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Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour

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 investi...

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Autores principales: Zhang, Yufeng, Shuang, Yang, Fu, Hang, Zhou, Wei, Qian, Li, Dai, Jing, Miron, Richard J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4696268/
https://www.ncbi.nlm.nih.gov/pubmed/26715589
http://dx.doi.org/10.1186/s12903-015-0154-z
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author Zhang, Yufeng
Shuang, Yang
Fu, Hang
Zhou, Wei
Qian, Li
Dai, Jing
Miron, Richard J.
author_facet Zhang, Yufeng
Shuang, Yang
Fu, Hang
Zhou, Wei
Qian, Li
Dai, Jing
Miron, Richard J.
author_sort Zhang, Yufeng
collection PubMed
description 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. CONCLUSION: 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.
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spelling pubmed-46962682015-12-31 Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour Zhang, Yufeng Shuang, Yang Fu, Hang Zhou, Wei Qian, Li Dai, Jing Miron, Richard J. BMC Oral Health Research Article 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. CONCLUSION: 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. BioMed Central 2015-12-30 /pmc/articles/PMC4696268/ /pubmed/26715589 http://dx.doi.org/10.1186/s12903-015-0154-z Text en © Zhang et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Zhang, Yufeng
Shuang, Yang
Fu, Hang
Zhou, Wei
Qian, Li
Dai, Jing
Miron, Richard J.
Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title_full Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title_fullStr Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title_full_unstemmed Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title_short Characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
title_sort characterization of a shorter recombinant polypeptide chain of bone morphogenetic protein 2 on osteoblast behaviour
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4696268/
https://www.ncbi.nlm.nih.gov/pubmed/26715589
http://dx.doi.org/10.1186/s12903-015-0154-z
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