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Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicine for reducing scar formation in skin repair

A protein and trauma technology, applied in the field of biopharmaceuticals, can solve problems such as safety issues, long time required for onset of effects, and long course of treatment

Active Publication Date: 2022-06-17
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]Traditional wound repair products have certain limitations, such as common wound silicone gel, wound spray and other products, when used, have a greater impact on activities Restrictive, and the use process is more complicated; most products only play the role of disinfection and isolation of external infections, and inhibit the growth of bacteria, and it takes a long time to take effect; the price is high, the course of treatment is long, and the economic level is average or backward It is difficult for the public to afford it, and it is difficult to apply to various groups of people. The allergy rate of the population is relatively high, and some are even higher than 0.005%. It is easy to cause safety problems, and it is difficult to effectively treat skin wounds on various wounds. The scope of application is limited to a certain type or several wound types
Most importantly, in the process of skin wound repair, scars are often formed when the repair reaches deep into the dermis or subcutaneous tissue, which seriously affects the appearance of the repaired skin wound

Method used

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  • Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicine for reducing scar formation in skin repair
  • Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicine for reducing scar formation in skin repair
  • Wound protein sponge, preparation method thereof and application of wound protein sponge in preparation of medicine for reducing scar formation in skin repair

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preparation example Construction

[0025] In the present invention, the mass ratio of the fibroblast growth factor bFGF and collagen is preferably 1:10. The preparation method of the fibroblast growth factor bFGF preferably includes the following steps: constructing a recombinant strain expressing the fibroblast growth factor bFGF; isolating and purifying the bFGF protein after the recombinant strain is fermented and cultured. The nucleotide sequence of the fibroblast growth factor bFGF gene in the recombinant strain is as SEQ ID NO: 1; the separation and purification include Bio-Rex 70 ion exchange column chromatography, Heparin HyperD affinity column chromatography and Sephacryl S in turn -100 gel filtration chromatography. The bFGF prepared by the present invention greatly improves the purity of the fibroblast growth factor bFGF. Compared with the traditional method for preparing bFGF, the purity of the bFGF prepared by the present invention is increased by 58 times. The purity of the bFGF prepared by the p...

Embodiment 1

[0034] A kind of collagen extraction method

[0035] 1. Reagent preparation

[0036] 36% acetic acid solution: 39.18 mL of glacial acetic acid and distilled water to 100 mL.

[0037] NaHSO 3 Solution: 4g NaHSO 3 Dissolve in 125mL distilled water.

[0038] with NaHSO 3 The solutions were prepared with acetate buffers with pH 2.5, 3.0, 3.5, 4.0 and 4.5, and were measured with a pH meter. The pigskin was treated with 8.3 mL of the above 5 kinds of acetic acid buffers with different pH values, and placed in a constant temperature water bath at 60 °C for 1.5 h, stirring continuously during this period to make the pigskin evenly heated, and rinsed twice with 40 °C distilled water. Obtain collagen in pigskin.

[0039] The purity of collagen was determined by UV-Vis spectrophotometry as follows:

[0040]Add 5mg of sample to the capped test tube, add 5ml of 6mol / l hydrochloric acid solution, decompose at 110°C for 24h, dilute the volume to 25ml with distilled water, remove water...

Embodiment 2

[0046] Preparation and purification method of fibroblast factor bFGF

[0047] 1. Materials and methods:

[0048] 1. Engineering bacteria: the host bacteria is E.coliJM103, which contains the recombinant plasmid pUC-bFGF; the promoter is LacZ, followed by the cDNA sequence of human bFGF, which can express fibroblast growth factor under the induction of IPTC, purchased from Shanghai Zeye Biotechnology Co., Ltd.

[0049] 2. Cells: BALB / c 3T3 cells, provided by the China Pharmaceutical and Biological Products Laboratory.

[0050] 3. Instruments and equipment: Fermentation tank, NBS (40L) in the United States; high-speed refrigerated centrifuge, beckman J2-CM, ultrasonic mill, SLS-70 in the United States, protein monitoring recorder, ISCOUA6 in the United States

[0051] 4. Seed culture: the medium is 1.0% tryptone, 1.0% yeast extract, 0.5% molasses, 0.4% NaCl, 0.25% K 2 HPO 4 , 0.1%KH 2 PO 4 , pH 7.2~7.4, Amp final concentration 100μg / mL. Inoculate 0.1% bacteria at 37°C, 20...

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Abstract

The invention provides a wound protein sponge, a preparation method thereof and an application of the wound protein sponge in preparation of a medicine for reducing scar formation in skin repair, and belongs to the technical field of biological medicines. The wound protein sponge for skin repair is prepared by taking collagen as a carrier and taking a fibroblast growth factor bFGF as a main active ingredient. The trauma protein sponge not only can repair damaged skin, but also can avoid formation of scar tissues, and has higher market competitiveness compared with similar trauma repair products.

Description

technical field [0001] The invention belongs to the technical field of biological drugs, and particularly relates to a wound protein sponge, a preparation method thereof, and an application in the preparation of a medicament for reducing scar formation in skin repair. Background technique [0002] Skin wounds and scalds are common clinical diseases. The epidermis, dermis and subcutaneous tissue are damaged due to external causes (cut wounds, blunt trauma, accidental injuries, water and fire burns, chronic skin ulcers, etc.) or internal causes (such as bedsores, etc.) The pain is unbearable for patients, and in severe cases, skin tissue necrosis can be caused. Improper handling can lead to severe skin ulceration, and the wound is difficult to heal. The skin is the largest organ of the human body, a physical barrier between the human body and the external environment, and undertakes many important physiological functions, including perspiration, heat sensation and pain sensati...

Claims

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Application Information

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IPC IPC(8): A61L15/32A61L15/44A61L15/42A61K38/18A61K38/01A61P17/02C07K14/78C07K1/12C07K1/14
CPCA61L15/32A61L15/325A61L15/44A61L15/425A61K38/1825A61K38/014A61P17/02C07K14/78A61L2300/412A61L2300/414A61L2300/252C08L89/00A61K2300/00
Inventor 詹姆斯·柯提斯·德林周蕊
Owner WENZHOU MEDICAL UNIV
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