The Effect of Chitosan Treatment on the Proliferation and Senescence of Anterior Cruciate Ligament Fibroblasts and Synovial Membrane Derived Cells

In this study, we isolated primary cells form human synovial membrane and human anterior cruciate ligament. First, we seed the cells on the chitosan substrate and culture for a few days. After this treatment, we studied the change of cell function. Chitosan is a natural, biodegradable, biocompatible, non-toxic and U.S. Food and Drug Administration (FDA) approved polysaccharide. In this study, chitosan was used as the coating substrates. When synovial cells and anterior cruciate ligament fibroblasts were cultured on chitosan, all cells became suspended and aggregated into spheroids. After three days, we seeded the cell spheroids on TCPS (Tissue culture polystyrene), and studied change of cell proliferation and cell senescence expression. Furthermore, we established the population doubling curve, used MTT ([3-[4,5-dimethylthiahiazo-2-yl]-2,4-dipheny- tetrazolium bromide]) assay and BrdU (5-bromo-2’-deoxiuridine) assay, to study the cell proliferation change. We used the inverted microscope to observe the cell morphology and used senescence-associated-β-galactosidase (SA-β-gal) to study the cell senescence expression. Finally, we collected cells from various patients to study the relationship between the effect of chitosan treatment and patient’s age.