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The effect of iloprost on cell proliferation and angiogenesis-related gene expression in human periodontal ligament cells

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Abstract

Periodontal ligament is considered a rich source of mesenchymal stem cells for pulp regeneration. This study investigated the effect of iloprost, a prostacyclin analog, on cell proliferation and the expression of angiogenesis-related genes in human periodontal ligament cells (hPDLs) in vitro. The hPDLs were treated with 10−9–10−6 M iloprost for 1, 6 and 24 h. Cell proliferation was determined by an MTT assay. Vascular endothelial growth factor (VEGF), alpha-1 type I collagen (COL1), and basic fibroblast growth factor (bFGF) mRNA expression were determined by semi-quantitative and qPCR. ELISA was employed for assessment of VEGF expression. Immunofluorescence staining for COL1 protein expression was performed. A prostacyclin receptor (IP) antagonist was used to verify the signaling pathway. The Kruskal–Wallis and Tukey significant difference tests were used to analyze the results. From the results, iloprost treatment did not affect cell morphology or proliferation. Iloprost induced the upregulation of VEGF and COL1 mRNA levels as shown by PCR. The effect of iloprost on bFGF mRNA expression was not observed. The immunofluorescence assay revealed that COL1 protein expression was increased in the iloprost groups. Pretreating the hPDLs with the IP antagonist significantly suppressed the enhancing effect of iloprost on VEGF and COL1 mRNA expression and suppressed COL1 protein expression. In conclusion, iloprost promoted mRNA and protein expression of VEGF and COL1, but not of bFGF in hPDL cells. The increased effect of iloprost was abolished by IP receptor antagonist pretreatment. Iloprost might be a promising agent in dentin-pulp regeneration.

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Acknowledgements

This study was supported by the Dental Research Fund Faculty of Dentistry [DRF57007], Thailand Research grant [TRG 5880218] and the Research Chair Grant 2012 of the National Science and Technology Development Agency (NSTDA).We thank Assoc. Prof. Thanaphum Osathanon for his valuable comments in manuscript revision. We thank Dr. Kevin Tompkins for manuscript editing and Dr. Sonntana Seang, PhD candidate in Oral Biology program, for his assistance in image analysis.

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Authors and Affiliations

  1. Department of Operative Dentistry, Faculty of Dentistry, Chulalongkorn University, 34 Henri Dunant Road, Pathumwan, Bangkok, 10330, Thailand

    Thanomsuk Jearanaiphaisarn & Teeranuch Sanharati

  2. Mineralized Tissue Research Unit and Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand

    Prasit Pavasant

  3. Department of Physiology, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand

    Chalida Nakalekha Limjeerajarus

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  1. Thanomsuk Jearanaiphaisarn
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  2. Teeranuch Sanharati
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  3. Prasit Pavasant
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  4. Chalida Nakalekha Limjeerajarus
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Correspondence to Thanomsuk Jearanaiphaisarn.

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Ethical approval

The study’s protocol was approved by the Human Ethics Committee of the Faculty of Dentistry, Chulalongkorn University (HREC-DCU 2013-035).

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Jearanaiphaisarn, T., Sanharati, T., Pavasant, P. et al. The effect of iloprost on cell proliferation and angiogenesis-related gene expression in human periodontal ligament cells. Odontology 106, 11–18 (2018). https://doi.org/10.1007/s10266-017-0307-4

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  • DOI: https://doi.org/10.1007/s10266-017-0307-4

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