Effects of Osteogenic Growth Peptide and Insulin-like Growth Factor-I on Biological Function in Cultured Human Periodontal Ligament Cell
|School||Dalian Medical University|
|Keywords||Human periodontal ligament cells OGP Insulin -like growth factor-1 Proliferation ALP Total protein content|
Objective: In this study, cultured human periodontal ligament cells (periodontal ligament cells, PDLCs), then become osteogenic growth peptide (osteogenic growth peptide, OGP) and insulin-like growth factor -1 (insulin-like growth factor-I, IGF- a) alone or combined effect on the human PDLCs, observe people PDLCs proliferation, intracellular ALP (alkaline phosphatase, ALP) activity changes and changes in total protein content, in order to investigate the two growth factors alone or in combination on Cultured human PDLCs effect of its clinical treatment of periodontal disease provide a theoretical basis. Method: 1. PDLCs in vitro and human identification: Take 12 to 18 years clinical need for extraction for orthodontic periodontal health, caries fresh premolars, scraping with a razor blade root in 1/3 of the periodontal ligament tissue , tissue explant cultured PDLCs. The third passage cells were sABC method vimentin (Vimentin) and keratin (Kerati) immunohistochemical staining, cell source to be identified. (2) the OGP with IGF-1 alone in the human PDLCs, cultured for 3 days with a tetrazolium salt colorimetric (MTT) proliferation observed people PDLCs case, statistical analysis and selected IGF-1 对 OGP person acting PDLCs maximum and minimum concentrations markedly markedly concentration. 3 would be measured OGP and IGF-1 concentrations markedly maximum and minimum concentrations were markedly or combined effect on the human PDLCs, in the culture of the first 1,3,6 days, with four MTT assay ( MTT) assay to detect cell proliferation, measured absorbance value (A value), were compared between groups; alkaline phosphatase detection kit intracellular ALP activity, measured absorbance value (A value), were compared between groups; with Coomassie Brilliant Blue colorimetric detection of intracellular total protein content, measured absorbance value (A value), were compared between groups. Results: 1. Using tissue block culture first 7-9 days, cells from the tissue blocks are freed, immunohistochemical staining showed positive staining cells vimentin, cytokeratin negative. 2 day 3 of culture, OGP cells with different concentrations absorbance experimental group compared with control group all have differences (p lt; 0.05) markedly maximum concentration of 10 -9 sup> mol / L, the minimum concentration markedly to 10 -11 sup> mol / L. 3 day 3 of culture, IGF-1 cells with different concentrations of the experimental group compared with control group absorbance values ??with differences (p lt; 0.05) the maximum concentration was markedly 100ng/ml, the minimum concentration is markedly 1ng/ml. 4.OGP with the combined effects of IGF-1 in human PDLCs, promoting proliferation than the control group were significantly different (P lt; 0.05), and with two growth factors alone at the same concentration also increased significantly when compared difference (P lt; 0.05 ). Promoting proliferation from day 1, has been continued until the sixth day. 5.OGP the combined effects of IGF-1 in human PDLCs, both in cell culture medium or in cell lysates, the measured value of A in ALP activity compared with the control group were significantly different (P lt; 0.05), and with same concentration difference alone group was also significantly (P lt; 0.05). Promote enhancement of ALP activity from day 1, has been continued until the sixth day. 6.OGP with the combined effects of IGF-1 in human PDLCs, measured intracellular A value of the total protein content compared with the control group were significantly different (P lt; 0.05), but also with the same concentration difference alone group was also significantly ( P lt; 0.05). Promote the role of the total protein content increased from day 1, has been continued until the sixth day. Conclusion: The experimental results show that OGP alone can promote the proliferation of human PDLCs intracellular ALP activity increased, total protein content increased. (2) The experimental results show that IGF-1 alone can promote human PDLCs proliferation, ALP activity enhancement of intracellular, total protein content increased. 3 OGP The experimental results show that the combined effects of the IGF-1 can not only promote human PDLCs proliferation, ALP activity enhancement of intracellular, total protein content increased, and the two growth factors combination has a synergistic effect.