Transcriptome sequencing analysis reveals the effect of combinative treatment with low‑intensity pulsed ultrasound and magnesium ions on hFOB1.19 human osteoblast cells

Abstract

Biodegradable magnesium (Mg) materials are considered ideal as osteosynthesis implants. However, clinical application has proven complex. This is primarily associated with the issue of reducing the extent of implant degradation to a range acceptable for the human body, while simultaneously enhancing osteogenesis or osteoinduction. In the present study, a combination of Mg ions and low-intensity pulsed ultrasound (LIPUS) treatment was applied in hFOB 1.19 human osteoblast cells as a potential strategy to resolve this issue. A total of 7,314 differentially expressed genes (DEGs) and 826 shared DEGs in hFOB1.19 osteoblast cells were identified by microarray analysis following treatment with Mg and/or LIPUS. Gene Ontology analysis demonstrated that among cells treated with a combination of Mg and LIPUS, DEGs were significantly enriched in various functional annotations, including ‘wound healing’, ‘transforming growth factor beta receptor signaling pathway’, ‘transcription, DNA-templated’, ‘receptor complex’, ‘nucleus’, ‘SMAD protein complex’, ‘DNA binding’, ‘metal ion binding’ and ‘GTPase activator activity’. Notably, the transforming growth factor (TGF)-β, mitogen-activated protein kinase (MAPK) and tumor necrosis factor (TNF) signaling pathways were preferentially overrepresented in the Mg and LIPUS combination group, which was subsequently confirmed by reverse transcription-quantitative polymerase chain reaction. Furthermore, genes involved in osteoblast mineralization promotion, including bone morphogenetic protein 6, noggin, bone morphogenetic protein receptor (BMPR)1A, BMPR2 and SMAD 5/8, were significantly upregulated following combination treatment compared with the control group. Genes involved in the promotion of migration, including c-Jun N-terminal kinase, doublecortin, paxillin and Jun proto-oncogene AP-1 transcription factor subunit, were also upregulated in the combination treatment group compared with the control group. The DEG data were supported by morphological observations of the osteoblasts using alizarin red S staining and wound healing assays, which indicated that Mg and LIPUS combinative treatment had a synergistic effect on osteoblast mineralization and migration. Additionally, the combined treatment significantly upregulated metal transporter genes associated with Mg entry, including ATPase Na+/K+-transporting subunit α1, cyclin and CBS domain divalent metal cation transport mediator 2, K+ voltage-gated channel subfamily J member 14, transient receptor potential cation channel (TRP) subfamily M member 7 and TRP subfamily V member 2. In summary, the findings of the present study revealed that combined stimulation with Mg and LIPUS may exhibit a synergistic effect on human osteoblast bone formation through the TGF-β, MAPK and TNF signaling pathways, while also facilitating Mg influx. The present study demonstrated the potential of combinative LIPUS and Mg treatment as a novel therapeutic strategy for enhancing the osteoinduction, biocompatibility and biosafety of biodegradable Mg implants.

LncRNA MALAT1 shuttled by bone marrow-derived mesenchymal stem cells-secreted exosomes alleviates osteoporosis through mediating microRNA-34c/SATB2 axis

“…Human osteoblasts (hFOB1.19) purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) were cultured with DMEM/F12 (Invitrogen, Carlsbad, CA, USA) containing 10% FBS (GE Healthcare Life Sciences, Hyclone, UT, USA) and 1% penicillin and streptomycin (Gibco, Invitrogen, NY, USA) [35].…”

Slow degrading Mg-based materials induce tumor cell dormancy on an osteosarcoma-fibroblast coculture model

“…However, tumor cells can evade this proliferation suppression by inactivating mutations of downstream proteins like SMAD or initiation of inhibitors of cyclin-dependent kinases (CDK) to keep the cell cycle running [ 24 ]. In theory, TGF-β signaling can be linked to Mg degradation through the cofactor activity of Mg 2+ [ 25 ], while another proliferation-associated protein, interleukin-8 (IL-8), is secreted by long-lasting tumor cell exposure to hypoxia and decreased pH [ 26 ]. Moreover, IL-8 was already reported as a potential autocrine growth factor, which was associated with increased tumor cell proliferation [ [27] , [28] , [29] , [30] , [31] ].…”

Identification of key microRNAs and target genes for the diagnosis of bone nonunion

“…These same researchers also determined that the gap junction structural protein pannexin-3 is a mir-367-5p target (8). indeed, there is robust evidence that the expression of specific miRNAs is linked to the occurrence of Bn (9)(10)(11). However, the exact molecular mechanisms by which miRNAs influence gene expression, and thereby regulate Bn pathogenesis, remain incompletely understood, and as such, must be further studied in order to better prevent, treat and diagnose Bn.…”