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ABSTRACT: SMSCs (synovial mesenchymal stem cells) isolated from TMJs (temporomandibular joints) were induced to proliferate and differentiate in vitro by bFGF (basic fibroblast growth factor) and explore the potential of SMSC differentiation into neuronal cells. In this study, the cultured SMSCs were derived from the TMJ synovial membrane of condylar hyperplasia patients and were amplified with the indicated concentration of FCS (fetal calf serum) and DMEM (Dulbecco's modified Eagle's medium) in vitro. bFGF (25 ng/ml) was applied to induced synovial cells differentiated into neuronal cells. Inverted microscopy, scanning electron microscopy, immunocytochemical and RT-PCR were used for checking the change of the induced cells. Morphology was mostly spindle; a small part was of a polygon. The undifferentiated SMSCs showed the fibroblast-like morphology; however, most of the differentiated cells were in the shape of a spindle and the rest were polygonal. Furthermore, being induced by bFGF, SMSCs can be found to be a unique long extension from the cell body under the scanning electron microscope. RT-PCR and immunocytochemical analysis was made to confirm nestin (neural stem cell marker) and NF-L (neurofilament-light or neurofilament 68-kDa mature nerve cell marker) expression in SMSCs. SMSCs can differentiate into neuronal cells when induced by bFGF. The bFGF-induced SMSCs not only changed into neural-like cells but also expressed specific markers.
Cell Biology International 10/2010; 35(1):87-91. · 1.48 Impact Factor
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ABSTRACT: The objective was to investigate synovium-derived stromal cells (SDSCs) coupled with chitosan/collagen type I (CS/COL-I) scaffolds for cartilage engineering. CS/COL-I scaffolds were fabricated through freeze-drying and cross-linked by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. SDSCs were isolated from synovium and cultured onto CS/COL-I scaffolds, constructs of which were incubated in serum-free chondrogenic medium with sequential application of TGF-β1 and bFGF for up to 21 days and then implanted into nude mice. The physical characteristics of the scaffolds were examined. The quality of the in vitro constructs was assessed in terms of DNA content by PicoGreen assay and cartilaginous matrix by histological examination. The implants of the constructs were evaluated by histological and immunohistochemical examinations and reverse transcription PCR. Results indicated that the CS/COL-I scaffold showed porous structures, and the DNA content of SDSCs in CS/COL-I scaffolds increased at 1 week culture time. Both of the constructs in vitro and the implants were examined with positive stained GAGs histologically and the implants with positive collagen type II immunohistochemically. RT-PCR of the implants indicated that aggrecan and collagen type II expressed. It suggested that SDSCs coupled with CS/COL-I scaffolds treated sequentially with TGF-β1 and bFGF in vitro were highly competent for engineered cartilage formation in vitro and in vivo.
Biomedical Materials 10/2010; 5(5):055005. · 2.16 Impact Factor
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ABSTRACT: The development of suitable bioactive three-dimensional scaffold for the promotion of cellular proliferation and differentiation is critical in periodontal tissue engineering. In this study,porous beta-tricalcium phosphate/chitosan composite scaffolds were prepared through a freeze-drying method. These scaffolds were evaluated by analysis of microscopic structure, porosity, and cytocompatibility. The gene expression of bone sialoprotein (BSP) and cementum attachment protein (CAP) was detected with RT-PCR after human periodontal ligament cells (HPLCs) were seeded in these scaffolds. Then cell-scaffold complexes were implanted subcutaneously into athymic mice. The protein expression of alkaline phosphatase (ALP) and osteopontin (OPN) was detected in vivo. Results indicated that composite scaffolds displayed a homogeneous three-dimensional microstructure; suitable pore size (120 microm) and high porosity (91.07%). The composite scaffold showed higher proliferation rate than the pure chitosan scaffold, and up-regulated the gene expression of BSP and CAP. In vivo, HPLCs in the composite scaffold not only proliferated but also recruited vascular tissue ingrowth. The protein expression of ALP and OPN was up-regulated in the composite scaffold. Therefore, it was suggested that the composite scaffold could promote the differentiation of HPLCs towards osteoblast and cementoblast phenotypes.
Journal of Materials Science Materials in Medicine 11/2009; 21(2):489-96. · 2.32 Impact Factor
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ABSTRACT: Rheumatoid arthritis (RA) is a common, systemic autoimmune disease which leads to destruction of the joint architecture and consequent disability. Although the aetiology of RA remains unknown, accumulating studies have established a strong association between RA and periodontitis (PD). Recently, anti-cyclic citrullinated peptide (anti-CCP) autoantibody and citrullinated peptide have been realized to be involved in the breaking of self-tolerance and development of autoimmune in RA. The citrullinated peptide is generated by post-translational modification (citrullination) of protein-bound arginine by peptidylarginine deiminase (PAD). Porphyromonas gingivalis(P. gingivalis), the major aetiological agent of PD and the only bacterium known to express a PAD enzyme, has been reported to be significantly associated with RA. The antibody titers to P. gingivalis are significantly increased in patients with RA and P. gingivalis antibody titers are significantly correlated with anti-CCP antibody isotypes that are specific to RA. Recent study indicates that the major synovial targets of the RA-specific anti-CCP autoantibodies are deiminated forms of the alpha- and beta- chains of fibrin. Meanwhile, it is also confirmed that bacterial PAD produced by P. gingivalis has the capacity of deiminating arginine in fibrin found in the periodontal lesion. What's more, it has been demonstrated that citrullination of HLA binding peptide causes a 100-fold increase in peptide-MHC affinity and leads to the activation CD4(+)T cells in HLA DRB1 0401 transgenic mice. Therefore, we postulate that P. gingivalis may play a crucial role in the pathogenesis of periodontitis-associated RA. P. gingivalis, which colonizes in the oral cavity, produces PAD enzyme continuously that leads to the citrullination of RA autoantigen such as fibrin in synovium joint. These PAD engendered antigens, presented in association with major histocompatibility complex (MHC) molecules by antigen-presenting cells (APC), ultimately lead to production of the anti-CCP antibody. The anti-CCP antibodies form immune complexes with citrullinated proteins, which can be bound by inflammatory cells via their Fc receptors. The roles of these immune complexes and inflammatory cells are mediated by a complex cascade involving complement activation. These mechanisms result in a release of mediators of inflammation and joint destruction ultimately leading to the onset of RA. This hypothesis reveals that oral bacterial infection may play a role in peptide citrullination which might be involved in loss of self-tolerance and development of autoimmune in RA.
Medical Hypotheses 03/2009; 72(6):732-5. · 1.39 Impact Factor
