Osteogenic and angiogenic lineage differentiated adipose-derived stem cells for bone regeneration of calvarial defects in rabbits
Osteogenic and angiogenic lineage differentiated adipose-derived stem cells for bone regeneration of calvarial defects in rabbits
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成骨和血管生成谱系分化的脂肪干细胞用于兔颅骨缺损的骨再生
DOI：
10.1002/jbm.a.37036
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发表时间：
--
影响因子：
4.9
通讯作者：
Wu Buling
中科院分区：
文献类型：
--
作者：
Wang Zhifa;Han Leng;Sun Tianyu;Wang Weijian;Li Xiao;Wu Buling
细胞片技术在骨工程中有着广泛的应用。然而，血管化仍然是构建血管化工程骨的一个挑战。本研究的目的是诱导脂肪干细胞(ADSC)向成骨和血管生成方向分化，并研究双向分化的ADSC在骨再生中的应用。ADSCs经体外培养形成成骨细胞片。其他ADSCs被诱导分化为内皮祖细胞，并通过形态观察和CD31免疫荧光染色对其进行鉴定。然后，将ADSC片材-EPC复合体植入裸鼠皮下，同时单独植入ADSC片材作为对照。移植8 周后，通过显微CT和组织学观察骨形成情况。然后将复合体植入兔颅骨缺损处，通过显微CT和组织学分析评价骨修复情况。ADSC膜由多层细胞和细胞外基质组成。所获得的内皮细胞形成毛细血管样结构，并表达特异性抗原标记CD31。成骨ADSCSheet-EPC复合体在植入后8 形成致密、血管化良好的新骨组织。对照组的骨密度显著低于复合体组(p<； .05)。此外，复合体组兔颅骨缺损的修复明显大于对照组(p<； .05)。这些结果表明，双向分化的ADSCs工程化骨组织的方法能够实现骨再生，从而为骨缺损的修复提供了一种有前景的策略。
Cell sheet techniques are widely used in bone engineering. However, vascularization remains a challenge in fabricating vascularized engineered bone. The goal of this study was to induce adipose‐derived stem cell (ADSC) osteogenic and angiogenic lineage differentiation and investigate the use of bidiretionally differentiated ADSCs for bone regeneration. ADSCs were cultured to form an osteogenic cell sheet. Other ADSCs were induced to differentiate into endothelial progenitor cells (EPCs), which were identified and characterized by morphological observation and CD31 immunofluorescent staining. Then, the ADSC sheet‐EPC complexes were implanted subcutaneously into nude mice, while ADSC sheets alone were implanted as a control. After 8 weeks of transplantation, microcomputed tomography (micro‐CT) and histological observation were used to assess bone formation. We then implanted the complexes in calvarial defects in rabbits and assessed bone repair by micro‐CT and histological analysis. The ADSC sheets consisted of multiple layers of cells and extracellular matrix. The obtained EPCs formed capillary‐like structures and expressed the specific antigen marker CD31. The osteogenic ADSC sheet‐EPC complexes formed dense and well‐vascularized new bone tissue at 8 weeks after implantation. Bone density was significantly lower in the control group than in the complex group (p< .05). In addition, the reconstruction of calvarial defects in rabbits in complex group was obviously greater than that in the control group (p< .05). These results suggested that the approach of engineering bone tissue with bidiretionally differentiated ADSCs enabled bone regeneration, thus offering a promising strategy for repairing bone defects.