Chondrogenic differentiation of synovial fluid mesenchymal stem cells on human meniscus-derived decellularized matrix requires exogenous growth factors

Autor: Aillette Mulet-Sierra, Melanie Kunze, Adetola B Adesida, Nadr M. Jomha, Alexander R A Szojka, Stephen H. J. Andrews, Enaam Idrees, Yan Liang
Rok vydání: 2018
Předmět:
0301 basic medicine
Adult
Male
Adolescent
medicine.medical_treatment
Biomedical Engineering
02 engineering and technology
Cell Separation
Matrix (biology)
Meniscus (anatomy)
Biochemistry
Collagen Type I
Biomaterials
Extracellular matrix
Colony-Forming Units Assay
03 medical and health sciences
Tissue engineering
Synovial Fluid
medicine
Synovial fluid
Humans
Meniscus
Molecular Biology
Collagen Type II
Cell Proliferation
Glycosaminoglycans
Tissue Scaffolds
Chemistry
Growth factor
Mesenchymal stem cell
Cell Differentiation
Mesenchymal Stem Cells
General Medicine
DNA
Fibroblasts
Middle Aged
musculoskeletal system
021001 nanoscience & nanotechnology
Cell biology
Extracellular Matrix
030104 developmental biology
medicine.anatomical_structure
Intercellular Signaling Peptides and Proteins
Female
Stem cell
0210 nano-technology
Chondrogenesis
Biomarkers
Biotechnology
Zdroj: Acta biomaterialia. 80
ISSN: 1878-7568
Popis: The objective of this study was to investigate whether meniscus-derived decellularized matrix (DCM) has the capacity to induce differentiation of synovial fluid-derived mesenchymal stem cells (SF-MSCs) towards a meniscus fibrochondrocyte (MFC) phenotype. The potential roles of transforming growth factor beta-3 (TGF-β3) and insulin-like growth factor 1 (IGF-1) in the differentiation of SF-MSCs towards an MFC phenotype were also investigated. SF-MSCs were isolated via plastic adherence cell culture from the synovial fluid of five donors (5 male, average age 34 years). Porous DCM was generated by homogenizing and freeze-drying fresh normal human cadaveric meniscus tissue. SF-MSCs were seeded and cultured on the DCM scaffold in a defined serum-free media (SFM) supplemented with or without the combination of TGF-β3 and IGF-1. Cell pellets of SF-MSCs were cultured in SFM with either TGF-β3 or IGF-1 or their combination as controls. The duration of culture was 3 weeks for both experimental configurations. We assessed newly-formed tissues by biochemical assays, scanning electron microscopy (SEM), immunofluorescence and quantitative real-time PCR (qPCR). The combination of TGF-β3 and IGF-1 induced production of the cartilaginous matrix in DCM and upregulated the expression of aggrecan, collagens I and II. Moreover, the SF-MSCs exhibited a round morphology in the DCM scaffolds in the presence of the growth factors. In pellets, combined TGF-β3 and IGF-1 synergistically enhanced cartilaginous matrix production. In contrast to bone marrow mesenchymal stem cells (BM-MSCs), the differentiated SF-MSCs showed little evidence of the expression of the hypertrophic differentiation marker, collagen X. In conclusion, meniscus-derived DCM appears to require exogenous growth factor supplementation to direct differentiation of SF-MSCs. Statement of significance Meniscus tears are the most common injury of the knee joint. These tears pose a major risk factor for the early development of knee osteoarthritis. Unfortunately, the majority of these tears occur in the inner region of the meniscus and lacks blood supply with no reparative or regenerative capacity. The goal of this study was to determine if the native extracellular matrix (ECM) of human meniscus has the capacity to differentiate human knee synovial fluid resident mesenchymal stem cells (SF-MSCs) towards a meniscus phenotype as a potential strategy to repair avascular meniscal tears. Our findings show that the human meniscus-derived ECM without supplementation with growth factors (TGF-β3 and IGF-1) cannot differentiate SF-MSCs towards a meniscus phenotype. The use of meniscus-derived scaffolds as a material to stimulate endogenous repair of meniscus tears via differentiation of SF-MSCs may require supplementation with TGF-β3 and IGF-1.
Databáze: OpenAIRE
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