Objective To review the research appl ication and advance of synovium-derived mesenchymal stem cells (SMSCs) in tissue engineering. Methods The recent related l iterature was reviewed, concerning isolation method, characteristics of SMSCs, and its appl ication in tissue engineering. Results SMSCs are multi potent cell population with characteristics of easy isolation and high prol iferation, which have been appl icated in the cartilage, tendon, l igament, and bone tissue engineering. Conclusion SMSCs is a new member of mesenchymal stem cells family. It appears to be promising seedcells for tissue engineering, but further research is needed.
ObjectiveTo investigate the osteogenic differentiation potential and the biological features of synovium-derived mesenchymal stem cells (SMSCs) in vitro and to observe the osteogenic capability of the composite scaffolds constructed with SMSCs and hydroxylapatite/chitosan/poly L-latic acid (HA/CS/PLLA) in vivo. MethodSMSCs were separated and cultured with adherent method and enzymatic digestion method. Specific phenotypes of SMSCs were detected by flow cytometry after purification. Then, SMSCs were identified by oil red O staining, alkaline phosphatase (ALP) staining, and alizarin red staining after adipogenic and osteogenic induction, respectively. In vitro experiments:the expressions of osteogenic related genes[osteocalcin (OCN), collagen type I, ALP, and Runx-2] were detected by real-time fluorescent quantitative PCR at 1, 7, 14, 21, and 28 days after osteogenic induction; ALP activities were also determined by ELISA at 1, 3, 5, 7, 9, and 11 days after osteogenic induction; meanwhile, extracellular matrix calcium mineralization was detected by alizarin red S method at 7, 14, 21, and 28 days after osteogenic induction; the normal SMSCs were harvested as control group. In vivo experiments:Twenty-four Sprague Dawley (SD) rats were randomly divided into experimental group (n=12) and control group (n=12) . The 3rd passage SMSCs were seeded on HA/CS/PLLA to construct composite scaffolds, after adhesion for 72 hours in vitro, the composite scaffolds were implanted into the right thigh muscle of 12 SD rats as experimental group; HA/CS/PLLA was implanted into the right thigh muscle of the other 12 SD rats as control group. At 4 and 8 weeks after implantation, the scaffolds were harvested for X-ray film and histological examination to observe ectopic bone formation. ResultsThe positive rates of CD147, CD90, CD105, and CD44 were more than 95%, while the positive rates of CD117, CD34, CD14, and CD45 were less than 10%. Oil red O staining demonstrated red lipid droplets in the cytoplasm, and alizarin red staining showed flaky red calcifications, and cytoplasm was dyed brown by the ALP staining. The mRNA expressions of collagen type I, ALP, and Runx-2 were significantly increased at 7 days after osteogenic induction, and OCN mRNA expression was significantly increased at 14 days after osteogenic induction; ALP activity was significantly higher at 5, 7, 9, 11 days after osteogenic induction in the SMSC-induced group than control group and reached a maximum at 7 days (P<0.05) . Calcium mineralization was significantly enhanced at 14 days after osteogenic induction, and gradually increased with time (P<0.05) ; moreover, it was significantly higher in the SMSC-induced group than control group (P<0.05) . X-ray and histological examination demonstrated that the new bone tissues formed in 2 groups, but bone formation content of the experimental group was significantly more than that of the control group at 4 and 8 weeks after implantation (P<0.05) . ConclusionsSMSCs can be induced into osteoblasts both in vitro and in vivo, so SMSCs might be a promising seed cells for bone tissue engineering.
ObjectiveTo investigate the anti-apoptotic ability of synovium-derived mesenchymal stem cells (SMSCs) by comparing the apoptosis induced by tumor necrosis factor α (TNF-α) between SMSCs and bone marrow mesenchymal stem cells (BMSCs). MethodSMSCs and BMSCs were isolated with tissue adhering and density gradient centrifugation respectively, and cells at passages 3-5 were used in further experiments. After immunophenotype identification and differentiation induction, cells were divided into 4 groups. In the experimental groups, apoptosis of SMSCs and BMSCs were induced by 20 ng/mL TNF-α and 10 μg/mL cycloheximide, and cells were cultured in normal culture medium in the control groups. Cellular morphology were observed by inverted phase contrast microscope. After apoptosis induction for 24 hours, cell viability was determined by cell counting kit 8 assay and apoptotic index was detected by flow cytometer. Moreover, the level of Cleaved Caspase-8, 3 were determined by Western blot. ResultsBoth SMSCs and BMSCs accorded with the definition criteria of MSCs according to results of immunophenotype identification and differentiation induction. After apoptosis induction, cells became shrinking and partially floated and cellular morphologies became worse than those in the control groups. After apoptosis induction for 24 hours, cell viabilities of SMSCs and BMSCs in the control groups were both 100%, and no apoptotic cells were observed. However, cell viabilities of SMSCs and BMSCs in the experimental groups were 60.13%±8.63% and 46.55%±10.54% respectively, which were both significantly lower than those in the control groups (P<0.05) , and cell viability in the SMSCs experimental group was significantly higher than that in the BMSCs experimental group (t=3.152, P=0.006) . The apoptotic index was 36.54%±8.63% in the SMSCs experimental group and was 53.77%±11.52% in the BMSCs experimental group, both were significantly higher than the control groups (1.12%±0.24% and 1.35%±0.31%) (P<0.05) . What's more, it was significantly lower in SMSCs experimental group than that in BMSCs experimental group (t=3.785, P=0.001) . Moreover, no expression of Cleaved Caspase-8, 3 was detected in the control groups. But the levels of Cleaved Caspase-8, 3 were significantly enhanced in the experimental groups and they were lower in SMSCs than in BMSCs (t=13.870, P=0.000; t=7.309, P=0.000) . ConclusionsTNF-α induced apoptosis is lower in SMSCs than in BMSCs, which means that SMSCs may have stronger anti-apoptosis ability than BMSCs.