Functional Characterization of Detergent-Decellularized Equine Tendon Extracellular Matrix for Tissue Engineering Applications

Abstract

Natural extracellular matrix provides a number of distinct advantages for engineering replacement orthopedic tissue due to its intrinsic functional properties. The goal of this study was to optimize a biologically derived scaffold for tendon tissue engineering using equine flexor digitorum superficialis tendons. We investigated changes in scaffold composition and ultrastructure in response to several mechanical, detergent and enzymatic decellularization protocols using microscopic techniques and a panel of biochemical assays to evaluate total protein, collagen, glycosaminoglycan, and deoxyribonucleic acid content. Biocompatibility was also assessed with static mesenchymal stem cell (MSC) culture. Implementation of a combination of freeze/thaw cycles, incubation in 2% sodium dodecyl sulfate (SDS), trypsinization, treatment with DNase-I, and ethanol sterilization produced a non-cytotoxic biomaterial free of appreciable residual cellular debris with no significant modification of biomechanical properties. These decellularized tendon scaffolds (DTS) are suitable for complex tissue engineering applications, as they provide a clean slate for cell culture while maintaining native three-dimensional architecture.

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

“…Other research also showed that MSCs reseeded on 2% SDS-treated equine tendon ECM were successful in terms of plating efficiency, cell proliferation, and viability. 46 Therefore, the recellularization ability of ECM scaffolds derived from fibroblast cell sheets is independent of decellularization methods and the amount of cellular remnants in our study.…”

“…The hMSCs are often employed as a model cell line to evaluate the feasibility of using decellularized scaffolds for tissue engineering, 46 because hMSCs have wide applications in bone, tendon, skin, vascular, and neural tissue engineering. It was reported that when SDS was added to the decellularization protocol, the ability of porcine dermis to support in vitro cell growth significantly decreased due to the harsh effect of SDS.…”

Aligned Nanofiber Topography Directs the Tenogenic Differentiation of Mesenchymal Stem Cells

“…We found that MSCs adhered to and elongated on fibrous scaffolds of both parallel and perpendicular oriented fibers, maintained viability, and migrated along both individual and multiple fibers regardless of fiber orientation. Previous studies using aligned fibrous scaffolds have shown that MSCs elongate along aligned fibrous scaffolds to promote tenocyte-like morphologies and tendon differentiation, while random electrospun fibrous scaffolds promote polygonal cell morphology and random cell orientation [16,31,32]. Instead of randomly oriented fibers, we used aligned perpendicular fibers resulting in primarily two different morphologies.…”

Topical Collagen-Based Biomaterials for Chronic Wounds: Rationale and Clinical Application

“…Decellularization may be accomplished by acid, detergent, enzymatic, and mechanical methods, all of which can affect the biochemical and structural characteristics [47][48][49] ( Fig. 4).…”