Composition for preparing keratin gel dressings as well as preparation method and application thereof

A composition and keratin technology, applied in medical science, bandages, etc., can solve the problems of difficult to obtain and single keratin products, and achieve the effect of preventing secondary wound damage, easy preparation, and promoting wound healing

Active Publication Date: 2020-05-29
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is difficult to obtain a single keratin

Method used

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  • Composition for preparing keratin gel dressings as well as preparation method and application thereof
  • Composition for preparing keratin gel dressings as well as preparation method and application thereof
  • Composition for preparing keratin gel dressings as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0043] Example 1

[0044] Keratin preparation:

[0045] Construct a recombinant plasmid containing the target gene fragment of recombinant keratin, and transfer the recombinant plasmid to E. coli BL21(DE3). The transformed bacteria were inoculated into sterile LB-kana (50ug / mL) liquid medium at a ratio of 1:5000, and cultured overnight at 170 rpm and 37°C. When the OD value was 0.8-1.0, the temperature was reduced to 25°C, 170rpm, and IPTG with a final concentration of 1mM was used for induction. After 8 hours of induction, the bacteria were harvested by centrifugation at 4°C and 8000 rpm for 5 minutes. Resuspend the bacterial cells with 5 times the volume of Buffer A (50mM Tris pH8.0, 150mM NaCl, 5mM EDTA, 20mM β-mercaptoethanol) of the bacterial weight. Homogeneously break at 1000 bar, centrifuge at 20000 rpm at 4°C for 1 hour, and discard the supernatant. Resuspend the pellet with 10 times the volume of the pellet weight in Buffer A, ultrasonicate for 3 min, centrifuge at 10...

Example Embodiment

[0046] Example 2

[0047] Preparation of gel dressing:

[0048] Dissolve 1 part by weight of sodium carboxymethyl cellulose and 5 parts by weight of hydroxyethyl cellulose with 10 parts by weight of water, and let it stand overnight to allow it to swell naturally and stir evenly; take 0.05 parts by weight of keratin and suspend it In 5 parts by weight of water, add to the above liquid and stir evenly, take 50 parts by weight of sorbitol, add 50 parts by weight of water to dissolve, add to the upper liquid and stir, then add 50 parts by weight of glycerin and 1 part by weight of phenoxyethanol solution in turn, Finally, add 50 parts by weight of water to the final volume and mix well to obtain.

[0049] Dissolve 10 parts by weight of sodium carboxymethyl cellulose and 50 parts by weight of hydroxyethyl cellulose with 60 parts by weight of water, and let it stand overnight to allow it to swell naturally and stir evenly; take 2 parts by weight of keratin and suspend it In 5 parts by w...

Example Embodiment

[0050] Example 3

[0051] Cytotoxicity test: To detect the effect of different concentrations of keratin gel dressing on cell viability.

[0052] Keratin concentration: 0.05mg / ml-0.8mg / ml gradient setting (96well-100ul medium) after 24 treatments, add 20ul of MTT solution (5mg / ml prepared with PBS) to each well. Continue to incubate for 4 hours to terminate the culture, and carefully aspirate the culture supernatant in the well. Add 150ul DMSO to each hole and shake for 10 minutes to fully melt the crystals. Select 490nm wavelength, measure the light absorption value of each hole on the microplate reader, record the result and analyze, as attached figure 1 Shown. Cytotoxicity experiments showed that the recombinant keratin had no toxic effect on the growth of fibroblasts at 24h.

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Abstract

The invention relates to a composition for preparing keratin gel dressings as well as a preparation method and application thereof. The composition consists of 0.01 part to 5 parts of keratin, 80 parts to 500 parts of auxiliaries and water medium. The gel dressings prepared by adopting the method disclosed by the invention can effectively stop bleeding, quicken healing of wounds and isolate infection of bacteria, and have no toxic and side effects. The product can be volatilized and absorbed, and a step of disposal of the medicine again is avoided; in a practical repeated using process, an appropriate amount of dressings are only applied to the wounds, and therefore, the wounds cannot be hurt again due to disposal of residual medicines and too much pain is avoided. The invention also provides a method for preparing the gel dressings according to the components of the composition; and preparation or production procedures are not complicated, so that production processing of the medicines is facilitated.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a composition for preparing a keratin gel dressing, a preparation method and application thereof. Background technique [0002] As a non-toxic natural biomaterial, keratin has been successfully applied in tissue repair, because it contains abundant cysteine ​​residues, has the property of spontaneous assembly and polymerization, and can be made into various types of wound dressings. As a protein source for humans, its excellent biocompatibility and good biodegradability further broaden its application in regenerative medicine. Traditional human hair keratin is mostly extracted by chemical or enzymatic extraction methods. Therefore it has the problems of complex components and difficult quality control. Human hair contains not only keratin, but also abundant keratin-related proteins and intermediate filament proteins, etc. It is difficult to obtain a single kera...

Claims

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Application Information

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IPC IPC(8): A61L26/00
CPCA61L26/0095A61L26/0066C08L1/286
Inventor 黄玉钱王伯初郝石磊查国栋余钡
Owner CHONGQING UNIV
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