Effects Of Carboxymethyl Chitosan And PDGF-BB Combination On The Biological Activity Of Human Periodontal Ligament Cells

Objective:To study on effects of CM-CTS and PDGF-BB combination on the biological activity of human periodontal ligament cell, provide theoretical basis and research foundation for the regeneration of periodontal tissue.Methods:Human periodontal ligament cells were primarily cultured with coverslip covering tissue culture method in vitro. Inverted phase contrast microscope method was used to observe the morphology. Immunohistochemical methods were used to identify the source of human periodontal ligament cells. To investigate the influence of CM-CTS and PDGF-BB combination on the biological activity of human periodontal ligament cells by MTT assay, Coomassie brilliant blue assay, flow cytometry and spectrophotometric assay.Results:Shown by immunohistochemistry, the positive staining of vimentin and the negative staining of cytokeratin in cultured cells. MTT assay showed that, every group could promote the proliferation of HPDLCs after treatment for72hours, respectively (P<0.05), lOng/ml PDGF-BB and100μg/ml CM-CTS joint group showed the most significant effection on promoting cell proliferation at this time. Coomassie brilliant blue assay showed that, every group could increase the total content of protein of human periodontal ligament cells after treatment for72hours, respectively (P<0.05),10ng/ml PDGF-BB and100μg/ml CM-CTS joint group showed the most significant effection on promoting the total content of protein of human periodontal ligament cells at this time. FCM assay showed that, every group could enhance PI (S+G2M)%(P<0.05). lOng/ml PDGF-BB and100μg/ml CM-CTS joint group showed the most significant effection on promoting PI of human periodontal ligament cells. ALP activity assay showed that, after treatment for72hours, PDGF-BB separate group was inhibited ALP activity of human periodontal ligament cells,100μg/ml CM-CTS separate group、10ng/ml PDGF-BB and100μg/ml CM-CTS joint group could promote significantly the ALP activity of human periodontal ligament cells (P<0.05), and10ng/ml PDGF-BB and100μg/ml CM-CTS joint group showed the most significant effection on promoting of the ALP activity of human periodontal ligament cells.Conclusions:The experimental results have shown that concentration of CM-CTS could promote the proliferation of human periodontal ligament cells, and promote osteogenic differentiation of human periodontal ligament cells. Provide the experimental basis for clinical application of CM-CTS treatment of periodontal tissue regeneration. CM-CTS and PDGF-BB combination could significantly promote the proliferation of human periodontal ligament cells, increase the synthesis of cell protein, promote the DNA synthesis and osteogenic differentiation. CM-CTS and PDGF-BB combination have a synergistic effect, to provide theoretical basis and research foundation for the regeneration and repair of periodontal tissue.