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

Zhang Y, Shuang Y, Fu H, Zhou W, Qian L, Dai J, Miron RJ - BMC Oral Health (2015)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

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.

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.

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.

No MeSH data available.


Related in: MedlinePlus

Quantitative analysis of alizarin red staining. rhBMP2_108 demonstrate a concentration dependent increase in alizarin red staining (All experiments were conducted in triplicate with three independent experiments (n = 9). Data represents means +/− SEM. * denotes significant difference between two groups, p < 0.05; ** denotes significantly higher than all other modalities, p < 0.05, # denotes control samples significantly lower than all other treatment modalities, p < 0.05)
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Fig6: Quantitative analysis of alizarin red staining. rhBMP2_108 demonstrate a concentration dependent increase in alizarin red staining (All experiments were conducted in triplicate with three independent experiments (n = 9). Data represents means +/− SEM. * denotes significant difference between two groups, p < 0.05; ** denotes significantly higher than all other modalities, p < 0.05, # denotes control samples significantly lower than all other treatment modalities, p < 0.05)

Mentions: RhBMP2_108 was then assessed for its ability to speed up osteoblast differentiation at various concentrations (Fig. 3, Fig. 4, Fig. 5, Fig. 6). First it was found that rhBMP2_108 was able to significantly increase ALP activity at seven and 14 days post seeding for all concentrations of rhBMP2_108 (Fig. 3). Interestingly, it was observed at 7 days that ALP activity was significantly higher at rhBMP2_108 concentrations of 50, 100 and 200 ng/ml when compared to control samples (Fig. 3). At 14 days, all treatment modalities receiving rhBMP2_108 at various concentrations demonstrated a marked 16-fold increase in ALP activity when compared to control samples void of rhBMP2_108 (Fig. 3).Fig. 3


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

Zhang Y, Shuang Y, Fu H, Zhou W, Qian L, Dai J, Miron RJ - BMC Oral Health (2015)

Quantitative analysis of alizarin red staining. rhBMP2_108 demonstrate a concentration dependent increase in alizarin red staining (All experiments were conducted in triplicate with three independent experiments (n = 9). Data represents means +/− SEM. * denotes significant difference between two groups, p < 0.05; ** denotes significantly higher than all other modalities, p < 0.05, # denotes control samples significantly lower than all other treatment modalities, p < 0.05)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4696268&req=5

Fig6: Quantitative analysis of alizarin red staining. rhBMP2_108 demonstrate a concentration dependent increase in alizarin red staining (All experiments were conducted in triplicate with three independent experiments (n = 9). Data represents means +/− SEM. * denotes significant difference between two groups, p < 0.05; ** denotes significantly higher than all other modalities, p < 0.05, # denotes control samples significantly lower than all other treatment modalities, p < 0.05)
Mentions: RhBMP2_108 was then assessed for its ability to speed up osteoblast differentiation at various concentrations (Fig. 3, Fig. 4, Fig. 5, Fig. 6). First it was found that rhBMP2_108 was able to significantly increase ALP activity at seven and 14 days post seeding for all concentrations of rhBMP2_108 (Fig. 3). Interestingly, it was observed at 7 days that ALP activity was significantly higher at rhBMP2_108 concentrations of 50, 100 and 200 ng/ml when compared to control samples (Fig. 3). At 14 days, all treatment modalities receiving rhBMP2_108 at various concentrations demonstrated a marked 16-fold increase in ALP activity when compared to control samples void of rhBMP2_108 (Fig. 3).Fig. 3

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

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.

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.

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.

No MeSH data available.


Related in: MedlinePlus