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Chemokine SDF-1 on human periodontal ligament stem cell proliferation , migration and differentiation

Author: DuLingZuo
Tutor: YangZuoShan；GeShaoHua
School: Shandong University
Course: Clinical Stomatology
Keywords: Periodontal ligament stem cells SDF-1 Chemotactic effect Periodontal Tissue Regeneration
CLC: R781.4
Type: Master's thesis
Year: 2011
Downloads: 63
Quote: 0
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Abstract


Objective: chemokine SDF-1 by being able to raise mesenchymal stem cells (MSCs), regulate their proliferation and differentiation, and in tissue regeneration plays an important role. However, it human periodontal ligament stem cells (PDLSCs) role is still unclear. From human periodontal ligament tissue isolated PDLSCs, belonging adult mesenchymal stem cells in the appropriate culture conditions can differentiate into cementum-like cells, osteoblasts, adipocytes and collagen-forming cells, and in animal periodontal defect model can no longer generate cementum / periodontal ligament-like structure. Therefore PDLSCs periodontal tissue regeneration is considered to be seed cells and is used to stem cell-based tissue regeneration project. PDLSCs the human chemokine receptor expression of the chemokine from the PDLSCs chemotactic effects and tissue repair process in the subsequent regeneration of periodontal tissue effect may be a crucial process. The purpose of this study was to investigate the chemokine SDF-1 receptor CXCR4 in human PDLSCs on SDF-1 expression in human PDLSCs growth activity, proliferation, migration and differentiation potential impact. Methods: limiting dilution and removal from the need for orthodontic healthy premolars were isolated, cultured people PDLSCs. The use of immunohistochemical staining was used to detect vimentin and cytokeratin and mesenchymal stem cells tables flag STRO-1 expression on the PDLSCs. Using real-time quantitative PCR and immunohistochemical staining was used to detect the chemokine SDF-1 receptor CXCR4 expression on the PDLSCs. Using the MTT assay method and BrdU incorporation assay SDF-1 对 PDLSCs growth activity and proliferation of state. Using real-time quantitative PCR method to detect SDF-1 对 PDLSCs differentiation factor Ⅰ collagen (Col Ⅰ) and alkaline phosphatase (ALP) in rats. The use of a 24-well Transwell cell culture chamber to detect SDF-1 对 PDLSCs chemotactic effect. All data were statistically using SPSS13.0 package ANOVA. P values ​​less than 0.05 were considered statistically significant. Results: Immunocytochemistry demonstrated a broad spectrum of people PDLSCs negative expression of cytokeratin, vimentin expression, and mesenchymal stem cell markers STRO-1 positive expression. Real-time quantitative PCR analysis confirmed that people PDLSCs expression at the mRNA level of chemokine receptor CXCR4; qualitative immunocytochemistry show people PDLSCs receptor expression at the protein level of CXCR4. Compared with medium alone, SDF-1 enhances the growth of human PDLSCs activity and proliferation (P lt; 0.05). Real-time quantitative PCR showed that, SDF-1 can significantly increase Co1 Ⅰ expression levels (P lt; 0.01), for the expression of ALP levels were not significantly affected (P gt; 0.05). Compared with the control group, and 400ng/ml in 100ng/m1 between the chemokine SDF-1 could promote PDLSCs migration (P lt; 0.05), the largest concentration of promoting migration is 200ng/ml. Use CXCR4 neutralizing antibody treatment, PDLSCs proliferation and migration effect was significantly inhibited (P lt; 0.05). Conclusion: In this study, we demonstrated that expression of human PDLSCs chemokine receptor CXCR4, the chemokine SDF-1 对 PDLSCs a dose-dependent chemotactic effect, this chemotactic effect may be mediated by its specific receptor CXCR4 mediated. Chemokine SDF-1 may have raised the PDLSCs periodontal tissue destruction potential and by promoting the activity of these stem cells, proliferation and differentiation affect their ability to promote destruction of periodontal tissue regeneration.

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