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Application of bromide ions 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 secondary articular cartilage degeneration, inability to significantly improve joint pain, and increased subchondral bone density. Achieve broad application prospects, excellent in vitro biological activity, and the effect of promoting cartilage repair

Active Publication Date: 2019-05-10
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 ions in cartilage tissue engineering
  • Application of bromide ions in cartilage tissue engineering
  • Application of bromide ions 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 application of bromide ions in cartilage tissue engineering. It is put forward and proved for the first time that the bromide ions can promote the proliferation and differentiation of cartilage cells and the secretion of a cartilage cell epimatrix (such as glycosaminoglycan), have the functions of inducing cartilage cell regeneration and differentiation, and can effectivelyhave the effect of promoting cartilage repairing; meanwhile, the bromide ions have no obvious influences on the proliferating, spreading and adhering of the cells. The invention further provides microspheres loaded with the bromide ions. The microspheres can be directly injected to a defect part for in-situ repairing or can be combined with a stent material for preparing a biological stent material. The biological stent material can be used as a cartilage-subchondral bone defect implantation material, has the excellent in-vitro bioactivity, has the obvious bidirectional biological function characteristics of in-vitro cartilage-subchondral bone integral repairing, and has wide application prospects in the field of cartilage-subchondral bone defect repairing.

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 Applications(China)
IPC IPC(8): C12N5/077A61L27/18A61L27/54
Inventor 于涛刘旭周长忍
Owner JINAN UNIVERSITY
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