Advances in Clinical and Experimental Medicine
2014, vol. 23, nr 3, May-June, p. 463–467
Publication type: original article
Language: English
Effects of the Combination of Fibroblast Growth Factor-2 and Bone Morphogenetic Protein-2 on the Proliferation and Differentiation of Osteoprecursor Cells
1 Department of Periodontics, Seoul St Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea
Abstract
Objectives. Fibroblast growth factor (FGF) plays a critical role in bone growth; FGF-2 is known to be an important regulator of osteoblast activity because it stimulates osteoblast replication and decreases differentiation markers. Bone morphogenetic protein-2 (BMP-2) has been shown to be an active inducer of osteoblast differentiation and stimulates expression of mineralization-associated genes.
Material and Methods. The dose-dependent impact of FGF-2 and BMP-2 on the cellular proliferation and differentiation of osteoprecursor cells was evalauted. The alkaline phosphatase activity (ALP) test was performed to assess differentiation, and protein expressions related to bone formation were measured using the Western blot analysis.
Results. Cultures grown in the presence of FGF at 20 ng/mL showed significantly increased value when compared with control group and cultures loaded with FGF-2 at 20 ng/mL, and BMP-2 at 100 ng/mL showed significant decrease in cellular proliferation when compared with cultures loaded with FGF-2 at 20 ng/mL. The ALP activity increased when cells were treated with 10 and 100 ng/mL BMP-2, with relative ALP activity of 213.1% and 312.5%, respectively, when ALP activity of the uncontrolled control was considered 100%. However, when 100 ng/mL BMP-2 was combined with 20 ng/mL FGF-2, the relative increase reached up to 392.2%, but this did not reach a statistically significant increase when compared with 100 ng/mL BMP-2 alone.
Conclusion. Within the limits of this study, BMP-2 significantly enhanced osteoblast differentiation but combined delivery of FGF-2 and BMP-2 did not produce synergistic effects on osteoblast differentiation under the current experimental condition.
Key words
bone morphogenetic protein 2, differentiation, fibroblast growth factor 2, osteoblast, proliferation.
References (20)
- Mansukhani A, Bellosta P, Sahni M, Basilico C: Signaling by fibroblast growth factors (FGF) and fibroblast growth factor receptor 2 (FGFR2)-activating mutations blocks mineralization and induces apoptosis in osteoblasts. J Cell Biol 2000, 149, 1297–1308.
- Xiao G, Jiang D, Gopalakrishnan R, Franceschi RT: Fibroblast growth factor 2 induction of the osteocalcin gene requires MAPK activity and phosphorylation of the osteoblast transcription factor, Cbfa1/Runx2. J Biol Chem 2002, 277, 36181–36187.
- Sobue T, Naganawa T, Xiao L, Okada Y, Tanaka Y, Ito M, Okimoto N, Nakamura T, Coffin JD, Hurley MM: Over-expression of fibroblast growth factor-2 causes defective bone mineralization and osteopenia in transgenic mice. J Cell Biochem 2005, 95, 83–94.
- Kuhn LT, Ou G, Charles L, Hurley MM, Rodner CM, Gronowicz G: Fibroblast Growth Factor-2 and Bone Morphogenetic Protein-2 Have a Synergistic Stimulatory Effect on Bone Formation in Cell Cultures From Elderly Mouse and Human Bone. J Gerontol A Biol Sci Med Sci 2013.
- Hughes-Fulford M, Li CF: The role of FGF-2 and BMP-2 in regulation of gene induction, cell proliferation and mineralization. J Orthop Surg Res 2011, 6, 8.
- Park JB, Lee JY, Park HN, Seol YJ, Park YJ, Rhee SH, Lee SC, Kim KH, Kim TI, Lee YM, Ku Y, Rhyu IC, Han SB, Chung CP: Osteopromotion with synthetic oligopeptide-coated bovine bone mineral in vivo. J Periodontol 2007, 78, 157–163.
- Maegawa N, Kawamura K, Hirose M, Yajima H, Takakura Y, Ohgushi H: Enhancement of osteoblastic differentiation of mesenchymal stromal cells cultured by selective combination of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-2 (FGF-2). J Tissue Eng Regen Med 2007, 1, 306–313.
- Tanaka E, Ishino Y, Sasaki A, Hasegawa T, Watanabe M, Dalla-Bona DA, Yamano E, van Eijden TM, Tanne K: Fibroblast growth factor-2 augments recombinant human bone morphogenetic protein-2-induced osteoinductive activity. Ann Biomed Eng 2006, 34, 717–725.
- Wang H, Zou Q, Boerman OC, Nijhuis AW, Jansen JA, Li Y, Leeuwenburgh SC: Combined delivery of BMP-2 and bFGF from nanostructured colloidal gelatin gels and its effect on bone regeneration in vivo. J Control Release 2012, 166, 172–181.
- Park JB, Zhang H, Lin CY, Chung CP, Byun Y, Park YS, Yang VC: Simvastatin Maintains Osteoblastic Viability While Promoting Differentiation by Partially Regulating the Expressions of Estrogen Receptors alpha. J Surg Res 2012, 174, 278–283.
- Park JB: The effects of dexamethasone, ascorbic acid, and beta-glycerophosphate on osteoblastic differentiation by regulating estrogen receptor and osteopontin expression. J Surg Res 2012, 173, 99–104.
- Park JB: Combination of simvastatin and bone morphogenetic protein-2 enhances differentiation of osteoblastic cells by regulating the expressions of phospho-Smad1/5/8. Exp Ther Med 2012, 4, 303–306.
- Park JB: Effects of fibroblast growth factor 2 on osteoblastic proliferation and differentiation by regulating bone morphogenetic protein receptor expression. J Craniofac Surg 2011, 22, 1880–1882.
- Park JB: Effects of doxycycline, minocycline, and tetracycline on cell proliferation, differentiation, and protein expression in osteoprecursor cells. J Craniofac Surg 2011, 22, 1839–1842.
- Meleti Z, Shapiro IM, Adams CS: Inorganic phosphate induces apoptosis of osteoblast-like cells in culture. Bone 2000, 27, 359–366.
- Moursi AM, Winnard PL, Winnard AV, Rubenstrunk JM, Mooney MP: Fibroblast growth factor 2 induces increased calvarial osteoblast proliferation and cranial suture fusion. Cleft Palate Craniofac J 2002, 39, 487–496.
- Nakamura Y, Tensho K, Nakaya H, Nawata M, Okabe T, Wakitani S: Low dose fibroblast growth factor-2 (FGF-2) enhances bone morphogenetic protein-2 (BMP-2)-induced ectopic bone formation in mice. Bone 2005, 36, 399–407.
- Park JB: Effects of low dose of estrone on the proliferation, differentiation, and mineralization of the osteoprecursor cells. Exp Ther Med 2012, 4, 681–684.
- Huang W, Carlsen B, Rudkin G, Berry M, Ishida K, Yamaguchi DT, Miller TA: Osteopontin is a negative regulator of proliferation and differentiation in MC3T3-E1 pre-osteoblastic cells. Bone 2004, 34, 799–808.
- Quarles LD, Yohay DA, Lever LW, Caton R, Wenstrup RJ: Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner Res 1992, 7, 683–692.


