Secretoneurin, a Neuropeptide, Enhances Bone Regeneration in a Mouse Calvarial Bone Defect Model
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ORIGINAL ARTICLE
Secretoneurin, a Neuropeptide, Enhances Bone Regeneration in a Mouse Calvarial Bone Defect Model Freshet Assefa1 • Jiwon Lim1 • Ju-Ang Kim1 • Hye Jung Ihn2 • Soomin Lim1 Sang-Hyeon Nam1 • Yong Chul Bae3 • Eui Kyun Park1
•
Received: 11 August 2020 / Revised: 7 September 2020 / Accepted: 16 September 2020 Ó The Korean Tissue Engineering and Regenerative Medicine Society 2020
Abstract BACKGROUND: This study investigates the effects of a neuropeptide, secretoneurin (SN), on bone regeneration in an experimental mouse model. METHODS: The effects of SN on cell proliferation, osteoblast marker genes expression, and mineralization were evaluated using the CCK-8 assay, quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and alizarin red S staining, respectively. To examine the effects of SN on bone regeneration in vivo, bone defects were created in the calvaria of ICR mice, and 0.5 or 1 lg/ml SN was applied. New bone formation was analyzed by micro-computed tomography (micro-CT) and histology. New blood vessel formation was assessed by CD34 immunohistochemistry. RESULTS: SN had no significant effect on proliferation and mineralization of MC3T3-E1 cells. However, SN partially induced the gene expression of osteoblast differentiation markers such as runt-related transcription factor 2, alkaline phosphatase, collagen type I alpha 1, and osteopontin. A significant increase of bone regeneration was observed in SN treated calvarial defects. The bone volume (BV), BV/tissue volume, trabecular thickness and trabecular number values were significantly increased in the collagen sponge plus 0.5 or 1 lg/ml SN group (p \ 0.01) compared with the control group. Histologic analysis also revealed increased new bone formation in the SN-treated groups. Immunohistochemical staining of CD34 showed that the SN-treated groups contained more blood vessels compared with control in the calvarial defect area. CONCLUSION: SN increases new bone and blood vessel formation in a calvarial defect site. This study suggests that SN may enhance new bone formation through its potent angiogenic activity. Keywords Neuropeptide Secretoneurin Bone regeneration Bone
1 Introduction
& Eui Kyun Park [email protected] 1
Department of Oral Pathology and Regenerative Medicine, School of Dentistry, Kyungpook National University, 2177 Dalgubeol-daero, Jung-gu, Daegu 41940, Republic of Korea
2
Cell & Matrix Research Institute, Kyungpook National University, Daegu 41944, Republic of Korea
3
Department of Oral Anatomy and Neurobiology, School of Dentistry, Kyungpook National University, Daegu 41940, South Korea
Bone repair is a complex process that leads to new bone formation through cellular and molecular processes. It is regulated by multiple biological factors, such as the transforming growth factor-b (TGF-b) superfamily members and proinflammatory cytokines [1]. The bone repair and fracture healing processes consist of several overlapping stages including inflammation and angiogenesis [2]. Angiogensi
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