Effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation and expression of type II collagen and transforming growth factor-beta1 of rabbit knee articular chondrocytes / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals. OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage. METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis. RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.