Department of Orthopaedic Surgery, CHA Bundang Medical Center, CHA University School of Medicine, 59 Yatap-Ro, Seongnam-si 13496, Gyeonggi-do, Korea.
Department of Orthopaedic Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, 81 Irwon-Ro, Gangnam-gu, Seoul 06351, Korea.
Cells. 2021 Aug 30;10(9):2244. doi: 10.3390/cells10092244.
An important objective of vascularized tissue regeneration is to develop agents for osteonecrosis. We aimed to identify the pro-angiogenic and osteogenic efficacy of adipose tissue-derived (AD) pericytes combined with Nel-like protein-1 (NELL-1) to investigate the therapeutic effects on osteonecrosis. Tube formation and cell migration were assessed to determine the pro-angiogenic efficacy. Vessel formation was evaluated in vivo using the chorioallantoic membrane assay. A mouse model with a 2.5 mm necrotic bone fragment in the femoral shaft was used as a substitute for osteonecrosis in humans. Bone formation was assessed radiographically (plain radiographs, three-dimensional images, and quantitative analyses), and histomorphometric analyses were performed. To identify factors related to the effects of NELL-1, analysis using microarrays, qRT-PCR, and Western blotting was performed. The results for pro-angiogenic efficacy evaluation identified synergistic effects of pericytes and NELL-1 on tube formation, cell migration, and vessel formation. For osteogenic efficacy analysis, the mouse model for osteonecrosis was treated in combination with pericytes and NELL-1, and the results showed maximum bone formation using radiographic images and quantitative analyses, compared with other treatment groups and showed robust bone and vessel formation using histomorphometric analysis. We identified an association between FGF2 and the effects of NELL-1 using array-based analysis. Thus, combinatorial therapy using AD pericytes and NELL-1 may have potential as a novel treatment for osteonecrosis.
血管化组织再生的一个重要目标是开发治疗骨坏死的药物。我们旨在确定脂肪组织来源的（AD）周细胞与类似Nell 蛋白-1（NELL-1）联合应用的促血管生成和促骨生成效果，以研究其对骨坏死的治疗作用。通过管形成和细胞迁移评估来确定促血管生成效果。通过鸡胚尿囊膜试验评估体内血管形成。在股骨骨干中使用 2.5mm 坏死骨碎片的小鼠模型作为人类骨坏死的替代模型。通过影像学（普通 X 线片、三维图像和定量分析）和组织形态计量学分析评估骨形成。为了确定与 NELL-1 作用相关的因素，进行了微阵列分析、qRT-PCR 和 Western blot 分析。促血管生成功效评估的结果确定了周细胞和 NELL-1 对管形成、细胞迁移和血管形成的协同作用。对于成骨功效分析，用周细胞和 NELL-1 联合治疗骨坏死的小鼠模型，与其他治疗组相比，在影像学图像和定量分析中显示出最大的骨形成，并通过组织形态计量学分析显示出强大的骨和血管形成。我们使用基于阵列的分析确定了 FGF2 与 NELL-1 作用之间的关联。因此，AD 周细胞和 NELL-1 的组合疗法可能具有治疗骨坏死的新疗法的潜力。
