M Usui1,2, T Sato3, G Yamamoto4, Y Okamatsu5, T Hanatani1, Y Moritani1, K Sano1, M Yamamoto2, K Nakashima1. 1. Department of Cardiology and Periodontology, Division of Periodontology, Kyushu Dental University, Kitakyushu City, Fukuoka, Japan. 2. Department of Periodontology, Showa University School of Dentistry, Ohta-ku, Tokyo, Japan. 3. Department of Oral and Maxillofacial Surgery, Saitama Medical University, Iruma-gun, Saitama, Japan. 4. Department of Oral Pathology and Diagnosis, Showa University School of Dentistry, Shinagawa-ku, Tokyo, Japan. 5. Dental Clinic, Showa University Medical Hospital, Shinagawa-ku, Tokyo, Japan.
Abstract
BACKGROUND AND OBJECTIVE: Periodontal disease is dental plaque-induced inflammatory disease of the periodontal tissues that results in bone loss in the affected teeth. During bone resorption, receptor activator of nuclear factor kappa B ligand (RANKL) is an essential factor that regulates osteoclastogenesis. Recently, we found that gingival epithelial cells (GECs) in periodontal tissue produce RANKL, the expression of which is regulated by tumor necrosis factor-α and protein kinase A signaling. In this study, we asked whether RANKL-producing GECs induce bone marrow macrophages (BMMs) to form osteoclasts in a co-culture system. MATERIAL AND METHODS: Ca9-22 GECs and osteoclast precursor BMMs were co-cultured with or without the protein kinase A signaling activator forskolin or inhibitor H89 to examine whether the RANKL-producing GECs could be induced to form osteoclasts, as determined using a pit formation assay. RESULTS: Osteoclasts formed spontaneously in co-cultures of Ca9-22 cells and BMMs, even in the absence of RANKL. The cells were cultured on bone slices for 14 d, at which time resorption pits were observed. Forskolin treatment significantly increased osteoclast numbers in these co-cultures, but forskolin alone did not induce osteoclast formation by BMMs. CONCLUSION: GECs producing RANKL are able to support osteoclastogenesis in an in vitro co-culture system using GECs and BMMs, in a process promoted by forskolin.
BACKGROUND AND OBJECTIVE: Periodontal disease is dental plaque-induced inflammatory disease of the periodontal tissues that results in bone loss in the affected teeth. During bone resorption, receptor activator of nuclear factor kappa B ligand (RANKL) is an essential factor that regulates osteoclastogenesis. Recently, we found that gingival epithelial cells (GECs) in periodontal tissue produce RANKL, the expression of which is regulated by tumor necrosis factor-α and protein kinase A signaling. In this study, we asked whether RANKL-producing GECs induce bone marrow macrophages (BMMs) to form osteoclasts in a co-culture system. MATERIAL AND METHODS: Ca9-22 GECs and osteoclast precursor BMMs were co-cultured with or without the protein kinase A signaling activator forskolin or inhibitor H89 to examine whether the RANKL-producing GECs could be induced to form osteoclasts, as determined using a pit formation assay. RESULTS: Osteoclasts formed spontaneously in co-cultures of Ca9-22 cells and BMMs, even in the absence of RANKL. The cells were cultured on bone slices for 14 d, at which time resorption pits were observed. Forskolin treatment significantly increased osteoclast numbers in these co-cultures, but forskolin alone did not induce osteoclast formation by BMMs. CONCLUSION: GECs producing RANKL are able to support osteoclastogenesis in an in vitro co-culture system using GECs and BMMs, in a process promoted by forskolin.
Authors: Olivier Lapérine; Alexandra Cloitre; Jocelyne Caillon; Olivier Huck; Isaac Maximiliano Bugueno; Paul Pilet; Sophie Sourice; Elodie Le Tilly; Gaby Palmer; Jean-Luc Davideau; Valérie Geoffroy; Jérôme Guicheux; Sarah Beck-Cormier; Philippe Lesclous Journal: PLoS One Date: 2016-12-19 Impact factor: 3.240
Authors: Alexandra Cloitre; Boris Halgand; Sophie Sourice; Jocelyne Caillon; Olivier Huck; Isaac Maximiliano Bugueno; Fareeha Batool; Jérôme Guicheux; Valérie Geoffroy; Philippe Lesclous Journal: Sci Rep Date: 2019-12-17 Impact factor: 4.379