{"doi":"10.14814/phy2.14799","title":"A collagen extraction and deuterium oxide stable isotope tracer method for the quantification of bone collagen synthesis rates <i>in vivo</i>","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.","journal":"Physiological Reports","year":2021,"id":203446,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9584,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":784658,"name":"Matthew S. Brook","orcid":"0000-0002-9245-3095","position":1,"is_corresponding":false},{"id":784659,"name":"Kirsty J. Elliott‐Sale","orcid":"0000-0003-1122-5099","position":2,"is_corresponding":false},{"id":784660,"name":"Lívia Santos","orcid":"0000-0002-1915-6780","position":3,"is_corresponding":false},{"id":784661,"name":"Ian Varley","orcid":"0000-0002-3607-8921","position":4,"is_corresponding":false},{"id":777447,"name":"Sanna Lensu","orcid":"0000-0001-8197-7590","position":5,"is_corresponding":false},{"id":570893,"name":"Heikki Kainulainen","orcid":"0000-0001-5102-4350","position":6,"is_corresponding":false},{"id":252436,"name":"Lauren G. Koch","orcid":"0000-0002-2019-5939","position":7,"is_corresponding":false},{"id":348346,"name":"Steven L. Britton","orcid":"0000-0002-7372-8671","position":8,"is_corresponding":false},{"id":760735,"name":"Daniel J. Wilkinson","orcid":"0000-0001-8808-8243","position":9,"is_corresponding":false},{"id":513582,"name":"Kenneth Smith","orcid":"0000-0001-8971-6635","position":10,"is_corresponding":false},{"id":784662,"name":"Craig Sale","orcid":"0000-0002-5816-4169","position":11,"is_corresponding":false},{"id":760736,"name":"Philip J. Atherton","orcid":"0000-0002-7286-046X","position":12,"is_corresponding":false},{"id":785244,"name":"Rita Civil","orcid":null,"position":0,"is_corresponding":true}],"reference_count":33,"raw_metadata":null,"created_at":"2026-07-18T23:51:18.050175Z","pmid":"34042295","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}