富含血小板血纖維蛋白對於人類造骨母細胞之影響與臨床運用

富含血小板血纖維蛋白(Platelet Rich Fibrin簡稱PRF)，是第二代的血小板濃縮技術，採血過程完全不添加抗凝血劑便可以獲得富含血小板及生長因子的纖維蛋白薄膜。然而有關於PRF對於造骨母細胞的生物性作用的資料有限。本研究的目的在於討論PRF對於人類的造骨母細胞的生物性影響，以提供臨床使用PRF的參考。用PCO2桌上型離心機以2700 rpm離心12分鐘得到PRF凝塊。PRF薄膜經蘇木紫與伊紅染色法處理後，在光學顯微鏡下觀察PRF的結構，纖維蛋白呈現均質化的外觀，在PRF基質中散佈著白血球。在掃描式電子顯微鏡下觀察，其基質是由網狀的血纖維蛋白交織排列所構成，呈現薄膜狀的外觀。在基質表面有時候可以觀察到聚集的血小板。以倒立位相差顯微鏡觀察時，顯示造骨母細胞維持立方狀或扁平狀外觀，且可貼附在PRF的邊緣。將人類U2OS造骨母細胞株植入基礎培養基中培養，以MTT比色分析法觀察PRF對於細胞存活率與增殖的影響，結果顯示PRF不會影響U2OS細胞的存活率(p > 0.05)；在五天的細胞培養期間PRF對於造骨母細胞，具有促進細胞增生的作用（p < 0.05）。將U2OS細胞與PRF一起培養21天後，以von Kossa染色法進行礦化作用的測試，可以觀察到許多暗棕色到黑色點狀明顯的礦化結節，由此可知PRF可促進造骨母細胞的分化。本研究最後的部份，以三個病例以臨床觀察、病理組織檢查或放射線檢查，來評估PRF在臨床應用之成效。綜合本研究的結果可知，PRF可促進造骨母細胞的增殖與分化，所以在骨再生手術中使用PRF是相當有助益的。

關鍵字

富含血小板血纖維蛋白；造骨母細胞；增殖作用；臨床應用

並列摘要

Platelet-rich fibrin (PRF) is a second-generation platelet concentrate which allows one to obtain fibrin membranes enriched with platelets and growth factors, after an anticoagulant-free blood harvest. However, limited information is currently available concerning the biologic effects of PRF on osteoblasts. The aim of this study is to provide clear evidence for the clinical use of PRF, we investigated the biologic effects of PRF on human osteoblasts. PRF was obtained by centrifugation at 2700 rpm for 12 minutes with a PC-O2 table centrifuge. At first, light microscope examination of PRF reveals a homogenous fibrin appearance by hematoxylin and eosin stain. Leucocytes were located within the PRF matrix. According to SEM image of PRF, a matrix can be observed of membrane like appearance, arranged by fibrin network. Platelet aggregates are assembled on the matrix surface occasionally. Then primary cultured human U2OS osteoblast cell line were used to evaluate cell viability and proliferation resulting from PRF according to trypan blue and tetrazolium bromide reduction assays. The result shows that PRF did not interfere with cell viability of U2OS cells. PRF was found to increase osteoblast proliferation during 5 day incubation period (p < 0.05). Besides, osteoblasts were observed to attach at the margins of PRF by phase-contrast microscopy. Afterward, exam of U2OS cultures with PRF for 21 days. After Von Kossa staining, mineralisation nodules were easily detectable. This was a significant indication when considering the favorable effect of PRF on bone mineralization. At the end of this study, we present 3 clinical cases with clinical, histologic, and radiographic finding. All of these examinations revealed new bone formation. Taken together, PRF can stimulate osteoblasts proliferation. And PRF was suggested to with a potential role for new bone formation. The application of PRF may provide the benefit for the bone regeneration.

並列關鍵字

platelet-rich fibrin ； osteoblast ； proliferation ； clinical trial

參考文獻

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富含血小板血纖維蛋白對於人類造骨母細胞之影響與臨床運用

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