Jo Kwangrae, Kim Yongsun, Lee Seung Hoon, Yoon Yong Seok, Kim Wan Hee, Kweon Oh-Kyeong
BK21 PLUS Program for Creative Veterinary Science Research, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Gwanak-ro 1, Seoul 08826, Republic of Korea.
Heliyon. 2017 Aug 17;3(8):e00383. doi: 10.1016/j.heliyon.2017.e00383. eCollection 2017 Aug.
Demineralized bone allografts and mesenchymal stromal cells have been used to promote bone regeneration. However, the degree to which cortical bone should be demineralized for use in combination with adipose-derived mesenchymal stromal cells (Ad-MSCs) remains to be clarified. In this study, the osteogenic ability of Ad-MSCs on allografts was investigated in relation to the extent of demineralization. Three treatment groups were established by varying exposure time to 0.6 N HCL: partially demineralized (PDB; 12 h), fully demineralized (FDB; 48 h), and non-demineralized bone (NDB; 0 h, as a control). Allografts were prepared as discs 6 mm in diameter for evaluation, and their demineralization and structure were evaluated by micro-computed tomography and scanning electron microscopy. Ad-MSC adhesion and proliferation were measured by MTS assay, and osteogenesis-related gene expression was assessed by quantitative reverse transcription polymerase chain reaction. PDB and FDB demineralization rates were 57.13 and 92.30%, respectively. Moreover, Ad-MSC adhesion rates on NDB, PDB, and FDB were 53.41, 60.65, and 61.32%, respectively. Proliferation of these cells on FDB increased significantly after 2 days of culture compared to the other groups ( < 0.05). Furthermore, expression of the osteogenic genes , , and in the FDB group on culture day 3 was significantly elevated in comparison to the other treatments. Given its biocompatibility and promotion of the osteogenic differentiation of Ad-MSCs, our results suggest that FDB may be a suitable scaffold for use in the repair of bone defects.
脱矿骨同种异体移植物和间充质基质细胞已被用于促进骨再生。然而，与脂肪来源的间充质基质细胞（Ad-MSCs）联合使用时，皮质骨应脱矿的程度仍有待阐明。在本研究中，研究了Ad-MSCs在同种异体移植物上的成骨能力与脱矿程度的关系。通过改变对0.6 N盐酸的暴露时间建立了三个治疗组：部分脱矿（PDB；12小时）、完全脱矿（FDB；48小时）和未脱矿骨（NDB；0小时，作为对照）。将同种异体移植物制备成直径6 mm的圆盘用于评估，并通过显微计算机断层扫描和扫描电子显微镜评估其脱矿和结构。通过MTS测定法测量Ad-MSC的粘附和增殖，并通过定量逆转录聚合酶链反应评估成骨相关基因的表达。PDB和FDB的脱矿率分别为57.13%和92.30%。此外，Ad-MSC在NDB、PDB和FDB上的粘附率分别为53.41%、60.65%和61.32%。与其他组相比，培养2天后这些细胞在FDB上的增殖显著增加（<0.05）。此外，与其他处理相比，培养第3天FDB组中成骨基因、和的表达显著升高。鉴于其生物相容性和对Ad-MSCs成骨分化的促进作用，我们的结果表明FDB可能是用于修复骨缺损的合适支架。
