An integrated study of natural hydroxyapatite-induced osteogenic differentiation of mesenchymal stem cells using transcriptomics, proteomics and microRNA analyses

Abstract

This work combined transcriptomics, proteomics, and microRNA (miRNA) analyses to elucidate the mechanism of natural hydroxyapatite (NHA)-induced osteogenic differentiation of mesenchymal stem cells (MSCs). First, NHA powder was obtained from pig bones and fabricated into disc-shaped samples. Subsequently, the proliferation and osteogenic differentiation of MSCs cultured on NHA were investigated. Then, proteomics was employed to detect the protein expression profiles of MSCs cultured on NHA, and the effect of NHA on MSCs was analyzed through an integrated pathway analysis (including proteomics and previous transcriptomics data) in which specific NHA-induced differentiation pathways were analyzed. The pathway nodes with expression data at both the mRNA and protein levels (mRNA-protein pairs) were filtered in differentiation-related pathways. miRNAs corresponding to these target mRNA-protein pairs were predicted, screened and tested, and the regulatory effects of miRNAs on mRNA-protein pairs were analyzed. Finally, the NHA-induced osteogenic pathways were verified. The results of an MTT assay and alkaline phosphatase (ALP) staining showed that the cell proliferation rate decreased and the osteogenic performance improved in the presence of NHA. By integrating transcriptomics and proteomics, the genes and proteins involved in 89 pathways were shown to be differentially expressed. Among them, 5 differentiation-associated pathways, in which 9 miRNAs and 8 regulated-target mRNA-protein zby inhibiting the target mRNA-protein pair HSPA8 in the MAPK signaling pathway, and miR-26a and miR-26b might inhibit adipogenic differentiation by repressing the target mRNA-protein pair HMGA1 in the adipogenesis pathway. A verification experiment for the osteogenic pathway indicated that the ERK1/2 or JNK MAPK pathways might play an important role in NHA-induced osteogenic differentiation. In conclusion, NHA affected MSCs at both the transcriptional and translational levels, and MSC osteogenic differentiation eventually occurred through the MAPK and adipogenesis pathways, in which miRNAs and target mRNAs/proteins participated cooperatively.

Cellular and Genetic Determinants of the Sensitivity of Cancer to α-Particle Irradiation

“…The osteomimetic hydroxyapatite and the cellular response to a-particles Hydroxyapatite is a calcium phosphate similar to the human bone in morphology and composition insofar as it has a hexagonal structure and a similar stoichiometric calcium-to-phosphate ratio (37). Hydroxyapatite has previously been shown to induce osteogenic differentiation from cellular precursors (38) and alter the transcriptome and proteome of cocultured cells (39). Because 223 Ra exerts its effect by incorporation into the bone, we sought to assess the impact of hydroxyapatite on cellular sensitivity to a-particles using a coincubation method (Fig.…”

Transcriptomics of human multipotent mesenchymal stromal cells: Retrospective analysis and future prospects

“…MSC differentiation has been a focus of past comparative transcriptome approaches, which has included studies of functional differences in MSC populations (Jansen et al, 2010), characterization of MSC stemness (Gao et al, 2013;Steinert et al, 2015), and assessments of differentiation status (Brown et al, 2015;Granchi et al, 2010;Liu et al, 2014;Menssen et al, 2011;Modder et al, 2012;Satija et al, 2013;Skreti et al, 2014;Steinert et al, 2015;Wislet-Gendebien et al, 2012;Yu et al, 2013;Zhang et al, 2014). For example, temporal and spatial patterns of gene expression profiles were reported for chondrogenic differentiation (Skreti et al, 2014).…”

The effect of hydroxyapatite nanoparticles on adipogenic differentiation of human mesenchymal stem cells

“…According to the inverse relationship, HA has the potential to suppress adipogenesis. Several previous studies have shown that HA coating or HA discs can inhibit the adipogenesis of different cell types . For instance, Zhang et al found that HA discs composed of natural HA powder inhibited adipogenic differentiation of mouse bone marrow derived MSCs by regulating several microRNAs in the adipogenesis pathway.…”

“…Several previous studies have shown that HA coating or HA discs can inhibit the adipogenesis of different cell types . For instance, Zhang et al found that HA discs composed of natural HA powder inhibited adipogenic differentiation of mouse bone marrow derived MSCs by regulating several microRNAs in the adipogenesis pathway. Iijima et al have reported that HA coating and synthetic HA discs suppressed the adipogenic differentiation of hMSCs through decreasing the formation of lipid droplets and the expression of PPARγ and C/EBPα.…”