W J Rody1, O Krokhin2, V Spicer2, C A Chamberlain3, M Chamberlain4, K P McHugh5, S M Wallet4, A K Emory1, J D Crull6, L S Holliday1. 1. Department of Orthodontics, University of Florida, Gainesville, FL, USA. 2. Manitoba Center for Proteomics, Winnipeg, MB, Canada. 3. Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, FL, USA. 4. Department of Oral Biology, University of Florida, Gainesville, FL, USA. 5. Department of Periodontology, University of Florida, Gainesville, FL, USA. 6. Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, USA.
Abstract
OBJECTIVES: 1) To test the hypothesis that there would be proteomic differences in the composition of exosomes isolated from osteoclasts and odontoclasts and 2) to determine the clinical usefulness of these in vitro biomarker candidates. MATERIALS AND METHODS: Mouse bone marrow-derived precursors were cultured on either dentin or bone slices and allowed to mature and begin resorption. Exosomes were isolated from cell culture media and characterized by mass spectrometry. The proteomic data obtained from this in vitro study were compared with the data obtained from human samples in our previous work. RESULTS: There was a difference in the proteomic composition of exosomes from osteoclasts and odontoclasts. A total of 40 exosomal proteins were only present in osteoclast media, whereas six unique exosomal proteins were identified in odontoclast supernatants. Approximately 50% of exosomal proteins released by clastic cells in vitro can be found in oral fluids. CONCLUSION: Our data suggest that the mineralized matrix type plays a role in the final phenotypic characteristics of mouse clastic cells. Many in vitro biomarker candidates of bone and dentin resorption can also be found in human oral fluids, thus indicating that this approach may be a viable alternative in biomarker discovery.
OBJECTIVES: 1) To test the hypothesis that there would be proteomic differences in the composition of exosomes isolated from osteoclasts and odontoclasts and 2) to determine the clinical usefulness of these in vitro biomarker candidates. MATERIALS AND METHODS:Mouse bone marrow-derived precursors were cultured on either dentin or bone slices and allowed to mature and begin resorption. Exosomes were isolated from cell culture media and characterized by mass spectrometry. The proteomic data obtained from this in vitro study were compared with the data obtained from human samples in our previous work. RESULTS: There was a difference in the proteomic composition of exosomes from osteoclasts and odontoclasts. A total of 40 exosomal proteins were only present in osteoclast media, whereas six unique exosomal proteins were identified in odontoclast supernatants. Approximately 50% of exosomal proteins released by clastic cells in vitro can be found in oral fluids. CONCLUSION: Our data suggest that the mineralized matrix type plays a role in the final phenotypic characteristics of mouse clastic cells. Many in vitro biomarker candidates of bone and dentin resorption can also be found in human oral fluids, thus indicating that this approach may be a viable alternative in biomarker discovery.
Authors: Wellington J Rody; Casey A Chamberlain; Alyssa K Emory-Carter; Kevin P McHugh; Shannon M Wallet; Victor Spicer; Oleg Krokhin; L Shannon Holliday Journal: PLoS One Date: 2019-07-10 Impact factor: 3.240
Authors: Henrik Löfvall; Hannah Newbould; Morten A Karsdal; Morten H Dziegiel; Johan Richter; Kim Henriksen; Christian S Thudium Journal: Arthritis Res Ther Date: 2018-04-10 Impact factor: 5.156