富血小板纤维蛋白生物学特性鉴定及其对脂肪干细胞增殖和分化的影响
Identification of biological characteristics of platelet-rich fibrin and its effects on proliferation and differentiation of adipose derived stem cells

目的 ·探讨富血小板纤维蛋白（PRF）的生物学特性及其对脂肪干细胞（ADSCs）增殖和分化的影响。方法 ·取比格犬静脉血制备PRF膜，进行大体、组织学和超微结构观察。比格犬腹股沟处提取、分离培养ADSCs并进行多向分化潜能鉴定。体外将PRF作用于自体ADSCs，分为PRF组和对照组。CCK-8检测细胞增殖。对 2组细胞分别进行成骨诱导，于诱导前及诱导后第4、7日，采用RT-PCR检测成骨相关基因骨钙素（OCN）、骨桥素（OPN）和Ⅰ型胶原（Col-Ⅰ）的表达；诱导 7 d后进行碱性磷酸酶（ALP）活性检测。结果 · PRF为乳白色的纤维蛋白凝胶，具有弹性和韧性；胶原纤维构成疏松多孔的三维立体网络结构，大量的白细胞、血小板网络其间。PRF组细胞增殖活性显著高于对照组。成骨诱导后，PRF组OCN、OPN、Col-ⅠmRNA表达均显著升高，ALP活性表达增强。结论 · PRF是具有三维网络结构的纤维蛋白，其富含的血小板可以缓慢释放生长因子；PRF对ADSCs的增殖和成骨分化具有促进作用。
： Objective · To explore the biological characteristics of platelet-rich fibrin (PRF) and its effects on proliferation and differentiation of adipose derived stem cells (ADSCs). Methods · The whole blood was collected from the forelimb vein of healthy beagles to prepare the PRF membrane, which were observed with optical microscope and scanning electron microscope. ADSCs were collected from the inguinal adipose tissue and were isolated and cultured.Identification of multi-directional differentiation potential was performed. ADSCs were assigned to the PRF group and the control group, the former was treated with PRF in vitro. Cell proliferation was measured with CCK-8. Osteogenesis induction was performed for two groups and the expression of genes associated with osteogenesis, including osteocalcin (OCN), osteopontin (OPN) and collagen I (Col-Ⅰ), was measured with RT-PCR before induction and 4 and 7 days after induction. The alkaline phosphatase (ALP) activity was measured 7 days after induction. Results · PRF is a milk white fibrin glue with elasticity and toughness. PRF can form loose and porous three dimensional network structure, which harbors lots of platelets and leucocytes. The cell proliferation activity was significantly higher in the PRF group than in the control group. After osteogenesis induction, the ALP activity and the mRNA levels of OCN, OPN, and Col-Ⅰ were significantly increased. Conclusion · PRF is a fibrin glue with three dimension network structure and contains lots of platelets, which can slowly release growth factors. PRF can promote the proliferation and osteogenic differentiation of ADSCs