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Application of Bromide Ion in Cartilage Tissue Engineering

A technology of cartilage tissue and bromide ions, applied in bone/connective tissue cells, medical science, prosthesis, etc., can solve the problems of increased subchondral bone density, secondary articular cartilage degeneration, and inability to significantly improve joint pain, etc. Achieve the effects of promoting cartilage repair, broad application prospects, and excellent in vitro biological activity

Active Publication Date: 2021-07-27
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficacy of glucosamine and chondroitin sulfate in the treatment of osteoarthritis has been questioned: in 2006, Clegg DO of Utah State University School of Medicine in the United States found that glucosamine and chondroitin sulfate alone or in combination could not significantly reduce knee joint pain. Pain Symptoms in Patients with Inflammation
In 2010, Simon Wandel of the University of Bern in Sweden found that compared with placebo, glucosamine monotherapy can better reduce joint pain, but there is no clinically significant difference; while chondroitin sulfate monotherapy and glucosamine and sulfuric acid Neither combination therapy with chondroitin significantly improved joint pain
Increased subchondral bone density, sclerosis, or defect caused by abnormal bone remodeling greatly weakens the effect of subchondral bone absorption on stress shocks, making it lose the function of protecting articular cartilage and causing secondary articular cartilage degeneration

Method used

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  • Application of Bromide Ion in Cartilage Tissue Engineering
  • Application of Bromide Ion in Cartilage Tissue Engineering
  • Application of Bromide Ion in Cartilage Tissue Engineering

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 bromide ion (Br - ) on the differentiation of chondrocytes

[0035] Mouse embryonic tumor cells ATDC5 (purchased from Shanghai Lianmai Bioengineering Co., Ltd.) were inoculated in 96-well plates, and the cells were placed in DMEM / F12 medium containing different concentrations of bromide ions at 37°C and 95% humidity in carbon dioxide (CO 2 ) in an incubator. In examining the bromide ion (Br - ) on the influence of chondrocyte differentiation, the bromide ion solutions of 0.5mM, 1mM, 5mM, 10mM and 100mM which were prepared with NaBr were set.

[0036] 1. Cell Proliferation Assay

[0037] On the 2nd, 4th, 7th, 10th, and 14th day of ATDC5 cell culture, the proliferation of cells in different media was detected by using CCK-8 kit (purchased from DOJINDO, Japan). On the 2nd, 4th, 7th, 10th, and 14th day of cell culture, aspirate the original medium, and add 110 μL of medium mixed with CCK-8 reagent (per 100 mL of DMEM / F12 medium mixed with 10 mL of CCK-8 8 ...

Embodiment 2

[0052] Characterization of the influence of embodiment 2 bromide ions on cartilage calcification

[0053] The differentiation medium used in this example was prepared by adding the following final concentrations of the components in common DMEM (Gibco) medium: 100 nM dexamethasone (Sigma, USA), 50 μg / mL vitamin C (Sigma, USA), 100 μg Sodium pyruvate / mL (Aladdin, China), 40 μg / mL proline (Aladdin, China), 1 vol% ITS (West, USA), 1 vol% double antibody (penicillin / streptomycin, brand Gibco).

[0054] Mouse embryonic tumor cells ATDC5 were stored in DMEM medium supplemented with 5% fetal bovine serum at 37°C and 95% humidity in CO 2 cultured in an incubator. When the cell density in the culture flask reached 90-95%, the cells were trypsinized and centrifuged for passage. After the cells were expanded to the required number for the experiment, the cells were digested with trypsin and centrifuged to collect the cells. 5 Density per well The cells were seeded in 96-well culture p...

Embodiment 3

[0057] The preparation of embodiment 3 loaded bromide ion microspheres

[0058] This example utilizes polylactic acid-co-glycolic acid (PLGA) to prepare loaded Br - PLGA microspheres.

[0059] PLGA was dissolved in dichloromethane, containing Br - The aqueous solution (NaBr aqueous solution) was added dropwise therein to prepare colostrum; then the colostrum was added dropwise to 1.5% polyvinyl alcohol aqueous solution, stirred in an ice bath for 6 h, and then stirred at room temperature for 24 h to obtain the loaded Br - PLGA microspheres.

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Abstract

The invention provides the application of bromide ion in cartilage tissue engineering. The present invention proposes and proves for the first time that bromide ions can promote the proliferation and differentiation of chondrocytes and the secretion of cartilage extracellular matrix (such as glycosaminoglycans, etc.), have the function of inducing chondrocyte regeneration and differentiation, and can effectively promote cartilage repair At the same time, bromide ions had no significant effect on cell proliferation, spreading and adhesion. The present invention also provides microspheres loaded with bromide ions, which can be directly injected into the defect site for in situ repair or the microspheres can be compounded on the scaffold material to make a bio-scaffold material; the bio-scaffold material can be used as a cartilage- The subchondral bone defect implant material has excellent in vitro biological activity and remarkable bidirectional biological functional characteristics of cartilage-subchondral bone integrated repair in vivo, and has broad application prospects in the field of cartilage-subchondral bone defect repair.

Description

technical field [0001] The invention belongs to the field of biomaterials, in particular to the application of bromide ions in cartilage tissue engineering. Background technique [0002] Osteoarthritis (OA) is the most common chronic autoimmune or autoinflammatory mechanism dysfunction disease, which occurs in major weight-bearing joints, such as hip and knee. According to recent statistics, the prevalence of OA in the world has exceeded 4%, and the prevalence is especially high in the elderly population, even exceeding 18%, which has seriously affected human health. At present, the treatment of OA is still in a relatively conservative stage, usually including moderate exercise, weight loss or assisting in reducing joint pressure, pain relief, etc., there is no effective targeted treatment plan; if the bone joint is seriously deteriorated, artificial joint replacement Surgery is imperative, which greatly affects the patient's later health activities. Bone joints mainly inc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077A61L27/18A61L27/54
Inventor 于涛刘旭周长忍
Owner JINAN UNIVERSITY
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