Bone morphogenetic protein 7 mediates stem cells migration and angiogenesis: therapeutic potential for endogenous pulp regeneration

Abstract

Pulp loss is accompanied by the functional impairment of defense, sensory, and nutrition supply. The approach based on endogenous stem cells is a potential strategy for pulp regeneration. However, endogenous stem cell sources, exogenous regenerative signals, and neovascularization are major difficulties for pulp regeneration based on endogenous stem cells. Therefore, the purpose of our research is to seek an effective cytokines delivery strategy and bioactive materials to reestablish an ideal regenerative microenvironment for pulp regeneration. In in vitro study, we investigated the effects of Wnt3a, transforming growth factor-beta 1, and bone morphogenetic protein 7 (BMP7) on human dental pulp stem cells (h-DPSCs) and human umbilical vein endothelial cells. 2D and 3D culture systems based on collagen gel, matrigel, and gelatin methacryloyl were fabricated to evaluate the morphology and viability of h-DPSCs. In in vivo study, an ectopic nude mouse model and an in situ beagle dog model were established to investigate the possibility of pulp regeneration by implanting collagen gel loading BMP7. We concluded that BMP7 promoted the migration and odontogenic differentiation of h-DPSCs and vessel formation. Collagen gel maintained the cell adhesion, cell spreading, and cell viability of h-DPSCs in 2D or 3D culture. The transplantation of collagen gel loading BMP7 induced vascularized pulp-like tissue regeneration in vivo. The injectable approach based on collagen gel loading BMP7 might exert promising therapeutic application in endogenous pulp regeneration.

References

[1] Zafar MS, Khurshid Z, Almas K. Oral tissue engineering progress and challenges. Tissue Eng. Regenerative Med., 2015, 12: 387-397.

[2] Javed, F. et al. Growth factors and guided bone regeneration, in Dental Implants (eds Zafar, M. S. & Khurshid, Z.) 133–143 (Woodhead Publishing, Elsevier, United Kingdom, 2020).

[3] Torabinejad M, . Outcomes of nonsurgical retreatment and endodontic surgery: a systematic review. J. Endod., 2009, 35: 930-937.

[4] Wang X, . Histologic characterization of regenerated tissues in canal space after the revitalization/revascularization procedure of immature dog teeth with apical periodontitis. J. Endod., 2010, 36: 56-63.

[5] Shi X, Mao J, Liu Y. Pulp stem cells derived from human permanent and deciduous teeth: Biological characteristics and therapeutic applications. Stem Cells Transl. Med., 2020, 9: 445-464.

[6] Huang GT, Al-Habib M, Gauthier P. Challenges of stem cell-based pulp and dentin regeneration: a clinical perspective. Endod. Top., 2013, 28: 51-60.

[7] Eramo S, . Dental pulp regeneration via cell homing. Int Endod. J., 2018, 51: 405-419.

[8] He L, . Parenchymal and stromal tissue regeneration of tooth organ by pivotal signals reinstated in decellularized matrix. Nat. Mater., 2019, 18: 627-637.

[9] Li J, Parada C, Chai Y. Cellular and molecular mechanisms of tooth root development. Development, 2017, 144: 374-384.

[10] Tamura M, Nemoto E. Role of the Wnt signaling molecules in the tooth. Jpn Dent. Sci. Rev., 2016, 52: 75-83.

[11] Yin X, . Wnt signaling and its contribution to craniofacial tissue homeostasis. J. Dent. Res., 2015, 94: 1487-1494.

[12] Lu X, . Advances of Wnt signalling pathway in dental development and potential clinical application. Organogenesis, 2019, 15: 101-110.

[13] Boland GM, . Wnt 3a promotes proliferation and suppresses osteogenic differentiation of adult human mesenchymal stem cells. J. Cell Biochem, 2004, 93: 1210-1230.

[14] Zhao Y, . Wnt-responsive odontoblasts secrete new dentin after superficial tooth injury. J. Dent. Res., 2018, 97: 1047-1054.

[15] Peng L, . WNT5A inhibits human dental papilla cell proliferation and migration. Biochem. Biophys. Res. Commun., 2009, 390: 1072-1078.

[16] He F, . Wnt5a regulates directional cell migration and cell proliferation via Ror2-mediated noncanonical pathway in mammalian palate development. Development, 2008, 135: 3871-3879.

[17] Zhang Z, . Effects of WNT10A on proliferation and differentiation of human dental pulp cells. J. Endod., 2014, 40: 1593-1599.

[18] Oka S, . Cell autonomous requirement for TGF-beta signaling during odontoblast differentiation and dentin matrix formation. Mech. Dev., 2007, 124: 409-415.

[19] da Rosa WLO, Piva E, da Silva AF. Disclosing the physiology of pulp tissue for vital pulp therapy. Int. Endod. J., 2018, 51: 829-846.

[20] Niwa T, . The dynamics of TGF-beta in dental pulp, odontoblasts and dentin. Sci. Rep., 2018, 8

[21] Nakamura M, . TGF-beta1 signaling is essential for tissue regeneration in the Xenopus tadpole tail. Biochem. Biophys. Res. Commun., 2021, 565: 91-96.

[22] Wang C, . Cryogenic 3D printing of heterogeneous scaffolds with gradient mechanical strengths and spatial delivery of osteogenic peptide/TGF-beta1 for osteochondral tissue regeneration. Biofabrication, 2020, 12: 025030.

[23] Langenfeld EM, Langenfeld J. Bone morphogenetic protein-2 stimulates angiogenesis in developing tumors. Mol. Cancer Res., 2004, 2: 141-149.

[24] Fiedler J, . BMP-2, BMP-4, and PDGF-bb stimulate chemotactic migration of primary human mesenchymal progenitor cells. J. Cell Biochem., 2002, 87: 305-312.

[25] Shi C, . BMP signaling in regulating mesenchymal stem cells in incisor homeostasis. J. Dent. Res., 2019, 98: 904-911.

[26] Atalayin C, . Medium modification with bone morphogenetic protein 2 addition for odontogenic differentiation. Braz. Oral. Res., 2016, 30: 0020.

[27] Tannoury CA, An HS. Complications with the use of bone morphogenetic protein 2 (BMP-2) in spine surgery. Spine J., 2014, 14: 552-559.

[28] James AW, . A review of the clinical side effects of bone morphogenetic protein-2. Tissue Eng. Part B Rev., 2016, 22: 284-297.

[29] Saburi E, . Bone morphogenetic protein-7 incorporated polycaprolactone scaffold has a great potential to improve survival and proliferation rate of the human embryonic kidney cells. J. Cell Biochem., 2019, 120: 9859-9868.

[30] Malik Z, . Mesenchymal Bmp7 controls onset of tooth mineralization: a novel way to regulate molar cusp shape. Front. Physiol., 2020, 11: 698.

[31] Akiyama I, . Bone morphogenetic protein 7 increased vascular endothelial growth factor (VEGF)-a expression in human granulosa cells and VEGF receptor expression in endothelial cells. Reprod. Sci., 2014, 21: 477-482.

[32] Liang C, Liao L, Tian W. Stem cell-based dental pulp regeneration: insights from signaling pathways. Stem Cell Rev. Rep., 2021, 17: 1251-1263.

[33] Patel M, . Injectable thermogel for 3D culture of stem cells. Biomaterials, 2018, 159: 91-107.

[34] Gong T, . Current advance and future prospects of tissue engineering approach to dentin/pulp regenerative therapy. Stem Cells Int., 2016, 2016: 9204574.

[35] Rosa V, . Dental pulp tissue engineering in full-length human root canals. J. Dent. Res., 2013, 92: 970-975.

[36] Xu X, . Adipose tissue-derived microvascular fragments as vascularization units for dental pulp regeneration. J. Endod., 2021, 47: 1092-1100.

[37] Monteiro N, . Dental cell sheet biomimetic tooth bud model. Biomaterials, 2016, 106: 167-179.

[38] Khayat A, . GelMA-encapsulated hDPSCs and HUVECs for dental pulp regeneration. J. Dent. Res., 2017, 96: 192-199.

[39] Fukushima KA, . Screening of hydrogel-based scaffolds for dental pulp regeneration—a systematic review. Arch. Oral. Biol., 2019, 98: 182-194.

[40] Zhao H, . Secretion of shh by a neurovascular bundle niche supports mesenchymal stem cell homeostasis in the adult mouse incisor. Cell Stem Cell, 2014, 14: 160-173.

[41] Dissanayaka WL, Zhang C. The role of vasculature engineering in dental pulp regeneration. J. Endod., 2017, 43(9S): S102-S106.

[42] Zhang S, . Cotransplantation of human umbilical cord mesenchymal stem cells and endothelial cells for angiogenesis and pulp regeneration in vivo. Life Sci., 2020, 255: 117763.

[43] Saghiri MA, . Role of angiogenesis in endodontics: contributions of stem cells and proangiogenic and antiangiogenic factors to dental pulp regeneration. J. Endod., 2015, 41: 797-803.

[44] Widbiller M, . Cell homing for pulp tissue engineering with endogenous dentin matrix proteins. J. Endod., 2018, 44: 956-962.e2.

[45] Kim JY, . Regeneration of dental-pulp-like tissue by chemotaxis-induced cell homing. Tissue Eng. Part A, 2010, 16: 3023-3031.

[46] Zhai Q, . Dental stem cell and dental tissue regeneration. Front. Med., 2019, 13: 152-159.

[47] Li S, Pan Y. Differential expression of transforming growth factor-beta1, connective tissue growth factor, phosphorylated-SMAD2/3 and phosphorylated-ERK1/2 during mouse tooth development. J. Mol. Histol., 2017, 48: 347-355.

[48] Hu CC, . Reparative dentin formation in rat molars after direct pulp capping with growth factors. J. Endod., 1998, 24: 744-751.

[49] Zhang Y, . Effects of transforming growth factor-beta1 on odontoblastic differentiation in dental papilla cells is determined by IPO7 expression level. Biochem. Biophys. Res. Commun., 2021, 545: 105-111.

[50] Maeda S, . Endogenous TGF-beta signaling suppresses maturation of osteoblastic mesenchymal cells. EMBO J., 2004, 23: 552-563.

[51] Chang HH, . Regulation of the regenerative activity of dental pulp stem cells from exfoliated deciduous teeth (SHED) of children by TGF-beta1 is associated with ALK5/Smad2, TAK1, p38 and MEK/ERK signaling. Aging (Albany NY), 2020, 12: 21253-21272.

[52] Lin PS, . Transforming growth factor beta1 down-regulates Runx-2 and alkaline phosphatase activity of human dental pulp cells via ALK5/Smad2/3 signaling. Oral. Surg. Oral. Med Oral. Pathol. Oral. Radio. Endod., 2011, 111: 394-400.

[53] Huang GT, Gronthos S, Shi S. Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J. Dent. Res., 2009, 88: 792-806.

[54] Zhang L, . Transcriptomic landscape regulated by the 14 types of bone morphogenetic proteins (BMPs) in lineage commitment and differentiation of mesenchymal stem cells (MSCs). Genes Dis., 2019, 6: 258-275.

[55] Phimphilai M, . BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J. Bone Min. Res., 2006, 21: 637-646.

[56] Toth, F., Tozser, J. & Hegedus, C. Effect of inducible BMP-7 expression on the osteogenic differentiation of human dental pulp stem cells. Int. J. Mol. Sci. 22, 6182, https://doi.org/10.3390/ijms22126182 (2021).

[57] Qin W, . Dental pulp cells that express adeno-associated virus serotype 2-mediated BMP-7 gene enhanced odontoblastic differentiation. Dent. Mater. J., 2014, 33: 656-662.

[58] Bai M, . Microenvironmental stiffness regulates dental papilla cell differentiation: implications for the importance of fibronectin-paxillin-beta-catenin axis. ACS Appl Mater. Interfaces, 2018, 10: 26917-26927.

[59] Alarmo EL, . BMP7 influences proliferation, migration, and invasion of breast cancer cells. Cancer Lett., 2009, 275: 35-43.

[60] Hu M, . BMP signaling pathways affect differently migration and invasion of esophageal squamous cancer cells. Int J. Oncol., 2017, 50: 193-202.

[61] Wang, T. C., Luo, S. J. & Chang, S. F. Bone morphogenetic protein 7 effect on human glioblastoma cell transmigration and migration. Life (Basel) 11, 708, https://doi.org/10.3390/life11070708 (2021).

[62] Perron, J.C. et al. Chemotropic signaling by BMP7 requires selective interaction at a key residue in ActRIIA. Biol. Open 8, bio042283, https://doi.org/10.1242/bio.042283 (2019).

[63] David L, Feige JJ, Bailly S. Emerging role of bone morphogenetic proteins in angiogenesis. Cytokine Growth Factor Rev., 2009, 20: 203-212.

[64] Yeh LC, Lee JC. Osteogenic protein-1 increases gene expression of vascular endothelial growth factor in primary cultures of fetal rat calvaria cells. Mol. Cell Endocrinol., 1999, 153: 113-124.

[65] Gronthos S, . Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc. Natl Acad. Sci. USA, 2000, 97: 13625-13630.

[66] Li R, . Human treated dentin matrix as a natural scaffold for complete human dentin tissue regeneration. Biomaterials, 2011, 32: 4525-4538.

[67] Chrepa V, . Survival of the apical papilla and its resident stem cells in a case of advanced pulpal necrosis and apical periodontitis. J. Endod., 2017, 43: 561-567.

[68] Estrela C, . Mesenchymal stem cell marker expression in periapical abscess. J. Endod., 2019, 45: 716-723.

[69] Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J. Bone Min. Res., 2003, 18: 696-704.

[70] Xu F, . The potential application of concentrated growth factor in pulp regeneration: an in vitro and in vivo study. Stem Cell Res. Ther., 2019, 10: 134.

Funding

the Nature Science Foundation of China (82071092), the Fundamental Research Funds for the Central Universities (YJ201878), Key Project of Sichuan province (2019YFS0311, 2019YFS0515), Technology Innovation Research and Development Project of Chengdu (2019-YF05-00705-SN).

the National Key Research and Development Program of China (2017YFA0104800)