Secretoneurin, a Neuropeptide, Enhances Bone Regeneration in a Mouse Calvarial Bone Defect Model

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.

The potential bone regeneration effects of leptin- and osteolectin-coated 3D-printed PCL scaffolds: an in vivo study

“…The rat calvarial defect models were immersed in the Calci-Clear Rapid (National Diagnostics, Atlanta, GA) decalcification solution, embedded in paraffin wax, and then sectioned coronally at a thickness of 4 µm onto tissue slides. To assess bone regeneration, the sections were stained with hematoxylin and eosin (H&E, Sigma-Aldrich, Germany) and Masson's trichrome (Sigma-Aldrich, Germany) according to manufacturer instructions [61][62][63][64][65] and then scanned using a digital slide scanner (Motic Easy Scan Pro digital slide scanner, Motic Asia Corp., Kowloon, Hong Kong). Finally, bone regeneration and histological changes were observed and analyzed using motic DSAssistant software (Motic VM V1 Viewer 2.0, Motic Asia Corp., Kowloon, Hong Kong).…”

“…They were then incubated with biotinylated goat anti-rabbit IgG (Vector ABC Elite Kit) at room temperature for one hour and then incubated with avidin-biotin-peroxidase complex (Vector ABC Elite Kit) at room temperature for one hour. Peroxidase signal was then developed using the diaminobenzidine substrate (DAB kit, Vector Laboratories) [64][65][66], followed by scanning of the stained tissue slides using a digital slide scanner. Finally, the immunohistochemistry data were observed and analyzed using motic DSAssistant software.…”

Secretoneurin as a Novel Biomarker of Cardiovascular Episodes: Are We There Yet? A Narrative Review

“…Of note, in animal models SN also enhances bone regeneration by increasing blood vessel and bone marrow formation [ 20 ].…”

“…These effects are known to be mediated by adenosine-monophosphate or the extracellular-regulated kinase (ERK)/MAPK pathway. It is less probable that SN directly affects myocardial cell hypertrophy [ 20 ]. Nevertheless, SN gene therapy (in an animal model of isoproterenol-induced hypertrophy) led to significant reductions in natriuretic peptide levels, the heart-weight to body-weight ratio, and surprisingly, interstitial fibrosis [ 25 ].…”