Exploration of the role of the subodontoblastic layer in odontoblast-like cell differentiation after tooth drilling using Nestin-enhanced green fluorescent protein transgenic mice |
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| Institution: | 1. Faculty of Dentistry, Niigata University, Niigata, Japan;2. Division of Clinical Chemistry, Department of Medical Technology, Niigata University Graduate School of Health Sciences, Niigata, Japan;3. Department of Pathology, The Nippon Dental University School of Life Dentistry at Niigata, Niigata, Japan;4. Division of Anatomy and Cell Biology of the Hard Tissue, Department of Tissue Regeneration and Reconstruction, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan;5. Department of Human, Information and Life Sciences, School of Health Sciences, University of Occupational and Environmental Health, Kitakyushu, Japan;6. Department of Physiology, Keio University School of Medicine, Tokyo, Japan;1. Monell Chemical Senses Center, 3500 Market Street, Philadelphia, PA 19104, USA;2. Department of Oral Physiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan;3. Obesity Institute, Geisinger Medical Center, 100 North Academy Ave, Danville, PA 17822, USA;4. Graduate School of Dental Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 Japan;5. Oral Health Science Center, Tokyo Dental College, 2-9-18 Kandamisaki-cho, Chiyoda-ku, Tokyo 101-0061 Japan;6. Department of Oral Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1, Shikata-cho, Kita-ku, Okayama, 700-8558 Japan;1. Division of Forensic Odontology, Department of Diagnostic ＆ Therapeutic Sciences, Meikai University School of Dentistry, Saitama, Japan;2. Division of Dental Radiology, Department of Diagnostic ＆ Therapeutic Sciences, Meikai University School of Dentistry, Saitama, Japan;3. Forensic Odontology Center, Meikai University School of Dentistry, Saitama, Japan;4. Department of Legal Medicine, Nihon University School of Dentistry, Tokyo, Japan;1. Oral Biology and Biochemistry Research Group, Unidade de Investigação Em Ciências Orais e Biomédicas (UICOB), Faculdade de Medicina Dentária, Universidade de Lisboa, Lisboa, Portugal;2. Center for Microelectromechanical Systems (CMEMS), Dept. of Mechanical Engineering, University of Minho, Guimarães, Portugal;3. Oral Biology and Biochemistry Research Group, Unidade de Investigação Em Ciências Orais e Biomédicas (UICOB), LIBPhys-FCT UID/FIS/04559/2013, Faculdade de Medicina Dentária, Universidade de Lisboa, Lisboa, Portugal;4. CEMDBE - Cochrane Portugal, Faculdade de Medicina Dentária, Universidade de Lisboa, Lisboa, Portugal;5. Implant & Tissue Regeneration Group, Unidade de Investigação Em Ciências Orais e Biomédicas (UICOB), LIBPhys-FCT UID/FIS/04559/2013, Universidade de Lisboa, Faculdade de Medicina Dentária, Lisboa, Portugal;1. Division of Oral and Maxillofacial Surgery, Department of Oral Surgery, Iwate Medical University, Iwate, Japan;2. Division of Molecular Microbiology, Department of Microbiology, Iwate Medical University, Iwate, Japan;3. Division of Bioregulatory Pharmacology, Department of Pharmacology, Iwate Medical University, Iwate, Japan;1. Department of Basic Sciences, Faculty of Dental Sciences, University of Peradeniya, 20400, Sri Lanka;2. Dental Hospital (Teaching), University of Peradeniya, 20400, Sri Lanka |
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| Abstract: | ObjectivesOriginal odontoblasts and regenerated odontoblast-like cells (OBLCs) may differently regulate Nestin expression. This study aimed to investigate the role of the subodontoblastic layer (SOBL) using green fluorescent protein (GFP) reactivity in the process of OBLC differentiation after tooth drilling in Nestin-enhanced GFP transgenic mice.MethodsA groove-shaped cavity was prepared on the mesial surface of the maxillary first molars of 5- or 6-week-old mice under deep anesthesia. Immunohistochemical staining for Nestin and GFP and Nestin in situ hybridization were conducted on the sections obtained at 1–14 days postoperative.ResultsOdontoblasts showed intense endogenous Nestin protein and mRNA expression, whereas the coronal SOBL cells showed a Nestin-GFP–positive reaction in the control groups. The injured odontoblasts had significantly decreased Nestin immunoreactivity as well as decreased expression of Nestin mRNA 1–2 days after the injury; subsequently, newly differentiated OBLCs were arranged along the pulp–dentin border, with significantly increased Nestin expression as well as increased expression of Nestin mRNA on days 3–5 to form reparative dentin. Nestin-GFP–positive cells at the pulp–dentin border significantly increased in number on days 1 and 2. GFP(+)/Nestin(+) and GFP(?)/Nestin(+) cells were intermingled in the newly differentiated OBLCs.ConclusionsThe commitment of Nestin-GFP–positive cells into Nestin-positive OBLCs suggests that the restriction of endogenous Nestin protein and mRNA expression in the static SOBL cells was removed by exogenous stimuli, resulting in their migration along the pulp–dentin border and their differentiation into OBLCs. |
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| Keywords: | Cell differentiation Dental cavity preparation Dental pulp Odontoblasts Transgenic mice |
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