RhBMP-2 and -7 combined with absorbable collagen sponge carrier enhance ectopic bone formation: An in vivo bioassay

Preativatanyou, Kanok; Honsawek, Sittisak

doi:10.5372/1905-7415.0501.010

Abstract

Background: Recombinant human bone morphogenetic proteins (rhBMPs) have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies. However, delivery of more than one growth factor by sustained release carrier to orthopedic site has yet been questionable in terms of efficacy and synergism.

Objective: Evaluate osteoinductivity and synergistic effect of rhBMP-2 and -7 using absorbable collagen sponge (ACS) carrier system in vivo.

Methods: cDNA of BMP-2 and -7 active domains were cloned and expressed in Escherichia coli BL21 Star^TM (DE3) using pRSETc expression system. Then, the purified rhBMPs were loaded onto ACS and evaluated by in vivo rat subcutaneous bioassay. Two and eight weeks postoperatively, all treated groups were histologically verified for evidence of new bone formation and neovascularization by hematoxylin-eosin staining and light microscopy.

Results: The Wistar rat treated with rhBMP-2 or -7/ACS exhibited new bone formation, compared to ACS control. The group treated with ACS supplemented with both rhBMP-2 and -7 significantly showed the osteoid matrix very well-organized into trabeculae-like structure with significant blood vessel invasion.

Conclusion: The osteogenic induction of rhBMPs was combined with ACS carrier in the in vivo bioassay. In addition, the combination of both two potent recombinant osteoinductive cytokines, rhBMP-2 and -7, with ACS carrier demonstrated synergistic effect and might be a more promising and effective choice for therapeutic applications.

References

1. Hogan BL. Bone morphogenetic proteins in development. Curr Opin Genet Dev. 1996; 6:432-8.10.1016/S0959-437X(96)80064-5
Open DOI Search in Google Scholar Back to article
2. Nakase T, Yoshikawa H. Potential roles of bone morphogenetic proteins (BMPs) in skeletal repair and regeneration. J Bone Miner Metab. 2006; 24:425-33.10.1007/s00774-006-0718-8
Open DOI Search in Google Scholar Back to article
3. Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T, et al. Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol. 1994; 6:1755-66.10.1083/jcb.127.6.1755
Open DOI Search in Google Scholar Back to article
4. Wang EA, Rosen V, D’Alessandro JS, Bauduy M, Cordes P, Harada T, et al. Recombinant human bone morphogenetic protein induces bone formation. Proc Natl Acad Sci USA. 1990; 87:2220-4.10.1073/pnas.87.6.2220
Open DOI Search in Google Scholar Back to article
5. Pecina M, Giltaij LR, Vukicevic S. Orthopaedic applications of osteogenic protein-1 (BMP-7). Int Orthop. 2001; 25:203-8.10.1007/s002640100262
Open DOI Search in Google Scholar Back to article
6. Geiger M, Li RH, Friess W. Collagen sponges for bone regeneration with rhBMP-2. Adv Drug Deliv Rev. 2003; 55:1613-29.10.1016/j.addr.2003.08.010
Open DOI Search in Google Scholar Back to article
7. Friess W, Uludag H, Foskett S, Biron R, Sargeant C. Characterization of absorbable collagen sponges as rhBMP-2 carriers. Int J Pharm. 1999; 187:91-9.10.1016/S0378-5173(99)00174-X
Search in Google Scholar Back to article
8. Scheufler C, Sebald W, Hulsmeyer M. Crystal structure of human bone morphogenetic protein-2 at 2.7 angstrom resolution. J Mol Biol. 1999; 287:103-15.10.1006/jmbi.1999.259010074410
Search in Google Scholar Back to article
9. Griffith DL, Keck PC, Sampath TK, Rueger DC, Carlson WD. Three-dimensional structure of recombinant human osteogenic 1: Structural paradigm for the transforming growth factor-β superfamily. Proc Natl Acad Sci USA. 1996; 93:878-83.10.1073/pnas.93.2.878401518570652
Open DOI Search in Google Scholar Back to article
10. Israel DI, Nove J, Kerns KM, Moutsatsos IK, Kaufman RJ. Expression and characterization of bone morphogenetic protein-2 in Chinese hamster ovary cells. Growth Factors. 1992; 7:139-50.10.3109/089771992090464031419071
Open DOI Search in Google Scholar Back to article
11. Long S, Truong L, Bennett K, Phillips A, Wong-Staal F, Ma H. Expression, purification, and renaturation of bone morphogenetic protein-2 from Escherichia coli. Protein Expr Purif. 2006; 46:374-8.10.1016/j.pep.2005.09.025
Search in Google Scholar Back to article
12. Vallejo LF, Brokelmann M, Marten S, Trappe S, Cabrera- Crespo J, HoVmann A, et al. Renaturation and purification of bone morphogenetic protein-2 produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli. J Biotechnol. 2002; 94: 185-94.10.1016/S0168-1656(01)00425-4
Open DOI Search in Google Scholar Back to article
13. Bessa PC, Pedro AJ, Klosch B, Nobre A, van Griensven M, Reis RL, et al. Osteoinduction in human fat-derived stem cells by recombinant human bone morphogenetic protein-2 produced in Escherichia coli. Biotechnol Lett. 2008; 30:15-21.10.1007/s10529-007-9501-8
Open DOI Search in Google Scholar Back to article
14. Visser R, Arrabal PM, Becerra J, Rinas U, Cifuentes M. The effect of an rhBMP-2 absorbable collagen spongetargeted system on bone formation in vivo. Biomaterials. 2009; 30:2032-7.10.1016/j.biomaterials.2008.12.046
Open DOI Search in Google Scholar Back to article
15. Kubler NR, Reuther JF, Faller G, Kirchner T, Ruppert R, Sebald W. Inductive properties of recombinant human BMP-2 produced in a bacterial expression system. Int J Oral Maxillofac Surg. 1998; 27: 305-9.10.1016/S0901-5027(05)80621-6
Search in Google Scholar Back to article
16. Ten Dijke P, Yamashita H, Sampath TK, Reddi AH, Estevez M, Riddle DL, et al. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J Biol Chem. 1994; 269: 16985-8.10.1016/S0021-9258(17)32506-1
Search in Google Scholar Back to article
17. Ten Dijke P, Yamashita H, Ichijo H, Franzen P, Laiho M, Miyazono K, et al. Characterization of type I receptors for transforming growth factor-β and activin. Science. 1994; 264:101-4.10.1126/science.81404128140412
Search in Google Scholar Back to article
18. Lyons KM, Hogan BL, Robertson EJ. Colocalization of BMP 7 and BMP 2 RNAs suggests that these factors cooperatively mediate tissue interactions during murine development Mech Dev. 1995; 50:71-83.
Search in Google Scholar Back to article
19. Knosp WM, Scott V, Bachinger HP, Stadler HS. HOXA13 regulates the expression of bone morphogenetic proteins 2 and 7 to control distal limb morphogenesis. Development. 2004; 131:4581-92.10.1242/dev.0132715342482
Search in Google Scholar Back to article
20. Israel DI, Nove J, Kerns KM, Kaufman RJ, Rosen V, Cox KA, et al. Heterodimeric bone morphogenetic proteins show enhanced activity in vitro and in vivo. Growth Factors. 1996; 13:291-300.10.3109/089771996090032298919035
Open DOI Search in Google Scholar Back to article
21. Aoki H, Fujii M, Imamura T, Yagi K, Takehara K, Kato M, et al. Synergistic effects of different bone morphogenetic protein type I receptors on alkaline phosphatase induction. J Cell Sci. 2001; 114:1483-9.10.1242/jcs.114.8.148311282024
Search in Google Scholar Back to article
22. Lee JH, Kim CS, Choi KH, Jung UW, Yun JH, Choi SH, et al. The induction of bone formation in rat calvarial defects and subcutaneous tissues by recombinant human BMP-2, produced in Escherichia coli. Biomaterials. 2010; 31:3512-9.10.1016/j.biomaterials.2010.01.07520149447
Open DOI Search in Google Scholar Back to article
23. Kim CS, Kim JI, Kim J, Choi SH, Chai JK, Kim CK, et al. Ectopic bone formation associated with recombinant human bone morphogenetic proteins-2 using absorbable collagen sponge and beta tricalcium phosphate as carriers. Biomaterials. 2006; 26:2501-7.10.1016/j.biomaterials.2004.07.01515585252
Open DOI Search in Google Scholar Back to article
24. Seeherman H, Wozney JM. Delivery of bone morphogenetic proteins for orthopedic tissue regeneration. Cytokine Growth Factor Rev. 2005; 16: 329-45. 10.1016/j.cytogfr.2005.05.00115936978
Open DOI Search in Google Scholar Back to article

RhBMP-2 and -7 combined with absorbable collagen sponge carrier enhance ectopic bone formation: An in vivo bioassay

Abstract

Paradigm

My account