Interleukin-35 Promotes The Osteogenic Differentiation Of Dental Pulp Stem Cells In Vitro

Objective: Bone defects caused by chronic inflammation after trauma cases or burns,tumors and congenital defect can bring restricted mobility and abnormal appearance but there has not been any world recognized clinical method,providing bone graft in regardless of short supply.Dental pulp stem cells(DPSCs)are placed on increasingly great expectations to repair bone defects due to their ability of large expansion in laboratory cell cultures,stable osteogenic capability and the non-invasive way to be harvested.Interleukin-35(IL-35)is a novel anti-inflammatory cytokine and plays an important role in regulation of T cell mediated inflammation,which is reported to be functional in bone metabolic activities and bone diseases.In this study,I induced human DPSCs differentiation into osteoblasts with or without recombinant human IL-35 to explore the effects of IL-35 on DPSCs osteogenic differentiation.Methods: DPSCs were isolated from the third molar teeth of patients undergoing orthodontic extraction,followed by being cultured under alpha minimum essential medium to Passage 3.Osteogenic,adipogenic and neurogenesis conditional medium were used to induce DPSCs in order to identify them.DPSCs were cultivated in osteogenic conditioned medium with or without recombinant IL-35 for 3 weeks to drive them into osteoblasts differentiation.After 3-week differentiation,cells were stained with Alizarin Red S followed by quantitative analysis with Cetylpyridinium Chloride(CPC)colorimetry.Quantitative real-time polymerase chain reaction(RT-q PCR)was carried out to examine the expression of the receptors of IL-35 and osteogenesis-related gene markers,such as runt-related transcription factors(Runx2)and collagen type ?(COL-?).PCR specificity was evaluated by Agarose gel electrophoresis of PCR products.Results: DPSCs differentiation into osteoblast cells was demonstrated by Alizarin Red S staining and osteogenic gene markers,along with which DPSCs expressed glycoprotein 130(gp130)increasingly.Recombinant IL-35 promoted the osteogenic differentiation from DPSCs with increased expression of Runx2,COL-1 and the CPC quantitative analysis.Conclusion: IL-35 is able to facilitate the DPSC osteogenic differentiation for bone repair and regeneration from DPSCs.