CN101618232A - Silver-containing pig acellular dermis and method for preparing same - Google Patents
Silver-containing pig acellular dermis and method for preparing same Download PDFInfo
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- CN101618232A CN101618232A CN200810115916A CN200810115916A CN101618232A CN 101618232 A CN101618232 A CN 101618232A CN 200810115916 A CN200810115916 A CN 200810115916A CN 200810115916 A CN200810115916 A CN 200810115916A CN 101618232 A CN101618232 A CN 101618232A
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- sus domestica
- acellular dermis
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Abstract
The invention relates to silver-containing pig acellular dermis and a method for preparing the same. The method for preparing the silver-containing pig acellular dermis comprises steps of pretreating pigskin; splitting the pigskin; sterilizing a skin sheet; performing cell removing treatment on the skin sheet; and performing active silver solution treatment on the skin sheet. The invention relates to silver-containing pig acellular dermis prepared by the method for preparing the silver-containing pig acellular dermis. Because of the treatment of the active silver ion solution, the silver-containing pig acellular dermis of the invention has active bacteria inhibiting effect , and reduces infection occurrence rate after transplantation; meanwhile, the silver-containing pig acellular dermis of the invention not only removes the prior immunogenicity cell components in the matrix, but also maintains a basic structure of skin tissue and an integral collagen fiber support, contains silver ions with bacteria inhibiting effect at the same time, and provides a fine dermis substituent with active bacteria inhibiting function in clinic application.
Description
Technical field
The present invention relates to field of tissue engineering technology, particularly relate to a kind of silver-containing pig acellular dermis and preparation method thereof.
Background technology
In treatment large-area burns patient process, how to utilize limited skin source as early as possible wound closure be one of more scabrous problem.Sometimes adopt microparticle skin or epidermis cell to cultivate the area coverage that the method for transplanting enlarges the skin source clinically, so that wound surface seals as early as possible.Though these skin sources can wound closure,, cause healing rate low, even additional diseases such as serious vesicle, ulceration and cicatricial contracture occur owing to lack the support of corium composition.
At present a large amount of research worker have been launched many-sided research work at aspects such as providing support, improve the wound healing quality for epidermis cell.Through discovering for many years, the Corii Sus domestica wide material sources, biological character and fell are close, a series of processing removal epidermises of process and intradermal can be brought out the cell component of host's rejection, keep both non-immunogenicities of the acellular dermis made behind the extracellular matrix compositions such as complete fibrous tissue and basement membrane, the dermis scaffold that is similar to normal configuration is provided again, can have promoted the formation of the Growth and Differentiation and the epidermis-corium junctional area normal configuration of epidermis cell.Pig acellular dermis is to provide good support for fibroblast as the advantage in skin source, help the reconstruction of fibroblast to acellular dermis collagen composition, and form morphosis and the normal collagen fiber of arranged distribution, promote the high-quality healing of wound surface.But pig acellular dermis also has the following disadvantages aspect clinical practice: the no anti-infection ability of acellular dermis itself, clean-up performance to wound surface is had relatively high expectations, and that severe burn, wound and wound surface itself infect is general relatively more serious, therefore transplant and often occur situation about infecting behind the pig acellular dermis, the effect that influence is transplanted.
Summary of the invention
The purpose of this invention is to provide a kind of silver-containing pig acellular dermis and preparation method thereof, effectively solve existing pig acellular dermis and transplant defectives such as postoperative infection incidence rate height.
For achieving the above object, the invention provides a kind of preparation method of silver-containing pig acellular dermis, comprising:
The Corii Sus domestica pretreatment;
Corii Sus domestica cuts open layer;
The skin graft sterilization;
Skin graft goes cell to handle;
The active silver-colored solution-treated of skin graft.
Described Corii Sus domestica is the fragrant Corii Sus domestica skin of the crust horse at 4-6 monthly age.
In technique scheme, described Corii Sus domestica pretreatment is specially:
The visual examination of raw material Corii Sus domestica;
Scrub described Corii Sus domestica;
Corii Sus domestica after scrubbing is scraped hair.
Describedly scrub described Corii Sus domestica and be specially: the tow sides of repeatedly scrubbing described Corii Sus domestica with soap powder water are until cleaning, and the reuse percentage by weight is scrubbed described Corii Sus domestica for the 3-8% povidone iodine.
Described Corii Sus domestica cuts open layer and is specially: pretreated Corii Sus domestica is cutd open epidermis and hypodermis layer, obtain the skin graft that thickness is the 0.1-1.0 millimeter.
Described skin graft sterilization is specially: with the skin graft percentage by weight is 0.05-0.2% benzalkonium bromide rinsing 12-18 minute.
Described skin graft goes the cell processing to be specially: with the skin graft percentage by weight is that 0.1-0.2% triton x-100 solution was handled 22-26 hour down in temperature 22-28 ℃.
The active silver-colored solution-treated of described skin graft is specially: with skin graft in being to soak 12-18 minute in the active silver-colored solution of 0.02-0.1% at percentage by weight under temperature 35-40 ℃.
For achieving the above object, the present invention also provides a kind of silver-containing pig acellular dermis that adopts the preparation method preparation of described silver-containing pig acellular dermis.
The invention provides a kind of silver-containing pig acellular dermis and preparation method thereof, have the following advantages:
1, since silver-containing pig acellular dermis of the present invention through the processing of active silver ion solution, so pig acellular dermis has initiatively bacteriostatic effect, the infection rate after having reduced it and transplanting.
2, silver-containing pig acellular dermis of the present invention in original immunogenicity cell component, has also kept the basic structure of skin histology and complete collagen fiber support in having removed substrate.
3, silver-containing pig acellular dermis of the present invention contains the silver ion with bacteriostasis, for clinical practice provides a kind of initiatively good dermal substitute of bacteria resistance function that has.
Below by drawings and Examples, technical scheme of the present invention is described in further detail.
Description of drawings
Fig. 1 is the preparation method flow chart of silver-containing pig acellular dermis in silver-containing pig acellular dermis of the present invention and preparation method thereof;
Fig. 2 is Corii Sus domestica pretreatment process figure in silver-containing pig acellular dermis of the present invention and preparation method thereof.
The specific embodiment
Fig. 1 is the preparation method flow chart of silver-containing pig acellular dermis in silver-containing pig acellular dermis of the present invention and preparation method thereof, is specially:
Fig. 2 is Corii Sus domestica pretreatment process figure in silver-containing pig acellular dermis of the present invention and preparation method thereof, and in technical scheme shown in Figure 1, step 1 is specially:
Further specify technical scheme of the present invention below by specific embodiment.
First embodiment
The fragrant Corii Sus domestica skin of the crust horse at step 101,4 monthly ages of selection carries out visual examination as the raw material Corii Sus domestica to the raw material Corii Sus domestica, and the raw material Corii Sus domestica presents normal white, and skin surface is smooth smooth, and fur does not have blood stasis, do not have common dermatosis, and Pilus Sus domestica carefully lacks, and pore is little;
Step 102, scrub Corii Sus domestica tow sides three times after the inspection until cleaning with soap powder water, remove unholiness parts such as leftover bits and bulk fat, the reuse percentage by weight is that 3% povidone iodine is scrubbed Corii Sus domestica, and is clean with a large amount of clear water at last;
Step 103, the Corii Sus domestica after will scrubbing are slightly scraped hair with sharpening earlier, with clear water Corii Sus domestica are rinsed well, and the little razor of reuse is carefully scraped hair, washes by rubbing with the hands totally with clear water;
Step 104, the Corii Sus domestica that will scrape behind the hair thin, remove subcutaneous fat, obtain skin graft, after with drum dermatome skin graft being cutd open epidermis and subcutaneous tissue, only remaining corium part, thickness are 0.1 millimeter, and skin graft is cut into required specification, wash by rubbing with the hands totally with clear water, and the Rhizoma Imperatae in the cleaning pore;
Step 105, the skin graft percentage by weight that will cut open behind the layer are that 0.05% benzalkonium bromide rinsing was sterilized in 18 minutes, and a large amount of clear water of reuse clean 3 times, until clarification of water, free from admixture;
Step 106, the skin graft after will sterilizing go cell to handle, with percentage by weight is 0.1% triton x-100 (TritonX-100) solution in 22 ℃ of following oscillation treatment skin grafts of temperature 26 hours, the a large amount of aquesterilisa of reuse clean 3 times, clean 15 minutes, until clarification of water, free from admixture at every turn;
Step 107, will carry out active silver-colored solution-treated through the pig acellular skin graft after going cell to handle, be in 0.02% active silver-colored solution to soak 18 minute at percentage by weight with skin graft under 35 ℃ of temperature, takes out, and is silver-containing pig acellular dermis.
A kind of silver-containing pig acellular dermis that adopts the preparation method preparation of above-mentioned silver-containing pig acellular dermis.
Second embodiment
The fragrant Corii Sus domestica skin of the crust horse at step 201,5 monthly ages of selection carries out visual examination as the raw material Corii Sus domestica to the raw material Corii Sus domestica, and the raw material Corii Sus domestica presents normal white, and skin surface is smooth smooth, and fur does not have blood stasis, do not have common dermatosis, and Pilus Sus domestica carefully lacks, and pore is little;
Step 202, scrub Corii Sus domestica tow sides four times after the inspection until cleaning with soap powder water, remove unholiness parts such as leftover bits and bulk fat, the reuse percentage by weight is that 5% povidone iodine is scrubbed Corii Sus domestica, and is clean with a large amount of clear water at last;
Step 203, the Corii Sus domestica after will scrubbing are slightly scraped hair with sharpening earlier, with clear water Corii Sus domestica are rinsed well, and the little razor of reuse is carefully scraped hair, washes by rubbing with the hands totally with clear water;
Step 204, the Corii Sus domestica that will scrape behind the hair thin, remove subcutaneous fat, obtain skin graft, after with drum dermatome skin graft being cutd open epidermis and subcutaneous tissue, only remaining corium part, thickness are 0.5 millimeter, and skin graft is cut into required specification, wash by rubbing with the hands totally with clear water, and the Rhizoma Imperatae in the cleaning pore;
Step 205, the skin graft percentage by weight that will cut open behind the layer are that 0.1% benzalkonium bromide rinsing was sterilized in 15 minutes, and a large amount of clear water of reuse clean 4 times, until clarification of water, free from admixture;
Step 206, the skin graft after will sterilizing go cell to handle, with percentage by weight is 0.15% triton x-100 (TritonX-100) solution in 25 ℃ of following oscillation treatment skin grafts of temperature 24 hours, the a large amount of aquesterilisa of reuse clean 4 times, clean 15 minutes, until clarification of water, free from admixture at every turn;
Step 207, will carry out active silver-colored solution-treated through the pig acellular skin graft after going cell to handle, be in 0.05% active silver-colored solution to soak 15 minute at percentage by weight with skin graft under 37 ℃ of temperature, takes out, and is silver-containing pig acellular dermis.
A kind of silver-containing pig acellular dermis that adopts the preparation method preparation of above-mentioned silver-containing pig acellular dermis.
The 3rd embodiment
The fragrant Corii Sus domestica skin of the crust horse at step 301,6 monthly ages of selection carries out visual examination as the raw material Corii Sus domestica to the raw material Corii Sus domestica, and the raw material Corii Sus domestica presents normal white, and skin surface is smooth smooth, and fur does not have blood stasis, do not have common dermatosis, and Pilus Sus domestica carefully lacks, and pore is little;
Step 302, scrub Corii Sus domestica tow sides five times after the inspection until cleaning with soap powder water, remove unholiness parts such as leftover bits and bulk fat, the reuse percentage by weight is that 8% povidone iodine is scrubbed Corii Sus domestica, and is clean with a large amount of clear water at last;
Step 303, the Corii Sus domestica after will scrubbing are slightly scraped hair with sharpening earlier, with clear water Corii Sus domestica are rinsed well, and the little razor of reuse is carefully scraped hair, washes by rubbing with the hands totally with clear water;
Step 304, the Corii Sus domestica that will scrape behind the hair thin, remove subcutaneous fat, obtain skin graft, after with drum dermatome skin graft being cutd open epidermis and subcutaneous tissue, only remaining corium part, thickness are 1.0 millimeters, and skin graft is cut into required specification, wash by rubbing with the hands totally with clear water, and the Rhizoma Imperatae in the cleaning pore;
Step 305, the skin graft percentage by weight that will cut open behind the layer are that 0.2% benzalkonium bromide rinsing was sterilized in 12 minutes, and a large amount of clear water of reuse clean 5 times, until clarification of water, free from admixture;
Step 306, the skin graft after will sterilizing go cell to handle, with percentage by weight is 0.2% triton x-100 (TritonX-100) solution in 28 ℃ of following oscillation treatment skin grafts of temperature 22 hours, the a large amount of aquesterilisa of reuse clean 5 times, clean 15 minutes, until clarification of water, free from admixture at every turn;
Step 307, will carry out active silver-colored solution-treated through the pig acellular skin graft after going cell to handle, be in 0.1% active silver-colored solution to soak 12 minute at percentage by weight with skin graft under 40 ℃ of temperature, takes out, and is silver-containing pig acellular dermis.
A kind of silver-containing pig acellular dermis that adopts the preparation method preparation of above-mentioned silver-containing pig acellular dermis.
The fungistatic effect that has compared silver-containing pig acellular dermis and pig acellular dermis below by concrete bacteriostatic experiment.
Strain is selected traumatology department's patient's wound surface common staphylococcus aureus, escherichia coli, bacillus cloacae, klepsiella pneumoniae and bacillus pyocyaneus for use, and above-mentioned pure bacterium is moved into meat soup or normal saline, and to be mixed with concentration respectively be 1.5 * 10
8The bacterium liquid of pfu/ml, the bacterium liquid for preparing are coated agar plate (90 millimeters of diameters) respectively equably.The skin graft selected for use of experiment is silver-containing pig acellular dermis and common pig acellular dermis, and the dinking that two kinds of skin grafts are all crossed with sterilization treatment is washed into diameter under aseptic condition be 6 millimeters circular skin graft.Two kinds of circular skin grafts that prepare are placed above-mentioned each culture of strains ware simultaneously, and the culture dish of every kind of bacterium is divided into two sub-districts, places two kinds of skin grafts respectively, and then each culture of strains ware all being put into temperature is that 35 ℃ of incubators are cultivated.Cultivate after 24 hours, each culture dish is taken out, observe the inhibition zone of each strain, accurately measure the diameter of inhibition zone, and vertically take pictures in 0.5 meter height, every kind of bacterium reprocessing 20 times, every results averaged with digital camera.Experimental result shows: silver-containing pig acellular dermis is respectively 19 millimeters at the antibacterial circle diameter of staphylococcus aureus, escherichia coli, bacillus cloacae, klepsiella pneumoniae and bacillus pyocyaneus, 24 millimeters, 25 millimeters, 28 millimeters, 24 millimeters, and common pig acellular dermis is respectively 6 millimeters at the antibacterial circle diameter of each strain, 7 millimeters, 6 millimeters, 8 millimeters, 7 millimeters.This experimental result shows that fully the fungistatic effect of silver-containing pig acellular dermis is apparently higher than common pig acellular dermis.
Therefore, silver-containing pig acellular dermis of the present invention is owing to through the processing of active silver ion solution, have initiatively bacteriostatic effect, the infection rate after having reduced it and transplanting; Simultaneously, silver-containing pig acellular dermis of the present invention has not only removed original immunogenicity cell component in the substrate, the basic structure of skin histology and complete collagen fiber support have also been kept, the present invention simultaneously contains the silver ion with bacteriostasis, for clinical practice provides a kind of initiatively good dermal substitute of bacteria resistance function that has.
It should be noted last that, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can make amendment or be equal to replacement technical scheme of the present invention, and not break away from the spirit and scope of technical solution of the present invention.
Claims (9)
1, a kind of preparation method of silver-containing pig acellular dermis is characterized in that, comprising:
The Corii Sus domestica pretreatment;
Corii Sus domestica cuts open layer;
The skin graft sterilization;
Skin graft goes cell to handle;
The active silver-colored solution-treated of skin graft.
2, the preparation method of silver-containing pig acellular dermis according to claim 1 is characterized in that, described Corii Sus domestica is the fragrant Corii Sus domestica skin of the crust horse at 4-6 monthly age.
3, the preparation method of silver-containing pig acellular dermis according to claim 1 is characterized in that, described Corii Sus domestica pretreatment is specially:
The visual examination of raw material Corii Sus domestica;
Scrub described Corii Sus domestica;
Corii Sus domestica after scrubbing is scraped hair.
4, the preparation method of silver-containing pig acellular dermis according to claim 3, it is characterized in that, describedly scrub described Corii Sus domestica and be specially: the tow sides of repeatedly scrubbing described Corii Sus domestica with soap powder water are until cleaning, and the reuse percentage by weight is scrubbed described Corii Sus domestica for the 3-8% povidone iodine.
5, the preparation method of silver-containing pig acellular dermis according to claim 1 is characterized in that, described Corii Sus domestica cuts open layer and is specially: pretreated Corii Sus domestica is cutd open epidermis and hypodermis layer, obtain the skin graft that thickness is the 0.1-1.0 millimeter.
6, the preparation method of silver-containing pig acellular dermis according to claim 1 is characterized in that, described skin graft sterilization is specially: with the skin graft percentage by weight is 0.05-0.2% benzalkonium bromide rinsing 12-18 minute.
7, the preparation method of silver-containing pig acellular dermis according to claim 1 is characterized in that, described skin graft goes the cell processing to be specially: with the skin graft percentage by weight is that 0.1-0.2% triton x-100 solution was handled 22-26 hour down in temperature 22-28 ℃.
8, the preparation method of silver-containing pig acellular dermis according to claim 1, it is characterized in that the active silver-colored solution-treated of described skin graft is specially: with skin graft in being to soak 12-18 minute in the active silver-colored solution of 0.02-0.1% at percentage by weight under temperature 35-40 ℃.
9, a kind of silver-containing pig acellular dermis that adopts the preparation method preparation of the described silver-containing pig acellular dermis of arbitrary claim among the claim 1-8.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106474534A (en) * | 2016-09-22 | 2017-03-08 | 烟台正海生物科技股份有限公司 | A kind of collagen protein microfibre and preparation method thereof |
| EP3693029A1 (en) * | 2010-08-26 | 2020-08-12 | LifeCell Corporation | Passive methods for anti-microbial biological meshes |
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2008
- 2008-06-30 CN CN200810115916A patent/CN101618232A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3693029A1 (en) * | 2010-08-26 | 2020-08-12 | LifeCell Corporation | Passive methods for anti-microbial biological meshes |
| CN106474534A (en) * | 2016-09-22 | 2017-03-08 | 烟台正海生物科技股份有限公司 | A kind of collagen protein microfibre and preparation method thereof |
| CN106474534B (en) * | 2016-09-22 | 2019-10-25 | 烟台正海生物科技股份有限公司 | A kind of collagen microfibre and preparation method thereof |
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Application publication date: 20100106 |