Nucleus Pulposus Mesenchymal Stem Cells in Acidic Conditions Mimicking Degenerative Intervertebral Discs Give Better Performance than Adipose Tissue-Derived Mesenchymal Stem Cells

Abstract

The microenvironment of the intervertebral disc (IVD) is characterized by matrix acidity, hypoxia, hyperosmolarity and limited nutrition, which are major obstacles to stem cell-based regeneration. Our recent work showed that nucleus pulposus mesenchymal stem cells (NPMSCs) had advantages over traditional sources of cell therapy under IVD-like hypoxic and hyperosmotic conditions. Here, we examined the viability, proliferation and matrix metabolism of NPMSCs compared with adipose tissue-derived mesenchymal stem cells (ADMSCs) under IVD-like acidic conditions in vitro. ADMSCs and NPMSCs from Sprague-Dawley rats were cultured at four different pH levels representing the standard condition (pH 7.4) and the normal, mildly degenerated and severely degenerated IVD (pH 7.1, 6.8 and 6.5, respectively). Cell viability was examined by annexin-V-fluorescein isothiocyanate/propidium iodide staining. Cell proliferation was measured using a cell counting kit cell proliferation assay. The expression of aggrecan, collagen-I, collagen-II, matrix metalloproteinase-2 (MMP-2), a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS4) and the tissue inhibitor of metalloproteinase-3 (TIMP-3) was measured at mRNA and protein levels by RT-PCR and Western blotting. In both cell types, acidic pH inhibited cell viability and proliferation, downregulated the expression of aggrecan, collagen-I, collagen-II and TIMP-3, and upregulated the expression of MMP-2 and ADAMTS4. Compared with ADMSCs, NPMSCs were significantly less inhibited in viability and proliferation; they expressed significantly higher levels of aggrecan and collagen-II, and lower levels of MMP-2 and ADAMTS4. Thus, an acidic environment is a major obstacle for IVD regeneration by ADMSCs or NPMSCs. NPMSCs appeared less sensitive to inhibition by acidic pH and might be promising candidates for cell-based IVD regeneration.