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Application of procyanidin SA hydrogel in preparation of medicine for treating tendon ectopic calcification

A proanthocyanidin and hydrogel technology, used in drug combinations, pharmaceutical formulations, muscular system diseases, etc., can solve the problems of high incidence of re-rupture, hinder the research and development of effective drugs for tendinopathy, and inability to rebuild mechanical structure, and achieve sustained release. Effects of drugs, inhibition of tendon ectopic calcification, and economic cost reduction

Inactive Publication Date: 2020-04-03
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After long-term excessive load-bearing and tendon micro-tears, the tendon can only be repaired with scars, and the mechanical structure cannot be reconstructed, and the incidence of re-rupture is high
The current lack of effective treatments and unclear pathogenesis hampers the development of effective drugs for tendinopathy

Method used

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  • Application of procyanidin SA hydrogel in preparation of medicine for treating tendon ectopic calcification
  • Application of procyanidin SA hydrogel in preparation of medicine for treating tendon ectopic calcification
  • Application of procyanidin SA hydrogel in preparation of medicine for treating tendon ectopic calcification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Screening and isolation of human tendon stem progenitor cells (hTSPCs).

[0022] Extract hTSPCs from human embryonic tendon samples, soak the tissue samples in 5x-PS and 2x-PS for 2 minutes, cut the tissue with sterile instruments, digest with 4ml 0.2% collagenase, after 2 hours, only two strands remain, 3 Hours later, it was basically digested. Centrifuge at 2000rpm for 5 minutes, resuspend in 1ml, culture medium with 10% fetal bovine serum, 1% penicillin and streptomycin, at 37°C, 5% CO 2 cultured in an incubator. like figure 1 Shown are hTSPCs isolated from successful screening.

Embodiment 2

[0023] Example 2: CCK-8 detects the effects of proanthocyanidins (PC) on the cytotoxicity and proliferation of tendon stem / progenitor cells.

[0024] After hTSPCs were resuscitated and passaged, the logarithmic phase cells were inoculated in a 96-well plate with cell suspension (100ul / well) at a density of 10^4-10^5 / ml. The specific grouping is as follows: 1. Normal control group (Control): Cell culture is carried out with the low-sugar DMEM medium containing sodium alginate PBS solution of the same amount as the experimental group interleukin 1 (IL-1β); 2. The experimental group : (1) Interleukin 1 experimental group (IL group): add 5ng / ml IL-1β low-sugar DMEM medium for cell culture to simulate the inflammatory environment of tendinopathy; (2) Add different concentrations of proanthocyanidin group : Add 5ng / ml IL-1β under the low-sugar DMEM medium culture condition, add 2, 4, 6, 8ug / ml proanthocyanidins respectively for cell culture, named as IP2, IP4, IP6, IP8 group respect...

Embodiment 3

[0025] Example 3: The optimal concentration of proanthocyanidins to inhibit inflammation of tendon stem / progenitor cells (hTSPCs) was screened by alkaline phosphatase (ALP) and alizarin red (ARS) staining experiments.

[0026] After recovery, hTSPCs were inoculated in low-glucose DMEM medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, at 37°C, 5% CO 2 Place the culture in the incubator, and replace the culture once every 3 days to restore vitality.

[0027] 10,000 cells / well for ALP staining and 5,000 cells / well for ARS staining were seeded in a 24-well plate and 500ul of low-sugar DMEM medium was added. After most cells adhered to the wall, the medium was replaced with an osteogenic induction system every 3 days. ALP induced culture for 7 days, ARS induced culture for 15 days, stained with alkaline phosphatase kit and alizarin red.

[0028] The specific experimental plan for alkaline phosphatase staining: 1. Aspirate the medium, fix with 500ul of 4% pa...

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Abstract

The invention discloses an application of procyanidin in regulation and control of stem cell functions in a pathological microenvironment of tendon diseases, and provides a preparation method of procyanidin SA hydrogel and an application of the procyanidin SA hydrogel in treatment of tendon ectopic calcification. Based on the application, the invention proves that: the addition of the procyanidininto tendon stem / progenitor cells does not influence cell proliferation, and can effectively inhibit the calcium deposition of the tendon stem / progenitor cells and the subsequent potential for treating tendon diseases. Through Micro CT, hematoxylin-eosin staining, crocus sativus-solid green staining, collagen II and osteocalcin (OCN) immunohistochemical staining, the situation of the procyanidin SA hydrogel for treating tendon ectopic calcification is detected. The invention finds and proves that the procyanidin SA hydrogel can effectively inhibit tendon ectopic calcification. Meanwhile, the administration route of sustained-release administration can reduce corresponding side effects generated by systemic administration while improving the local drug concentration, and has a clinical application prospect.

Description

technical field [0001] The invention relates to a new application of proanthocyanidins, in particular to the application of proanthocyanidins to regulate stem cell function in the pathological microenvironment of tendinopathy. The preparation method of the proanthocyanidin SA hydrogel is provided, and the application of the proanthocyanidin SA hydrogel in the preparation of drugs for treating ectopic calcification of tendon is clarified. Background technique [0002] Tendon is an important tissue in the human body, and tendinopathy is a common disease of the musculoskeletal system. After long-term excessive weight-bearing and micro-tears of the tendon, the tendon can only be repaired with scars, and the mechanical structure cannot be reconstructed, and the incidence of re-rupture is high. Currently, there is a lack of effective treatments, and its pathogenesis is still unclear, which hinders the development of effective drugs for the treatment of tendinopathy. In the infla...

Claims

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Application Information

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IPC IPC(8): A61K31/353A61K9/06A61K47/02A61K47/18A61K47/36A61P19/00A61P21/00
CPCA61K9/06A61K31/353A61K47/02A61K47/183A61K47/36A61P19/00A61P21/00
Inventor 茵梓刘日纯陈晓张鸿常江
Owner ZHEJIANG UNIV
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