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Optimized anti-oxidative accellular protective solution

A decellularization and protection solution technology, applied in the field of optimized antioxidant decellularization protection solution and its preparation, can solve the problems of low self-degradation performance and poor histocompatibility, etc.

Active Publication Date: 2016-12-07
拜欧迪赛尔成都生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, artificially synthesized polymer materials are mostly used in clinical tissue transplantation and repair, which often have the problems of low self-degradation performance and poor tissue compatibility. urgent

Method used

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  • Optimized anti-oxidative accellular protective solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Preparation of anti-oxidant decellularization protection solution:

[0019] 1. Take 10g of DMEM powder and add 500ml of deionized water, dissolve and autoclave;

[0020] 2. Take 25g of chondroitin sulfate and add 275ml of deionized water to heat to dissolve and autoclave to make a chondroitin sulfate solution; add 20g of low molecular dextran to 100ml of water for injection to dissolve and autoclave to make a low molecular dextran solution; L -2.87g histidine hydrochloride, add 100ml water for injection to dissolve and autoclave to make L-histidine hydrochloride solution;

[0021] 3. Take a sterile container and add 500ml of DMEM culture solution, autoclave chondroitin sulfate 275ml, low molecular dextran solution 100ml, L-histidine hydrochloride solution 100ml; add 0.5g allopurinol, hydroxypropyl methyl fiber Sodium 5g, reduced glutathione 2g, dexamethasone injection 2mg, levofloxacin injection 0.1g and mix well;

[0022] 4. Adjust the PH value of Hepes buffer to 7.4;

[0023]...

Embodiment 2

[0026] Observation and detection of decellularization effect and tissue structure of porcine tendon tissue with decellularization protective liquid

[0027] Take out the tendon tissue within 3 hours after the fresh pig is slaughtered, and treat it aseptically, cut the aponeurosis to make 1cm×2cm tissue pieces, take 5 pieces and seal them in a plastic bag containing 10ml of the preservation solution prepared in Example 1; In the other control group, 5 tissue pieces were sealed in a plastic bag containing 10ml BSS. Under the condition of 600MP high static pressure, treat 8 times, each time is 3 minutes; after taking out the ligament, place it in a protection solution containing 0.2% sodium lauryl sulfate + 1000U / ml DNA enzyme at a temperature of 25 ℃, set the shaker speed to 100 revolutions / min, and process for 2 hours; take out the decellularized tendon and rinse in the protective solution for 2 hours. The enzymes and detergents of the control group and the final rinsing treatmen...

Embodiment 3

[0030] Observation and detection of decellularization effect and tissue structure of conjunctival tissue using decellularization protective liquid

[0031] Take out the conjunctival tissue of fresh pigs within 2 hours after slaughter, remove the subconjunctival tissue as much as possible, and perform aseptic processing. Cut the conjunctival tissue into 0.5cm×1cm tissue pieces, of which 5 pieces of conjunctival tissue are sealed in 10ml In the plastic bag of the preservation solution prepared in Example 1; in the control group, 5 pieces of conjunctival tissue were sealed in a plastic bag containing 10 ml of BSS. Under the condition of 300MP high static pressure, treat 3 times, each time is 1 minute; after taking out the conjunctival tissue, place it in the protective solution containing 0.2% sodium lauryl sulfate + 1000U / ml DNA enzyme, temperature Set the shaker speed to 100 revolutions per minute at 25°C, and treat for 1 hour; take out the decellularized conjunctiva and rinse it ...

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Abstract

The invention relates to an optimized anti-oxidative accelular protective solution, which has a critical effect on protection of structural completeness and biological nature of acellular tissues in the whole course of the decellularizing process. The protective solution is prepared from DMEM cellular culture medium L-histidine hydrochloride, allopurinol, chondroitin sulfate, low molecular dextran, hydroxypropyl methyl cellulose, HEPES buffer solution, dexamethason hydrochloride, reduced glutathione and levofloxacin. The protective solution is a light-red liquid with specific pH value, specific crystal and colloid osmotic pressure. The optimized anti-oxidative accelular protective solution is easily available in raw materials and economical in price, has strong applicability, wide application range and excellent anti-oxidation performance, and has a protective effect in the process of decellularizing biological tissues.

Description

Technical field [0001] The invention belongs to the technical field of chemical compositions, and specifically relates to an optimized anti-oxidation decellularization protection solution and a preparation method and application thereof. Background technique [0002] The 21st century is a century of new developments in biological sciences. As a rising sun in the field of biomedicine in the past 10 years, TERM (Tissue engineering & Regenerative Medicine) has attracted worldwide attention. In the research and application of TERM, related decellularization technology occupies a pivotal position. Decellularization refers to the process of separating extracellular mesenchyme from cells in the field of biomedical engineering. The separated extracellular mesenchymal scaffold can be used for artificial organ and tissue regeneration. The use of physical, chemical, enzymatic treatment and a mixture of the three is the currently widely used decellularization method worldwide. Its purpose i...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 史真史伟云
Owner 拜欧迪赛尔成都生物科技有限公司
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