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The comparative viability of hTERT periodontal ligament cell Line mediated by adenovirus and lentivirus.

OBJECTIVE: The aim of this study was to construct the periodontal ligament cells overexpressing Human telomerase reverse transcriptase(hTERT) mediated by adenovirus vectors and lentivirus, and compare the osteogenic and proliferative abilities of the two cell lines, in order to establish an efficient and stable cell model that will be more appropriate for applying to the periodontal ligament regeneration.

METHODS: After the construction of the recombinant adenovirus plasmid pAd-pshuttle-cmv-hTERT, then human periodontal ligament cells were infected by the successfully packaged adenovirus and lentivirus particles, by which the two periodontal ligament cell lines were established. The expression levels of hTERT and the mRNA of the alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, core-binding factor and type I collagen were test. After cultured in osteoinductive culture medium for 14 days, periodontal ligament cells were stained with alizarin red(pH=4.1) for 10 minutes and then observed the formation of mineralized nodules and proliferation ability by CCK-8. Results The qPCR results suggested that the hTERT was highly expressed in the infected two cell lines, and there was a significant difference compared with the normal periodontal ligament cells;The osteogenic gene was highly expressed in the adenovirus-mediated cell line, and there was a significant difference compared with the lentivirus-mediated cell line; Alizarin red staining showed that the mineralized nodules in adenovirus-mediated cell line were more than lentivirus-mediated cell line, and CCK-8 test showed that the adenovirus-mediated cell line had the more powerful proliferation ability. Conclusion Periodontal ligament cell line established by adenovirus has the better osteogenic differentiation and proliferative ability than that by lentivirus, which means the former one is more appropriate as the model of cell studying for the periodontal ligament regeneration.

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