Hamahashi K, Sato M, Yamato M, Kokubo M, Mitani G, Ito S, Nagai T, Ebihara G, Kutsuna T, Okano T, Mochida J
Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Kanagawa, Japan.
J Tissue Eng Regen Med. 2015 Jan;9(1):24-30. doi: 10.1002/term.1610. Epub 2012 Nov 19.
The authors aimed to repair and regenerate articular cartilage with layered chondrocyte sheets, produced using temperature-responsive culture dishes. The purpose of this study was to investigate the humoral factors produced by layered chondrocyte sheets. Articular chondrocytes and synovial cells were harvested during total knee arthroplasty. After co-culture, the samples were divided into three groups: a monolayer, 7 day culture sheet group (group M); a triple-layered, 7 day culture sheet group (group L); and a monolayer culture group with a cell count identical to that of group L (group C). The secretion of collagen type 1 (COL1), collagen type 2 (COL2), matrix metalloproteinase-13 (MMP13), transforming growth factor-β (TGFβ), melanoma inhibitory activity (MIA) and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assay (ELISA). Layered chondrocyte sheets produced the most humoral factors. PGE2 expression declined over time in group C but was significantly higher in groups M and L. TGFβ expression was low in group C but was significantly higher in groups M and L (p<0.05). Our results suggest that the humoral factors produced by layered chondrocyte sheets may contribute to cartilaginous tissue repair and regeneration.
作者旨在使用温度响应培养皿制备分层软骨细胞片来修复和再生关节软骨。本研究的目的是调查分层软骨细胞片产生的体液因子。在全膝关节置换术中采集关节软骨细胞和滑膜细胞。共培养后,将样本分为三组:单层7天培养片组(M组);三层7天培养片组(L组);以及细胞计数与L组相同的单层培养组(C组)。通过酶联免疫吸附测定(ELISA)测量1型胶原(COL1)、2型胶原(COL2)、基质金属蛋白酶-13(MMP13)、转化生长因子-β(TGFβ)、黑色素瘤抑制活性(MIA)和前列腺素E2(PGE2)的分泌。分层软骨细胞片产生的体液因子最多。C组中PGE2表达随时间下降,但在M组和L组中显著更高。C组中TGFβ表达较低,但在M组和L组中显著更高(p<0.05)。我们的结果表明,分层软骨细胞片产生的体液因子可能有助于软骨组织的修复和再生。