title: Paracrine effect of inflammatory cytokine-activated bone marrow mesenchymal stem cells and its role in osteoblast function.
abstract: Mesenchymal stem cells (MSCs) have a crucial function in bone regeneration. Inflammation is a well-documented component of the osteogenic microenvironment. In the present study, we investigated whether stimulation of MSCs with inflammatory cytokines promotes osteogenesis through a paracrine mediator. MSCs were pre-stimulated with the inflammatory factors IFN-γ and TNF-α. After pre-stimulation, the MSC secretion levels of IL-6, HGF, VEGF, and TGF-β were significantly elevated (p < 0.01); however, the production of IL-2, IL-4, and IL-10 was not changed (p > 0.05). MG63, an osteoblast-like cell line, was cultured in different MSC-conditioned media. After treatment with conditioned media collected from MSCs pre-treated with cytokines, the proliferation and migration of MG63 cells were significantly improved, and the expression levels of the osteoblast differentiation markers ALP, COLI, OCN and OPN were significantly increased as revealed by a quantitative PCR analysis (p < 0.05). Furthermore, an immunofluorescence staining assay showed that more MG63 cells were OPN-positive, while an Alizarin red staining indicated the increased formation of calcium nodules in the IFN-γ and TNF-α combined pretreatment group. The results indicated that conditioned medium from inflammatory cytokine-activated MSCs can significantly promote osteoblast proliferation, migration, differentiation, and mineralization and ultimately enhance osteogenesis through paracrine mechanisms. These findings present a new direction for the clinical application of MSCs in the repair of bone defects.
authors: Department of Orthopedics, The Second Affiliated Hospital, Soochow University, Suzhou 215004, China; Cyrus Tang Hematology Center, Soochow University, Suzhou 215123, China.
authors: Cheng
authors: C
authors: Li
authors: Department of Orthopedics, The Second Affiliated Hospital, Soochow University, Suzhou 215004, China; Cyrus Tang Hematology Center, Soochow University, Suzhou 215123, China.
authors: Guoqiang
authors: G
authors: Li
authors: Cyrus Tang Hematology Center, Soochow University, Suzhou 215123, China.
authors: Meng
authors: M
authors: Liu
authors: Cyrus Tang Hematology Center, Soochow University, Suzhou 215123, China.
authors: Tiantian
authors: T
authors: Zhou
authors: Department of Orthopedics, The Second Affiliated Hospital, Soochow University, Suzhou 215004, China. Electronic address: 13913133393@163.com.
authors: Haibin
authors: H
authors: Zhou
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D001861:Bone Regeneration / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D019899:Paracrine Communication / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D018836:Inflammation Mediators; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma
keywords: Bone marrow mesenchymal stem cells; Bone regeneration; Inflammatory cytokine; Osteogenesis; Paracrine