Characterization of the distributions of collagen and PGs content in the decellularized book-shaped enthesis scaffolds by SR-FTIR

BackgroundBone-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.MethodsNative enthesis tissues (NET) harvested from rabbit rotator cuff were sectioned into cuboid (about 30mmx1.2mmx10mm) 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.ResultsThe 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.ConclusionSR-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.

Automated summary

The study successfully obtained decellularized book-shaped enthesis scaffolds from rabbit rotator cuff, evaluated the effect of decellularization on extracellular matrix structure through histological staining and SEM, and found that the content of collagen and PGs in the decellularized scaffolds decreased significantly. This innovative use of SR-FTIR can provide quantitative mapping of collagen and PGs distribution in enthesis scaffolds.