Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology

Experimental Study on Intervention of Weilingxian on Three-dimensional Cultured of Injectable Chitosan/β-glyceroPHosPHate Gel Encapsulating Allograft Chondrocytes and Its Role in Repair of Articular Cartilage Defects

Author JinXiang
Tutor XuJianAn
School Nanjing University of Traditional Chinese Medicine
Course TCM orthopedics and traumatology
Keywords Articular cartilage defects Repair Clematis Tissue Engineering
CLC R285
Type PhD thesis
Year 2010
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Background and purpose of articular cartilage defects difficult to repair, has always been the difficulty of the medical profession and research focus. Currently, tissue engineering has become a major research direction, and has made some progress, tissue engineering seed cells, scaffolds, growth factors. In addition, traditional Chinese medicine in the field, has been proved to have good clinical efficacy. We screened single herb - Clematis as research subjects, studies have shown Clematis able to maintain and promote proteoglycan synthesis of chondrocytes with type II collagen, and possibly through inhibition of interleukin-1 levels increased protection of joint cartilage. The subject of Clematis and tissue engineering technologies to observe the Clematis promote chondrocyte proliferation, intervention C / beta-GP gel composite allogenic chondrocytes in vitro three-dimensional culture, and to repair cartilage defects, explore Chinese medicine and modern medicine the way and value. Method in vitro culture of rabbit articular chondrocytes, toluidine blue staining, take in a randomized the 3rd generation of the logarithmic growth phase cells were cultured, to inverted microscope cartilage changes in cell morphology, MTT assay different concentrations of Clematis proliferation of chondrocytes, RT-PCR method to detect the expression of TGF-beta1. 2 7:1 scale preparation C / beta-GP composite cartilage cell gel, were randomly divided into experimental and control groups were given the appropriate treatment, MTT assay different concentrations of Clematis gel chondrocyte proliferation HE, TB and type II collagen immunohistochemical staining the 1,4 weeks gel chondrocytes histological changes. Copy articular cartilage defect model, 24 New Zealand white rabbits were randomly divided into three groups, the experimental group (C / beta-GP chondrocytes Clematis medium intra-articular injection group), control group (C / beta-GP cartilage intra-articular injection of cells in normal medium group), model group (model group), were given the appropriate treatment, after 6 and 12 weeks, respectively, after line general observation and HE, TB, type II collagen immunohistochemical staining and Wakitani ratings . The 1 different concentrations of Clematis can promote the proliferation of chondrocytes, especially 0.5mg/ml proliferation effect is obvious, and the proliferation peak in the first three days of the Clematis intervention. The 0.05,0.1,0.5,1.0 mg / ml Clematis can promote cartilage cell expression of TGF-beta1 mRNA 4-group difference was not significant, but the peak of the role of 0.5mg/ml group. 2 chondrocytes good C / beta-GP gel stretching, adhesion, growth and proliferation. Various concentrations of Clematis can promote the proliferation of chondrocytes in the gel, which 0.5mg/ml Weilingxian, the most obvious effect, and can promote chondrocyte type II collagen, extracellular matrix secretion. 3, the experimental group and control group visible defect of the articular surface is preferably filled, and the formation of hyaline cartilage-like structure, the surface flatness of the experimental group and the surrounding tissue and the degree of integration compared with the control group, histological visible class cartilage formation and secretion cartilage matrix and specificity of type II collagen, model group no repair the visible tissue degeneration. Degree of integration of experimental group repair tissue and the surrounding tissue, histology, and type II collagen secretion in the control group. The Rating: experimental and control groups score significantly lower than the model group scores (P lt; 0.01), and the scores of the experimental group was significantly lower than the scores of the control group (P lt; 0.05) Wakitani. Conclusion 1. Clematis can promote chondrocyte proliferation and TGF-beta1 mRNA expression, this type of cell growth factor role may be one of the mechanisms to promote cartilage repair; 2.C/β-GP gel chondrocytes good compatibility, Clematis can promote the proliferation of chondrocytes in the C / ss-GP gel type II collagen, proteoglycan secretion into cartilage development. Clematis can promote injectable C / beta-GP gel composite allograft cartilage cells to repair articular cartilage defects, which is a combination of Chinese medicine and tissue engineering technology to provide new ideas, new methods.

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