Preparation method and application of decellularized fishskin substrates

A decellularized dermis and decellularized technology, applied in medical science, bandages, absorbent pads, etc., can solve the problems of poor thermal stability of collagen, limited source of raw materials, cumbersome production process, etc., to achieve low production costs, reduce medical costs, Effects that improve clinical utility and safety

Inactive Publication Date: 2020-05-01
烟台蓝创生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, at present, cod has not yet achieved large-scale artificial breeding, and the raw materials of wild cod have poor traceability, so the source of raw materials for medical products prepared from cod is very limited. In addition, cod is a deep-sea fish, and its collagen poor thermal stability
In addition, the production process described in patent WO 2011 / 042794 is cumbersome and does not include virus inactivation process steps. Therefore, the development of a clean fish collagen product with a large supply of raw materials, thermal stability and close to mammals has become the current fish collagen industry. problems that urgently need to be solved

Method used

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  • Preparation method and application of decellularized fishskin substrates
  • Preparation method and application of decellularized fishskin substrates
  • Preparation method and application of decellularized fishskin substrates

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 A kind of preparation method of decellularized fish skin matrix

[0035] 1. Pretreatment

[0036] Take 100g of fresh tilapia skin to remove scales, residual meat and fat, soak it in 1% NaCl at 15°C for 6 hours, the ratio of solid to liquid is 1:10 (w / v), repeat once, and drain. This step is mainly to remove the stains on the surface of the fish skin mechanically, which is beneficial to the subsequent operation.

[0037] 2. Scrape off the scales

[0038] First, take a piece of filter paper to fully soak it in 0.1% acetic acid solution, then spread the fish skin on the filter paper, make the side containing the scales fully contact with the filter paper, take it out after 15 minutes, scrape off the black scales with a knife , until the scales are basically scraped off, rinsed with pure water for 3 times, and then temporarily stored in a phosphate buffer solution with a pH of 7.4.

[0039] 3. Surfactant washing

[0040] Drain the fish skin in the previous ...

Embodiment 2

[0047] Embodiment 2 A kind of preparation method of decellularized fish skin matrix

[0048] 1. Pretreatment

[0049] Thaw 100 g of frozen tilapia skin, soak it with 5% NaCl at 10°C for 0.5 hour, the ratio of solid to liquid is 1:30 (w / v), repeat once, and drain.

[0050] 2. Scrape off the scales

[0051] First, take a piece of filter paper to fully soak it in 3% citric acid solution, then spread the fish skin on the filter paper, make the side containing the scales fully contact with the filter paper, take it out after 1 minute, and scrape off the black scales with a knife until the scales are basically scraped off, rinsed with pure water for 3 times, and then temporarily stored in MES buffer at pH 6.0.

[0052] 3. Surfactant washing

[0053] Add 1% Tween 80 (w / w) solution to the fish skin, the ratio of material to liquid is 1:30 (w / v), shake at 10°C and 250rpm for 2 hours, drain the water, rinse 3 times with pure water, and drain .

[0054] 4. Alkaline washing

[0055]...

Embodiment 3

[0060] Embodiment 3 A kind of preparation method of decellularized fish skin matrix

[0061] 1. Pretreatment

[0062] Take 100g of new tilapia skin to remove scales, residual meat and fat, soak in 3% KCl for 12 hours at 20°C, the ratio of solid to liquid is 1:20 (w / v), repeat once, and drain.

[0063] 2. Scrape off the scales

[0064] First, take a piece of filter paper to fully soak it in 0.6% acetic acid solution, then spread the fish skin on the filter paper, make the side containing the scales fully contact with the filter paper, take it out after 5 minutes, scrape off the black scales with a knife , until the scales are basically scraped off, rinsed with pure water for 3 times, and temporarily stored in Tris-Hcl buffer solution with pH 8.0.

[0065] 3. Surfactant washing

[0066] Add 0.3% sodium deoxycholate solution to the fish skin, the ratio of solid to liquid is 1:20 (w / v), shake at 20°C and 175rpm for 48 hours, drain the water, rinse with pure water 3 times, and d...

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Abstract

The invention discloses a preparation method of novel decellularized fishskin substrates. The method comprises the following steps of (1) performing pretreatment of washing Tilapia skin, removing impurities and the like; (2) after acid treatment, scraping scales; (3) performing washing with a surfactant for decellularization; (4) performing washing with alkali liquor; (5) performing treatment witha chemical bleaching agent; and (6) performing post-treatment of drying, sterilization and the like. The decellularized fishskin substrates prepared by the preparation method disclosed by the invention have the advantages of being low in production cost, high in heat stability, free from zoonoses virus propagation risk and the like, and besides, a scale removing technique is also innovatively provided, so that the clinical practicality of products is greatly increased; and various techniques are comprehensively applied for virus inactivation, so that the biological safety of the products is increased. Besides, the invention discloses an application of the decellularized fishskin substrates as wound surface dressing, so that wound healing and regenerated tissue epithelization can be promoted.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and relates to a preparation method of a novel decellularized fish skin matrix material and its application as a wound dressing. Background technique [0002] The correct treatment of surgical wounds is one of the keys to the success of surgical treatment, and good wound healing is the prerequisite for the recovery of body functions after trauma. Therefore, research on accelerating wound healing is very important, and the development of wound dressings is one of the hotspots of research. one. With the in-depth research on wound healing, people realize that the purpose of using dressings is far more than just to cover the wound surface, and the dressing must also help the wound heal. Therefore, the development of functional dressings with wound healing or hemostatic activity has become a current research hotspot. At present, there are many brands of wound dressings used clinically in China, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L15/40A61L15/42
CPCA61L15/40A61L15/42
Inventor 李东升谢则平张淑敏李诚博宫世周曲承蕾申胜标宋国梁
Owner 烟台蓝创生物技术有限公司
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