplication factor (Mag)：
973 times, operating distance (WD)：12.7mm, operating pressure (Pressure)：515.7Pa, operating temperature (Tem)：0 DEG C, ADM the 6th
Week.Eardrum is rebuild please refer to cavy ear is implanted into shown in Figure 11, after acellular dermal matrix of the invention processing postoperative 8 weeks,
Visible surface is attached with a large amount of cells, forms multilayer (→), scale：20 μm, operating voltage：15.0kv, beam spot size control ginseng
Number：5.0, multiplication factor：2200 times, operating distance：11.5mm operating pressure：556.1Pa, 0 DEG C of operating temperature, ADM the 8th week.
Scanning electron microscopic observation visible surface is smooth, locally there is nipple-like elevation, the collagenous fibres of a large amount of interleaved arrangements be present, has no cell
Spline structure.Normal guinea pig eardrum scanning electron microscopic observation can be by the squamous cell in external auditory meatus face, middle fibrous layer and tympanum
The flat epithelial cell in face.After acellular dermal matrix eardrum repair materials progress eardrum Reconstruction is implanted into after the cavy tympanic membrane perforation
8 weeks, it is seen that there is substantial amounts of squamous cell in collagenous fibres, external auditory meatus face, and facies tympanica takes off with the presence of a large amount of flat epithelial cells
Visible a large amount of immature flat epithelial cells in the middle part of acellular dermal matrix eardrum repair materials.
Embodiment 2
Please refer to shown in Figure 12, it is smooth that external ear tissue renovation material of the invention transplants preceding surface, it is seen that great Liang Jiao
The collagenous fibres (→) of arrangement are knitted, are adhered to without any cell, scale：30μm.Please refer to shown in Figure 13, external ear of the invention
Local surfaces have a small amount of nipple spline structure (→) before tissue renovation material transplanting, it is seen that the collagenous fibres of a large amount of interleaved arrangements, nothing
Any cell attachment, scale：30μm.Please refer to shown in Figure 14, normal human tympanic membrane external auditory meatus face, it is seen that scaly epithelium furfur
The squamous cell (▲) of (→) and demyelinate, scale：30μm.Please refer to shown in Figure 15, outer ear tissue of the invention
8 weeks external auditory meatus faces after repair materials transplanting, it is seen that there are a large amount of squamous cells (→), scale in collagenous fibres, surface：30μm.
Please refer to shown in Figure 16, normal human tympanic membrane facies tympanica, it is seen that there are a large amount of flat epithelial cells (→) on radial fiber, surface,
Scale：30μm.Please refer to 8 weeks facies tympanicas after the external ear tissue renovation material transplanting that shown in Figure 17, prepared by the present invention, it is seen that
There are the flat epithelial cell (→) of maturation, scale in collagenous fibres, surface：30μm.Please refer to shown in Figure 18, of the invention is outer
8 weeks facies tympanicas after the transplanting of ear tissue repair materials, it is seen that there are a large amount of immature flat epithelial cells on collagenous fibres, surface
(→), scale：30μm.
The acellular dermal matrix eardrum and external ear tissue renovation material obtained using the preparation method of the present invention, by facing
Bed experiment proves that tissue integrity reparation can be completed completely, and the feature of eardrum can obtain basic recovery, clinical
Effect is greatly improved, and solves the problems, such as the scarcity of clinically ear drum membrane repair materials, has widened ear drum membrane and outer
The source of ear tissue repair materials.
It is described above, it is presently preferred embodiments of the present invention, not the present invention is imposed any restrictions, those skilled in the art's profit
A little simple modification, equivalent variations or modification are made with the content of the disclosure above, is all fallen within protection scope of the present invention.
Claims (6)
1. the preparation method of a kind of acellular dermal matrix eardrum and external ear tissue renovation material, it is characterised in that including following step
Suddenly：
Step A：Choose the dermal tissue of mammalian body or human body；
Step B：By contrasting the structure of dermal tissue and the eardrum tissue, determine in the dermal tissue structure with the drum
The structure of membrane tissue is close and adjacent 3 layers of dermal tissue α Rotating fields；
Step C:Other layers in addition to dermal tissue α Rotating fields in the dermal tissue are removed, at the beginning of obtaining dermal tissue α layers
Level raw material；
Step D:De- cell processing is carried out to the dermal tissue α layers primary raw materials, it is primary to obtain acellular dermal tissue α layers
Raw material；
Step E：It is former in balance by the acellular dermal tissue α layers primary raw materials handled through step D under sterile, constant temperature
In liquid mix shake certain time, restore collagen in the acellular dermal tissue α layer primary raw materials, active factors it is biological
Energy；
Step F：By the α layer primary raw materials of the acellular dermal tissue after being handled in step E, handle, lead in stoste is balanced
The mode of the permeability for changing albuminous membranae and supplement ion balance is crossed, closes the protein of dermal tissue α layer primary raw materials
Potential point, obtain acellular dermal matrix eardrum and external ear tissue renovation material；
Wherein, in the step F, the dermal tissue α layers after cell are taken off by combined ionic or single ionic described in leveling
The potential difference of the protein of primary raw materials, reach the purpose of the dermal tissue α layer primary raw materials potential points after the de- cell of closing, make
All potential points for obtaining the α layer primary raw materials albumen of dermal tissue are 0.
2. the preparation method of acellular dermal matrix eardrum according to claim 1 and external ear tissue renovation material, it is special
Sign is,
In the step D, de- cell processing during, by I, V, VI collagen type, basic fibroblast growth because
Son, transforming growth factor-β, fibronectin are fixed.
3. the preparation method of acellular dermal matrix eardrum according to claim 1 and external ear tissue renovation material, it is special
Sign is,
In the step C, dermal tissue α layer primary raw materials is fixed using fixer.
4. the preparation method of acellular dermal matrix eardrum according to claim 1 and external ear tissue renovation material, it is special
Sign is,
In the step D, dermal tissue α layer primary raw materials immunitys are removed, retain the collagen of natural tissues space framework
With elastic fibers material, and the frame structure of its natural material is maintained.
5. the preparation method of acellular dermal matrix eardrum according to claim 1 and external ear tissue renovation material, it is special
Sign is,
In the step E, in the constant temperature, temperature range of choice is 16 ~ 37 degrees Celsius.
6. the preparation method of acellular dermal matrix eardrum according to claim 1 and external ear tissue renovation material, it is special
Sign is,
It it is 4 ~ 24 hours in balancing the mixed time shaken in stoste in the step E.
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| 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 |
| CN106983909B (en) * | 2017-03-03 | 2018-01-30 | 北京博辉瑞进生物科技有限公司 | A kind of otology repair materials, preparation method and application |
| 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 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1324567A (en) * | 2000-07-13 | 2001-12-05 | 胡杰 | Method of preparing cartilage graft |
| CN102038564A (en) * | 2009-10-14 | 2011-05-04 | 周星 | Supporting frame capable of being taken out |
| RU2013114451A (en) * | 2013-04-02 | 2013-08-27 | Государственное бюджетное учреждение здравоохранения города Москвы Научно-исследовательский институт скорой помощи имени Н.В. Склифосовского Департамента здравоохранения г. Москвы | METHOD FOR PRODUCING A DERMAL MATRIX |
| CN103764071A (en) * | 2011-07-11 | 2014-04-30 | 澳大利亚耳科学研究所 | Device for ear drum repair |
| CN104288837A (en) * | 2014-09-11 | 2015-01-21 | 杨淑珍 | Acellular dermal matrix and preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050013870A1 (en) * | 2003-07-17 | 2005-01-20 | Toby Freyman | Decellularized extracellular matrix of conditioned body tissues and uses thereof |
-
2015
- 2015-11-24 CN CN201510825102.1A patent/CN105233343B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1324567A (en) * | 2000-07-13 | 2001-12-05 | 胡杰 | Method of preparing cartilage graft |
| CN102038564A (en) * | 2009-10-14 | 2011-05-04 | 周星 | Supporting frame capable of being taken out |
| CN103764071A (en) * | 2011-07-11 | 2014-04-30 | 澳大利亚耳科学研究所 | Device for ear drum repair |
| RU2013114451A (en) * | 2013-04-02 | 2013-08-27 | Государственное бюджетное учреждение здравоохранения города Москвы Научно-исследовательский институт скорой помощи имени Н.В. Склифосовского Департамента здравоохранения г. Москвы | METHOD FOR PRODUCING A DERMAL MATRIX |
| CN104288837A (en) * | 2014-09-11 | 2015-01-21 | 杨淑珍 | Acellular dermal matrix and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 脱细胞真皮基质黏膜组织补片在内外植法鼓膜修补术中的应用;李特等;《检验医学与临床》;20150331;第12卷(第5期);第614-615页 * |
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