Donor-Matched Comparison of Chondrogenic Potential of Equine Bone Marrow- and Synovial Fluid-Derived Mesenchymal Stem Cells: Implications for Cartilage Tissue Regeneration.

Mohammed Zayed,Nabil Misk,Madhu S Dhar,Christopher Caniglia

doi:10.3389/fvets.2016.00121

Abstract

Mesenchymal stem cells (MSCs) have been demonstrated to be useful for cartilage tissue regeneration. Bone marrow (BM) and synovial fluid (SF) are promising sources for MSCs to be used in cartilage regeneration. In order to improve the clinical outcomes, it is recommended that prior to clinical use, the cellular properties and, specifically, their chondrogenic potential must be investigated. The purpose of this study is to compare and better understand the in vitro chondrogenic potential of equine bone marrow-derived mesenchymal stem cells (BMMSCs) and synovial fluid-derived mesenchymal stem cells (SFMSCs) populated from the same equine donor. BM- and SF-derived MSCs cultures were generated from five equine donors, and the MSCs were evaluated in vitro for their morphology, proliferation, trilineage differentiation, and immunophenotyping. Differences in their chondrogenic potentials were further evaluated quantitatively using glycosaminoglycan (GAG) content and via immunofluorescence of chondrogenic differentiation protein markers, SRY-type HMG box9, Aggrecan, and collagen II. The BMMSCs and SFMSCs were similar in cellular morphology, viability, and immunophenotype, but, varied in their chondrogenic potential, and expression of the key chondrogenic proteins. The SFMSCs exhibited a significant increase in GAG content compared to the BMMSCs (P < 0.0001) in three donors, suggesting increased levels of chondrogenesis. The expression of the key chondrogenic proteins correlated positively with the GAG content, suggesting that the differentiation process is dependent on the expression of the target proteins in these three donors. Our findings suggest that even though SFMSCs were hypothesized to be more chondrogenic relative to BMMSCs, there was considerable donor-to-donor variation in the primary cultures of MSCs which can significantly affect their downstream application.

Highlights

Osteoarthritis (OA) is a chronic disease pertaining to progressive deterioration of the articular cartilage and subchondral bone [1]
Cellular proliferation and viability of bone marrow-derived mesenchymal stem cells (BMMSCs) and synovial fluid-derived mesenchymal stem cells (SFMSCs) were assessed through a period of 8 days using MTS assay (Figure 2)
To alleviate some of the technical challenges associated with the tissue harvest and properties of mesenchymal stem cells (MSCs) cultured ex vivo from bone marrow (BM) and adipose tissue (AD), recently, synovial fluid (SF) and the synovium membrane have been identified as alternative sources

Summary

Introduction

Osteoarthritis (OA) is a chronic disease pertaining to progressive deterioration of the articular cartilage and subchondral bone [1]. Attempts have been made to treat OA and prevent joint degeneration, which primarily includes injection of pain relief medications to reduce inflammation and treat pain [5]. Cartilage degeneration is a sequel to OA, aggressive surgical approaches such as chondrocyte implantation have been used in the treatment of massive chondral injuries in human and equine patients [6, 7]. No therapies are available to effectively regenerate the affected tissue, posing OA as a major health concern. Mesenchymal stem cells (MSCs) have shown promise in human and equine regenerative medicine and have been used in tissue regeneration and treatment of many diseases including those affecting the joint [8,9,10,11,12]

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