Neuropeptide Y stimulates osteoblastic differentiation and VEGF expression of bone marrow mesenchymal stem cells related to canonical Wnt signaling activating in vitro

Liu, Song; Jin, Dayong; Wu, Jianxin; Xu, Ziyi; Fu, Su; Mei, Gang; Zou, Zhenlv; Ma, Shenghui

doi:10.1016/j.npep.2015.12.008

Cited by 48 publications

(47 citation statements)

References 32 publications

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“…Classically, this has been performed for individual proteins by western blotting, which is still widely reported in the current literature (Bayerl et al, 2016;Liu et al, 2016a). Owing to the typically small size of neuropeptides, western blotting is normally used to assess other, related protein changes or expression of neuropeptide receptors (Bayerl et al, 2016;Liu et al, 2016b). Other complementary examples of a targeted technique are enzyme-linked immunosorbent assay (ELISA) (Bilgic et al, 2016;Javadian et al, 2016) and use of radioactively labeled ligand and a γ-counter (Dhuria et al, 2016).…”

Section: Measuring Neuropeptide Levelsmentioning

confidence: 99%

New techniques, applications and perspectives in neuropeptide research

DeLaney

Buchberger

Atkinson

et al. 2018

Journal of Experimental Biology

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Neuropeptides are one of the most diverse classes of signaling molecules and have attracted great interest over the years owing to their roles in regulation of a wide range of physiological processes. However, there are unique challenges associated with neuropeptide studies stemming from the highly variable molecular sizes of the peptides, low concentrations, high degree of structural diversity and large number of isoforms. As a result, much effort has been focused on developing new techniques for studying neuropeptides, as well as novel applications directed towards learning more about these endogenous peptides. The areas of importance for neuropeptide studies include structure, localization within tissues, interaction with their receptors, including ion channels, and physiological function. Here, we discuss these aspects and the associated techniques, focusing on technologies that have demonstrated potential in advancing the field in recent years. Most identification and structural information has been gained by mass spectrometry, either alone or with confirmations from other techniques, such as nuclear magnetic resonance spectroscopy and other spectroscopic tools. While mass spectrometry and bioinformatic tools have proven to be the most powerful for large-scale analyses, they still rely heavily on complementary methods for confirmation. Localization within tissues, for example, can be probed by mass spectrometry imaging, immunohistochemistry and radioimmunoassays. Functional information has been gained primarily from behavioral studies coupled with tissue-specific assays, electrophysiology, mass spectrometry and optogenetic tools. Concerning the receptors for neuropeptides, the discovery of ion channels that are directly gated by neuropeptides opens up the possibility of developing a new generation of tools for neuroscience, which could be used to monitor neuropeptide release or to specifically change the membrane potential of neurons. It is expected that future neuropeptide research will involve the integration of complementary bioanalytical technologies and functional assays.

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Section: Measuring Neuropeptide Levelsmentioning

confidence: 99%

New techniques, applications and perspectives in neuropeptide research

DeLaney

Buchberger

Atkinson

et al. 2018

Journal of Experimental Biology

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“…Bone is innervated by sensory and sympathetic nerve bers that, in addition to skeletal pain transmission, play a role in bone metabolism [3]. Many neuropeptides secreted by sensory and autonomic nerves within bone tissues, including substance P, calcitonin gene-related peptide from sensory nerves, vasoactive intestinal peptide from parasympathetic nerves, and neuropeptide Y from sympathetic nerves, positively regulate the function of bone cells [11][12][13]. Similarly, neurotrophins, including nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), directly stimulate bone cells [14,15].…”

Section: Introductionmentioning

confidence: 99%

Schwann Cells Promote Prevascularization and Osteogenesis of Tissue-engineered Bone via Bone Marrow Mesenchymal Stem Cell-derived Endothelial Cells

Zhang

Jiang

et al. 2020

Preprint

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Background: Tissue-engineered bone grafts (TEBGs) that undergo vascularization and neurotization evolve into functioning bone tissue. Previously, we verified that implanting sensory nerve tracts into TEBGs promoted osteogenesis. However, the precise mechanisms and interaction between seed cells were not explored. In this study, we hypothesized that neurotization may influence the osteogenesis of TEBGs through vascularization. Methods: We cultured rat Schwann cells (SCs), aortic endothelial cells (AECs) and bone marrow-derived mesenchymal stem cells (BM-MSCs) and then obtained BM-MSC-derived induced endothelial cells (IECs) and induced osteoblasts (IOBs). IECs and AECs were cultured in SC-conditioned medium (SC-CM) to assess proliferation, migration, capillary-like tube formation and angiogenesis, and the vascular endothelial growth factor (VEGF) levels in the supernatants were detected. We established an indirect coculture model to detect the expression of nestin and VEGF receptors in IECs and tissue inhibitor of metalloproteinase (TIMP)-2 in SCs. Then, SCs, IECs and IOBs were labeled and loaded into a β-tricalcium phosphate scaffold to induce prevascularization, and the scaffold was implanted into a 6-mm-long defect of rat femurs. Three groups were set up according to the loaded cells: I, SCs and IECs (coculture for 3 days) plus IOBs; II, IECs (culture for 3 days) plus IOBs; III, IOBs. Nestin and TIMP-2 expression and osteogenesis of TEBGs were evaluated at 12 weeks post implantation through histological and radiological assessment.Results: We found that SC-CM promoted IEC proliferation, migration, capillary-like tube formation and angiogenesis, but no similar effects were observed for AECs. IECs expressed nestin extensively, while AECs barely expressed nestin, and SC-CM promoted the VEGF secretion of IECs. In the coculture model, SCs promoted nestin and VEGF receptor expression in IECs, and IECs inhibited TIMP-2 expression in SCs. The promotion of prevascularized TEBGs by SCs and IECs in group I augmented new bone formation at 6 and 12 weeks. Nestin expression was higher in group I than in the other groups, while TIMP-2 expression was lower at 12 weeks. Conclusions: This study demonstrated that SCs can promote TEBG osteogenesis via IECs and further revealed the mechanisms related to specific characteristics of IECs, providing preliminary cytological evidence for neurotization of TEBGs.

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“…Recently, increasing evidence showed that neural signaling within the central nervous system was involved in bone remodeling through various pathways. Previous studies demonstrated that neuropeptide Y (NPY), a 36-amino acid peptide that exerts its function in the local bone by its Y1 receptor (NPY1R), enhanced migration and osteoblastic differentiation of MSCs and facilitated fracture healing [14,15]. However, the association of MLT with NPY/NPY1R in osteoblastic differentiation of MSCs and fracture healing has not been elaborated yet.…”

Section: Introductionmentioning

confidence: 99%

Melatonin Induces Osteoblastic Differentiation of Mesenchymal Stem Cells and Promotes Fracture Healing in a Rat Model of Femoral Fracture via Neuropeptide Y/Neuropeptide Y Receptor Y1 Signaling

Dong

Zhu

et al. 2018

Pharmacology

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The function of melatonin (MLT) in promoting fracture healing has been demonstrated in previous studies. However, the molecular mechanism underlying therapeutic effects of MLT is not entirely clear. In this study, mesenchymal stem cells (MSCs) were isolated from rat bone marrow and identified by flow cytometry. We found that MLT treatment upregulated the neuropeptide Y (NPY) and NPY receptor Y1 (NPY1R) expression, and promoted the proliferation and migration of MSCs, which was suppressed by BIBP3226, an inhibitor of NPY1R. Moreover, the levels of NPY and NPY1R in MSCs undergoing osteoblastic differentiation were upregulated after MLT administration. MLT-induced osteoblastic differentiation of MSCs was suppressed by BIBP3226 treatment, as evidenced by decreased levels of alkaline phosphatase (ALP), collagen type I α1 chain, osteocalcin, and runt-related transcription factor 2, downregulated activity of ALP, as well as reduced calcium nodule formation. Furthermore, we demonstrated that MLT could promote fracture healing in a rat model of femoral fracture, which was accompanied by the elevated expression of NPY and NPY1R. The administration of BIBP3226 inhibited fracture healing mediated by MLT. To sum up, our results show that MLT promotes osteoblastic differentiation of MSCs and fracture healing by NPY/NPY1R signaling.

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Neuropeptide Y stimulates osteoblastic differentiation and VEGF expression of bone marrow mesenchymal stem cells related to canonical Wnt signaling activating in vitro

Cited by 48 publications

References 32 publications

New techniques, applications and perspectives in neuropeptide research

New techniques, applications and perspectives in neuropeptide research

Schwann Cells Promote Prevascularization and Osteogenesis of Tissue-engineered Bone via Bone Marrow Mesenchymal Stem Cell-derived Endothelial Cells

Melatonin Induces Osteoblastic Differentiation of Mesenchymal Stem Cells and Promotes Fracture Healing in a Rat Model of Femoral Fracture via Neuropeptide Y/Neuropeptide Y Receptor Y1 Signaling

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