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人類骨髓間葉幹細胞在不同植體表面進行骨生成分化之體外研究
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| 作者 |
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温明璇 |
| 學校系所 |
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國防醫學院生物及解剖學研究所 |
| 地點 |
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全臺 全部
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| 研究內容 |
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[中文摘要]
人類骨髓間葉幹細胞為多重分化潛能細胞，可以分化為造骨細胞系列和其他細胞系列。鈦是常見的金屬製人工植體廣泛用於骨科和口腔頜面部重建手術。鈦人工植體依靠表面之化學及物理特性，可直接與骨組織結合。然而，純鈦表面與骨組織之結合與新骨形成是緩慢的過程及不完整的骨整合。經過表面改造的鈦人工植體，增加表面的粗糙性及模擬生物活性的材質，能夠增進細胞的貼附、增生及分化。然而以往的研究集中在纖維母細胞、造骨細胞及骨髓間質細胞與植體表面之間的整合。目前為止，人類骨髓間葉幹細胞或骨髓間質細胞與植體之不同表面改造之間的關係/交互作用仍然不清楚。因此，我們利用掃描式電子顯微鏡、溴化去氧尿甘分析法、反轉錄聚合酶連鎖反應及鹼性磷酸酶活性分析法評估人類骨髓間葉幹細胞在經過不同改造方式的鈦植體表面上骨生成分化潛能的變化。相較於純鈦表面，人類骨髓間葉幹細胞的貼附被漿噴塗製造出的粗糙表面改變。根據溴化去氧尿甘分析法，三種表面對於細胞增生沒有顯著的影響。鹼性磷酸酶活性分析與反轉錄聚合酶連鎖反應顯示人類骨髓間葉幹細胞在鈦漿噴塗及羥基磷灰石漿噴塗表面的骨生成潛能比磨光的鈦人工植體低。本研究結果顯示漿噴塗製造出的粗糙性和羥基磷灰石塗層會影響人類骨髓間葉幹細胞的骨生成分化。
中文關鍵詞：人類骨髓間葉幹細胞、鈦金屬人工植體、骨生成分化、鈦漿噴塗、羥基磷灰石漿噴塗、掃描式電子顯微鏡、溴化去氧尿甘分析法、反轉錄聚合酶連鎖反應、鹼性磷酸酶活性分析法
[英文摘要Abstract]
Human bone marrow mesenchymal stem cells (hBMSCs) are multipotential cells which can differentiate into osteogenic and other cell lineages. Titanium (Ti) implants are common metal implant widely used in orthopedic and oral maxillofacial reconstructive surgery. Ti implant directly bond to bone depended on chemical and physical properties of the surface. However, the bonding and bone ingrowth between pure Ti implant and bone are slow process and incomplete osseointegration. Ti implant with specific surface modifications which increase surface roughness or mimic bioactive materials can improve the cell adhesion, proliferation and differentiation. Nevertheless, previous studies have been focused on integration between fibroblasts, osteoblasts or bone marrow stromal cells and implant surface. Till now, interrelationship/interaction between hBMSCs or marrow stromal cells and different surface modification of implant are still unclear. Therefore, we investigated the changes in osteogenic potential of hBMSCs on different modified surfaces of Ti implant using scanning electron microscopy, bromodeoxyuridine (BrdU) assay, alkaline phosphatase (ALP) activity, and reverse transcriptase polymerase chain reaction (RT-PCR). Compared to the pure Ti, hBMSCs adhesion was changed due to roughness produced by plasma spraying. With BrdU assay, there was no significant difference on cell proliferation between three modified surfaces. The results of ALP activity and RT-PCR indicated that the osteogenic potential of hBMSC was lower on the Ti plasma spraying and plasma sprayed hydroxyapatite (HA) coating surfaces than on polished Ti surface. The results of this study demonstrated that roughness and HA coating produced by plasma spraying interfered with osteogenic differentiation of hBMSCs.
Keywords：human bone marrow mesenchymal stem cell、titanium implant、osteogenesis、titanium plasma spraying、plasma sprayed hydroxyapatite、scanning electron microscopy、bromodeoxyuridine assay、alkaline phosphatase activity、reverse transcriptase polymerase chain reaction |
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