骨膜微环境下脂肪脱细胞基质再生和分化的研究

Regeneration and differentiation of decellularized adipose matrix in periosteal microenvironment

目的观察骨膜微环境下,脂肪脱细胞基质(DAM)在体内向成脂或成骨方向分化的情况。方法2020年6—12月,将人体吸脂术获取的脂肪组织在全军整形外科研究所制备为DAM,选取6只4~6周雄性SD大鼠,按照同等初始体积植入到大鼠顶骨骨膜下。术后84 d取材,通过大体观察、组织学染色、免疫荧光染色观察DAM支架材料成脂和成骨分化情况;非标记定量蛋白质谱检测DAM中成骨相关蛋白。结果大体观察显示,DAM支架材料周围血管化丰富。HE和Masson染色观察到DAM脂肪再生和血管化现象,同时OCN免疫荧光染色证实,DAM少部分向成骨方向分化。质谱筛选出了成骨分化相关蛋白。结论骨膜微环境下,DAM主要向脂肪组织方向分化,但少部分具有向成骨方向分化的潜能,可能与DAM中含有部分成骨相关蛋白有关。

Objective To observe the lipogenic or osteogenic differentiation of decellularized adipose matrix(DAM)in the periosteal microenvironment.Methods From June 2020 to December 2020,adipose tissue obtained by human liposuction was prepared as DAM at the National Institute of Plastic Surgery.Six male SD rats of 4 to 6 weeks were selected and implanted into the subperiosteum of the rat parietal bone according to the same initial volume.After 12 weeks,adipogenesis and osteogenic differentiation of DAM were observed by gross specimens,histological staining,and immunofluorescence staining.Label-free quantitative protein mass spectrometry was used for detection of osteogenic-associated proteins in DAM.Results Vascularization around the DAM was evident.Adipogenesis and angiogenesis were observed in DAM by H&E and Masson staining,while OCN immunofluorescence staining confirmed osteogenic differentiation of DAM.The osteogenic differentiation related proteins were screened by mass spectrometry.Conclusions In the microenvironment of periosteum,DAM mainly differentiates towards adipose tissue,but a few of them havs the potential to differentiate towards osteogenesis,which might be related to some of the osteogenesis-related proteins contained in DAM.