富含血小板纖維蛋白可促進人類造骨細胞的細胞附著、增殖及膠原相關的蛋白質之表現

Platelet-rich fibrin increases cell attachment, proliferation, and collagen-related proteins expression of human osteoblasts

吳志隆(Chih-Lung Wu)

指導教授 ： 呂克修

中山醫學大學/醫學院/醫學研究所/博士(2012年)

摘要

研究背景:藉由法國休葛蘭醫師 (Choukroun, 2001, Nice, France) 所創新的血小板濃縮技術，不用任何特殊處理即可將富含血小板纖維蛋白從自體的血液萃取出來，此富含血小板纖維蛋白被發現可以增進造骨細胞的生長與增殖，然而為何富含血小板纖維蛋白具有這種功能，其機轉至今尚未被確切了解，本研究是在確立富含血小板纖維蛋白對於人類造骨細胞的細胞附著、增生及phosphorylated Akt、heat shock protein 47 (HSP47) 與 lysyl oxidase (LOX) 等蛋白質表現的效應。
研究方法:收集10位健康的自願者的血液，分別使用細胞增生分析試劑WST-1和螢光偵測染劑alamlar blue方法，經由比色法測定來估量人類造骨細胞 (U2OS) 的細胞吸附與增生效應，用西方墨點法 (Western blot) 來評估p-Akt、 HSP47及LOX等蛋白的表現。
研究結果:在與對照組比較下，富含血小板纖維蛋白被發現可獨自刺激U2OS的細胞附著，且在5天的細胞培養期間可促進造骨細胞的增生。除此之外，富含血小板纖維蛋白以隨時間增加而增加的方式作用(time-dependent manner) 來促進Akt調控蛋白質的磷酸化作用，諸如HSP47與LOX等有相關的膠原蛋白濃度，也因富含血小板纖維蛋白的刺激而提升。
結論: 富含血小板的纖維蛋白能夠增進造骨細胞的細胞附著、增生及同時調升相關膠原蛋白質產物，這些作用可有效促進骨頭新生。

關鍵字

富含血小板的纖維蛋白 (PRF) ； 造骨細胞 ； 附著 ； 增生 ； Akt ； HSP47 ； LOX

並列摘要

Background: Platelet-rich fibrin (PRF) by Choukroun’s technique is derived from anautogenous preparation of concentrated platelets without any manipulation. PRF was found to increase osteoblast growth and proliferation. However, the underlying mechanisms are not yet completely understood. This study was to determine the effects of PRF on cell attachment, proliferation, phosphorylated Akt, heat shock protein 47 (HSP47), and lysyl oxidase (LOX) expression on human osteoblasts.
Methods: Blood collection was carried out from ten healthy volunteers. Cell attachment and proliferation were measured by colorimetric assay with WST-1 and alamlar blue in human osteoblast cell line U2OS cells, respectively. Western blot was employed to evaluate the expression of p-Akt, HSP47, and LOX.
Results: PRF alone was found to stimulate U2OS cell attachment as compared with untreated controls (p<0.05). PRF was found to increase osteoblast proliferation during 5 day incubation period (p<0.05). PRF was found to increase Akt phosphorylation in a time-dependent manner (p<0.05). Collagen-related proteins HSP47 and LOX were significantly elevated by the stimulation with PRF as compared with untreated controls (p<0.05).
Conclusions: It is suggested that PRF is capable of increasing osteoblast attachment, proliferation and simultaneously upregulating collagen- related proteins production. These actions in combination would effectively promote bone regeneration.

並列關鍵字

platelet-rich fibrin ； osteoblasts ； attachment ； proliferation ； Akt ； heat shock protein 47 ； lysyl oxidase

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