Cell Adhesion of Human Bone Marrow-derived Mesenchymal Stem (HMS0014) Cells on Titanium Discs with Different Surface Modifications

HMS0014骨髓衍生間質細胞在不同表面處理的鈦金屬板上的細胞附著

中塚美智子(Michiko Nakatsuka) ； 岩井康智(Yasutomo Iwai) ； 黃純德(Shun-Te Huang) ； 紺井拡隆(Hirotaka Kon-I) ； 郭蓮(Lian Guo) ； 森下愛子(Aiko Morishita) ； 黃宏智(Hung-Chin Huang) ； 隈部俊二(Shunji Kumabe)

《Taiwan Journal of Oral Medicine Sciences》 28卷2期 (2013/08) Pp. 71-81+83-84

摘要

人工牙根的表面處理可以增加人工牙根與周圍骨組織的結合，以利於人工牙根的初期穩定性及骨整合（osseointegration）。我們先前對人類骨髓出來的間葉幹細胞（HMS0014）的單層培養，細胞的繁衍及分化，與細胞外物質（ECM）的礦化先作了初步研討。接著，我們以骨再生誘導法（GBR），將HMS0014細胞培養在不同表面處理的鈦或鈦合金的圓板上，並啓始其骨化作用。然後，以免疫螢光組織學的方法及掃瞄電子顯微鏡探討在inducing condition下，附著、貼附於鈦金屬圓板的表面基質上的HMS0014細胞，並觀察細胞的伸展以及細胞間鍵結的發生。本研究得到以下三個結果：1.掃瞄電子顯微鏡顯觀察到，許多圓形或多角形（直徑約10－40μm）的HMS0014細胞分佈於鈦金屬圓板上。在60－180分鐘的培養後，繁衍並分化為扁平多角形（大小約30x90－100x20μm2）的細胞；此細胞具有板狀偽足及髮絲樹突狀偽足，用以作為細胞間以及細胞與鈦金屬圓板表面基質間的結合。2. 免疫螢光組織學顯示，附著於鈦金屬板的HMS0014細胞有F-actin（actin filament）和CD51（αV integrin）的共同存在。3.從第一天的培養後，成熟的類骨母細胞樣（osteoblast-like）HMS0014細胞的ECM已有礦化現象，而到第7至第14天，在ECM內可明顯地觀察到點狀、塊狀的鈣化物的分佈。我們得到一個結論，GBR可促使HMS0014細胞在經過不同表面處理的鈦或鈦合金表層上產生骨化現象。

關鍵字

細胞附著 ； 鈦金屬板 ； HMS0014細胞

並列摘要

The modification of surface microtextures on the implant fixture increases direct bone-to-implant contact compatibility to benift primary stability and osseointegration of dental implant therapy. Cell proliferation and differentiation of HMS0014 cells, and mineralisation of the ECM in monolayer cell culture were preliminarily studied. Subsequently, the HMS0014 cells were GBR-engineered to initiate osteogenesis on either titanium (Ti) or Ti alloy discs modified with different surface substrates. Attachment onto the substratum, extension and intercellular contact of the HMS0014 cells under inducing condition were studied with fluorescent immunohistology and conventional scanning electron microscopy (SEM). The present study obtained results as follows: (i) The SEM demonstrated that the spherical-to-polygonal (d= 10-40μm) HMS0014 cells proliferated and differentiated into flat polygonal (30x90-100x200 μm2) cells, showing prominent lamellipodia and dendritic filopodia, to employ cell-to-substrate and intercellular attachments on the Ti disc surface between 60 to 180 minutes of culture. (ii)The fluorescent immunohistochemistry demonstrated the co-expression of F-actin (actin filament, cytoskeleton protein) and CD51 (α V integrin, interfacing specific receptor protein) in the attached HMS0014 cells. (iii) The matured osteoblast-like HMS0014 cells initiated mineralisation on day 1 of culture; distribution of calcification loci in the ECM was prominently observed between days 7 and 14 of culture. We concluded that the present GBR method enhanced HMS0014 cells to initiate contact osteogenesis on Ti and Ti alloy discs subject to different surface modifications.

並列關鍵字

cell adhesion ； surface substrate ； HMS0014 cells