Li S.-T.,Chongqing Medical University |
Liu Y.,Chongqing Medical University |
Zhou Q.,Chongqing Medical University |
Lue R.-F.,No. 184 hospital of PLA |
And 4 more authors.
Tissue Engineering - Part C: Methods | Year: 2014
This study introduced a prototype of an axial-stress bioreactor system that supports long-term growth and development of engineered tissues. The main features of this bioreactor are an integrated substance exchanger and feedback control of pH and PO2. A 21-day study was conducted to validate the system's ability to maintain a stable environment, while remaining sterile. Our results showed that the pH, PO2, and nutrient (glucose) remained balanced at appropriate levels, while metabolic waste (lactic acid) was removed. No bacteria or fungi were detected in the system or tissue; thus, demonstrating that it was sterile. These data indicate the bioreactor's strong potential for long-term tissue culture. To explore this idea, the effect of dynamic culture, including cyclic compression and automatic substance exchange, on mouse bone-marrow mesenchymal stem cells (BMSCs) seeded in decalcified bone matrix was studied using the bioreactor prototype. Histological sections of the engineered tissues showed higher cell densities in scaffolds in dynamic culture compared to those in static culture, while cell cycle analysis showed that dynamic culture promoted BMSC proliferation (proliferation index, PI=34.02±1.77) more effectively than static culture (PI=26.66±1.81). The results from a methyl thiazolyl tetrazolium assay were consistent with the loading experimental data. Furthermore, elevated alkaline phosphatase activity and calcium content were observed in dynamic condition compared to static culture. In conclusion, this bioreactor system supplies a method of modulating the pH and PO2 in defined ranges with only small fluctuations; it can be used as a physiological or pathological analog. Automatic control of the environment is a practical solution for long-term, steady-state culture for future commercialization. © Copyright 2014, Mary Ann Liebert, Inc. 2014.
Mo J.,No. 184 hospital of PLA |
Yanke Z.,No. 184 hospital of PLA |
Jiaming Y.,No. 184 hospital of PLA |
Peng'an X.,No. 184 hospital of PLA |
Shanhua T.,No. 184 hospital of PLA
Chinese Journal of Cancer Biotherapy | Year: 2015
Objective: To explore the influence and the mechanism of matrix metalloproteinase-3 (MMP-3) on invasion and metastasis of osteosarcoma. Methods: 40 cases of patients with osteosarcoma in No. 184 Hospital of PLA from January 1, 2013 to November 1, 2013 and 40 cases of healthy people were collected, and the serum of the patients with osteosarcoma and the healthy people were collected and the content of MMP-3 was detected with ELISA. And, the correlation between MMP-3 and the pathogenesis of osteosarcoma was also analyzed by SPSS. The mRNA levels of MMP-3 were detected by the method of RT-PCR in different osteosarcoma cell lines such as U2-0S,H0S-143b,MG-63cell lines. The proliferation of MG-63 after given different concentration of MMP-3 or knockdown MMP-3 was detected using MTT assay. And the invasion ability was detected by Transwell. Finally, the MMP-3 promotes the invasion and metastasis related proteins of osteosarcoma such as p-AKT, NF-κB were detected by Western blotting. Results: MMP-3 was highly expression in the serum of the patients with osteosarcoma (F=186. 4, P = 0. 000), and there was a strong positive correlation between MMP-3 exepression and Enneking stage in osteosarcoma(r =0. 736, P =0. 043). MMP-3 was highly expression in the cell lines of MG-63 (P <0. 05). The overexpression of MMP-3 stimulated by MMP-3 cultivation increased the expression of p-AKT and the intranuclear NF-κB, then promoted the proliferation and invasion ability of MG-63 cells (P < 0. 05); In contrast, MMP-3-shRNA transfection knockdown the MMP-3, result in reduction of the expression of p-AKT and the intranuclear NF-κB, then suppressed the proliferation and invasion ability of MG-63 cells (P < 0. 05). Conclusion: MMP-3 promotes the invasion and metastasis of osteosarcoma may via increasing the phosphorylation of AKT and promoting the translocation of NF-κB 1. © 2015, Editorial office of Chinese Journal of Cancer Biotherapy. All rights reserved.