Naoki Edanami1, Nagako Yoshiba1, Naoto Ohkura1, Ryosuke Takeuchi1, Aiko Tohma1, Yuichiro Noiri1, Kunihiko Yoshiba2. 1. Division of Cariology, Operative Dentistry and Endodontics, Department of Oral Health Science, Course for Oral Life Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan. 2. Division of Cariology, Operative Dentistry and Endodontics, Department of Oral Health Science, Course for Oral Life Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan. Electronic address: yoshiba@dent.niigata-u.ac.jp.
Abstract
INTRODUCTION: Myofibroblasts express alpha smooth muscle actin (α-SMA) and play a critical role in wound healing. Myofibroblast differentiation is controlled by the joint actions of transforming growth factor beta 1 (TGF-β1) and the extradomain A fibronectin splice variant (EDA-FN). Currently, the contribution of myofibroblasts to dental pulp healing is unknown. Therefore, we analyzed expressional characteristics of α-SMA-positive cells and investigated TGF-β1, EDA-FN, and α-SMA expression levels after pulpotomy to better understand dental pulp healing. METHODS: The maxillary first molars of 8-week-old Wistar rats were pulpotomized with mineral trioxide aggregate. After 1 to 14 days, localization and colocalization of α-SMA, rat endothelial cell antigen-1 (as a marker of endothelial cells), neuron-glial antigen 2 (as a marker of perivascular cells), prolyl-4-hydroxylase (P4H, as an additional marker of myofibroblasts), and EDA-FN were analyzed using immunohistochemistry and double immunofluorescence. Time-course changes in the messenger RNA expression levels of TGF-β1, EDA-FN, and α-SMA were evaluated using quantitative real-time polymerase chain reaction analysis. RESULTS: Spindle-shaped α-SMA-positive cells transiently appeared after pulpotomy. These cells initially emerged in the pulp core on day 3 and then accumulated at the wound site by day 5. These cells were isolated from rat endothelial cell antigen-1 positive cells and did not express neuron-glial antigen 2 but did express P4H. The messenger RNA levels of TGF-β1, EDA-FN, and α-SMA were significantly up-regulated after pulpotomy. EDA-FN and α-SMA were colocalized at the wound sites on day 5. CONCLUSIONS: In association with up-regulation of TGF-β1 and EDA-FN expression, α-SMA and P4H double-positive cells accumulated at the wound sites after pulpotomy. This suggests that myofibroblasts participate in dental pulp healing.
INTRODUCTION: Myofibroblasts express alpha smooth muscle actin (α-SMA) and play a critical role in wound healing. Myofibroblast differentiation is controlled by the joint actions of transforming growth factor beta 1 (TGF-β1) and the extradomain A fibronectin splice variant (EDA-FN). Currently, the contribution of myofibroblasts to dental pulp healing is unknown. Therefore, we analyzed expressional characteristics of α-SMA-positive cells and investigated TGF-β1, EDA-FN, and α-SMA expression levels after pulpotomy to better understand dental pulp healing. METHODS: The maxillary first molars of 8-week-old Wistar rats were pulpotomized with mineral trioxide aggregate. After 1 to 14 days, localization and colocalization of α-SMA, rat endothelial cell antigen-1 (as a marker of endothelial cells), neuron-glial antigen 2 (as a marker of perivascular cells), prolyl-4-hydroxylase (P4H, as an additional marker of myofibroblasts), and EDA-FN were analyzed using immunohistochemistry and double immunofluorescence. Time-course changes in the messenger RNA expression levels of TGF-β1, EDA-FN, and α-SMA were evaluated using quantitative real-time polymerase chain reaction analysis. RESULTS: Spindle-shaped α-SMA-positive cells transiently appeared after pulpotomy. These cells initially emerged in the pulp core on day 3 and then accumulated at the wound site by day 5. These cells were isolated from rat endothelial cell antigen-1 positive cells and did not express neuron-glial antigen 2 but did express P4H. The messenger RNA levels of TGF-β1, EDA-FN, and α-SMA were significantly up-regulated after pulpotomy. EDA-FN and α-SMA were colocalized at the wound sites on day 5. CONCLUSIONS: In association with up-regulation of TGF-β1 and EDA-FN expression, α-SMA and P4H double-positive cells accumulated at the wound sites after pulpotomy. This suggests that myofibroblasts participate in dental pulp healing.