Department of Biomaterials, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands; Department of Oral and Maxillofacial Surgery, Beijing Stomatological Hospital, Capital Medical University, Beijing, 100050, China.
J Biomed Mater Res A. 2014 Apr;102(4):1026-36. doi: 10.1002/jbm.a.34775. Epub 2013 Jun 4.
The aim of this study was to compare the osteogenic capacity between human adipose tissue-derived mesenchymal stem cells (AT-MSCs) and their cocultures with human umbilical vein endothelial cells (HUVECs) in vitro and their biological performance in vivo. First, the optimal cell ratio in cocultures for osteogenic differentiation was determined by seeding AT-MSCs and HUVECs in ratios varying from 100:0 to 0:100 on tissue culture plates. Afterward, AT-MSCs and AT-MSCs/HUVECs (50:50) were seeded on porous titanium fiber mesh scaffolds (Ti) for both in vitro and in vivo osteogenic evaluation. For in vitro evaluation, cell osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity and calcium assay. For in vivo evaluation, the scaffolds were implanted bilaterally into rat cranial defects (5 mm diameter) and bone formation was assessed histologically and histomorphometrically after 8 weeks. The ratio of 50:50 was chosen in the cocultures because this coculture condition retained similar amount of calcium deposition while using the least amount of AT-MSCs. Moreover, AT-MSCs showed higher osteogenic differentiation in comparison to AT-MSCs/HUVECs on Ti in vitro. Furthermore, superior bone formation was observed in AT-MSCs compared to AT-MSCs/HUVECs in rat cranial defects. In conclusion, AT-MSCs showed significantly higher osteogenic potential compared to AT-MSCs/HUVECs both in vitro and in vivo.
本研究旨在比较人脂肪组织来源间充质干细胞(AT-MSCs)及其与人脐静脉内皮细胞(HUVECs)共培养物的体外成骨能力及其体内生物学性能。首先,通过将 AT-MSCs 和 HUVECs 以 100:0 至 0:100 的比例接种于细胞培养板上,确定共培养物中用于成骨分化的最佳细胞比例。然后,将 AT-MSCs 和 AT-MSCs/HUVECs(50:50)接种于多孔钛纤维网支架(Ti)上,用于体外和体内成骨评价。体外评价时,通过碱性磷酸酶(ALP)活性和钙测定评估细胞成骨分化。体内评价时,将支架双侧植入大鼠颅缺损(5mm 直径)中,8 周后进行组织学和组织形态计量学评估骨形成。选择 50:50 的共培养比例是因为这种共培养条件在使用最少数量的 AT-MSCs 的情况下保留了相似数量的钙沉积。此外,与 Ti 上的 AT-MSCs/HUVECs 相比,AT-MSCs 的体外成骨分化更高。此外,在大鼠颅缺损中,与 AT-MSCs/HUVECs 相比,AT-MSCs 观察到更好的骨形成。总之,与 AT-MSCs/HUVECs 相比,AT-MSCs 在体外和体内均显示出更高的成骨潜力。