^{Department of Pediatrics and Developmental Biology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Department of Obstetrics and Gynecology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Section of Periodontology, Department of Hard Tissue Engineering, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Department of Cellular Physiological Chemistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Behavioral Dentistry, Department of Comprehensive Oral Health Care, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Global COE Program, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.}

^{Corresponding author.}

Address correspondence to:, Motohiro Komaki, DDS, PhD, Department of Nanomedicine (DNP), Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan,
E-mail:Email: pj.ca.dmt@irep.ikamok

Received 2013 Jan 11; Accepted 2013 Sep 10.

Abstract

Periodontal disease is characterized by the destruction of tooth supporting tissues. Regeneration of periodontal tissues using ex vivo expanded cells has been introduced and studied, although appropriate methodology has not yet been established. We developed a novel cell transplant method for periodontal regeneration using periodontal ligament stem cell (PDLSC)-transferred amniotic membrane (PDLSC-amnion). The aim of this study was to investigate the regenerative potential of PDLSC-amnion in a rat periodontal defect model. Cultured PDLSCs were transferred onto amniotic membranes using a glass substrate treated with polyethylene glycol and photolithography. The properties of PDLSCs were investigated by flow cytometry and in vitro differentiation. PDLSC-amnion was transplanted into surgically created periodontal defects in rat maxillary molars. Periodontal regeneration was evaluated by microcomputed tomography (micro-CT) and histological analysis. PDLSCs showed mesenchymal stem cell-like characteristics such as cell surface marker expression (CD90, CD44, CD73, CD105, CD146, and STRO-1) and trilineage differentiation ability (i.e., into osteoblasts, adipocytes, and chondrocytes). PDLSC-amnion exhibited a single layer of PDLSCs on the amniotic membrane and stability of the sheet even with movement and deformation caused by surgical instruments. We observed that the PDLSC-amnion enhanced periodontal tissue regeneration as determined by micro-CT and histology by 4 weeks after transplantation. These data suggest that PDLSC-amnion has therapeutic potential as a novel cell-based regenerative periodontal therapy.

Abstract

References

1. Page R.C., Offenbacher S., Schroeder H.E., Seymour G.J., and Kornman K.S. Advances in the pathogenesis of periodontitis: summary of developments, clinical implications and future directions. Periodontol 200014, 216, 1997 [[PubMed]
2. Pihlstrom B.L., and Michalawiz B.S. Periodontal diseases. Lancet366, 1809, 2005 [[PubMed]
3. Ankrum J., and Karp J.M. Mesenchymal stem cell therapy: two steps forward, one step back. Trends Mol Med16, 203, 2010
4. Pittenger M.F., Mackay A.M., Beck S.C., Jaiswal R.K., Douglas R., Mosca J.D., Moorman M.A., Simonetti D.W., Craig S., and Marshak D.R. Multilineage potential of adult human mesenchymal stem cells. Science284, 143, 1999 [[PubMed]
5. Liu Z.J., Zhuge Y., and Velazquez O.C. Trafficking and differentiation of mesenchymal stem cells. J Cell Biochem.106, 984, 2009 [[PubMed]
6. Iwata T., Yamato M., Tsuchioka H., Takagi R., Mukobata S., Washio K., Okano T., and Ishikawa I. Periodontal regeneration with multi-layered periodontal ligament-derived cell sheets in a canine model. Biomaterials30, 2716, 2009 [[PubMed]
7. Park J.Y., Jeon S.H., and Choung P.H. Efficacy of periodontal stem cell transplantation in the treatment of advanced periodontitis. Cell Transplant20, 271, 2011 [[PubMed]
8. Ding G., Liu Y., Wang W., Wei F., Liu D., Fan Z., An Y., Zhang C., and Wang S. Allogeneic periodontal ligament stem cell therapy for periodontitis in swine. Stem Cells28, 1829, 2010
9. Yang Y., Rossi FM., and Putnins EE. Periodontal regeneration using engineered bone marrow mesenchymal stromal cells. Biomaterials31, 8574, 2010 [[PubMed]
10. Li H., Yan F., Lei L., Li Y., and Xiao Y. Application of autologous cryopreserved bone marrow mesenchymal stem cells for periodontal regeneration in dogs. Cells Tissues Organs190, 94, 2009 [[PubMed]
11. Kawaguchi H., Hirachi A., Hasegawa N., Iwata T., Hamaguchi H., Shiba H., Takata T., Kato Y., and Kurihara H. Enhancement of periodontal tissue regeneration by transplantation of bone marrow mesenchymal stem cells. J Periodontol75, 1281, 2004 [[PubMed]
12. Tobita M., Uysal AC., Ogawa R., Hyakusoku H., and Mizuno H. Periodontal tissue regeneration with adipose-derived stem cells. Tissue Eng Part A14, 945, 2008 [[PubMed]
13. Tsumanuma Y., Iwata T., Washio K., Yoshida T., Yamada A., Takagi R., Ohno T., Lin K., Yamato M., Ishikawa I., Okano T., and Izumi Y. Comparison of different tissue-derived stem cell sheets for periodontal regeneration in a canine 1-wall defect model. Biomaterials32, 5819, 2011 [[PubMed]
14. Jiang J., Wu X., Lin M., Doan N., Xiao Y., and Yan F. Application of autologous periosteal cells for the regeneration of class III furcation defects in Beagle dogs. Cytotechnology62, 235, 2010
15. Kawase T., Okuda K., Kogami H., Nakayama H., Nagata M., Sato T., Wolff L.F., and Yoshie H. Human periosteum-derived cells combined with superporous hydroxyapatite blocks used as an osteogenic bone substitute for periodontal regenerative therapy: an animal implantation study using nude mice. J Periodontol81, 420, 2010 [[PubMed]
16. Seo B.M., Miura M., Gronthos S., Bartold P.M, Batouli S., Brahim J., Young M., Robey P.G., Wang C.Y, and Shi S. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet364, 149, 2004 [[PubMed]
17. Wada N., Menicanin D., Shi S., Bartold P.M., and Gronthos S. Immunomodulatory properties of human periodontal ligament stem cells. J Cell Physiol219, 667, 2009 [[PubMed]
18. Gronthos S., Mrozik K., Shi S., and Bartold P.M. Ovine periodontal ligament stem cells: isolation, characterization, and differentiation potential. Calcif Tissue Int79, 310, 2006 [[PubMed]
19. Mrozik K.M., Zilm P.S., Bagley C.J., Hack S., Hoffmann P., Gronthos, S, and Bartold P.M. Proteomic characterization of mesenchymal stem cell-like populations derived from ovine periodontal ligament, dental pulp, and bone marrow: analysis of differentially expressed proteins. Stem Cells Dev19, 1485, 2010 [[PubMed]
20. Niknejad H., Peirovi H., Jorjani M., Ahmadiani A., Ghanavi J., and Seifalian A.M. Properties of the amniotic membrane for potential use in tissue engineering. Eur Cell Mater15, 88, 2008 [[PubMed]
21. Díaz-Prado S., Muiños-López E., Hermida-Gómez T., Cicione C., Rendal-Vázquez M.E., Fuentes-Boquete I., de Toro F.J., and Blanco F.J. Human amniotic membrane as an alternative source of stem cells for regenerative medicine. Differentiation81, 162, 2011 [[PubMed]
22. Lo V., and Pope E. Amniotic membrane use in dermatology. Int J Dermatol48, 935, 2009 [[PubMed]
23. Altan-Yaycioglu R., Akova Y.A., and Oto S. Amniotic membrane transplantation for treatment of symblepharon in a patient with recessive dystrophic epidermolysis bullosa. Cornea25, 971, 2006 [[PubMed]
24. Akahori T., Kobayashi A., Komaki M., Hattori H., Nakahama K., Ichinose S., Abe M., Takeda S., and Morita I. Implantation of capillary structure engineered by optical lithography improves hind limb ischemia in mice. Tissue Eng Part A16, 953, 2010 [[PubMed]
25. Yoshida T., Komaki M., Hattori H., Negishi J., Kishida A., Morita I., and Abe M. Therapeutic angiogenesis by implantation of a capillary structure constituted of human adipose tissue microvascular endothelial cells. Arterioscler Thromb Vasc Biol30, 1300, 2010 [[PubMed]
26. Tsugawa J., Komaki M., Yoshida T., Nakahama K., Amagasa T., and Morita I. Cell-printing and transfer technology applications for bone defects in mice. J Tissue Eng Regen Med5, 695, 2011 [[PubMed]
27. Fujisato T., Minatoya K., Yamazaki S., Meng Y., Niwaya K., Kishida A., Nakatani T., and Kitamura S. Preparation and recellularization of tissue engineered bioscaffold for feat valve replacement. In: Mori H., editor; , and Matsuda H., editor. , eds. Cardiovascular Regeneration Therapies Using Tissue Engineering Approaches. Tokyo: Springer, 2005, p. 83 [PubMed]
28. Wilshaw S.P., Kearney J.N., Fisher J., and Ingham E. Production of an acellular amniotic membrane matrix for use in tissue engineering. Tissue Eng12, 2117, 2006 [[PubMed]
29. Inukai T., Katagiri W., Yoshimi R., Osugi M., Kawai T., Hibi H., and Ueda M. Novel application of stem cell-derived factors for periodontal regeneration. Biochem Biophys Res Commun430, 763, 2013 [[PubMed]
30. Zhang J., An Y., Gao L.N., Zhang Y.J., Jin Yand Chen F.M. The effect of aging on the pluripotential capacity and regenerative potential of human periodontal ligament stem cells. Biomaterials33, 6974, 2012 [[PubMed][Google Scholar]
31. Liu O., Xu J., Ding G., Liu D., Fan Z., Zhang C., Chen W., Ding Y., Tang Z., and Wang S. Periodontal ligament stem cells regulate B lymphocyte function via programmed cell death protein 1. Stem Cells31, 1371, 2013 [[PubMed]
32. Bosshardt D.D. Are cementoblasts a subpopulation of osteoblasts or a unique phenotype? J Dent Res84, 390, 2005 [[PubMed]
33. Lekic P., and McCulloch C.A. Periodontal ligament cell population: the central role of fibroblasts in creating a unique tissue. Anat Rec245, 327, 1996 [[PubMed]
34. Komaki M., Iwasaki K., Arzate H., Narayanan A.S., Izumi Y., and Morita I. Cementum protein 1 (CEMP1) induces a cementoblastic phenotype and reduces osteoblastic differentiation in periodontal ligament cells. J Cell Physiol227, 649, 2012 [[PubMed]
35. Paula-Silva FW., Ghosh A., Arzate H., Kapila S., da Silva L.A., and Kapila Y.L. Calcium hydroxide promotes cementogenesis and induces cementoblastic differentiation of mesenchymal periodontal ligament cells in a CEMP1- and ERK-dependent manner. Calcif Tissue Int87, 144, 2010 [[PubMed]
36. Zeichner-David M. Regeneration of periodontal tissues: cementogenesis revisited. Periodontol 200041, 196, 2006 [[PubMed]
37. Grzesik W.J., and Narayanan A.S. Cementum and periodontal wound healing and regeneration. Crit Rev Oral Biol Med13, 474, 2002 [[PubMed]
38. Ma Z., Li S., Song Y., Tang L., Ma D., Liu B., and Jin Y. The biological effect of dentin noncollagenous proteins (DNCPs) on the human periodontal ligament stem cells (HPDLSCs) in vitro and in vivo. Tissue Eng Part A. 14, 2059, 2008 [[PubMed]