RESUMO
This study aimed to investigate the hub genes and miRNA-mRNA regulatory network around periodontal ligament stem cells (PDLSC) for osteogenic differentiation through bioinformatic analysis. The dataset with osteogenic differentiation of human PDLSC was downloaded from the GEO database. The Weighted gene coexpression network analysis (WGCNA) was performed to identify key modules and hub genes. In addition, differentially expressed genes (DEGs) analysis was conducted with limma. The functional enrichment of differentially expressed hub genes was implemented with KEGG and GSEA analysis. The targeted genes of differentially expressed miRNA were predicted based on miRWalk database. The miRNA-mRNA interaction network of osteogenic differentiation of PDLSC was constructed and visualized. The WGNCA results showed that the light-cyan module was positively correlated with osteogenic differentiation (r=0.98, P<0.05). A total of 3125 hub genes and 1426 differentially expressed hub genes were detected in OG group. Innate immune-related signaling pathways and metabolic pathways were involved in the osteogenic differentiation. In addition, total of 2 upregulated miRNAs with 63 targeted DEGs and 6 downregulated miRNAs with 214 targeted DEGs were detected, which contributed to osteogenic differentiation by regulating amino acid metabolism signaling pathway. We identified hub genes and miRNA-mRNA regulatory network contributing to osteogenic differentiation of human PDLSC, which will provide novel strategy for periodontal disease therapy.
Assuntos
Diferenciação Celular , Redes Reguladoras de Genes , MicroRNAs , Osteogênese , Ligamento Periodontal , RNA Mensageiro , Células-Tronco , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Osteogênese/genética , Células-Tronco/metabolismo , Células-Tronco/citologia , Diferenciação Celular/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Perfilação da Expressão Gênica , Biologia Computacional/métodos , Regulação da Expressão Gênica , Transdução de Sinais/genéticaRESUMO
Osteoinductive bone filling biomaterials are in high demand for effective bone defect reconstruction. In this study, we aimed to design both organic and inorganic substances containing strontium-doped hydroxyapatite/silk fibroin (SrHA/SF) biocomposite nanospheres as an osteoinductive bone defect-filling biomaterial. SrHA/SF nanospheres were prepared with different concentration of Sr using ultrasonic coprecipitation method. The nanospheres were characterized using XRD, FTIR, SEM, TEM, ICP-AES and TGA. Solid and dense SrHA/SF nanospheres with 500-700â¯nm size and rough surfaces were synthesized successfully. Higher crystallinity and HA/SF phase were observed with the increase in Sr-concentration. The doping of different concentration of Sr did not affect the size and surface characteristics of the nanospheres. ICP-AES data showed that Sr/Ca ratio in SrHA/SF is very close to the nominal value. Nanospheres with higher concentration of Sr did not negatively affect the biocompatibility, but enhanced viability of mesenchymal stem cells (MSCs). Moreover, SrHA/SF nanospheres showed higher osteogenic differentiation potential compared to HA/SF nanospheres as indicated by the results from ALP staining, ALP activity, and Runx2, Alp, Col-1 and Opn gene expression assay in MSCs culture. Our findings suggest this novel design of biocompatible and osteoinductive SrHA/SF biocomposite nanospheres as a potential bone defect-filling biomaterial for bone regenerative applications.
Assuntos
Fibroínas/química , Hidroxiapatitas/química , Nanosferas/química , Seda/química , Estrôncio/química , Engenharia Tecidual , Alicerces Teciduais/química , Animais , Materiais Biocompatíveis/química , Biomarcadores , Diferenciação Celular , Células Cultivadas , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Osteogênese , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
Extending the release cycle of growth factors to match the cycle of bone remodeling is difficult. When using concentrated growth factors (CGFs), the release of growth factors is excessively rapid. In the present study, CGF samples were prepared by centrifugation. CGF samples were then lyophilized and grinded into a powder, which was termed freezedried CGF. The freezedried CGF samples were mixed with chitosanalginate composite hydrogels, and the mixture was lyophilized. The result was a chitosanalginate composite CGF membrane, which was called sustainedrelease CGF. This study investigated whether freezedried CGF in a chitosanalginate composite gel can release CGF steadily to achieve effective osteogenesis. The proliferation and osteogenic expression of MC3T3E1 cells induced by the supernatants from incubation with freezedried CGF and sustainedrelease CGF were evaluated. The concentrations of the growth factors, transforming growth factor ß1 (TGFß1), insulinlike growth factor1 (IGF1), plateletderived growth factorAB (PDGFAB) and vascular endothelial growth factor (VEGF), in these two experimental groups at different times were determined by ELISA kits. The freezedried CGF showed better osteogenic performance than the sustainedrelease CGF in the early stages. At later stages, the sustainedrelease CGF had significant advantages over freezedried CGF in terms of promoting osteogenic mineralization. By characterizing the biologic properties of the CGF in the two different forms in vitro, we obtained a better understanding of their clinical effects.
