Preparation of Recombinant Bone Morphogenetic Protein 2 at Laboratory Scale

Mahmoud Kazemi, Ali Hassani, Mojgan Bandehpour, Bahram Kazemi

Abstract


356

Introduction: Millions cases of bone fracture are reported annually worldwide. The conventional methods for the repair often fail. The complications of using routine methods include stimulation of bone repair which should be performed without infection risk and immunologic responses. The aim of this study was cloning and expression of bone morphogenenic protein 2 (BMP2), a key growth factor used in bone repair. Materials and Methods: mRNA was extracted from human maxillary osteosarcoma and reverse transcribed. The BMP2 gene was amplified by PCR and cloned into pETDuet vector and was expressed. Results: The BMP2 gene was expressed successfully and expression was confirmed by western blotting using anti- His tag antibody. Conclusion: Recombinant protein preparation method has been recognized to be accessible and the production is cost effective.


Keywords


BMP2; Bone regeneration; Bone fracture

Full Text:

PDF

154

References


Pressley JC, Kendig TD, Frencher SK, Barlow B, Quitel L, Waqar F. Epidemiology of bone fracture across the age span in blacks and whites. J Trauma. 2011;71(5 Suppl 2):S541-8.

Dhanwal DK, Dennison EM, Harvey NC, Cooper C. Epidemiology of hip fracture: Worldwide geographic variation. Indian J Orthop. 2011;45(1):15-22.

Filipov O. Epidemiology and social burden of the femoral neck fractures. Journal of IMAB–Annual Proceeding Scientific Papers. 2014;20(4):516-8.

Allareddy V, Allareddy V, Nalliah RP. Epidemiology of facial fracture injuries. J Oral Maxillofac Surg. 2011;69(10):2613-8.

Chee W, Jivraj S. Failures in implant dentistry. Br Dent J. 2007;202(3):123-9.

Park J, Ries J, Gelse K, Kloss F, von der Mark K, Wiltfang J, et al. Bone regeneration in critical size defects by cell-mediated BMP-2 gene transfer: a comparison of adenoviral vectors and liposomes. Gene Ther. 2003;10(13):1089-98.

Rao SM, Ugale GM, Warad SB. Bone morphogenetic proteins: periodontal regeneration. N Am J Med Sci. 2013;5(3):161-8.

Erickson DM, Harris SE, Dean DD, Harris MA, Wozney JM, Boyan BD, et al. Recombinant bone morphogenetic protein (BMP)-2 regulates costochondral growth plate chondrocytes and induces expression of BMP-2 and BMP-4 in a cell maturation-dependent manner. J Orthop Res. 1997;15(3):371-80.

Carragee EJ, Hurwitz EL, Weiner BK. A critical review of recombinant human bone morphogenetic protein-2 trials in spinal surgery: emerging safety concerns and lessons learned. Spine J. 2011;11(6):471-91.

Clemson M, Kelly WJ. Optimizing alkaline lysis for DNA plasmid recovery. Biotechnol Appl Biochem. 2003;37(Pt 3):235-44.

Kram KE, Finkel SE. Rich Medium Composition Affects Escherichia coli Survival, Glycation, and Mutation Frequency during Long-Term Batch Culture. Appl Environ Microbiol. 2015;81(13):4442-50.

Hansen LH, Knudsen S, Sorensen SJ. The effect of the lacY gene on the induction of IPTG inducible promoters, studied in Escherichia coli and Pseudomonas fluorescens. Curr Microbiol. 1998;36(6):341-7.

Burnette WN. Western blotting: remembrance of things past. Methods Mol Biol. 2015;1312:9-12.

Chen D, Zhao M, Mundy GR. Bone morphogenetic proteins. Growth Factors. 2004;22(4):233-41.

Carreira AC, Zambuzzi WF, Rossi MC, Astorino Filho R, Sogayar MC, Granjeiro JM. Bone Morphogenetic Proteins: Promising Molecules for Bone Healing, Bioengineering, and Regenerative Medicine. Vitam Horm. 2015;99:293-322.

Khan SN, Lane JM. The use of recombinant human bone morphogenetic protein-2 (rhBMP-2) in orthopaedic applications. Expert Opin Biol Ther. 2004;4(5):741-8.

Xiong S, Yu L, Tan H, Huang T, Wang C, Liu D, et al. Expression and purification of recombinant bone morphogenetic protein-2 in E. coli. Di 1 jun yi da xue xue bao= Academic journal of the first medical college of PLA. 2002;22(5):413-6.

Ma Q, Dang G, Ma D. Human bone morphogenetic protein-2: recombinant expression in E. coli. Zhonghua Yi Xue Za Zhi. 1998;78(4):308-10.

Ihm HJ, Yang SJ, Huh JW, Choi SY, Cho SW. Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli. BMB Rep. 2008;41(5):404-7.




DOI: https://doi.org/10.22037/rrr.v1i3.11968

Refbacks

  • There are currently no refbacks.


ISSN:2476-5163 (Print); 2476-5171 (Online)

 
Creative Commons LicenseThe Journal of "Regeneration, Reconstruction, & Restoration" is licensed under a Creative Commons Attribution 4.0 International License.