Abstract

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 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.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.

Highlights

  • Bone-tendon interface plays a pivotal role in relaxing load transfer between otherwise structurally and functionally distinct tissue types

  • Hematoxylin and eosin (H&E) and Toluidine blue fast green staining showed that the cellular components of the decellularized bookshaped enthesis scaffolds were absolutely removed, while the structure and morphology of the native enthesis extracellular matrix were well preserved (Fig. 2b, c)

  • Dapi-positive cell nuclei were rarely shown in the decellularized book-shaped enthesis scaffolds (Fig. 2d)

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Summary

Introduction

Bone-tendon interface (enthesis) plays a pivotal role in relaxing load transfer between otherwise structurally and functionally distinct tissue types. Bone-tendon interface (BTI), which is named as enthesis, serves as an interface for force transmission from bone to tendon that consists of four transitional tissues: tendon, uncalcified fibrocartilage, calcified fibrocartilage, and bone [1, 2] This transitional enthesis allows smooth transmission of forces derived from muscle contraction and minimizes formation of stress peaks [3, 4]. Enthesis injury, those in the rotator cuff, are prevalent conditions that often lead to disability and persistent pain [5]. As a result, decellularized extracellular matrix from enthesis may provide a natural three-dimensional scaffold with tissue-specific orientations of extracellular matrix molecules for enthesis regeneration. In this study, bone was treated together with fibrocartilage and tendons with different solutions in the same composite

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