EVALUATION OF REGENERATIVE THERAPY USING CELL SHEET THROUGH CEMENTUM PROTEIN-1 EXPRESSION ON MACACA NEMESTRINA

  • Syanti Wahyu Astuty Department of Periodontology, Faculty of Dentistry, Jakarta, Universitas Indonesia, Indonesia
  • Hari Sunarto Department of Periodontology, Faculty of Dentistry, Jakarta, Universitas Indonesia, Indonesia
  • Lisa Amir Department of Oral Biology, Faculty of Dentistry, Jakarta, Universitas Indonesia, Indonesia
  • Erik Idrus Department of Oral Biology, Faculty of Dentistry, Jakarta, Universitas Indonesia, Indonesia

Abstract

Objective: The main objective of periodontal therapy is tissue regeneration. Previous studies have identified the potential of mesenchymal stem cells to improve major periodontal defect reconstruction in bone tissue engineering. Cell sheet technology (CST), in which a cell culture is obtained from a material coated with a temperature-sensitive substrate, has been developed for the reconstruction of various tissues, including periodontal tissue. Cementum protein-1 (CEMP-1) is a 50-kDa protein that plays a crucial role in cementogenesis by enhancing the combining of cells formed by cell cementoblast. Evaluate periodontal tissue regeneration with CST, and the application of chitosan, chitosan cell sheet or arginineglycineaspartic acid (RGD)-modified chitosan cell sheet on Macaca nemestrina one-wall defect model.

Methods: The CEMP-1 was analyzed expression in a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel electrophoresis assay.

Results: CEMP-1 expression in gingival crevicular fluid was observed using the SDS-PAGE method every week for 3 weeks. Protein band expressions on SDS-PAGE gel were identified at around 50 kDa with different thicknesses between groups. The chitosan, chitosan cell sheet, and RGD-modified chitosan cell sheet groups showed protein bands of CEMP-1 between 50 and 70 kDa at weeks 1, 2, and 3; weeks 2 and 3; and weeks 1 and 2, respectively.

Conclusion: Our results demonstrated that the application of chitosan and RGD-modified chitosan cell sheets could enhance bone regeneration, as evidenced by CEMP-1 protein expression levels.

Keywords: Periodontitis, Bone defect, Cell sheet, Cementum protein-1

References

1. Craig R, Kamer A, editors. A Clinician’s Guide to Systemic Effects of Periodontal Diseases. New York: Springer; 2016.
2. Sculean A, Nikolidakis D, Nikou G, Ivanovic A, Chapple IL, Stavropoulos A. Biomaterials for promoting periodontal regeneration in human intrabony defects: A systematic review. Periodontol 2000 2015;68:182-216.
3. Lee J, Stavropoulos A, Susin C, Wikesjö UM. Periodontal regeneration: Focus on growth and differentiation factors. Dent Clin North Am 2010;54:93-111.
4. Bachtiar EW, Amir LR, Suhardi P, Abas B. Scaffold degradation during bone tissue reconstruction in Macaca nemestrina mandible. Interv Med Appl Sci 2016;8:77-81.
5. Tsumanuma Y, Iwata T, Kinoshita A, Washio K, Yoshida T, Yamada A, et al. Allogeneic transplantation of periodontal ligament-derived multipotent mesenchymal stromal cell sheets in canine critical-size supra-alveolar periodontal defect model. Biores Open Access 2016;5:22-36.
6. Ivanovski S, Vaquette C, Gronthos S, Hutmacher DW, Bartold PM. Multiphasic scaffolds for periodontal tissue engineering. J Dent Res 2014;93:1212-21.
7. Yang J, Yamato M, Shimizu T, Sekine H, Ohashi K, Kanzaki M, et al. Reconstruction of functional tissues with cell sheet engineering. Biomaterials 2007;28:5033-43.
8. Matsuda N, Shimizu T, Yamato M, Okano T. Tissue engineering based on cell sheet technology. Adv Mater 2007;19:3089-99.
9. Mohammadi R, Amini K. Guided bone regeneration of mandibles using chitosan scaffold seeded with characterized uncultured omental adipose-derived stromal vascular fraction: An animal study. Int J Oral Maxillofac Implants 2015;30:216-22.
10. Tsai W, Chen Y, Liu H. RGD-conjugated crosslinked chitosan scaffolds for culture and osteogenic differentiation of mesenchymal stem cells. J Taiwan Inst Chem Eng 2013;44:1-7.
11. Torii D, Konishi K, Watanabe N, Goto S, Tsutsui T. Cementogenic potential of multipotential mesenchymal stem cells purified from the human periodontal ligament. Odontology 2015;103:27-35.
12. Carmona-Rodríguez B, Alvarez-Pérez MA, Narayanan AS, Zeichner-David M, Reyes-Gasga J, Molina-Guarneros J, et al. Human Cementum Protein 1 induces expression of bone and cementum proteins by human gingival fibroblasts. Biochem Biophys Res Commun 2007;358:763-9.
13. Tsumanuma Y, Iwata T, Washio K, Yoshida T, Yamada A, Takagi R, et al. Comparison of different tissue-derived stem cell sheets for periodontal regeneration in a canine 1-wall defect model. Biomaterials 2011;32:5819-25.
14. Iwata T, Yamato M, Tsuchioka H, Takagi R, Mukobata S, Washio K, et al. Periodontal regeneration with multi-layered periodontal ligament-derived cell sheets in a canine model. Biomaterials 2009;30:2716-23.
15. Akizuki T, Oda S, Komaki M, Tsuchioka H, Kawakatsu N, Kikuchi A, et al. Application of periodontal ligament cell sheet for periodontal regeneration: A pilot study in beagle dogs. J Periodontal Res 2005;40:245-51.
16. Hansson A, Hashom N, Falson F, Rousselle P, Jordan O, Borchard G. In vitro evaluation of an RGD-functionalized chitosan derivative for enhanced cell adhesion. Carbohydr Polym 2012;90:1494-500.
Statistics
244 Views | 219 Downloads
How to Cite
Astuty, S., Sunarto, H., Amir, L., & Idrus, E. (2018). EVALUATION OF REGENERATIVE THERAPY USING CELL SHEET THROUGH CEMENTUM PROTEIN-1 EXPRESSION ON MACACA NEMESTRINA. International Journal of Applied Pharmaceutics, 9, 107-109. https://doi.org/10.22159/ijap.2017.v9s2.26
Section
Original Article(s)