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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 product using traditional extraction methods

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

Embodiment 1

[0044] Keratin preparation:

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

Embodiment 2

[0047] Preparation of gel dressing:

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

[0049] Dissolve 10 parts by weight of sodium carboxymethylcellulose and 50 parts by weight of hydroxyethyl cellulose with 60 parts by weight of water, let it stand overnight, let it swell naturally, and stir evenly; take 2 parts by weight of keratin to suspend Add 5 parts by weight of water to the above liquid an...

Embodiment 3

[0051] Cytotoxicity experiment: 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 in PBS) to each well. Continue to incubate for 4 hours, terminate the culture, and carefully aspirate and discard the culture supernatant in the well. Add 150ul DMSO to each well and shake for 10 minutes to fully melt the crystals. Select a wavelength of 490nm, measure the light absorption value of each well on a microplate reader, record and analyze the results, as shown in the attached figure 1 shown. Cytotoxicity experiments showed that the recombinant keratin had no toxic effect on the growth of fibroblasts at 24 hours.

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PUM

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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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