A1 Journal article (refereed)
A collagen extraction and deuterium oxide stable isotope tracer method for the quantification of bone collagen synthesis rates in vivo (2021)

Civil, R., Brook, M. S., Elliott‐Sale, K. J., Santos, L., Varley, I., Lensu, S., Kainulainen, H., Koch, L. G., Britton, S. L., Wilkinson, D. J., Smith, K., Sale, C., & Atherton, P. J. (2021). A collagen extraction and deuterium oxide stable isotope tracer method for the quantification of bone collagen synthesis rates in vivo. Physiological Reports, 9(10), Article e14799. https://doi.org/10.14814/phy2.14799

JYU authors or editors

Publication details

All authors or editors: Civil, Rita; Brook, Matthew S.; Elliott‐Sale, Kirsty J.; Santos, Lívia; Varley, Ian; Lensu, Sanna; Kainulainen, Heikki; Koch, Lauren G.; Britton, Steven L.; Wilkinson, Daniel J.; et al.

Journal or series: Physiological Reports

ISSN: 2051-817X

eISSN: 2051-817X

Publication year: 2021

Volume: 9

Issue number: 10

Article number: e14799

Publisher: John Wiley & Sons

Publication country: United Kingdom

Publication language: English

DOI: https://doi.org/10.14814/phy2.14799

Publication open access: Openly available

Publication channel open access: Open Access channel

Publication is parallel published (JYX): https://jyx.jyu.fi/handle/123456789/76082

Abstract

The development of safe and practical strategies to prevent weakening of bone tissue is vital, yet attempts to achieve this have been hindered by a lack of understanding of the short-term (days-weeks) physiology of bone collagen turnover. To address this, we have developed a method to quantify bone collagen synthesis in vivo, using deuterium oxide (D2O) tracer incorporation techniques combined with gas chromatography pyrolysis isotope-ratio mass spectrometry (GC-pyrolysis-IRMS). Forty-six male and female rats from a selectively bred model ingested D2O for 3 weeks. Femur diaphyses (FEM), tibia proximal (T-PRO), and distal (T-DIS) epiphyses-metaphyses and tibia mid-shaft diaphyses (T-MID) were obtained from all rats after necropsy. After demineralisation, collagen proteins were isolated and hydrolysed and collagen fractional synthetic rates (FSRs) determined by incorporation of deuterium into protein-bound alanine via GC-pyrolysis-IRMS. The collagen FSR for the FEM (0.131 ± 0.078%/day; 95% CI [0.106–0.156]) was greater than the FSR at T-MID (0.055 ± 0.049%/day; 95% CI [0.040–0.070]; p < 0.001). The T-PRO site had the highest FSR (0.203 ± 0.123%/day; 95% CI [0.166–0.241]) and T-DIS the lowest (0.027 ± 0.015%/day; 95% CI [0.022–0.031]). The three tibial sites exhibited different FSRs (p < 0.001). Herein, we have developed a sensitive method to quantify in vivo bone collagen synthesis and identified site-specific rates of synthesis, which could be applicable to studies of human bone collagen turnover.

Keywords: bone; bony tissues; regeneration (biology); collagens; biomarkers; isotope analysis

Free keywords: bone turnover; collagen synthesis; deuterium oxide; GC-pyrolysis-IRMS; stable isotopes

Contributing organizations

Ministry reporting: Yes

VIRTA submission year: 2021

JUFO rating: 1