title: Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour.
abstract: BACKGROUND
  Recombinant bone morphogenetic protein two (rhBMP2) has been utilised for a variety of clinical applications in orthopaedic surgery and dental procedures. Despite its widespread use, concerns have been raised regarding its short half-life and transient bioactivity in vivo. Recent investigation aimed at developing rhBMP2 synthesized from a shorter polypeptide chain (108 amino acids) has been undertaken.
  
  
  METHODS
  The osteopromotive properties of BMP2 were investigated on cell behaviour. Five concentrations of rhBMP2_108 including 10, 50, 100, 200 and 500 ng/ml were compared to a commercially available rhBMP2 (100 ng/ml). Each of the working concentrations of rhBMP2_108 were investigated on MC3T3-E1 osteoblasts for their ability to induce osteoblast recruitment, proliferation and differentiation as assessed by alkaline phosphatase (ALP) staining, alizarin red staining, and real-time PCR for genes encoding ALP, osteocalcin (OCN), collagen-1 (COL-1) and Runx2.
  
  
  RESULTS
  The results demonstrate that all concentrations of rhBMP2_108 significantly improved cell recruitment and proliferation of osteoblasts at 5 days post seeding. Furthermore, rhBMP2_108 had the most pronounced effects on osteoblast differentiation. It was found that rhBMP2_108 had over a four fold significant increase in ALP activity at seven and 14 days post-seeding and the concentrations ranging from 50 to 200 ng/ml demonstrated the most pronounced effects. Analysis of real-time PCR for genes encoding ALP, OCN, COL-1 and Runx2 further confirmed dose-dependant increases at 14 days post-seeding. Furthermore, alizarin red staining demonstrated a concentration dependant increase in staining at 14 days.
  
  
  CONCLUSIONS
  The results from the present study demonstrate that this shorter polypeptide chain of rhBMP2_108 is equally as bioactive as commercially available rhBMP2 for the recruitment of progenitor cells by facilitating their differentiation towards the osteoblast lineage. Future in vivo study are necessary to investigate its bioactivity.
authors.affiliation: The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, People's Republic of China. zyf@whu.edu.cn.|Department of Oral Implantology, School of Stomatology, Wuhan University, Wuhan, 430079, China. zyf@whu.edu.cn.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, People's Republic of China. 2009302220042@whu.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shuang
authors.affiliation: Hangzhou JIuyuan Gene Engineering Co, Ltd;East of No.8 Street, Hangzhou Econ. and Tech. Development Zone, Hangzhou, China Hangzhou China, 310018, China. 584946138@qq.com.
authors.forename: Hang
authors.initials: H
authors.lastname: Fu
authors.affiliation: Hangzhou JIuyuan Gene Engineering Co, Ltd;East of No.8 Street, Hangzhou Econ. and Tech. Development Zone, Hangzhou, China Hangzhou China, 310018, China. 380497633@qq.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Hangzhou JIuyuan Gene Engineering Co, Ltd;East of No.8 Street, Hangzhou Econ. and Tech. Development Zone, Hangzhou, China Hangzhou China, 310018, China. 271107294@qq.com.
authors.forename: Li
authors.initials: L
authors.lastname: Qian
authors.affiliation: The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, People's Republic of China. daijing0508@qq.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Dai
authors.affiliation: The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, People's Republic of China. richard.miron@zmk.unibe.ch.|Department of Oral Surgery and Stomatology, University of Bern, Freiburgstrasse 7, Bern, 3010, Switzerland. richard.miron@zmk.unibe.ch.|Department of Periodontology, School of Dental Medicine, University of Bern, Freiburgstrasse 7, Bern, 3010, Switzerland. richard.miron@zmk.unibe.ch.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Miron
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D024042:Collagen Type I / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects*; D015675:Osteocalcin / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; D010455:Peptides; D011994:Recombinant Proteins; C493716:Runx2 protein, mouse; D015675:Osteocalcin; D000469:Alkaline Phosphatase