利用雙光子螢光及二倍頻顯微術觀測外來作用力對於由幹細胞分化的軟骨母細胞分泌膠原蛋白的影響

Abstract

軟骨的損傷一直是人體一項不可避免的疾病。而且對於運動員而言，長期持續的運動使的軟骨更容易受傷。一開始的作法是從身體其他部分取出不會影響到正常生理機能的軟骨去填補受傷的部份。不過漸漸的，所需要的軟骨組織越來越多，於是開始試著製作軟骨，這就是組織工程中一個很大的題目。在眾多細胞來源中，利用幹細胞配合適當的鷹架跟生長因子來培養軟骨組織一直受到很大的重視，而怎樣適當的參數會影響到軟骨分泌膠原蛋白也廣被研究。 本實驗即是利用雙光子及倍頻顯微術，觀察受力 ( 0G, 10000G, 20000G，每天施力15分鐘)的軟骨母細胞，其膠原蛋白生長的型態和情形。(一共培養四個禮拜。) 我們觀察到有施力的細胞，膠原蛋白有成束狀的發展，且有局部生長的情形發生。

Rheumatoid arthritis (RA) is a major disease for human. For a sportsman, longtime motion makes articular cartilage hurt easily. Initially we take some cartilage from other part in body to repair the hurt part. Because the more cartilage needed, we try to make the cartilage from cells. This is a big term in tissue engineering. Between all the cell’s sources, we pay much attention to use the stem cells with proper scaffolds and transfer growth factors to cultivate cartilage. In my experiment, I utilize the auto-fluorescence and second harmonic generation to observe the forced chondrocytes producing collagen.We cultivate mesenchymal stem cells (MSCs) in a medium with TGF-β3 added. TGF-β3 induces the MSCs to differentiate into chondrocytes and the chondrocytes subsequently produce collagen. During culture, we impart force on the cells by using a centrifuge. We choose three force-0G,10000G,200000G and exerted the force on the cells for 15 minutes a day for 4 weeks. We observed that the cells exerted with greater force produce collagen that tend to accumulate in lumps at early stage, and generate stronger second harmonic signal.