Abstract
Meniscus fibrochondrocytes (MFCs) may be the optimal cell source to repair non-healing meniscus injuries using tissue engineering strategies. In this study, we investigated the effects of mitotic divisions and oxygen tension on the plasticity of adult human MFCs. Our assessment techniques included gene expression, biochemical, histological, and immunofluorescence assays. MFCs were expanded in monolayer culture with combined growth factors TGFβ1 and FGF-2 (T1F2) under normoxia (21% O2). Trilineage (adipogenesis, chondrogenesis and osteogenesis) differentiation was performed under both normoxic (21% O2) and hypoxic (3% O2) conditions. The data demonstrated that MFCs with a mean total population doubling of 10 can undergo adipogenesis and chondrogenesis. This capability was enhanced under hypoxic conditions. The MFCs did not undergo osteogenesis. In conclusion, our findings suggest that extensively expanded human MFCs have the capacity to generate tissues with the functional matrix characteristics of avascular meniscus. To this end, expanded MFCs may be an ideal cell source for engineering functional constructs for the replacement or repair of avascular meniscus.
Highlights
Due to the unsatisfactory outcomes of current treatments, cell-based tissue engineering (TE) strategies have been an area of interest for meniscus repair or replacement[15]
Isolated meniscus fibrochondrocytes (MFCs) were cultured in monolayer with transforming growth factor β1 (TGFβ1) and fibroblast growth factor-2 (FGF-2) (T1F2) under normal oxygen tension (21% O2)
The primary goals of this study were to: 1) investigate the proliferation rates and subsequent chondrogenic differentiation of TGFβ1 and FGF-2 (T1F2)-expanded MFCs under normal oxygen (21% O2) or low oxygen tension (3%); 2) identify the number of population doublings (PD) these cells can undergo while still maintaining the capacity to form meniscus-like extracellular matrix; 3) to characterize the plasticity of these cells in terms of multilineage differentiation in chondrogenesis, adipogenesis and osteogenesis
Summary
Due to the unsatisfactory outcomes of current treatments, cell-based tissue engineering (TE) strategies have been an area of interest for meniscus repair or replacement[15]. Current work focuses on MFCs as a preferred cell source for two main reasons They are derived from native meniscus tissue and are conditioned to synthesize the functional ECM of meniscus[8]. After expansion in TGFβ1 and FGF-2 both periosteal cells and articular chondrocytes demonstrated enhanced chondrogenic differentiation and restoration of the matrix-forming capacity, respectively. Little is known about the effect of combined TGFβ1 and FGF-2 (T1F2) on MFC proliferation and subsequent redifferentiation capacity under different oxygen tensions To this end, our objectives were to characterize the maximal population doublings (PD) for T1F2-expanded MFCs while retaining their functional matrix-forming capacity. The effect of oxygen tension (normoxia 21% O2, hypoxia 3% O2) on chondrogenic differentiation and matrix-forming phenotype of T1F2-expanded MFCs was tested. We characterized the adipogenic and osteogenic differentiation potential of these T1F2-expanded MFCs
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