Enhanced osteocalcin expression by osteoblast-like cells (MC3T3-E1) exposed to bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system) ions

V. G. Varanasi; E. Saiz; P. M. Loomer; B. Ancheta; N. Uritani; S. P. Ho; A. P. Tomsia; S. J. Marshall; G. W. Marshall

doi:10.1016/j.actbio.2009.05.035

Enhanced osteocalcin expression by osteoblast-like cells (MC3T3-E1) exposed to bioactive coating glass (SiO₂-CaO-P₂O₅-MgO-K₂O-Na₂O system) ions

V. G. Varanasi, E. Saiz, P. M. Loomer, B. Ancheta, N. Uritani, S. P. Ho, A. P. Tomsia, S. J. Marshall, G. W. Marshall

Research output: Contribution to journal › Article › peer-review

105 Scopus citations

Abstract

This study tested the hypothesis that bioactive coating glass (SiO₂-CaO-P₂O₅-MgO-K₂O-Na₂O system), used for implant coatings, enhanced the induction of collagen type 1 synthesis and in turn enhanced the expression of downstream markers alkaline phosphatase, Runx2 and osteocalcin during osteoblast differentiation. The ions from experimental bioactive glass (6P53-b) and commercial Bioglass^TM (45S5) were added to osteoblast-like MC3T3-E1 subclone 4 cultures as a supplemented ion extract (glass conditioned medium (GCM)). Ion extracts contained significantly higher concentrations of Si and Ca (Si, 47.9 ± 10.4 ppm; Ca, 69.8 ± 14.0 for 45S5; Si, 33.4 ± 3.8 ppm; Ca, 57.1 ± 2.8 ppm for 6P53-b) compared with the control extract (Si < 0.1 ppm, Ca 49.0 ppm in α-MEM) (ANOVA, p < 0.05). Cell proliferation rate was enhanced (1.5× control) within the first 3 days after adding 45S5 and 6P53-b GCM. MC3T3-E1 subclone 4 cultures were then studied for their response to the addition of test media (GCM and control medium along with ascorbic acid (AA; 50 ppm)). Each GCM + AA treatment enhanced collagen type 1 synthesis as observed in both gene expression results (day 1, Col1α1, 45S5 GCM + AA: 3× control + AA; 6P53-b GCM + AA: 4× control + AA; day 5, Col1α2, 45S5 GCM + AA: 3.15× control + AA; 6P53-b GCM + AA: 2.35× control + AA) and in histological studies (Picrosirius stain) throughout the time course of early differentiation. Continued addition of each GCM and AA treatment led to enhanced expression of alkaline phosphatase (1.4× control + AA after 5 days, 2× control + AA after 10 days), Runx2 (2× control + AA after 7 days) and osteocalcin gene (day 3, 45S5 GCM + AA: 14× control + AA; day 5, 6P53-b GCM + AA: 19× control + AA) and protein expression (40×-70× control + AA after 6 days). These results indicated the enhanced effect of bioactive glass ions on key osteogenic markers important for the bone healing process.

Original language	English (US)
Pages (from-to)	3536-3547
Number of pages	12
Journal	Acta Biomaterialia
Volume	5
Issue number	9
DOIs	https://doi.org/10.1016/j.actbio.2009.05.035
State	Published - Nov 2009
Externally published	Yes

Keywords

Bioactive glass ions
Calcium
Osteoblasts
Osteogenesis
Silicon

ASJC Scopus subject areas

Biotechnology
Biomaterials
Biochemistry
Biomedical Engineering
Molecular Biology

Access to Document

10.1016/j.actbio.2009.05.035

Cite this

@article{199a20ce043947029497866a197d371c,

title = "Enhanced osteocalcin expression by osteoblast-like cells (MC3T3-E1) exposed to bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system) ions",

abstract = "This study tested the hypothesis that bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system), used for implant coatings, enhanced the induction of collagen type 1 synthesis and in turn enhanced the expression of downstream markers alkaline phosphatase, Runx2 and osteocalcin during osteoblast differentiation. The ions from experimental bioactive glass (6P53-b) and commercial BioglassTM (45S5) were added to osteoblast-like MC3T3-E1 subclone 4 cultures as a supplemented ion extract (glass conditioned medium (GCM)). Ion extracts contained significantly higher concentrations of Si and Ca (Si, 47.9 ± 10.4 ppm; Ca, 69.8 ± 14.0 for 45S5; Si, 33.4 ± 3.8 ppm; Ca, 57.1 ± 2.8 ppm for 6P53-b) compared with the control extract (Si < 0.1 ppm, Ca 49.0 ppm in α-MEM) (ANOVA, p < 0.05). Cell proliferation rate was enhanced (1.5× control) within the first 3 days after adding 45S5 and 6P53-b GCM. MC3T3-E1 subclone 4 cultures were then studied for their response to the addition of test media (GCM and control medium along with ascorbic acid (AA; 50 ppm)). Each GCM + AA treatment enhanced collagen type 1 synthesis as observed in both gene expression results (day 1, Col1α1, 45S5 GCM + AA: 3× control + AA; 6P53-b GCM + AA: 4× control + AA; day 5, Col1α2, 45S5 GCM + AA: 3.15× control + AA; 6P53-b GCM + AA: 2.35× control + AA) and in histological studies (Picrosirius stain) throughout the time course of early differentiation. Continued addition of each GCM and AA treatment led to enhanced expression of alkaline phosphatase (1.4× control + AA after 5 days, 2× control + AA after 10 days), Runx2 (2× control + AA after 7 days) and osteocalcin gene (day 3, 45S5 GCM + AA: 14× control + AA; day 5, 6P53-b GCM + AA: 19× control + AA) and protein expression (40×-70× control + AA after 6 days). These results indicated the enhanced effect of bioactive glass ions on key osteogenic markers important for the bone healing process.",

keywords = "Bioactive glass ions, Calcium, Osteoblasts, Osteogenesis, Silicon",

author = "Varanasi, {V. G.} and E. Saiz and Loomer, {P. M.} and B. Ancheta and N. Uritani and Ho, {S. P.} and Tomsia, {A. P.} and Marshall, {S. J.} and Marshall, {G. W.}",

note = "Funding Information: The authors would like to thank the following contributors to this paper: Tiffany Vallortigara, Janet Wong, Kelly Leong and Garrett Porteous. The authors would also like to thank the following for their kind advice related to the above work: Linda Prentice, Larry Watanabe, Grace Nonomura, Steven Lee, Michael Carillo, Dr. Stuart Gansky, Professor Pamela DenBeston, Dr. Yuan Zhang, Dr. James Chen, Dr. Stefan Habelitz, Dr. Kuniko Saeki and Dr. Huynh Tri. In addition, the authors thank UC Davis ICP–MS facility for solution and GCM analyses. Finally, the authors appreciate the financial support by the National Institutes of Health/National Institute of Dental and Craniofacial Research Grants K25 DE018230 Varanasi (PI) and R01 DE11289 Tomsia (PI). ",

year = "2009",

month = nov,

doi = "10.1016/j.actbio.2009.05.035",

language = "English (US)",

volume = "5",

pages = "3536--3547",

journal = "Acta Biomaterialia",

issn = "1742-7061",

publisher = "Elsevier BV",

number = "9",

}

TY - JOUR

T1 - Enhanced osteocalcin expression by osteoblast-like cells (MC3T3-E1) exposed to bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system) ions

AU - Varanasi, V. G.

AU - Saiz, E.

AU - Loomer, P. M.

AU - Ancheta, B.

AU - Uritani, N.

AU - Ho, S. P.

AU - Tomsia, A. P.

AU - Marshall, S. J.

AU - Marshall, G. W.

N1 - Funding Information: The authors would like to thank the following contributors to this paper: Tiffany Vallortigara, Janet Wong, Kelly Leong and Garrett Porteous. The authors would also like to thank the following for their kind advice related to the above work: Linda Prentice, Larry Watanabe, Grace Nonomura, Steven Lee, Michael Carillo, Dr. Stuart Gansky, Professor Pamela DenBeston, Dr. Yuan Zhang, Dr. James Chen, Dr. Stefan Habelitz, Dr. Kuniko Saeki and Dr. Huynh Tri. In addition, the authors thank UC Davis ICP–MS facility for solution and GCM analyses. Finally, the authors appreciate the financial support by the National Institutes of Health/National Institute of Dental and Craniofacial Research Grants K25 DE018230 Varanasi (PI) and R01 DE11289 Tomsia (PI).

PY - 2009/11

Y1 - 2009/11

N2 - This study tested the hypothesis that bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system), used for implant coatings, enhanced the induction of collagen type 1 synthesis and in turn enhanced the expression of downstream markers alkaline phosphatase, Runx2 and osteocalcin during osteoblast differentiation. The ions from experimental bioactive glass (6P53-b) and commercial BioglassTM (45S5) were added to osteoblast-like MC3T3-E1 subclone 4 cultures as a supplemented ion extract (glass conditioned medium (GCM)). Ion extracts contained significantly higher concentrations of Si and Ca (Si, 47.9 ± 10.4 ppm; Ca, 69.8 ± 14.0 for 45S5; Si, 33.4 ± 3.8 ppm; Ca, 57.1 ± 2.8 ppm for 6P53-b) compared with the control extract (Si < 0.1 ppm, Ca 49.0 ppm in α-MEM) (ANOVA, p < 0.05). Cell proliferation rate was enhanced (1.5× control) within the first 3 days after adding 45S5 and 6P53-b GCM. MC3T3-E1 subclone 4 cultures were then studied for their response to the addition of test media (GCM and control medium along with ascorbic acid (AA; 50 ppm)). Each GCM + AA treatment enhanced collagen type 1 synthesis as observed in both gene expression results (day 1, Col1α1, 45S5 GCM + AA: 3× control + AA; 6P53-b GCM + AA: 4× control + AA; day 5, Col1α2, 45S5 GCM + AA: 3.15× control + AA; 6P53-b GCM + AA: 2.35× control + AA) and in histological studies (Picrosirius stain) throughout the time course of early differentiation. Continued addition of each GCM and AA treatment led to enhanced expression of alkaline phosphatase (1.4× control + AA after 5 days, 2× control + AA after 10 days), Runx2 (2× control + AA after 7 days) and osteocalcin gene (day 3, 45S5 GCM + AA: 14× control + AA; day 5, 6P53-b GCM + AA: 19× control + AA) and protein expression (40×-70× control + AA after 6 days). These results indicated the enhanced effect of bioactive glass ions on key osteogenic markers important for the bone healing process.

AB - This study tested the hypothesis that bioactive coating glass (SiO2-CaO-P2O5-MgO-K2O-Na2O system), used for implant coatings, enhanced the induction of collagen type 1 synthesis and in turn enhanced the expression of downstream markers alkaline phosphatase, Runx2 and osteocalcin during osteoblast differentiation. The ions from experimental bioactive glass (6P53-b) and commercial BioglassTM (45S5) were added to osteoblast-like MC3T3-E1 subclone 4 cultures as a supplemented ion extract (glass conditioned medium (GCM)). Ion extracts contained significantly higher concentrations of Si and Ca (Si, 47.9 ± 10.4 ppm; Ca, 69.8 ± 14.0 for 45S5; Si, 33.4 ± 3.8 ppm; Ca, 57.1 ± 2.8 ppm for 6P53-b) compared with the control extract (Si < 0.1 ppm, Ca 49.0 ppm in α-MEM) (ANOVA, p < 0.05). Cell proliferation rate was enhanced (1.5× control) within the first 3 days after adding 45S5 and 6P53-b GCM. MC3T3-E1 subclone 4 cultures were then studied for their response to the addition of test media (GCM and control medium along with ascorbic acid (AA; 50 ppm)). Each GCM + AA treatment enhanced collagen type 1 synthesis as observed in both gene expression results (day 1, Col1α1, 45S5 GCM + AA: 3× control + AA; 6P53-b GCM + AA: 4× control + AA; day 5, Col1α2, 45S5 GCM + AA: 3.15× control + AA; 6P53-b GCM + AA: 2.35× control + AA) and in histological studies (Picrosirius stain) throughout the time course of early differentiation. Continued addition of each GCM and AA treatment led to enhanced expression of alkaline phosphatase (1.4× control + AA after 5 days, 2× control + AA after 10 days), Runx2 (2× control + AA after 7 days) and osteocalcin gene (day 3, 45S5 GCM + AA: 14× control + AA; day 5, 6P53-b GCM + AA: 19× control + AA) and protein expression (40×-70× control + AA after 6 days). These results indicated the enhanced effect of bioactive glass ions on key osteogenic markers important for the bone healing process.

KW - Bioactive glass ions

KW - Calcium

KW - Osteoblasts

KW - Osteogenesis

KW - Silicon

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