Pulp-dentin regeneration in the apical region of immature permanent teeth represents a significant clinical challenge. Tissue engineering approaches using bioactive molecules and scaffolds may have the potential to regenerate the natural apical structure of these teeth, representing a superior alternative to existing treatment regimens. The aims of this study are (i) to evaluate the VitroGel 3D system, an animal origin-free polysaccharide hydrogel, as a possible injectable scaffold for pulp-dentin regeneration and (ii) to investigate the effects of stromal cell-derived factor-1α (SDF-1α) and bone morphogenetic protein-2(BMP-2) cotreatment on odontogenic differentiation of human stem cells from apical papilla (SCAP) cultured in the VitroGel 3D system. The morphology, viability and proliferation of SCAP cultured in the VitroGel 3D system were measured via scanning electron microscopy (SEM), live and dead cell staining and CCK-8 assays. Alkaline phosphatase (ALP) activity, real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR) and Western blot analysis were further used to evaluate the odontogenic differentiation of SCAP cultured in the VitroGel 3D system in vitro. Finally, the odontogenic differentiation was assessed in vivo through ectopic subcutaneous injection. The results showed that SCAP cultured in 3D hydrogel demonstrated favorable viability and proliferation. SDF-1α and BMP-2 cotreatment enhanced odontogenic differentiation-related gene and protein expression in vitro and promoted odontogenic differentiation of SCAP in vivo. In conclusion, the present study demonstrated that the VitroGel 3D system promoted SCAP proliferation and differentiation. Moreover, SDF-1α cotreatment had synergistic effects on BMP-2-induced odontogenic differentiation of human SCAP cultured in the VitroGel 3D system both in vitro and in vivo.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Bessa PC, Casal M, Reis RL (2008) Bone morphogenetic proteins in tissue engineering: the road from laboratory to clinic, part II (BMP delivery). J Tissue Eng Regen Med 2:81–96
Bruderer M, Richards RG, Alini M, Stoddart MJ (2014) Role and regulation of RUNX2 in osteogenesis. Eur Cells Mater 28:269–286
Carmen L, Asuncion M, Beatriz S, Rosa YV (2017) Revascularization in immature permanent teeth with necrotic pulp and apical pathology: case series. Case Rep Dent 2017:3540159
Cavalcanti BN, Zeitlin BD, Nor JE (2013) A hydrogel scaffold that maintains viability and supports differentiation of dental pulp stem cells. Dent Mater 29:97–102
Chen YP, Jovani-Sancho Mdel M, Sheth CC (2015) Is revascularization of immature permanent teeth an effective and reproducible technique? Dental traumatology : official publication of international association for. Dent Traumatol 31:429–436
Chrepa V, Austah O, Diogenes A (2017) Evaluation of a commercially available hyaluronic acid hydrogel (Restylane) as injectable scaffold for dental pulp regeneration: an in vitro evaluation. J Endod 43:257–262
De-Colle C, Monnich D, Welz S, Boeke S, Sipos B, Fend F, Mauz PS, Tinhofer I, Budach V, Jawad JA, Stuschke M, Balermpas P, Rodel C, Grosu AL, Abdollahi A, Debus J, Bayer C, Belka C, Pigorsch S, Combs SE, Lohaus F, Linge A, Krause M, Baumann M, Zips D, Menegakis A (2017) SDF-1/CXCR4 expression in head and neck cancer and outcome after postoperative radiochemotherapy. Clin Transl Radiat Oncol 5:28–36
Dissanayaka WL, Hargreaves KM, Jin L, Samaranayake LP, Zhang C (2015) The interplay of dental pulp stem cells and endothelial cells in an injectable peptide hydrogel on angiogenesis and pulp regeneration in vivo. Tissue Eng A 21:550–563
Edmondson R, Broglie JJ, Adcock AF, Yang L (2014) Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol 12:207–218
Ferreira SA, Faull PA, Seymour AJ, Yu TTL, Loaiza S, Auner HW, Snijders AP, Gentleman E (2018) Neighboring cells override 3D hydrogel matrix cues to drive human MSC quiescence. Biomaterials 176:13–23
Garzon I, Martin-Piedra MA, Carriel V, Alaminos M, Liu X, D'Souza RN (2018) Bioactive injectable aggregates with nanofibrous microspheres and human dental pulp stem cells: a translational strategy in dental endodontics. J Tissue Eng Regen Med 12:204–216
Gazitt Y (2004) Homing and mobilization of hematopoietic stem cells and hematopoietic cancer cells are mirror image processes, utilizing similar signaling pathways and occurring concurrently: circulating cancer cells constitute an ideal target for concurrent treatment with chemotherapy and antilineage-specific antibodies. Leukemia 18:1–10
Gibson MP, Zhu Q, Wang S, Liu Q, Liu Y, Wang X, Yuan B, Ruest LB, Feng JQ, D'Souza RN, Qin C, Lu Y (2013) The rescue of dentin matrix protein 1 (DMP1)-deficient tooth defects by the transgenic expression of dentin sialophosphoprotein (DSPP) indicates that DSPP is a downstream effector molecule of DMP1 in dentinogenesis. J Biol Chem 288:7204–7214
Gu S, Liang J, Wang J, Liu B (2013) Histone acetylation regulates osteodifferentiation of human dental pulp stem cells via DSPP. Front Biosci (Landmark Ed) 18:1072–1079
Harris H (1990) The human alkaline phosphatases: what we know and what we don’t know. Clin Chim Acta 186:133–150
Hosogane N, Huang Z, Rawlins BA, Liu X, Boachie-Adjei O, Boskey AL, Zhu W (2010) Stromal derived factor-1 regulates bone morphogenetic protein 2-induced osteogenic differentiation of primary mesenchymal stem cells. Int J Biochem Cell Biol 42:1132–1141
Huang GT, Gronthos S, Shi S (2009) Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res 88:792–806
Huang L, Xiao L, Jung Poudel A, Li J, Zhou P, Gauthier M, Liu H, Wu Z, Yang G (2018) Porous chitosan microspheres as microcarriers for 3D cell culture. Carbohydr Polym 202:611–620
Jiang HW, Ling JQ, Gong QM (2008a) The expression of stromal cell-derived factor 1 (SDF-1) in inflamed human dental pulp. J Endod 34:1351–1354
Jiang L, Zhu YQ, Du R, Gu YX, Xia L, Qin F, Ritchie HH (2008b) The expression and role of stromal cell-derived factor-1alpha-CXCR4 axis in human dental pulp. J Endod 34:939–944
Kim DS, Kim YS, Bae WJ, Lee HJ, Chang SW, Kim WS, Kim EC (2014) The role of SDF-1 and CXCR4 on odontoblastic differentiation in human dental pulp cells. Int Endod J 47:534–541
Langer R, Vacanti JP (1993) Tissue engineering. Science (New York, NY) 260:920–926
Lee CH, Jin MU, Jung HM, Lee JT, Kwon TG (2015) Effect of dual treatment with SDF-1 and BMP-2 on ectopic and orthotopic bone formation. PLoS One 10:e0120051
Lin J, Zeng Q, Wei X, Zhao W, Cui M, Gu J, Lu J, Yang M, Ling J (2017) Regenerative endodontics versus apexification in immature permanent teeth with apical periodontitis: a prospective randomized controlled study. J Endod 43:1821–1827
Liu W, Lee BS, Mieler WF, Kang-Mieler JJ (2018) Biodegradable microsphere-hydrogel ocular drug delivery system for controlled and extended release of bioactive aflibercept in vitro. Curr Eye Res
Lovelace TW, Henry MA, Hargreaves KM, Diogenes A (2011) Evaluation of the delivery of mesenchymal stem cells into the root canal space of necrotic immature teeth after clinical regenerative endodontic procedure. J Endod 37:133–138
Mahauad-Fernandez WD, Okeoma CM (2018) B49, a BST-2-based peptide, inhibits adhesion and growth of breast cancer cells. Sci Rep 8:4305
Muller AS, Artner M, Janjic K, Edelmayer M, Kurzmann C, Moritz A, Agis H (2018) Synthetic clay-based hypoxia mimetic hydrogel for pulp regeneration: the impact on cell activity and release kinetics based on dental pulp-derived cells in vitro. J Endod 44:1263–1269
Nam H, Kim GH, Bae YK, Jeong DE, Joo KM, Lee K, Lee SH (2017) Angiogenic capacity of dental pulp stem cell regulated by SDF-1alpha-CXCR4 axis. Stem Cells Int 2017:8085462
Park HJ, Lee WY, Kim JH, Park C, Song H (2018) Expression patterns and role of SDF-1/CXCR4 axis in boar spermatogonial stem cells. Theriogenology 113:221–228
Razzouk S, Sarkis R (2012) BMP-2: biological challenges to its clinical use. N Y State Dent J 78:37–39
Rosa V, Zhang Z, Grande RH, Nor JE (2013) Dental pulp tissue engineering in full-length human root canals. J Dent Res 92:970–975
Seo BB, Choi H, Koh JT, Song SC (2015) Sustained BMP-2 delivery and injectable bone regeneration using thermosensitive polymeric nanoparticle hydrogel bearing dual interactions with BMP-2. J Control Release 209:67–76
Shen X, Zhang Y, Gu Y, Xu Y, Liu Y, Li B, Chen L (2016) Sequential and sustained release of SDF-1 and BMP-2 from silk fibroin-nanohydroxyapatite scaffold for the enhancement of bone regeneration. Biomaterials 106:205–216
Togel F, Isaac J, Hu Z, Weiss K, Westenfelder C (2005) Renal SDF-1 signals mobilization and homing of CXCR4-positive cells to the kidney after ischemic injury. Kidney Int 67:1772–1784
Tsesis I, Rosen E, Tamse A, Taschieri S, Kfir A (2010) Diagnosis of vertical root fractures in endodontically treated teeth based on clinical and radiographic indices: a systematic review. J Endod 36:1455–1458
Wang X, Thibodeau B, Trope M, Lin LM, Huang GT (2010) Histologic characterization of regenerated tissues in canal space after the revitalization/revascularization procedure of immature dog teeth with apical periodontitis. J Endod 36:56–63
Wang W, Dang M, Zhang Z, Hu J, Eyster TW, Ni L, Ma PX (2016) Dentin regeneration by stem cells of apical papilla on injectable nanofibrous microspheres and stimulated by controlled BMP-2 release. Acta Biomater 36:63–72
Yang G, Yuan G, MacDougall M, Zhi C, Chen S (2017) BMP-2 induced Dspp transcription is mediated by Dlx3/Osx signaling pathway in odontoblasts. Sci Rep 7:10775
Yu J, Li M, Qu Z, Yan D, Li D, Ruan Q (2010) SDF-1/CXCR4-mediated migration of transplanted bone marrow stromal cells toward areas of heart myocardial infarction through activation of PI3K/Akt. J Cardiovasc Pharmacol 55:496–505
Zaruba MM, Franz WM (2010) Role of the SDF-1-CXCR4 axis in stem cell-based therapies for ischemic cardiomyopathy. Expert Opin Biol Ther 10:321–335
Zhu W, Boachie-Adjei O, Rawlins BA, Frenkel B, Boskey AL, Ivashkiv LB, Blobel CP (2007) A novel regulatory role for stromal-derived factor-1 signaling in bone morphogenic protein-2 osteogenic differentiation of mesenchymal C2C12 cells. J Biol Chem 282:18676–18685
Zwingenberger S, Yao Z, Jacobi A, Vater C, Valladares RD, Li C, Nich C, Rao AJ, Christman JE, Antonios JK, Gibon E, Schambach A, Maetzig T, Goodman SB, Stiehler M (2014) Enhancement of BMP-2 induced bone regeneration by SDF-1alpha mediated stem cell recruitment. Tissue Eng A 20:810–818
Zwingenberger S, Langanke R, Vater C, Lee G, Niederlohmann E, Sensenschmidt M, Jacobi A, Bernhardt R, Muders M, Rammelt S, Knaack S, Gelinsky M, Gunther KP, Goodman SB, Stiehler M (2016) The effect of SDF-1alpha on low dose BMP-2 mediated bone regeneration by release from heparinized mineralized collagen type I matrix scaffolds in a murine critical size bone defect model. J Biomed Mater Res A 104:2126–2134
We appreciate Beidi Zhang for technical assistance of cell culture and animal experimental and Rong Kuang and Wei Wang for intellectual help on the project.
This study was funded by the National Natural Science Foundation of China (grant numbers 81670975, 31500786, 31600786).
Ethical conduct of research
All animal procedures were performed and experimental protocols were approved by the guidelines of the Animal Care Committee of the Fourth Military Medical University, Xi’an, China (SCXK (Military) 2007-007), which was in compliance with the NIH Guide for the Care and Use of Laboratory Animals.
Conflict of interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
About this article
Cite this article
Xiao, M., Qiu, J., Kuang, R. et al. Synergistic effects of stromal cell-derived factor-1α and bone morphogenetic protein-2 treatment on odontogenic differentiation of human stem cells from apical papilla cultured in the VitroGel 3D system. Cell Tissue Res 378, 207–220 (2019). https://doi.org/10.1007/s00441-019-03045-3
- Stem cells from apical papilla
- Stromal cell-derived factor-1α
- Bone morphogenetic protein-2
- Odontoblastic differentiation