Odontogenic differentiation of vascular endothelial growth factor‑transfected human dental pulp stem cells in vitro

Zhang,Wen; Liu,Wei; Ling,Junqi; Lin,Zhengmei; Gao,Yan; Mao,Xueli; Jian,Yutao

doi:10.3892/mmr.2014.2481

Odontogenic differentiation of vascular endothelial growth factor‑transfected human dental pulp stem cells in vitro

Authors:
- Wen Zhang
- Wei Liu
- Junqi Ling
- Zhengmei Lin
- Yan Gao
- Xueli Mao
- Yutao Jian
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Affiliations: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Province Key Laboratory of Stomatology, Sun Yat‑sen University, Guangzhou, Guangdong 510055, P.R. China
Published online on: August 11, 2014 https://doi.org/10.3892/mmr.2014.2481
Pages: 1899-1906

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Abstract

Dental pulp stem cells (DPSCs) can be induced towards odontogenic differentiation. Previous studies have shown that vascular endothelial growth factor (VEGF) is able to induce the osteogenic differentiation of cells, but the effectiveness of VEGF in the odontogenic differentiation of DPSCs remains unclear. This study aimed to investigate the effects of lentivirus‑mediated human VEGF gene transfection on the proliferation and odontogenic differentiation of human DPSCs in vitro. DPSCs were transfected with either lentiviral pCDH‑CMV‑MCS‑EFI‑copGFP (pCDH) vector or recombinant pCDH‑VEGF vector, and the growth characteristics of the resulting DPSCs/Vector and DPSCs/VEGF were subsequently assessed. The odontogenic differentiation genes of the two groups of cells, including alkaline phosphatase, osteocalcin, dentin sialophosphoprotein and dentin matrix protein 1 (DMP1), were evaluated by quantitative polymerase chain reaction (qPCR). The specific proteins of odontogenic differentiation, including dentin sialoprotein and DMP1, were analyzed by western blotting. DPSCs/VEGF showed similar proliferation characteristics to DPSCs/Vector during the observation period. qPCR results showed that the relative VEGF gene expression was significantly higher in DPSCs/VEGF than that in DPSCs/Vector two days after transfection (P<0.01). Similarly, western blot analysis showed that the protein expression levels of VEGF were higher in DPSCs/VEGF than those in DPSCs/Vector. On the first, fourth, eighth and 16th days after lentivirus‑mediated transfection, the expression of odontogenic differentiation‑specific genes and proteins was higher in DPSCs/VEGF than that in DPSCs/Vector. These results indicated that lentivirus‑mediated VEGF gene transfection promoted the odontogenic differentiation of human DPSCs in vitro.

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1	Gronthos S, Mankani M, Brahim J, Robey PG and Shi S: Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci USA. 97:13625–13630. 2000. View Article : Google Scholar : PubMed/NCBI
2	Kawashima N: Characterisation of dental pulp stem cells: a new horizon for tissue regeneration? Arch Oral Biol. 57:1439–1458. 2012. View Article : Google Scholar : PubMed/NCBI
3	Rodríguez-Lozano FJ, Bueno C, Insausti CL, et al: Mesenchymal stem cells derived from dental tissues. Int Endod J. 44:800–806. 2011.PubMed/NCBI
4	Huang GT, Gronthos S and Shi S: 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. 2009.PubMed/NCBI
5	Yang X, van der Kraan PM, van den Dolder J, et al: STRO-1 selected rat dental pulp stem cells transfected with adenoviral-mediated human bone morphogenetic protein 2 gene show enhanced odontogenic differentiation. Tissue Eng. 13:2803–2812. 2007. View Article : Google Scholar
6	Matsushita K, Motani R, Sakuta T, et al: The role of vascular endothelial growth factor in human dental pulp cells: induction of chemotaxis, proliferation, and differentiation and activation of the AP-1-dependent signaling pathway. J Dent Res. 79:1596–1603. 2000. View Article : Google Scholar
7	Tan YY, Yang YQ, Chai L, Wong RW and Rabie AB: Effects of vascular endothelial growth factor (VEGF) on MC3T3-E1. Orthod Craniofac Res. 13:223–228. 2010. View Article : Google Scholar : PubMed/NCBI
8	Lee JH, Um S, Jang JH and Seo BM: Effects of VEGF and FGF-2 on proliferation and differentiation of human periodontal ligament stem cells. Cell Tissue Res. 348:475–484. 2012. View Article : Google Scholar : PubMed/NCBI
9	D’Alimonte I, Nargi E, Mastrangelo F, et al: Vascular endothelial growth factor enhances in vitro proliferation and osteogenic differentiation of human dental pulp stem cells. J Biol Regul Homeost Agents. 25:57–69. 2011.
10	Wozney JM and Rosen V: Bone morphogenetic protein and bone morphogenetic protein gene family in bone formation and repair. Clin Orthop Relat Res. 26–37. 1998.PubMed/NCBI
11	Samee M, Kasugai S, Kondo H, Ohya K, Shimokawa H and Kuroda S: Bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) transfection to human periosteal cells enhances osteoblast differentiation and bone formation. J Pharmacol Sci. 108:18–31. 2008. View Article : Google Scholar
12	Liu B, Li X, Liang G and Liu X: VEGF expression in mesenchymal stem cells promotes bone formation of tissue-engineered bones. Mol Med Rep. 4:1121–1126. 2011.PubMed/NCBI
13	Jiang J, Fan CY and Zeng BF: Osteogenic differentiation effects on rat bone marrow-derived mesenchymal stromal cells by lentivirus-mediated co-transfection of human BMP2 gene and VEGF165 gene. Biotechnol Lett. 30:197–203. 2008. View Article : Google Scholar
14	Bakopoulou A, Leyhausen G, Volk J, et al: Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells (DPSCs) and stem cells from the apical papilla (SCAP). Arch Oral Biol. 56:709–721. 2011. View Article : Google Scholar
15	Lahmy R, Soleimani M, Sanati MH, Behmanesh M, Kouhkan F and Mobarra N: Pancreatic islet differentiation of human embryonic stem cells by microRNA overexpression. J Tissue Eng Regen Med. Jul 30–2013.(Epud ahead of print).
16	Hwang SY, Foley J, Numaga-Tomita T, Petranka JG, Bird GS and Putney JW Jr: Deletion of Orai1 alters expression of multiple genes during osteoclast and osteoblast maturation. Cell Calcium. 52:488–500. 2012. View Article : Google Scholar
17	Pereira LO, Rubini MR, Silva JR, et al: Comparison of stem cell properties of cells isolated from normal and inflamed dental pulps. Int Endod J. 45:1080–1090. 2012. View Article : Google Scholar : PubMed/NCBI
18	Shi S and Gronthos S: Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res. 18:696–704. 2003. View Article : Google Scholar : PubMed/NCBI
19	Jiang L, Peng WW, Li LF, Yang Y and Zhu YQ: Isolation and identification of CXCR4-positive cells from human dental pulp cells. J Endod. 38:791–795. 2012. View Article : Google Scholar : PubMed/NCBI
20	Lee UL, Jeon SH, Park JY and Choung PH: Effect of platelet-rich plasma on dental stem cells derived from human impacted third molars. Regen Med. 6:67–79. 2011. View Article : Google Scholar : PubMed/NCBI
21	Trubiani O, Tripodi D, Delle Fratte T, Caputi S and Di Primio R: Human dental pulp vasculogenesis evaluated by CD34 antigen expression and morphological arrangement. J Dent Res. 82:742–747. 2003. View Article : Google Scholar : PubMed/NCBI
22	Hata R and Senoo H: L-ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts. J Cell Physiol. 138:8–16. 1989. View Article : Google Scholar : PubMed/NCBI
23	Hitomi K, Torii Y and Tsukagoshi N: Increase in the activity of alkaline phosphatase by L-ascorbic acid 2-phosphate in a human osteoblast cell line, HuO-3N1. J Nutr Sci Vitaminol (Tokyo). 38:535–544. 1992. View Article : Google Scholar : PubMed/NCBI
24	Torii Y, Hitomi K and Tsukagoshi N: L-ascorbic acid 2-phosphate promotes osteoblastic differentiation of MC3T3-E1 mediated by accumulation of type I collagen. J Nutr Sci Vitaminol (Tokyo). 40:229–238. 1994. View Article : Google Scholar : PubMed/NCBI
25	Shima N, Kimoto M, Yamaguchi M and Yamagami S: Increased proliferation and replicative lifespan of isolated human corneal endothelial cells with L-ascorbic acid 2-phosphate. Invest Ophthalmol Vis Sci. 52:8711–8717. 2011. View Article : Google Scholar
26	MacDougall M, Gu TT and Simmons D: Dentin matrix protein-1, a candidate gene for dentinogenesis imperfecta. Connect Tissue Res. 35:267–272. 1996. View Article : Google Scholar : PubMed/NCBI
27	Ritchie HH, Berry JE, Somerman MJ, et al: Dentin sialoprotein (DSP) transcripts: developmentally-sustained expression in odontoblasts and transient expression in pre-ameloblasts. Eur J Oral Sci. 105:405–413. 1997. View Article : Google Scholar : PubMed/NCBI
28	Lee SY, Kim SY, Park SH, Kim JJ, Jang JH and Kim EC: Effects of recombinant dentin sialoprotein in dental pulp cells. J Dent Res. 91:407–412. 2012. View Article : Google Scholar : PubMed/NCBI
29	Suzuki S, Sreenath T, Haruyama N, et al: Dentin sialoprotein and dentin phosphoprotein have distinct roles in dentin mineralization. Matrix Biol. 28:221–229. 2009. View Article : Google Scholar : PubMed/NCBI