Research The Biology Charateristics And Clinical Application Of The Human Articular Cartilage Cells

Background:Articular cartilage defected is a common clinical disease. Because the chondrocytes are trapped by Extracellular Matrix(ECM), with much limited the ability of migration, so the spontaneous repair of damaged cartilage is limited. Therefore, we should use surgical operation to treat the articular cartilage defects. Early therapeutic methods include Abrasion, Micro fracture, Mosaicplasty, Osteochondral transplantation and so on, but none of these have been successful in regeneration a normal and functional articular cartilage layer. Resently, Autologous Chondrocyte Implantation(ACI) technique treat the articular cartilage defected as a new method, which has been widely used in USA, Germany, Australia, Sweden, Singapore and so on, but less in our Country. Objective:To meet the clinical needs, reseach the biology characteristics about human articular cartilage cells, this experiment establish a set of system about the autologous chondrocyte culturing, expanding and detecting in vitro, provide a method to repair the defects of articular cartilage, take technical support and protection for autologous chondrocytes implantation on clinical applying.Research content:1.Chondrocytes was cultured in monolayer culture, observed the morphological characteristics; made cell slide and analyzed though Safranin O; detected the proliferation ability of P1, P3and P5generation of chondrocytes by MTT, and the expression of collagen Ⅱ gene(COL-Ⅱ) by Reverse Transcription-Polymerase Chain Reaction(RT-PCR).2.Articular cartilage cells, carried in an alginate polyer, were injected into the dorsa subcutaneous of node mice and developed in vivo. The cartilage tissue was harvested after3months and analyzed through histological and immunohistochemical.3.Cultured and proliferated patients’chondrocytes, implanted the cells to the injured parts, evaluated the effect of the repaired parts by preoperation and postoperation clinical symptom and the magnetic resonance imaging(MRI).Results:1.Observed chondrocytes by microscope, the morphological characteristics was polygon when the cells adherence, the morphological was slabs as chondrocytes expanded over the flask bottom; the chondrocytes showed positive by safranin O analyzed; the expression level of COL-Ⅱ in P1generation of chondrocytes is significantly higher than in the P3and P5. The results is concordant with MTT. The results showed that chondrocytes exited dedifferentiation in vitro cultured.2.The histological and immunohistochemical results demonstrated that chondroytes-alginate formed a mature cartilage tissue at the dorsa subcutaneous of node mice.3.The MRI results revealed that the defects parts improved obviously after implanting autologous culture-expanded chondrocytes2and6months. The clinical symptom were also improved. Conclusions:1.We suggest that using P1generation chondrocytes treat the cartilage defects by compared the proliferation ability and the expression level of COL-Ⅱ in P1, P3and P5generation.2. ACI has a effective to treat the defects of cartilage.