Fabrication and characterization of a decellularized bovine tendon sheet for tendon reconstruction

Abstract

Obtaining a performing decellularized tendon scaffold with proper dimensions and adequate availability is highly desirable. However, the combined study of complete decellularization and detailed characterization of native tendon extracellular matrix (ECM) from large animals is still lacking. In the present study, we developed a new decellularization protocol, including physical methods and enzymatic solutions for processing bovine Achilles tendons, and produced a decellularized bovine tendon sheet (DBTS) scaffold for tendon reconstruction. The decellularization effectiveness was demonstrated by DNA quantification and histological qualification. The removal of the alpha-gal epitopes was confirmed by ELISA analysis and immunohistochemical staining. After decellularization, there were no significant alterations of the native tendon extracellular matrix (ECM) properties, including the internal ultrastructure, biochemical compositions such as collagen, glycosaminoglycans (GAGs), basic fibroblast growth factor (bFGF) and transforming growth factor-β1 (TGF-β1), fibronectin and decorin, as well as substantial mechanical strength. Furthermore, the DBTS scaffold showed no cytotoxic and promoted the proliferation of NIH-3T3 fibroblasts in vitro. When implanted into rat subcutaneous tissue, the DBTS scaffold displayed excellent histocompatibility in vivo. Our results, while offering a new decellularization protocol for large tendons, can provide a promising biologic scaffold with a combination of mechanical strength and tendon ECM bioactive factors that may have many potential applications in tendon reconstruction. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2299-2311, 2017.

Improved tendon healing using bFGF, BMP-12 and TGFβ1 in a rat model

“…Other approaches use nanostructural coatings to prolong the release of bFGF and VEGF for several weeks (Yu et al, 2017) or exploit new protocols to decellularise bovine Achilles tendons for use as scaffolds retaining high concentrations of bFGF and TGFβ 1 (Ning et al, 2017). These developments indicate current interest in combining GFs and extending their release profiles.…”

Tenogenesis of Decellularized Porcine Achilles Tendon Matrix Reseeded with Human Tenocytes in the Nude Mice Xenograft Model

“…However, since tendon contains only few cells (5–10% of the tissue volume), no major volume loss could be detected in the present study. The biomechanical properties of the decellularized xECM are dependent on the protocol and are in most cases not severely compromised [ 23 , 25 , 35 , 36 ]; hence, they were not investigated here. Nevertheless, decent tissue clefts are of some advantage to favor cell penetration and distribution during and after reseeding [ 24 ].…”

Evaluation of Decellularized Bovine Tendon Sheets for Achilles Tendon Defect Reconstruction in a Rabbit Model

“…DBTSs were fabricated with a previously published protocol. 31 In brief, preparation of DBTSs included the following procedures: The tendon midsubstances were compressed to ~25% of the initial diameter of the tendon, repetitively frozen in liquid nitrogen and thawed at 37°C a total of 5 times, and treated with nuclease solution (including RNase and DNase) and α-galactose. Then, the sheets were cut longitudinally into pieces 2-cm length × 1.5-cm width, lyophilized, and sterilized with ethylene oxide for further use.…”

“…12,30,38 To prepare the bioscaffolds, the tenocytes and xenoantigens should be removed from xenogenic tendons to minimize inflammatory responses. These bioscaffolds partially retain the structural and functional proteins 31,33 and possess good biocompatibility, thereby offering tendon repair the advantages of recellularization, revascularization, 39 and specific mechanical properties. 41…”

“…Most recently, a study showed that decellularized bovine tendon sheets (DBTSs) prepared from compressed AT segments possessed the inherent ultrastructure, fundamental mechanical strength, multiple ECM components, and good biocompatibility of native tendon. 31 However, the efficacy of the DBTS grafts to reconstruct tendon defects in vivo has not been evaluated. The purpose of this study was to investigate the feasibility of using a DBTS graft as a bioscaffold to repair a large AT defect in a rabbit model.…”