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Quantification of Bone Formation on Calcium Phosphate Ceramic Thin Film, in Vitro by Tetracycline Labelling

Published online by Cambridge University Press:  02 July 2020

J.E. Davies ,
F. Krasnoshtein ,
L. Hryhorenko  and
D. Sindrey
J.E. Davies
Affiliation:
Centre for Biomaterials, University of Toronto, Toronto, Ont, CanadaM5S 1A1
F. Krasnoshtein
Affiliation:
Centre for Biomaterials, University of Toronto, Toronto, Ont, CanadaM5S 1A1
L. Hryhorenko
Affiliation:
Allelix Biopharmaceuticals Inc., Mississauga, Ont, CanadaL4V 1V7
D. Sindrey
Affiliation:
Allelix Biopharmaceuticals Inc., Mississauga, Ont, CanadaL4V 1V7
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Introduction

We have previously described a quantitative in vitro method for the assessment of bone matrix deposition, employing the fluorescent antibiotic tetracycline1. In a present study we demonstrate the relationship of bone matrix formation by osteoblasts as a function of cell seeding density, and time, on calcium phosphate ceramic thin film substrates.

Materials and Methods

Substrata. Commercially available 3x1 inch quartz Osteologic™ chamber slides (Millenium Biologix Inc., Kingston, Ont.) previously shown to support bone growth, were used as substrates for osteoblastic cell growth2. These chamber slides are coated with a sintered sub-micron calcium phosphate ceramic thin film, and are fitted with a removable 16 well superstructure.

Rat Bone Marrow Cell Culture. Bone marrow cells from young adult male Wistar rat (115-120g) femora were cultured according to a previously described protocol. Each well of each Osteologic™ chamber slide (14/16 wells; 2 wells contained media, but without cells, to provide background for tetracycline fluorescence) was inoculated with a 250 μl of cell suspension.

Type
Biomaterials
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 936 - 937
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Todescan, R. et al., J. Biomed. Mat. Res. (1998).(submitted)Google Scholar
2.Qui, Q. et al., Cells and Materials 3 (1993)351.Google Scholar
3.Davies, et al., Cells and Materials l(1991a)3.Google Scholar
4. Acknowledgements. The Osteologic™ slides were kindly donated by Millenium Biologix Inc. Financial Support from both the Canadian Space Agency and the Natural Sciences and Engineering Research Council Canada) to JED are greatfully acknowledged.Google Scholar