Bone defect regeneration is a complex process that involves the coordination of a variety of different type of cells. As bone tissues are innervated and rich in nerve fibers, the neuropeptides released from various never fibers could regulate bone development, metabolism, and remodeling. Among all the neuropeptides, vasoactive intestinal peptide (VIP) could modulate the functions of both osteoblasts and osteoclasts, and may play a vital role in bone marrow mesenchymal stem cell (BMSC) osteogenesis during bone repair. In this study, we investigated the role of VIP in bone formation and the mechanisms of VIP in mediating BMSC osteogenic differentiation, and its possibility in clinical application of bone defect reconstruction. Our in vitro study results indicated that VIP promoted BMSC osteogenic differentiation by activating Wnt/β-catenin signaling pathway in BMSCs. VIP could also stimulate tube formation of EA.hy926 endothelial cell and increase vascular endothelial growth factor (VEGF) expression in BMSCs. Furthermore, in the rat skull defect model, VIP-conjugated functionalized hydrogel significantly enhanced cranial bone defect repair compared with the control group, with increased bone formation and angiogenesis. Taken together, as a member of neuropeptides, VIP could promote the BMSCs osteogenesis and angiogenesis differentiation in vitro and stimulate bone repair in vivo by activating Wnt/β-catenin signaling pathway. The knowledge obtained from this study emphasized the close association between innervation and bone repair process, and VIP may be a potential therapeutic agent for augmenting bone repair.
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References
1.
CraneJL and CaoX. (2014). Bone marrow mesenchymal stem cells and TGF-beta signaling in bone remodeling. J Clin Invest, 124:466–472.
2.
KularJ, TicknerJ, ChimSM and XuJ. (2012). An overview of the regulation of bone remodelling at the cellular level. Clin Biochem, 45:863–873.
3.
MuschterD, SchaferN, StanglH, StraubRH and GrasselS. (2015). Sympathetic neurotransmitters modulate osteoclastogenesis and osteoclast activity in the context of collagen-induced arthritis. PLoS One, 10:e0139726.
4.
NiedermairT, KuhnV, DoranehgardF, StangeR, WieskotterB, BeckmannJ, SalmenP, SpringorumHR, StraubRH, et al. (2014). Absence of substance P and the sympathetic nervous system impact on bone structure and chondrocyte differentiation in an adult model of endochondral ossification. Matrix Biol, 38:22–35.
5.
KonttinenY, ImaiS and SudaA. (1996). Neuropeptides and the puzzle of bone remodeling. State of the art. Acta Orthop Scand, 67:632–639.
6.
LiuX, LiuH, XiongY, YangL, WangC, ZhangR and ZhuX. (2018). Postmenopausal osteoporosis is associated with the regulation of SP, CGRP, VIP, and NPY. Biomed Pharmacother, 104:742–750.
7.
JiangW, GaoSG, ChenXG, XuXC, XuM, LuoW, TuM, ZhangFJ, ZengC and LeiGH. (2012). Expression of synovial fluid and articular cartilage VIP in human osteoarthritic knee: a new indicator of disease severity?. Clin Biochem, 45:1607–1612.
8.
HuangS, XuL, ZhangY, SunY and LiG. (2015). Systemic and local administration of allogeneic bone marrow-derived mesenchymal stem cells promotes fracture healing in rats. Cell Transplant, 24:2643–2655.
9.
Lerner UH. (2000). Osteoclast formation and resorption. Matrix Biol, 19:107–120.
10.
SassoliC, PiniA, ChelliniF, MazzantiB, NistriS, NosiD, SaccardiR, QuercioliF, Zecchi-OrlandiniS and FormigliL. (2012). Bone marrow mesenchymal stromal cells stimulate skeletal myoblast proliferation through the paracrine release of VEGF. PLoS One, 7:e37512.
11.
RameshwarP, GasconP, OhHS, DennyTN, ZhuG and GaneaD. (2002). Vasoactive intestinal peptide (VIP) inhibits the proliferation of bone marrow progenitors through the VPAC1 receptor. Exp Hematol, 30:1001–1009.
12.
WangL, XiaoQ, WangCH, LiX, LuoSQ and TangCW. (2014). Vasoactive intestinal polypeptide suppresses proliferation of human cord blood-derived hematopoietic progenitor cells by increasing TNF-alpha and TGF-beta production in the liver. Genet Mol Res, 13:9032–9043.
13.
MartiniL, StaffaG, GiavaresiG, SalamannaF, ParrilliA, SerchiE, PressatoD, ArcangeliE and FiniM. (2012). Long-term results following cranial hydroxyapatite prosthesis implantation in a large skull defect model. Plast Reconstr Surg, 129:625e–635e.
14.
ZouL, ZouX, ChenL, LiH, MygindT, KassemM and BungerC. (2008). Effect of hyaluronan on osteogenic differentiation of porcine bone marrow stromal cells in vitro. J Orthop Res, 26:713–720.
15.
ZhuM, LinS, SunY, FengQ, LiG and BianL. (2016). Hydrogels functionalized with N-cadherin mimetic peptide enhance osteogenesis of hMSCs by emulating the osteogenic niche. Biomaterials, 77:44–52.
16.
XuL, SongC, NiM, MengF, XieH and LiG. (2012). Cellular retinol-binding protein 1 (CRBP-1) regulates osteogenenesis and adipogenesis of mesenchymal stem cells through inhibiting RXRalpha-induced beta-catenin degradation. Int J Biochem Cell Biol, 44:612–619.
17.
SunY, XuL, HuangS, HouY, LiuY, ChanKM, PanXH and LiG. (2015). mir-21 overexpressing mesenchymal stem cells accelerate fracture healing in a rat closed femur fracture model. Biomed Res Int, 2015:412327.
18.
SunY, XuJ, XuL, ZhangJ, ChanK, PanX and LiG. (2017). MiR-503 promotes bone formation in distraction osteogenesis through suppressing Smurf1 expression. Sci Rep, 7:409.
19.
ShiL, FengL, LiuY, DuanJQ, LinWP, ZhangJF and LiG. (2018). MicroRNA-218 promotes osteogenic differentiation of mesenchymal stem cells and accelerates bone fracture healing. Calcif Tissue Int, 103:227–236.
20.
ArandaE and OwenGI. (2009). A semi-quantitative assay to screen for angiogenic compounds and compounds with angiogenic potential using the EA.hy926 endothelial cell line. Biol Res, 42:377–389.
21.
FengL, ShiL, LuYF, WangB, TangT, FuWM, HeW, LiG and ZhangJF. (2018). Linc-ROR promotes osteogenic differentiation of mesenchymal stem cells by functioning as a competing endogenous RNA for miR-138 and miR-145. Mol Ther Nucleic Acids, 11:345–353.
22.
ChenS, YangP, JiangF, WeiY, MaZ and KangL. (2010). De novo analysis of transcriptome dynamics in the migratory locust during the development of phase traits. PLoS One, 5:e15633.
23.
BianL, HouC, TousE, RaiR, MauckRL and BurdickJA. (2013). The influence of hyaluronic acid hydrogel crosslinking density and macromolecular diffusivity on human MSC chondrogenesis and hypertrophy. Biomaterials, 34:413–421.
24.
VandermeersA, VandenborreS, HouX, de NeefP, RobberechtP, Vandermeers-PiretMC and ChristopheJ. (1992). Antagonistic properties are shifted back to agonistic properties by further N-terminal shortening of pituitary adenylate-cyclase-activating peptides in human neuroblastoma NB-OK-1 cell membranes. Eur J Biochem, 208:815–819.
25.
XuJ, WuT, SunY, WangB, ZhangJ, LeeWY, ChaiY and LiG. (2017). Staphylococcal enterotoxin C2 expedites bone consolidation in distraction osteogenesis. J Orthop Res, 35:1215–1225.
26.
OrimoH. (2010). The mechanism of mineralization and the role of alkaline phosphatase in health and disease. J Nippon Med Sch, 77:4–12.
27.
GordonJA, TyeCE, SampaioAV, UnderhillTM, HunterGK and GoldbergHA. (2007). Bone sialoprotein expression enhances osteoblast differentiation and matrix mineralization in vitro. Bone, 41:462–473.
28.
OrimoH and ShimadaT. (2008). The role of tissue-nonspecific alkaline phosphatase in the phosphate-induced activation of alkaline phosphatase and mineralization in SaOS-2 human osteoblast-like cells. Mol Cell Biochem, 315:51–60.
29.
Hughes-FulfordM and LiCF. (2011). The role of FGF-2 and BMP-2 in regulation of gene induction, cell proliferation and mineralization. J Orthop Surg Res, 6:8.
30.
ZhaoZ, ZhaoM, XiaoG and FranceschiRT. (2005). Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo. Mol Ther, 12:247–253.
31.
BertramH, MayerH and SchliephakeH. (2005). Effect of donor characteristics, technique of harvesting and in vitro processing on culturing of human marrow stroma cells for tissue engineered growth of bone. Clin Oral Implants Res, 16:524–531.
32.
MaW, ZhangX, ShiS and ZhangY. (2013). Neuropeptides stimulate human osteoblast activity and promote gap junctional intercellular communication. Neuropeptides, 47:179–186.
33.
LundbergP, BostromI, MukohyamaH, BjurholmA, SmansK and LernerUH. (1999). Neuro-hormonal control of bone metabolism: vasoactive intestinal peptide stimulates alkaline phosphatase activity and mRNA expression in mouse calvarial osteoblasts as well as calcium accumulation mineralized bone nodules. Regul Pept, 85:47–58.
34.
NatsumeH, TokudaH, MizutaniJ, AdachiS, Matsushima-NishiwakiR, MinamitaniC, KatoK, KozawaO and OtsukaT. (2010). Synergistic effect of vasoactive intestinal peptides on TNF-alpha-induced IL-6 synthesis in osteoblasts: amplification of p44/p42 MAP kinase activation. Int J Mol Med, 25:813–817.
35.
PerssonE, VoznesenskyOS, HuangYF and LernerUH. (2005). Increased expression of interleukin-6 by vasoactive intestinal peptide is associated with regulation of CREB, AP-1 and C/EBP, but not NF-kappaB, in mouse calvarial osteoblasts. Bone, 37:513–529.
36.
LescherB, HaenigB and KispertA. (1998). sFRP-2 is a target of the Wnt-4 signaling pathway in the developing metanephric kidney. Dev Dyn, 213:440–451.
37.
DavoodiS, CookeRF, FernandesAC, CappellozzaBI, VasconcelosJL and CerriRL. (2016). Expression of estrus modifies the gene expression profile in reproductive tissues on day 19 of gestation in beef cows. Theriogenology, 85:645–655.
38.
CarrollTJ, ParkJS, HayashiS, MajumdarA and McMahonAP. (2005). Wnt9b plays a central role in the regulation of mesenchymal to epithelial transitions underlying organogenesis of the mammalian urogenital system. Dev Cell, 9:283–292.
39.
CawthornWP, BreeAJ, YaoY, DuB, HematiN, Martinez-SantibanezG and MacDougaldOA. (2012). Wnt6, Wnt10a and Wnt10b inhibit adipogenesis and stimulate osteoblastogenesis through a beta-catenin-dependent mechanism. Bone, 50:477–489.
40.
JinL, CaoY, YuG, WangJ, LinX, GeL, DuJ, WangL, DiaoS, et al. (2017). SFRP2 enhances the osteogenic differentiation of apical papilla stem cells by antagonizing the canonical WNT pathway. Cell Mol Biol Lett, 22:14.
41.
KressE, RezzaA, NadjarJ, SamarutJ and PlaterotiM. (2009). The frizzled-related sFRP2 gene is a target of thyroid hormone receptor alpha1 and activates beta-catenin signaling in mouse intestine. J Biol Chem, 284:1234–1241.
42.
MastriM, ShahZ, HsiehK, WangX, WooldridgeB, MartinS, SuzukiG and LeeT. (2014). Secreted Frizzled-related protein 2 as a target in antifibrotic therapeutic intervention. Am J Physiol Cell Physiol, 306:C531–C539.
43.
LinH, AngeliM, ChungKJ, EjimaduC, RosaAR and LeeT. (2016). sFRP2 activates Wnt/beta-catenin signaling in cardiac fibroblasts: differential roles in cell growth, energy metabolism, and extracellular matrix remodeling. Am J Physiol Cell Physiol, 311:C710–C719.
44.
LanY, RyanRC, ZhangZ, BullardSA, BushJO, MaltbyKM, LidralAC and JiangR. (2006). Expression of Wnt9b and activation of canonical Wnt signaling during midfacial morphogenesis in mice. Dev Dyn, 235:1448–1454.
45.
WillisCM and KluppelM. (2012). Inhibition by chondroitin sulfate E can specify functional Wnt/beta-catenin signaling thresholds in NIH3T3 fibroblasts. J Biol Chem, 287:37042–37056.
46.
DayTF, GuoX, Garrett-BealL and YangY. (2005). Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell, 8:739–750.
47.
JacksonA, VayssiereB, GarciaT, NewellW, BaronR, Roman-RomanS and RawadiG. (2005). Gene array analysis of Wnt-regulated genes in C3H10T1/2 cells. Bone, 36:585–598.
48.
GlassDA, II and KarsentyG. (2007). In vivo analysis of Wnt signaling in bone. Endocrinology, 148:2630–2634.
49.
SzpalskiC, BarrJ, WetterauM, SaadehPB and WarrenSM. (2010). Cranial bone defects: current and future strategies. Neurosurg Focus, 29:E8.
50.
LiuX, HuangR, SuR, QiW, WangL and HeZ. (2014). Grafting hyaluronic acid onto gold surface to achieve low protein fouling in surface plasmon resonance biosensors. ACS Appl Mater Interfaces, 6:13034–13042.
51.
WangY, ChenZ, LuoG, HeW, XuK, XuR, LeiQ, TanJ, WuJ and XingM. (2016). In-situ-generated vasoactive intestinal peptide loaded microspheres in mussel-inspired polycaprolactone nanosheets creating spatiotemporal releasing microenvironment to promote wound healing and angiogenesis. ACS Appl Mater Interfaces, 8:7411–7421.
52.
ColladoB, Sanchez-ChapadoM, PrietoJC and CarmenaMJ. (2006). Hypoxia regulation of expression and angiogenic effects of vasoactive intestinal peptide (VIP) and VIP receptors in LNCaP prostate cancer cells. Mol Cell Endocrinol, 249:116–122.
53.
YuXJ, RenXH, XuYH, ChenLM, ZhouCL and LiCY. (2010). Vasoactive intestinal peptide induces vascular endothelial growth factor production in human HaCaT keratinocytes via MAPK pathway. Neuropeptides, 44:407–411.
54.
YangJ, ShiQD, SongTB, FengGF, ZangWJ, ZongCH and ChangL. (2013). Vasoactive intestinal peptide increases VEGF expression to promote proliferation of brain vascular endothelial cells via the cAMP/PKA pathway after ischemic insult in vitro. Peptides, 42:105–111.
55.
YangJ, ZongCH, ZhaoZH, HuXD, ShiQD, XiaoXL and LiuY. (2009). Vasoactive intestinal peptide in rats with focal cerebral ischemia enhances angiogenesis. Neuroscience, 161:413–421.
56.
HuangC, NessVP, YangX, ChenH, LuoJ, BrownEB and ZhangX. (2015). Spatiotemporal analyses of osteogenesis and angiogenesis via intravital imaging in cranial bone defect repair. J Bone Miner Res, 30:1217–1230.
57.
HolsteinJH, BeckerSC, FiedlerM, GarciaP, HistingT, KleinM, LaschkeMW, CorstenM, PohlemannT and MengerMD. (2011). Intravital microscopic studies of angiogenesis during bone defect healing in mice calvaria. Injury, 42:765–771.
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