Molecular Characterization and Differentiation of Mesenchymal Progenitor Cells from Human Rheumatoid Arthritis Cartilage.

The presence of mesenchymal progenitor cells (MPCs) in rheumatoid arthritis (RA) articular cartilage is sparsely investigated largely owing to the persistent pathogenic disease condition and lack of specific biomarkers. Considering the recent advancements for potential cell-based therapies in immunomodulatory diseases, such as RA, this in vitro study was aimed at investigating the cellular, molecular, and differentiation characteristics of human RA cartilage-derived MPCs. Articular cartilage fragments from RA patients were obtained for the isolation of MPCs and characterization of their cellular and biological properties, cytogenetic stability, pluripotency, and plasticity. Established MPCs were phenotypically identified using a panel of markers, and their differentiation ability into mesenchymal lineages was assessed by cytochemical staining and the expression of molecular markers. MPCs displayed a heterogenous population of cells with characteristic features of multipotent stem cells. Cells had higher viability, proliferative rate, and colony-forming ability. Further, MPCs showed the expression of pluripotency markers, cytogenetic stability, and minimal replicative senescence. In addition, MPCs differentiated into osteocytes, adipocytes, and chondrocytes, and modulated the expression of each lineage-specific gene markers. The results demonstrated the availability of a viable pool of MPCs residing in RA cartilage, which could serve as an ideal cell source for reinstating native homotypic cartilage.

Influence of human teeth matrix on the cellular and biological properties of dental pulp stem cells - An in vitro study.

BACKGROUND: A major challenge in bone tissue regeneration is the use of right combination of stem cells with osteoinductive biomaterials. Hence, the present in vitro study was aimed at evaluating the effect of mineralized teeth matrix (MTM) and demineralized teeth matrix (DTM) on the selected cellular and biological characteristics of human dental pulp stem cells (DPSCs). METHODS: Established DPSCs were cultured in conditioned media (CM) of MTM and DTM and analyzed on their morphology, proliferation rate, population doubling time (PDT), viability, migration ability, ploidy and expression of cell surface markers, Further, the effect of MTM and DTM on the biocompatibility and osteogenic differentiation ability of DPSCs was evaluated. RESULTS: The DPSCs exhibited a fibroblast-like morphology with >80% viability. Cells were highly proliferative with an average PDT of 61 â€‹± â€‹12 â€‹h. A greater proliferation of DPSCs in the scratched area was observed when cultured in CM of teeth matrix compared to the cells in basal media. Moreover, no chromosomal abnormalities were induced during the culture of DPSCs. Flow cytometry analysis showed that DPSCs in basal media and CM of MTM and DTM were positive for CD29, CD44, CD73, CD90 (>70%), and negative for CD34 and CD45 (<0.1%). Alizarin red staining showed the higher deposition of mineralized nodules in DPSCs cultured with DTM compared to MTM. CONCLUSION: MTM and DTM-derived CM enhanced the proliferation and selected phenotypic markers expression with no chromosomal abnormalities in DPSCs. In addition, both matrices were biocompatible with DPSCs and increased the osteogenic differentiation through higher nodule formation.

Effect of histone acetylation modification with sodium butyrate, a histone deacetylase inhibitor, on cell cycle, apoptosis, ploidy and gene expression in porcine fetal fibroblasts.

The present study evaluated the effective dose of sodium butyrate (NaB), a histone deacetylase (HDAC) inhibitor, for determination of the level of enhancement of histone acetylation in porcine fetal fibroblasts (PFFs) based on their morphology, growth, apoptosis and cell cycle status. Cells were analyzed for their histone acetylation levels at H3, H4 and H2A and expression of genes related to histone deacetylation (HDAC1, HDAC2 and HDAC3), pro-apoptosis (Bax and Bak) and anti-apoptosis (Bcl-2). PFFs at passage 3-4 were cultured with 0, 0.5, 1.0, 2.0 and 3.0 mM NaB for 96 h. NaB inhibited cell proliferation at all tested concentrations in a dose-dependent manner. However, there was slow cell growth for PFFs treated with 2.0 and 3.0 mM NaB compared with those of untreated PFFs and those treated with other lower concentrations (0.5 and 1.0 mM). More than 85% of the cells that were untreated or treated with 0.5 or 1.0 mM NaB had intact membranes, whereas, approximately 30% of the cells treated with 2.0 or 3.0 mM NaB had increased cell sizes and a more flattened and elongated appearance. NaB induced apoptosis in a dose-dependent manner; the rates of apoptosis were 2.5 +/- 0.4% for 1.0 mM NaB, 7.6 +/- 1.1% for 2.0 mM NaB and 11.2 +/- 1.4% for 3.0 mM NaB. The chromosomal sets of PFFs treated with 0.5 and 1.0 mM NaB were normal, whereas a lower proportion of PFFs treated with 2.0 and 3.0 mM were classified as normal. NaB at 0.5 and 1.0 mM showed little effect on cell cycle. However, 2.0 and 3.0 mM resulted in an increased cell population at the G(0)/G(1) phase. Increased NaB concentrations led to elevated acetylation of H3, H4 and H2A. NaB altered the expression of histone deacetylation and apoptosis-related genes. In conclusion, 1.0 mM NaB induced histone hyperacetylation in the PFFs and produced less deleterious effects than other concentrations; these PFFs might serve as suitable donors for porcine somatic cell nuclear transfer (SCNT).