通过同步辐射傅里叶变换红外光谱（SR-FTIR）对脱细胞书本状附着点支架中胶原蛋白和蛋白聚糖含量的分布进行表征。
Characterization of the distributions of collagen and PGs content in the decellularized book-shaped enthesis scaffolds by SR-FTIR.
作者信息
Shi Qiang, Chen Can, Li Muzhi, Chen Yang, Xu Yan, Hu Jianzhong, Liu Jun, Lu Hongbin
机构信息
Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China.
Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, 410008, Hunan, China.
出版信息
BMC Musculoskelet Disord. 2021 Mar 1;22(1):235. doi: 10.1186/s12891-021-04106-x.
BACKGROUND
Bone-tendon interface (enthesis) plays a pivotal role in relaxing load transfer between otherwise structurally and functionally distinct tissue types. Currently, decellularized extracellular matrix (DEM) from enthesis provide a natural three-dimensional scaffold with tissue-specific orientations of extracellular matrix molecules for enthesis regeneration, however, the distributions of collagen and PGs content in the decellularized book-shaped enthesis scaffolds from rabbit rotator cuff by SR-FTIR have not been reported.
METHODS
Native enthesis tissues (NET) harvested from rabbit rotator cuff were sectioned into cuboid (about 30 mm × 1.2 mm × 10 mm) for decalcification. The decellularized book-shaped enthesis scaffolds and intrinsic ultrastructure were evaluated by histological staining and scanning electron microscopy (SEM), respectively. The distributions of collagen and PGs content in the decellularized book-shaped enthesis scaffolds from rabbit rotator cuff were also measured innovatively by SR-FTIR.
RESULTS
The decellularized book-shaped enthesis scaffolds from rabbit rotator cuff were successfully obtained. Histomorphology and SEM evaluated the effect of decellularization and the structure of extracellular matrix during decellularization. After mechanical testing, the failure load in the NET group showed significantly higher than that in the DEM group (P < 0.05). Meanwhile, the stiffness of the DEM group was significantly lower than the NET group. Furthermore, the distributions of collagen and PGs content in the decellularized book-shaped enthesis scaffolds were decreased obviously after decellularization by SR-FTIR quantitative analysis.
CONCLUSION
SR-FTIR was applied innovatively to characterize the histological morphology of native enthesis tissues from rabbit rotator cuff. Moreover, this technology can be applied for quantitative mapping of the distribution of collagen and PGs content in the decellularized book-shaped enthesis scaffolds.
背景
骨 - 肌腱界面（附着点）在缓解结构和功能不同的组织类型之间的负荷传递中起关键作用。目前，来自附着点的脱细胞细胞外基质（DEM）为附着点再生提供了具有细胞外基质分子组织特异性取向的天然三维支架，然而，尚未有关于通过同步辐射傅里叶变换红外光谱（SR - FTIR）对兔肩袖脱细胞书本状附着点支架中胶原蛋白和蛋白聚糖（PGs）含量分布的报道。
方法
从兔肩袖获取的天然附着点组织（NET）切成长方体（约30mm×1.2mm×10mm）进行脱钙处理。分别通过组织学染色和扫描电子显微镜（SEM）评估脱细胞书本状附着点支架及其固有超微结构。还通过SR - FTIR创新性地测量了兔肩袖脱细胞书本状附着点支架中胶原蛋白和PGs的含量分布。
结果
成功获得了兔肩袖脱细胞书本状附着点支架。组织形态学和SEM评估了脱细胞效果以及脱细胞过程中细胞外基质的结构。力学测试后，NET组的破坏载荷显著高于DEM组（P < 0.05）。同时，DEM组的刚度明显低于NET组。此外，通过SR - FTIR定量分析表明，脱细胞后兔肩袖脱细胞书本状附着点支架中胶原蛋白和PGs的含量分布明显降低。
结论
创新性地应用SR - FTIR来表征兔肩袖天然附着点组织的组织形态学。此外，该技术可用于定量绘制脱细胞书本状附着点支架中胶原蛋白和PGs含量的分布图。
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