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Objective: To investigate the role of p38 MAPK pathway in IL-1β induced inflammatory secretory properties mesenchymal stem cells(SFMSCs) from human synovial fluid of temporomandibular joint. Methods: In vitro cultured hSFMSCs were divided into control group and IL-1β treated group(10 ng/ml IL-1β for 2 h), the activation of p38 MAPK was examined with western blot. Then, the gene expression and the secretion of IL-6 and IL-8 in control group, IL-1β treated group and p38 MAPK inhibitor pre-treated group(pretreatment with SB-203580 10 μmol/L prior to the addition of 10 ng/ml IL-1β) were examined by RT-PCR and cytometric bead array(CBA). Results: It was observed that the protein level of p-p38 MAPK, the secretion of IL-8 and IL-6 were increased in IL-1β treated group, and suppressed in p38 MAPK inhibitor pre-treated group. Conclusion: p38 MAPK pathway of SFMSCs might play a role in IL-6 and IL-8 secretion by SFMSCs in inflammatory environment.
ABSTRACT
Objective@#To analyze related factors on the number of mesenchymal stem cells in the synovial fluid of the temporomandibular joint (TMJ) and provide an research basis for understanding of the source and biological role of mesenchymal stem cells derived from synovial fluid in TMJ.@*Methods@#One hundred and twenty-two synovial fluid samples from 91 temporomandibular disorders (TMD) patients who visited in Department of TMJ Center, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University from March 2013 to December 2013 were collected in this study, and 6 TMJ synovial fluid samples from 6 normal volunteers who were studying in the North Campus of Sun Yat-sen University were also collected, so did their clinical information. Then the relation between the number of mesenchymal stem cells derived from synovial fluid and the health status of the joints, age of donor, disc perforation, condylar bony destruction, blood containing and visual analogue scale score of pain were investigated using Mann-Whitney U test and Spearman rank correlation test.@*Results@#The number of mesenchymal stem cells derived from synovial fluid had no significant relation with visual analogue scale score of pain (r=0.041, P=0.672), blood containing (P=0.063), condylar bony destruction (P= 0.371). Linear correlation between the number of mesenchymal stem cells derived from synovial fluid and age of donor was very week (r=0.186, P=0.043). The number of mesenchymal stem cells up-regulated when the joint was in a disease state (P=0.001). The disc perforation group had more mesenchymal stem cells in synovial fluid than without disc perforation group (P=0.042).@*Conclusions@#The number of mesenchymal stem cells derived from synovial fluid in TMJ has no correlation with peripheral blood circulation and condylar bony destruction, while has close relation with soft tissue structure damage of the joint.
ABSTRACT
BACKGROUND: Jaw defects are common clinically. It is desirable to find ideal seed cells combined with scaffolds to construct tissue engineered jaws for curing these diseases. OBJECTIVE: To investigate the growth characteristics of bone marrow mesenchymal stem cells (BMSCs) transfected with basic fibroblast growth factor (bFGF) gene after seeded on coral scaffold in vitro. DESIGN, TIME AND SETTING: An experimental study of bone tissue engineering was performed in the Research Institute of Stomatology, Sun Yat-sen University between March 2006 and June 2008. MATERIALS: Natural coral from China Hainan bench was made into pieces of 8 mm×8 mm×2 mm. METHODS: BMSCs were isolated from New England rabbits by density gradient centrifugation and then purified by adherent separation. bFGF-pcDNA3 gene was transfected into BMSCs using Lipofectamine TM 2000. bFGF gene-transfected (transfected group) or untransfected (untransfected group)BMSCs were seeded on different coral scaffolds. In addition, bFGF gene-transfected BMSCs were simply cultured but not on the coral scaffold for control (simple culture group). MAIN OUTCOME MEASURES: BMSC proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and BMSC growth on coral scaffold was observed under the scanning electron microscope. RESULTS: MTT assay showed that the BMSC proliferation rate was significantly higher in the transfected group than in the untransfected group (P < 0.05) and that there was no significant difference in BMSC proliferation between the transfected and simple culture groups (P > 0.05). Scanning electron microscope results displayed that BMSCs adhered to and spread over the coral scaffold, exhibiting various appearances, with some cells had grown into scaffold micropores or spanned micropore surface, and some extracellular matrix secreted by BMSCs were found. CONCLUSION: The transfected group exhibited better growth of BMSCs transfected by bFGF gene than the untransfected group. These findings indicate that coral skeleton does not influence BMSC proliferation and can be used as a scaffold of BMSCs to construct tissue-engineered bone.