Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (49): 9218-9222.doi: 10.3969/j.issn.1673-8225.2011.49.023

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Retinoic acid induces osteogenic differentiation of periodontal ligament stem cells from miniature swine in vitro

Zhang Peng-tao1, Zhong Liang-jun2, Zhang Yuan1, Zhang Yuan-ming3, Xu Yan4   

  1. 1First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
    2Clinical College of Hangzhou Normal University, Hangzhou  310015, Zhejiang Province, China
    3Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Uygur Autonomous Region, China
    4Stomatological Hospital of Nanjing Medical University, Nanjing  210000, Jiangsu Province, China
  • Received:2011-08-25 Revised:2011-09-13 Online:2011-12-03 Published:2011-12-03
  • Contact: Zhang Yuan-ming, Professor, Doctoral supervisor, Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Uygur Autonomous Region, China sparkwalker@qq.com
  • About author:Zhang Peng-tao★, Studying for master’s degree, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China sparkwalker@qq.com
  • Supported by:

    the Science and Technology Supported Project of Xinjiang Uygur Autonomous Region, No. 201091144*

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Abstract:

BACKGROUND: Recent studies have found that retinoic acid can induce the osteogenic differentiation of embryonic stem cells and multiple adult stem cells.
OBJECTIVE: To observe the effect of retinoic acid on osteogenic differentiation of porcine periodontal ligament stem cells (PDLSCs).
METHODS: Porcine PDLCs were harvested by using outgrowth method; PDLSCs were isolated by limited dilution of culture cells for single cell clone. Immunofluorescence was used to detect the expression of STRO-1 and immunocytochemistry to detect the expression of vimentin and pan-Cytokeratin (PCK) of porcine PDLSCs. Cell counting kit8 (CCK8) was applied to evaluated cell proliferation of PDLSCs. The colony formation of porcine PDLSCs ratio was assayed. Third passage PDLSCs were induced with mineralized conditional medium containing retinoic acid, ascorbic acid and β-glycerophosphate. Mineralized nodules were studied by Alizarin red S staining. Osteopontin, osteocalcin, collagen typeⅠ and collagen type Ⅲ were detected by immunocytochemistry.
RESULTS AND CONCLUSION: Porcine PDLSCs expressed STRO-1 and vimentin, and the result of pan-Cytokeratin was negative. Porcine PDLSCs colony formation ratio was 2.8%. PDLSCs induced by retinoic acid showed positive expression of alkaline phosphatase at 14 days and positive expression of Alizarin red S at 21 days. Osteopontin, osteocalcin, collagen typeⅠ were positive but collagen type Ⅲ was negative at 21 days. These findings indicate that retinoic acid can be an effective inducer of osteogenic differentiation of porcine PDLSCs.

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