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Published in: BMC Musculoskeletal Disorders 1/2011

Open Access 01-12-2011 | Research article

Cooperative effects in differentiation and proliferation between PDGF-BB and matrix derived synthetic peptides in human osteoblasts

Authors: Thomas Vordemvenne, Jürgen RJ Paletta, Rene Hartensuer, Thomas Pap, Michael J Raschke, Sabine Ochman

Published in: BMC Musculoskeletal Disorders | Issue 1/2011

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Abstract

Background

Enhancing osteogenic capabilities of bone matrix for the treatment of fractures and segmental defects using growth factors is an active area of research. Recently, synthetic peptides like AC- 100, TP508 or p-15 corresponding to biologically active sequences of matrix proteins have been proven to stimulate bone formation. The platelet-derived growth factor (PDGF) BB has been identified as an important paracrine factor in early bone healing. We hypothesized that the combined use of PDGF-BB with synthetic peptides could result in an increase in proliferation and calcification of osteoblast-like cells.

Methods

Osteoblast-like cell cultures were treated with PDGF and synthetic peptides, singly and as combinations, and compared to non-treated control cell cultures. The cultures were evaluated at days 2, 5, and 10 in terms of cell proliferation, calcification and gene expression of alkaline phosphate, collagen I and osteocalcin.

Results

Experimental findings revealed that the addition of PDGF, p-15 and TP508 and combinations of PDGF/AC-100, PDGF/p-15 and PDGF/TP508 resulted in an increase in proliferating osteoblasts, especially in the first 5 days of cultivation. Proliferation did not significantly differ between single factors and factor combinations (p > 0.05). The onset of calcification in osteoblasts occurred earlier and was more distinct compared to the corresponding control or PDGF stimulation alone. Significant difference was found for the combined use of PDGF/p-15 and PDGF/AC-100 (p < 0.05).

Conclusions

Our findings indicate that PDGF exhibits cooperative effects with synthetic peptides in differentiation and proliferation. These cooperative effects cause a significant early calcification of osteoblast-like cells (p < 0.05). We suggest the combination of synthetic peptides and PDGF as a potential clinical approach for accelerating bone healing or coating osteosynthesis materials.
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Literature
1.
go back to reference Beck LS, Wong RL, DeGuzman L, Lee WP, Ongpipattanakul B, Nguyen TH: Combination of bone marrow and TGF-beta1 augment the healing of critical-sized bone defects. J Pharm Sci. 1998, 87 (11): 1379-1386.CrossRefPubMed Beck LS, Wong RL, DeGuzman L, Lee WP, Ongpipattanakul B, Nguyen TH: Combination of bone marrow and TGF-beta1 augment the healing of critical-sized bone defects. J Pharm Sci. 1998, 87 (11): 1379-1386.CrossRefPubMed
2.
go back to reference Bostrom M, Lane JM, Tomin E, Browne M, Berberian W, Turek T, Smith J, Wozney J, Schildhauer T: Use of bone morphogenetic protein-2 in the rabbit ulnar nonunion model. Clin Orthop Relat Res. 1996, 272-282. 327 Bostrom M, Lane JM, Tomin E, Browne M, Berberian W, Turek T, Smith J, Wozney J, Schildhauer T: Use of bone morphogenetic protein-2 in the rabbit ulnar nonunion model. Clin Orthop Relat Res. 1996, 272-282. 327
3.
go back to reference Laflamme C, Rouabhia M: Effect of BMP-2 and BMP-7 homodimers and a mixture of BMP-2/BMP-7 homodimers on osteoblast adhesion and growth following culture on a collagen scaffold. Biomed Mater. 2008, 3 (1): 015008-CrossRefPubMed Laflamme C, Rouabhia M: Effect of BMP-2 and BMP-7 homodimers and a mixture of BMP-2/BMP-7 homodimers on osteoblast adhesion and growth following culture on a collagen scaffold. Biomed Mater. 2008, 3 (1): 015008-CrossRefPubMed
4.
go back to reference Liao J-C, Tzeng S-T, Keorochana G, Lee K-B, Johnson JS, Morishita Y, Murray SS, Wang JC: Enhancement of recombinant human BMP-7 bone formation with bmp binding peptide in a rodent femoral defect model. J Orthop Res. 2011, 29 (5): 753-759.CrossRefPubMed Liao J-C, Tzeng S-T, Keorochana G, Lee K-B, Johnson JS, Morishita Y, Murray SS, Wang JC: Enhancement of recombinant human BMP-7 bone formation with bmp binding peptide in a rodent femoral defect model. J Orthop Res. 2011, 29 (5): 753-759.CrossRefPubMed
6.
go back to reference Ranly D, McMillan J, Keller T, Lohmann C, Meunch T, Cochran D, Schwartz Z, Boyan B: Platelet-derived growth factor inhibits demineralized bone matrix-induced intramuscular cartilage and bone formation - A study of immunocompromised mice. J Bone Joint Surg Am. 2005, 87A (9): 2052-2064.CrossRef Ranly D, McMillan J, Keller T, Lohmann C, Meunch T, Cochran D, Schwartz Z, Boyan B: Platelet-derived growth factor inhibits demineralized bone matrix-induced intramuscular cartilage and bone formation - A study of immunocompromised mice. J Bone Joint Surg Am. 2005, 87A (9): 2052-2064.CrossRef
7.
go back to reference Vanhaesebroeck B, Claesson-Welsh L: The PI 3-kinase isoforms p110(alpha) and p110(beta) have differential roles in PDGF- and insulin-mediated signaling. J Cell Sci. 2000, 113 (Pt 2): 207-14.PubMed Vanhaesebroeck B, Claesson-Welsh L: The PI 3-kinase isoforms p110(alpha) and p110(beta) have differential roles in PDGF- and insulin-mediated signaling. J Cell Sci. 2000, 113 (Pt 2): 207-14.PubMed
8.
go back to reference Barnes GL, Kostenuik PJ, Gerstenfeld LC, Einhorn TA: Growth Factor Regulation of Fracture Repair. J Bone Miner Res. 1999, 14 (11): 1805-1815.CrossRefPubMed Barnes GL, Kostenuik PJ, Gerstenfeld LC, Einhorn TA: Growth Factor Regulation of Fracture Repair. J Bone Miner Res. 1999, 14 (11): 1805-1815.CrossRefPubMed
9.
go back to reference Mohan S: Bone growth factors. Clin Orthop Relat Res. 1991, 30-48. 263 Mohan S: Bone growth factors. Clin Orthop Relat Res. 1991, 30-48. 263
10.
go back to reference Siegbahn A, Hammacher A, Westermark B, Heldin CH: Differential effects of the various isoforms of platelet-derived growth factor on chemotaxis of fibroblasts, monocytes, and granulocytes. J Clin Invest. 1990, 85 (3): 916-920.CrossRefPubMedPubMedCentral Siegbahn A, Hammacher A, Westermark B, Heldin CH: Differential effects of the various isoforms of platelet-derived growth factor on chemotaxis of fibroblasts, monocytes, and granulocytes. J Clin Invest. 1990, 85 (3): 916-920.CrossRefPubMedPubMedCentral
11.
go back to reference Park YJ, Lee YM, Lee JY, Seol YJ, Chung CP, Lee SJ: Controlled release of platelet-derived growth factor-BB from chondroitin sulfate-chitosan sponge for guided bone regeneration. J Control Release. 2000, 67 (2-3): 385-394.CrossRefPubMed Park YJ, Lee YM, Lee JY, Seol YJ, Chung CP, Lee SJ: Controlled release of platelet-derived growth factor-BB from chondroitin sulfate-chitosan sponge for guided bone regeneration. J Control Release. 2000, 67 (2-3): 385-394.CrossRefPubMed
12.
go back to reference Walsh WR, Kim HD, Jong YS, Valentini RF: Controlled release of platelet-derived growth factor using ethylene vinyl acetate copolymer (EVAc) coated on stainless-steel wires. Biomaterials. 1995, 16 (17): 1319-1325.CrossRefPubMed Walsh WR, Kim HD, Jong YS, Valentini RF: Controlled release of platelet-derived growth factor using ethylene vinyl acetate copolymer (EVAc) coated on stainless-steel wires. Biomaterials. 1995, 16 (17): 1319-1325.CrossRefPubMed
13.
go back to reference Schmidmaier G, Lucke M, Schwabe P, Raschke M, Haas NP, Wildemann B: Collective review: bioactive implants coated with poly(D, L-lactide) and growth factors IGF-I, TGF-beta1, or BMP-2 for stimulation of fracture healing. J Long Term Eff Med Implants. 2006, 16 (1): 61-69.CrossRefPubMed Schmidmaier G, Lucke M, Schwabe P, Raschke M, Haas NP, Wildemann B: Collective review: bioactive implants coated with poly(D, L-lactide) and growth factors IGF-I, TGF-beta1, or BMP-2 for stimulation of fracture healing. J Long Term Eff Med Implants. 2006, 16 (1): 61-69.CrossRefPubMed
14.
go back to reference Schmidmaier G, Wildemann B, Bail H, Lucke M, Fuchs T, Stemberger A, Flyvbjerg A, Haas NP, Raschke M: Local application of growth factors (insulin-like growth factor-1 and transforming growth factor-beta1) from a biodegradable poly(D,L-lactide) coating of osteosynthetic implants accelerates fracture healing in rats. Bone. 2001, 28 (4): 341-350.CrossRefPubMed Schmidmaier G, Wildemann B, Bail H, Lucke M, Fuchs T, Stemberger A, Flyvbjerg A, Haas NP, Raschke M: Local application of growth factors (insulin-like growth factor-1 and transforming growth factor-beta1) from a biodegradable poly(D,L-lactide) coating of osteosynthetic implants accelerates fracture healing in rats. Bone. 2001, 28 (4): 341-350.CrossRefPubMed
15.
go back to reference Hayashibara T, Hiraga T, Yi B, Nomizu M, Kumagai Y, Nishimura R, Yoneda T: A synthetic peptide fragment of human MEPE stimulates new bone formation in vitro and in vivo. J Bone Miner Res. 2004, 19 (3): 455-462.CrossRefPubMed Hayashibara T, Hiraga T, Yi B, Nomizu M, Kumagai Y, Nishimura R, Yoneda T: A synthetic peptide fragment of human MEPE stimulates new bone formation in vitro and in vivo. J Bone Miner Res. 2004, 19 (3): 455-462.CrossRefPubMed
16.
go back to reference Six N, Septier D, Chaussain-Miller C, Blacher R, DenBesten P, Goldberg M: Dentonin, a MEPE fragment, initiates pulp-healing response to injury. J Dent Res. 2007, 86 (8): 780-785.CrossRefPubMed Six N, Septier D, Chaussain-Miller C, Blacher R, DenBesten P, Goldberg M: Dentonin, a MEPE fragment, initiates pulp-healing response to injury. J Dent Res. 2007, 86 (8): 780-785.CrossRefPubMed
17.
go back to reference Wang Y, Wan C, Szöke G, Ryaby JT, Li G: Local injection of thrombin-related peptide (TP508) in PPF/PLGA microparticles-enhanced bone formation during distraction osteogenesis. J Orthop Res. 2008, 26 (4): 539-546.CrossRefPubMed Wang Y, Wan C, Szöke G, Ryaby JT, Li G: Local injection of thrombin-related peptide (TP508) in PPF/PLGA microparticles-enhanced bone formation during distraction osteogenesis. J Orthop Res. 2008, 26 (4): 539-546.CrossRefPubMed
18.
go back to reference Hanratty BM, Ryaby JT, Pan X-H, Li G: Thrombin related peptide TP508 promoted fracture repair in a mouse high energy fracture model. J Orthop Surg Res. 2009, 4: 1-CrossRefPubMedPubMedCentral Hanratty BM, Ryaby JT, Pan X-H, Li G: Thrombin related peptide TP508 promoted fracture repair in a mouse high energy fracture model. J Orthop Surg Res. 2009, 4: 1-CrossRefPubMedPubMedCentral
19.
go back to reference Wang H, Li X, Tomin E, Doty SB, Lane JM, Carney DH, Ryaby JT: Thrombin peptide (TP508) promotes fracture repair by up-regulating inflammatory mediators, early growth factors, and increasing angiogenesis. J Orthop Res. 2005, 23 (3): 671-679.CrossRefPubMed Wang H, Li X, Tomin E, Doty SB, Lane JM, Carney DH, Ryaby JT: Thrombin peptide (TP508) promotes fracture repair by up-regulating inflammatory mediators, early growth factors, and increasing angiogenesis. J Orthop Res. 2005, 23 (3): 671-679.CrossRefPubMed
20.
go back to reference Qian JJ, Bhatnagar RS: Enhanced cell attachment to anorganic bone mineral in the presence of a synthetic peptide related to collagen. J Biomed Mater Res. 1996, 31 (4): 545-54.CrossRefPubMed Qian JJ, Bhatnagar RS: Enhanced cell attachment to anorganic bone mineral in the presence of a synthetic peptide related to collagen. J Biomed Mater Res. 1996, 31 (4): 545-54.CrossRefPubMed
21.
go back to reference Nguyen H, Qian JJ, Bhatnagar RS, Li S: Enhanced cell attachment and osteoblastic activity by P-15 peptide-coated matrix in hydrogels. Biochem Biophys Res Commun. 2003, 311 (1): 179-86.CrossRefPubMed Nguyen H, Qian JJ, Bhatnagar RS, Li S: Enhanced cell attachment and osteoblastic activity by P-15 peptide-coated matrix in hydrogels. Biochem Biophys Res Commun. 2003, 311 (1): 179-86.CrossRefPubMed
22.
go back to reference Siggelkow H, Niedhart C, Kurre W, Ihbe A, Schulz A, Atkinson MJ, Hüfner M: In vitro differentiation potential of a new human osteosarcoma cell line (HOS 58). Differentiation. 1998, 63 (2): 81-91.CrossRefPubMed Siggelkow H, Niedhart C, Kurre W, Ihbe A, Schulz A, Atkinson MJ, Hüfner M: In vitro differentiation potential of a new human osteosarcoma cell line (HOS 58). Differentiation. 1998, 63 (2): 81-91.CrossRefPubMed
23.
go back to reference Martin I, Jakob M, Schäfer D, Dick W, Spagnoli G, Heberer M: Quantitative analysis of gene expression in human articular cartilage from normal and osteoarthritic joints. Osteoarthritis Cartilage. 2001, 9 (2): 112-8.CrossRefPubMed Martin I, Jakob M, Schäfer D, Dick W, Spagnoli G, Heberer M: Quantitative analysis of gene expression in human articular cartilage from normal and osteoarthritic joints. Osteoarthritis Cartilage. 2001, 9 (2): 112-8.CrossRefPubMed
24.
go back to reference Frank O, Heim M, Jakob M, Barbero A, Schäfer D, Bendik I, Dick W, Heberer M, Martin I: Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem. 2002, 85 (4): 737-46.CrossRefPubMed Frank O, Heim M, Jakob M, Barbero A, Schäfer D, Bendik I, Dick W, Heberer M, Martin I: Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem. 2002, 85 (4): 737-46.CrossRefPubMed
25.
go back to reference Li G, Cui Y, McIlmurray L, Allen WE, Wang H: rhBMP-2, rhVEGF(165), rhPTN and thrombin-related peptide, TP508 induce chemotaxis of human osteoblasts and microvascular endothelial cells. J Orthop Res. 2005, 23 (3): 680-685.CrossRefPubMed Li G, Cui Y, McIlmurray L, Allen WE, Wang H: rhBMP-2, rhVEGF(165), rhPTN and thrombin-related peptide, TP508 induce chemotaxis of human osteoblasts and microvascular endothelial cells. J Orthop Res. 2005, 23 (3): 680-685.CrossRefPubMed
26.
go back to reference Kübler A, Neugebauer J, Oh J-H, Scheer M, Zöller JE: Growth and proliferation of human osteoblasts on different bone graft substitutes: an in vitro study. Implant Dent. 2004, 13 (2): 171-179.CrossRefPubMed Kübler A, Neugebauer J, Oh J-H, Scheer M, Zöller JE: Growth and proliferation of human osteoblasts on different bone graft substitutes: an in vitro study. Implant Dent. 2004, 13 (2): 171-179.CrossRefPubMed
27.
go back to reference Liu H, Li W, Gao C, Kumagai Y, Blacher RW, DenBesten PK: Dentonin, a fragment of MEPE, enhanced dental pulp stem cell proliferation. J Dent Res. 2004, 83 (6): 496-499.CrossRefPubMed Liu H, Li W, Gao C, Kumagai Y, Blacher RW, DenBesten PK: Dentonin, a fragment of MEPE, enhanced dental pulp stem cell proliferation. J Dent Res. 2004, 83 (6): 496-499.CrossRefPubMed
28.
go back to reference Miyamoto S, Teramoto H, Gutkind JS, Yamada KM: Integrins can collaborate with growth factors for phosphorylation of receptor tyrosine kinases and MAP kinase activation: roles of integrin aggregation and occupancy of receptors. J Cell Biol. 1996, 135 (6 Pt 1): 1633-1642.CrossRefPubMed Miyamoto S, Teramoto H, Gutkind JS, Yamada KM: Integrins can collaborate with growth factors for phosphorylation of receptor tyrosine kinases and MAP kinase activation: roles of integrin aggregation and occupancy of receptors. J Cell Biol. 1996, 135 (6 Pt 1): 1633-1642.CrossRefPubMed
29.
go back to reference Baron W, Shattil SJ, ffrench-Constant C: The oligodendrocyte precursor mitogen PDGF stimulates proliferation by activation of alpha(v)beta3 integrins. EMBO J. 2002, 21 (8): 1957-1966.CrossRefPubMedPubMedCentral Baron W, Shattil SJ, ffrench-Constant C: The oligodendrocyte precursor mitogen PDGF stimulates proliferation by activation of alpha(v)beta3 integrins. EMBO J. 2002, 21 (8): 1957-1966.CrossRefPubMedPubMedCentral
30.
go back to reference Bartold PM, Raben A: Growth factor modulation of fibroblasts in simulated wound healing. J Periodont Res. 1996, 31 (3): 205-216.CrossRefPubMed Bartold PM, Raben A: Growth factor modulation of fibroblasts in simulated wound healing. J Periodont Res. 1996, 31 (3): 205-216.CrossRefPubMed
31.
go back to reference Getachew R, Ballinger ML, Burch ML, Reid JJ, Khachigian LM, Wight TN, Little PJ, Osman N: PDGF beta-receptor kinase activity and ERK1/2 mediate glycosaminoglycan elongation on biglycan and increases binding to LDL. Endocrinology. 2010, 151 (9): 4356-4367.CrossRefPubMed Getachew R, Ballinger ML, Burch ML, Reid JJ, Khachigian LM, Wight TN, Little PJ, Osman N: PDGF beta-receptor kinase activity and ERK1/2 mediate glycosaminoglycan elongation on biglycan and increases binding to LDL. Endocrinology. 2010, 151 (9): 4356-4367.CrossRefPubMed
32.
go back to reference Wight T, Kinsella M: The role of proteoglycans in cell adhesion, migration and proliferation. Current opinion in cell biology. Curr Opin Cell Biol. 1992, 4 (5): 793-801.CrossRefPubMed Wight T, Kinsella M: The role of proteoglycans in cell adhesion, migration and proliferation. Current opinion in cell biology. Curr Opin Cell Biol. 1992, 4 (5): 793-801.CrossRefPubMed
33.
go back to reference Kim SJ, Kim SY, Kwon CH, Kim YK: Differential effect of FGF and PDGF on cell proliferation and migration in osteoblastic cells. Growth Factors. 2007, 25 (2): 77-86.CrossRefPubMed Kim SJ, Kim SY, Kwon CH, Kim YK: Differential effect of FGF and PDGF on cell proliferation and migration in osteoblastic cells. Growth Factors. 2007, 25 (2): 77-86.CrossRefPubMed
34.
go back to reference Wildemann B, Burkhardt N, Luebberstedt M, Vordemvenne T, Schmidmaier G: Proliferating and differentiating effects of three different growth factors on pluripotent mesenchymal cells and osteoblast like cells. J Orthop Surg Res. 2007, 2: 27-CrossRefPubMedPubMedCentral Wildemann B, Burkhardt N, Luebberstedt M, Vordemvenne T, Schmidmaier G: Proliferating and differentiating effects of three different growth factors on pluripotent mesenchymal cells and osteoblast like cells. J Orthop Surg Res. 2007, 2: 27-CrossRefPubMedPubMedCentral
35.
go back to reference Chaudhary LR, Hofmeister AM, Hruska KA: Differential growth factor control of bone formation through osteoprogenitor differentiation. Bone. 2004, 34 (3): 402-411.CrossRefPubMed Chaudhary LR, Hofmeister AM, Hruska KA: Differential growth factor control of bone formation through osteoprogenitor differentiation. Bone. 2004, 34 (3): 402-411.CrossRefPubMed
36.
go back to reference Yu X, Hsieh SC, Bao W, Graves DT: Temporal expression of PDGF receptors and PDGF regulatory effects on osteoblastic cells in mineralizing cultures. Am J Physiol. 1997, 272 (5 Pt 1): C1709-16.PubMed Yu X, Hsieh SC, Bao W, Graves DT: Temporal expression of PDGF receptors and PDGF regulatory effects on osteoblastic cells in mineralizing cultures. Am J Physiol. 1997, 272 (5 Pt 1): C1709-16.PubMed
37.
go back to reference Kono S-J, Oshima Y, Hoshi K, Bonewald LF, Oda H, Nakamura K, Kawaguchi H, Tanaka S: ERK pathways negatively regulate matrix mineralization. Bone. 2007, 40 (1): 68-74.CrossRefPubMed Kono S-J, Oshima Y, Hoshi K, Bonewald LF, Oda H, Nakamura K, Kawaguchi H, Tanaka S: ERK pathways negatively regulate matrix mineralization. Bone. 2007, 40 (1): 68-74.CrossRefPubMed
38.
go back to reference Zhang L, Leeman E, Carnes DC, Graves DT: Human osteoblasts synthesize and respond to platelet-derived growth factor. Am J Physiol. 1991, 261 (2 Pt 1): C348-54.PubMed Zhang L, Leeman E, Carnes DC, Graves DT: Human osteoblasts synthesize and respond to platelet-derived growth factor. Am J Physiol. 1991, 261 (2 Pt 1): C348-54.PubMed
Metadata
Title
Cooperative effects in differentiation and proliferation between PDGF-BB and matrix derived synthetic peptides in human osteoblasts
Authors
Thomas Vordemvenne
Jürgen RJ Paletta
Rene Hartensuer
Thomas Pap
Michael J Raschke
Sabine Ochman
Publication date
01-12-2011
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2011
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/1471-2474-12-263

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