Tajima Satoshi, Tobita Morikuni, Orbay Hakan, Hyakusoku Hiko, Mizuno Hiroshi
1 Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine , Tokyo, Japan .
Tissue Eng Part A. 2015 Mar;21(5-6):895-905. doi: 10.1089/ten.TEA.2014.0336. Epub 2014 Nov 12.
A key goal for successful bone regeneration is to bridge a bone defect using healing procedures that are stable and durable. Adipose-derived stem cells (ASCs) have the potential to differentiate into bone. Meanwhile, platelet-rich plasma (PRP) is an interesting biological means to repair tissue by inducing chemotactic, proliferative, and anabolic cellular responses. This study evaluated bone regeneration using a combination of ASCs and PRP in a rat calvarial defect model. ASCs were isolated from inguinal fat pads of F344 inbred rats, while PRP was prepared from these rats. ASCs were cultured in control medium supplemented with 10% fetal bovine serum or 5% PRP in vitro. After 1 week, levels of growth factors including insulin-like growth factor-1, transforming growth factor-β1, hepatocyte growth factor, and vascular endothelial growth factor in the culture supernatant were measured by enzyme-linked immunosorbent assays. Moreover, the ASC/PRP admixture was transplanted into the rat calvarial defect. Microcomputed tomography, histological, and immunohistochemical (osteopontin and osteocalcin) analyses were performed at 4 and 8 weeks after transplantation. The in vitro study showed that the levels of growth factors secreted by ASCs were significantly increased by the addition of PRP. Transplantation of the ASC/PRP admixture had dramatic effects on bone regeneration overtime in comparison with rats that received other transplants. Furthermore, some ASCs directly differentiated into osteogenic cells in vivo. These findings suggest that the combination of ASCs and PRP has augmentative effects on bone regeneration. The ASC/PRP admixture may be a promising source for the clinical treatment of cranial defects.
成功进行骨再生的一个关键目标是通过稳定且持久的愈合程序来桥接骨缺损。脂肪来源干细胞(ASC)具有分化为骨的潜力。同时,富血小板血浆(PRP)是一种通过诱导趋化、增殖和合成代谢细胞反应来修复组织的有趣生物学手段。本研究在大鼠颅骨缺损模型中评估了ASC与PRP联合应用对骨再生的作用。ASC从F344近交系大鼠的腹股沟脂肪垫中分离,而PRP则由这些大鼠制备。ASC在补充有10%胎牛血清的对照培养基或含5%PRP的培养基中体外培养。1周后,通过酶联免疫吸附测定法测量培养上清液中包括胰岛素样生长因子-1、转化生长因子-β1、肝细胞生长因子和血管内皮生长因子在内的生长因子水平。此外,将ASC/PRP混合物移植到大鼠颅骨缺损处。在移植后4周和8周进行微型计算机断层扫描、组织学和免疫组织化学(骨桥蛋白和骨钙素)分析。体外研究表明,添加PRP可显著提高ASC分泌的生长因子水平。与接受其他移植的大鼠相比,ASC/PRP混合物的移植对骨再生随时间推移具有显著影响。此外,一些ASC在体内直接分化为成骨细胞。这些发现表明,ASC与PRP联合应用对骨再生具有增强作用。ASC/PRP混合物可能是颅骨缺损临床治疗的一个有前景的来源。