CN114632183A - Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicines for reducing scar formation in skin repair - Google Patents
Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicines for reducing scar formation in skin repair Download PDFInfo
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- CN114632183A CN114632183A CN202210326732.4A CN202210326732A CN114632183A CN 114632183 A CN114632183 A CN 114632183A CN 202210326732 A CN202210326732 A CN 202210326732A CN 114632183 A CN114632183 A CN 114632183A
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- wound
- bfgf
- collagen
- protein sponge
- growth factor
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention provides a wound protein sponge, a preparation method thereof and application thereof in preparing a medicament for reducing scar formation in skin repair, and belongs to the technical field of biological medicaments. The invention relates to a wound protein sponge for skin repair, which is prepared by taking collagen as a carrier and a fibroblast growth factor (bFGF) as a main active ingredient. The wound protein sponge not only can repair damaged skin, but also can avoid the formation of scar tissues, and has stronger market competitiveness compared with similar wound repair products.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a wound protein sponge, a preparation method thereof and application thereof in preparing medicines for reducing scar formation in skin repair.
Background
Skin wounds and scalds are common clinical symptoms, and the skin, the dermis and the subcutaneous tissue are damaged due to external factors (cutting wounds, blunt injuries, accidental injuries, burns and scalds, chronic skin ulcers and the like) or internal factors (bedsores and the like), so that patients can feel hard to feel painful, severe patients can also cause skin tissue necrosis, severe skin ulceration can be caused by improper treatment, and wounds are difficult to heal. The skin is the largest organ of the human body, is the physical barrier between the human body and the external environment, and assumes many important physiological functions, including perspiration, heat sensation, and pain sensation. Once the skin is damaged, it causes a disturbance of the internal and external environment of the body, which is followed by malnutrition, multiple organ failure and even death.
The traditional wound repair products have certain limitations, for example, when the common products such as wound silica gel, wound spray and the like are used, the activity is greatly limited, and the use flow is complex; most products only have the functions of disinfecting, isolating external infection and inhibiting bacterial growth, and the time required for the effect is long; the price is high, the treatment course is long, the general economic level or the laggard people are difficult to bear, the traditional Chinese medicine composition is difficult to be suitable for various crowds, the use allergy rate of the crowds is high and is even higher than 0.005%; the method is easy to cause safety problems, difficult to effectively treat skin wounds of various wound surfaces and limited in application range to one or more wound surface types. Most importantly, in the process of repairing skin wounds, scars are often formed when the dermis or subcutaneous tissue is repaired, and the attractiveness of the repaired skin wound surface is seriously affected. Therefore, it is very important and necessary to find new materials for promoting the repair of skin wounds.
Disclosure of Invention
In view of the above, the present invention aims to provide an economical and safe wound protein sponge which can not only achieve skin repair but also reduce scar formation.
The invention provides a wound protein sponge, which comprises a fibroblast growth factor (bFGF) and collagen;
the mass ratio of the fibroblast growth factor bFGF to the collagen is (1-2): (8-12).
Preferably, the mass ratio of the fibroblast growth factor bFGF to the collagen is 1: 10.
preferably, the purity of the fibroblast growth factor bFGF is more than 93%.
Preferably, the preparation method of the fibroblast growth factor bFGF comprises the following steps: constructing a recombinant strain expressing a fibroblast growth factor (bFGF); and (3) separating and purifying bFGF protein after the recombinant strain is subjected to fermentation culture to obtain the recombinant protein bFGF.
Preferably, the purity of the collagen is 98.9 wt% or more.
Preferably, the preparation method of the collagen comprises the following steps:
and respectively treating the pigskin with acetic acid buffer solutions with the pH values of 2.5, 3.0, 3.5, 4.0 and 4.5, placing the pigskin in a water bath at the temperature of 55-65 ℃ for 1-2 h for each treatment, separating protein, and rinsing the protein with warm water at the temperature of 38-42 ℃ to obtain the collagen.
Preferably, the temperature of the water bath is 60 ℃, and the time of the water bath is 1.5 h.
The invention provides a preparation method of the wound protein sponge, which comprises the following steps:
adding a fibroblast growth factor (bFGF) solution into the collagen solution at the temperature of 4-8 ℃, mixing, and standing at the temperature of 1-3 ℃ to obtain the wound protein sponge.
The invention provides application of the wound protein sponge or the preparation method in preparing a medicine for reducing scar formation in skin repair.
The invention provides a wound protein sponge, which comprises a fibroblast growth factor (bFGF) and collagen; the mass ratio of the fibroblast growth factor bFGF to the collagen is (1-2): (8-12). The collagen can improve the metabolism of the skin, increase the blood circulation of the local skin, promote the inflammation absorption and dissipation of the traumatic skin, promote the generation of subcutaneous protein, fill the collapse of the damaged skin and repair the damaged skin; meanwhile, the fibroblast growth factor bFGF has the effects of promoting epidermal healing and reducing scar formation. In addition, compared with the single use of the fibroblast growth factor bFGF and the collagen, on one hand, the fibroblast growth factor can be uniformly adhered to the wound surface, and the healing time is shortened; on the other hand, the collagen can resist bacterial invasion and prevent infection, and the collagen play a positive feedback positive role mutually. Therefore, the wound protein sponge provided by the invention is wide in application range, and the raw materials are animal sources, so that the wound protein sponge has the characteristics of economy and safety.
Drawings
FIG. 1 is a standard curve of collagen, wherein the left graph shows a standard curve of collagen prepared by a conventional method, and the right graph shows a standard curve of collagen prepared in example 1;
FIG. 2 is an electrophoretogram of bFGF prepared by the present invention;
FIG. 3 is an SDS-PAGE electrophoresis of ion-exchanged purified bFGF prepared in the present invention;
FIG. 4 is a diagram showing the appearance of a wound protein sponge prepared according to the present invention;
FIG. 5 is a graph of wound healing after the use of sterilized water, collagen alone, fibroblast growth factor alone, and wound protein sponge, respectively;
FIG. 6 is a sectional view showing the growth of dermal tissue of damaged skin after sterilized water, collagen alone, fibroblast growth factor alone and a wound protein sponge, respectively.
Detailed Description
The invention provides a wound protein sponge, which comprises a fibroblast growth factor (bFGF) and collagen; the mass ratio of the fibroblast growth factor bFGF to the collagen is (1-2): (8-12).
In the present invention, the mass ratio of the fibroblast growth factor bFGF to the collagen is preferably 1: 10. the preparation method of the fiber-forming vitamin growth factor bFGF preferably comprises the following steps: constructing a recombinant strain expressing a fibroblast growth factor (bFGF); and (3) separating and purifying bFGF protein after fermentation culture of the recombinant strain. The nucleotide sequence of the fibroblast growth factor bFGF gene in the recombinant strain is shown as SEQ ID NO. 1; the separation and purification sequentially comprises Bio-Rex 70 ion exchange column chromatography, Heparin HyperD affinity column chromatography and Sephacryl S-100 gel filtration chromatography. The purity of the bFGF prepared by the method is greatly improved, and compared with the traditional bFGF preparation method, the purity of the bFGF prepared by the method is improved by 58 times. The purity of the bFGF prepared by the present invention is preferably 93% or more. The bFGF can promote the cells related to skin repair to divide, reproduce and regenerate rapidly, so that the active repair of the wound surface is promoted, the repair capability is in direct proportion to the purification rate of the bFGF, and the higher the purification rate of the bFGF is, the stronger the wound protein sponge has the effect of promoting the wound healing.
In the invention, the preparation method of the collagen comprises the following steps: and respectively treating the pigskin with acetic acid buffer solutions with the pH values of 2.5, 3.0, 3.5, 4.0 and 4.5, placing the pigskin in a water bath at the temperature of 55-65 ℃ for 1-2 h for each treatment, separating protein, and rinsing the protein with warm water at the temperature of 38-42 ℃ to obtain the collagen. The temperature of the water bath is preferably 60 ℃, and the time of the water bath is preferably 1.5 h. The method used by the invention can effectively reduce the loss of collagen, and the self-made glacial acetic acid is used for carrying out restrictive degradation on the collagen, namely, the terminal peptides are cut off, as the covalent bonds among the chains of the collagen peptides are formed by the interaction of lysine or hydroxylysine in the terminal peptides of the molecules, after the terminal peptides are cut off, the main part containing the triple-helix structure can be dissolved in dilute organic acid and extracted, the terminal peptides of collagen fiber can be hydrolyzed, and the yield of the collagen is improved; but also can not destroy the triple helix structure of the collagen and maintain the characteristics of the collagen. Meanwhile, compared with the traditional method for physically extracting collagen, the preparation method provided by the invention can obviously improve the purity, and the purity of the collagen prepared by the invention is preferably more than 98.9 wt%.
The invention provides a preparation method of the wound protein sponge, which comprises the following steps:
adding a fibroblast growth factor (bFGF) solution into the collagen solution at 4-8 ℃, mixing, and standing at 1-3 ℃ to obtain the wound protein sponge.
In view of the synergistic effect of the fibroblast growth factor (bFGF) and the collagen, which is helpful for repairing skin wounds, particularly reducing the formation of scars, the invention provides the application of the wound protein sponge or the preparation method in preparing the medicine for reducing the formation of scars in skin repair.
In the present invention, the medicament preferably further comprises a medically acceptable adjuvant. The mass percentage of the wound protein sponge in the medicine is 47%. The application method of the medicine is preferably to coat the wound surface with the wound protein sponge or the medicine prepared from the wound protein sponge, and replace the wound protein sponge or the medicine every two days until the wound surface is recovered to be healthy.
In the invention, the wound protein sponge and the medicine prepared from the same are suitable for various fresh wounds, preferably including wounds of mechanical injury of soft tissues, sharp injury, laser wound, wounds after medical cosmetology operation, operation wounds and residual cavities in parenchymal organ operation, filling of stumps, surrounding of anastomotic stoma of hollow visceral organs, nerve injury, and wounds of injury of cartilage and tendon; old wounds include diabetic ulcers, radiation ulcers, bed sores, fistula sinuses, mucosal ulcers and erosions and burn and frostbite.
The wound protein sponge provided by the present invention, its preparation method and its application in preparing a medicament for reducing scar formation in skin repair are described in detail below with reference to the examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Collagen extraction method
1. Reagent preparation
36% acetic acid solution: 39.18mL of glacial acetic acid and distilled water to 100 mL.
NaHSO3Solution: 4g NaHSO3Dissolved in 125mL of distilled water.
With NaHSO3The solutions were prepared into acetic acid buffers having pH values of 2.5, 3.0, 3.5, 4.0 and 4.5, and measured by a pH meter. Treating Corii Sus Domestica with 8.3mL of the above 5 kinds of acetic acid buffer solutions with different pH values, respectively, placing in a 60 deg.C constant temperature water bath kettle, water bathing for 1.5 hr, stirring to heat Corii Sus Domestica uniformly, rinsing with 40 deg.C distilled water twice to obtain Corii Sus DomesticaCollagen protein of (1).
The purity of the collagen is measured by an ultraviolet-visible spectrophotometry, and the method comprises the following steps:
5mg of the sample was added to the capped test tube, 5ml of 6mol/l hydrochloric acid solution was added, and after decomposing at 110 ℃ for 24 hours, distilled water was added to a volume of 25ml, water and hydrochloric acid were removed by rotary evaporation at 40 ℃, 30ml of distilled water was added to dissolve, 0.5ml of distilled water was absorbed to measure absorbance, and the concentration was calculated from a standard curve (right panel in FIG. 1).
Through purity measurement, the purity of the collagen extracted by the method is 98.9%.
Comparative example 1
The high pressure assisted physical method is used for extracting collagen from the pigskin, and the method comprises the following steps: cleaning pigskin, beating into homogenate, soaking with 50mmol/L ethanol solution for 1h, high pressure power 250MPa, processing time 10min, hot water treatment temperature 70 deg.C, hot water extraction time 4h, centrifuging (4000r/min) for 15min, concentrating, and drying.
The molecular structure of the collagen is destroyed under the high temperature condition, so that the collagen is denatured and precipitated, and the purity of the collagen extracted by the method is low (only 52 percent) and the conversion efficiency is low (only 43 percent).
Example 2
Preparation and purification method of fibroblast bFGF
Materials and methods:
1. engineering bacteria: the host bacterium is E.coliJM103 and contains a recombinant plasmid pUC-bFGF; the promoter is LacZ, and the cDNA sequence of human bFGF is behind the promoter, and the promoter can express fibroblast growth factor under IPTC induction and is purchased from Shanghai Zea leaf Biotech Co.
2. Cell: BALB/c 3T3 cells, provided by the Chinese drug biologicals assay.
3. Instruments and devices: fermentor, us NBS (40L); high speed refrigerated centrifuge, beckmann J2-CM, sonicator, US SLS-70, protein monitoring recorder, US ISCOIA 6
4. Seed culture: the culture medium comprises 1.0% tryptone, 1.0% yeast extract, 0.5% molasses, 0.4% NaCl, and 0.25% K2HPO4，0.1％KH2PO4The pH value is 7.2-7.4, and the final Amp concentration is 100 mu g/mL. Inoculating 0.1% of strain, shake culturing at 37 deg.C and 200r/min for 6 hr to obtain 1 generation strain, inoculating 5% of 1 generation strain, shake culturing at 37 deg.C and 200r/min for 8 hr to obtain 2 generation strain.
5. Fermentation culture: the culture medium is 1.6% tryptone, 1.6% yeast extract, and the other components are the same as the seed culture medium. 20L of culture medium is filled in a 40L tank, 10 percent of 2 generation bacteria are inoculated, and the culture is carried out at 37 ℃ and under the pH value of 7.0-7.5. Low-density culture: and adding IPTG (isopropyl thiogalactoside) at the beginning of fermentation until the final concentration of the IPTG is 50 mu g/mL, ventilating and stirring at the rotating speed of 400-500 r/min without adding sugar. High-density culture: continuously adding 5mmol/L glucose solution, adjusting the flow glucose adding speed according to the pH change to control the pH, adding IPTG after fermenting for 6 hours, controlling the ventilation stirring rotating speed at 500-600 r/min in the fermentation process, sampling every 1 hour, and adopting a BLISA method bFGF expression quantity, which specifically comprises the following steps: the bFGF antigen was diluted to 254g/mL with 0.01M carbonate buffer (pH 9.5), plated at 100. mu.L/well and incubated at 37 ℃ for 3 hours (or further incubated in a refrigerator at 4 ℃ overnight). Wash 3 times with heavy salt wash for 3 minutes each time. Serum diluent 1 for serum to be detected: 100 dilution, 100 u L/hole, 100L/hole, 37 degrees 1 hours incubation, washing 3 times, each time 3 minutes. Bio-RABG was diluted at a volume ratio of 1:400, added at a volume of 100. mu.L/well, incubated at 37 ℃ for 30 minutes, and washed 3 times for 3 minutes each. Diluting AVi-HRP at a volume ratio of 1:3000, adding 100 μ L/well, incubating at 37 deg.C for 30min, and washing for 3 times, each for 3 min; the substrate solution was added in an amount of 100. mu.L/well, and the reaction was carried out at 37 ℃ for 30 minutes with exclusion of light. Adding 2M H according to the amount of 50 mu 1/hole2SO4The reaction was terminated. OD value of each well is measured by DG-3022 type enzyme-linked immunosorbent assay instrument with wavelength of 450 nm. The OD values were recorded at the same time.
6. 10000r/min and 4 ℃ are used for continuous centrifugal separation, 8 times of buffer solution containing 0.1mol/L NaCl, 10mmol/L and 20mmol/L EDTA is used for suspending the harvested bacteria, after ultrasonic treatment at 4 ℃, 15000r/min and 30min are used for centrifugation, and the supernatant is the crude bFGF (target protein I).
7. And (3) purification:
7.1Bio-Rex 70 ion exchange column (XK50/30) chromatography:
and (2) balancing with 0.1mol/L NaC1 and 20mmol/L PB (pH 7.0) buffer solution, carrying out gradient elution (0.1-0.6 mol/L NaCl) with the buffer solution at the flow rate of 20mL/min, collecting samples of each section, and determining the target protein II through immunogenicity determination.
7.2Heparin HyperD affinity column (XK50/30) chromatography was equilibrated with 20mmol/LPB (pH9.0) buffer containing 0.6mol/L NaCl, gradient elution (0.6-2.0 mol/L NaCl) was performed with the same buffer at a flow rate of 15mL/min, and the target protein III was collected.
7.3Sephacryl S-100 gel filtration chromatography (2.6X 50 cm): and (3) balancing and eluting by using a buffer solution with the concentration of 20mmol/LPB (pH 7.0), and collecting the target protein IV, namely the pure bFGF fusion protein.
8. Detection of pure product
The method is carried out according to the requirements of the quality control key points of recombinant DNA products.
The results are shown in FIG. 2. The purity of bFGF prepared in this example was 93%.
9. Detection of biological Activity of bFGF Using MTT method
Fibroblast cell line NIH3T3 was cultured in complete medium in CO at 37 deg.C2Culture loop for controlling CO2The concentration was 5%. The cell concentration was controlled to 1ml containing 1.0X 105～5.0×105Culturing the individual cells for 24-36 hours, removing the culture solution in a culture bottle, digesting and collecting the cells, and preparing 1ml of the culture solution containing 5.0 multiplied by 104～1.0×105The cell suspension of each cell was inoculated in a 96-well cell culture medium in an amount of 100. mu.l per well at 37 ℃ with 5% CO2Culturing under the conditions of 24 hr, removing the complete culture medium by suction, and maintaining the culture medium at 37 deg.C and 5% CO2Starved culture under conditions for 24 hours.
The desalted hbFGF was prepared into different concentrations (2, 5, 10, 20, 50, 75, 100, 200ng/mL) with maintenance medium, replacing the original maintenance medium, 100. mu.L per well, 100. mu.L of complete medium cultured cells as positive control, and cells cultured in maintenance medium without hbFGF as negative control. Three parallel controls were performed for each concentration gradient at 37 ℃ with 5% CO2The cells were cultured under the conditions for 72 hours and the number of the cells was observed microscopically.
And (3) MTT method detection: mu.L of MTT solution was added to each well at 37 ℃ with 5% CO2The cell culture box is incubated for 4 h. Add 100. mu.L of formanzan lysis solution to each well and continue incubation in the cell incubator until formanzan is completely lysed under a normal light microscope and maintained at 37 ℃ for 4 h. The absorbance was measured at 570nm and the measurement was recorded.
The result of detecting the bioactivity of the hbFGF by using an MTT method shows that the very low concentration of hbFGF (2ng/mL) can promote the proliferation of fibroblasts, and the effect of promoting the proliferation of cells is improved in a certain range along with the increase of the concentration of hbFGF, when the concentration of hbFGF is 10ng/mL, the effect of promoting the proliferation of cells is the best, the total number of living cells is 54.1% higher than that of a negative control group without adding hbFGF, but the positive control group is lower than that of a positive control group, because the positive control group is added with 10% bovine serum complete culture solution, various nutritional factors are quite recombined, an experimental group only contains hbFGF single nutritional factor, but when the concentration of hbFGF is continuously increased, the effect of promoting the proliferation of cells is reduced, probably because the high concentration of hbFGF can generate the already-inhibited effect on the proliferation of cells, and a microscope can also show that the number of the experimental group added with hbFGF obviously has surplus cells only used for maintaining the culture solution, and less than cells cultured with complete medium, indicating that hbFGF did promote fibroblast proliferation.
Comparative example 2
The conventional method for preparing recombinant human bFGF is as follows:
(a) isolating RNA encoding human bFGF from the cells (SEQ ID NO:1, 5'-gaa ttc atc gaa ggt cgt cca gct ctg ccg gag gac gg-3');
(b) synthesizing a single-stranded Complementary DNA (CDNA) according to the RNA, and then synthesizing a corresponding double-stranded DNA;
(c) inserting the complementary DNA into a plasmid;
(d) transforming a host with the resulting recombinant plasmid;
(e) culturing the obtained transformant and isolating a plasmid containing the desired DNA;
(f) excising the desired cloned DNA from the plasmid;
(g) inserting the cloned DNA into a vector at a site downstream of a promoter to obtain an expression plasmid;
(h) transforming a host cell with the vector to obtain a transformant;
(i) culturing the transformant in a medium and recovering the produced and accumulated human bFGF in the culture medium.
The method has low specificity and low transformation efficiency. The purity of bFGF prepared by the method is 75%.
Therefore, the specific antibody of the fibroblast growth factor bFGF is adopted to purify the fusion protein expression vector and the expression strain, the specificity is high, and the purification rate of the bFGF is improved by 58 times compared with the traditional extraction method.
Example 3
Adding a fibroblast growth factor (bFGF) solution into the collagen solution at the temperature of 8 ℃, mixing until the mass ratio of the collagen to the bFGF is 10:1, and standing at the temperature of 2 ℃ to obtain the wound protein sponge.
The appearance of the wound protein sponge is shown in fig. 3. As can be seen from FIG. 3, the wound protein sponge is white in appearance, has a certain elasticity, and has a specification of 8cm × 10 cm.
Comparative example 3
A wound protein sponge was prepared according to the method of example 3 using the collagen prepared in comparative example 1 and the bFGF prepared in comparative example 2.
Example 4
Skin repairing method using wound protein sponge
The mouse simulation wound is treated by respectively using wound protein sponge, a fibroblast growth factor and collagen, the medicine is replaced every 2 days, and the wound growth condition after no treatment is set. After 3, 5, 7, 9, 11 days of application, the wound condition was observed and recorded. The skin tissues after 3, 5, 7, 9 and 11 days of drug coating are sliced and subjected to HE staining, and the growth condition of the skin dermal tissues is observed.
Wound healing was as shown in figure 4. As can be seen from fig. 4, compared with the group without any treatment, the wound sponge, the fibroblast growth factor bFGF alone and the collagen alone can effectively promote the repair of the wound, and the wound treated by the wound sponge has a faster healing speed than the wound treated by the fibroblast growth factor alone and the collagen alone, and no obvious scar is formed on the wound.
The section of the damaged dermal tissue is shown in FIG. 5. As can be seen from fig. 5, the growth recovery rate of dermal tissue of skin is ranked from fast to slow in the wound protein sponge group > bFGF group > collagen group > without any treatment group, and the overall recovery condition is consistent with the apparent recovery condition of wound.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Wenzhou university of medical science
<120> a wound protein sponge, a preparation method thereof and application thereof in preparing medicines for reducing scar formation in skin repair
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 38
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
gaattcatcg aaggtcgtcc agctctgccg gaggacgg 38
Claims (9)
1. A wound protein sponge comprising a fibroblast growth factor, bFGF, and collagen;
the mass ratio of the fiber-forming vitamin growth factor bFGF to the collagen is (1-2): (8-12).
2. The wound protein sponge according to claim 1, wherein the mass ratio of the fibroblast growth factor bFGF to the collagen is 1: 10.
3. the wound protein sponge according to claim 1 or 2, wherein the purity of the fibroblast growth factor, bFGF, is 93% or more.
4. The wound protein sponge according to claim 3, wherein the process for preparing the fibroblast growth factor, bFGF, comprises the steps of:
constructing a recombinant strain expressing a fibroblast growth factor (bFGF); and (3) separating and purifying bFGF protein after the recombinant strain is subjected to fermentation culture to obtain the recombinant protein bFGF.
5. The wound protein sponge according to claim 1 or 2, wherein the collagen has a purity of 98.9 wt% or more.
6. The wound protein sponge according to claim 5, wherein the collagen is prepared by a method comprising the steps of:
and respectively treating the pigskin with acetic acid buffer solutions with the pH values of 2.5, 3.0, 3.5, 4.0 and 4.5, placing the pigskin in a water bath at the temperature of 55-65 ℃ for 1-2 h for each treatment, separating protein, and rinsing the protein with warm water at the temperature of 38-42 ℃ to obtain the collagen.
7. The wound protein sponge according to claim 6, wherein the temperature of the water bath is 60 ℃ and the time of the water bath is 1.5 h.
8. A method of preparing a wound protein sponge according to any one of claims 1 to 7, comprising the steps of:
adding a fibroblast growth factor (bFGF) solution into the collagen solution at 4-8 ℃, mixing, and standing at 1-3 ℃ to obtain the wound protein sponge.
9. Use of a wound protein sponge according to any one of claims 1 to 8 in the manufacture of a medicament for reducing scarring in the repair of skin.
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| ZA2022/05323A ZA202205323B (en) | 2022-03-30 | 2022-05-13 | Collagen-based sponge for trauma, preparation method thereof, use thereof in preparation of medicament for reducing scar formation in skin repair |
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