저분자 키토산-TGF-β1 복합체가 사람 연골 세포의 시험관 내배양에 미치는 효과

Abstract

We aim to investigate the effects of LMC(Low Molecular weight Chitosan)- TGF-β1 conjugate on adult human chondrocytesin vitro. Primary chondrocytes were harvested from two patients who underwent total knee arthroplasty. The chondrocytes fromone patient were cultured in Mebiol gel??, 3 D culture system, for evaluation of cell proliferation and collagen type II expressionusing semi-quantitative RT-PCR. A monolayer culture for histological evaluation was performed with the Chondrocytesharvested from the other patient using H&E, safranin-O and immunohistochemistry stain(S-100 and type II collagen). Each culturewas divided into 4 experimental groups; group 1: control (PBS), group 2: chitosan(5.8g/mL), group 3: TGF-β1(15 ng/mL), group4: LMC-TGF-β1 conjugation(LMC 5.8g/mL, TGF-β1 15 ng/mL). In each group, conditioning was made only one time andsustained for 3days. Cell proliferation was sustained for 3 days and cell count declined slightly from day 6 in all groups.However, increased cell count was preserved well in the group 4, especially at day 3,6,9(p < 0.05) and in the group 3 and 4 attochemisty and RT-PCR revealed that Collagen type II synthesis was more enhanced with longerpreservation in the group 4 compared to other groups. The conjugation of TGF-β1 and LMC could have a more positive effecton the proliferation and production of type II collagen of human chondrocyte in vitro than using of single TGF-β1.