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Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells
OBJECTIVES: Untargeted metabolomics analyses by our group have shown that plasma linoleic acid (LA) was inversely associated with bone mineral density Z-score and that bone formation indices were associated with energy-generating metabolic pathways, including fatty acid b-oxidation, in adult cohorts...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9193374/ http://dx.doi.org/10.1093/cdn/nzac057.018 |
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author | Nesbeth, Paula-Dene Ziegler, Thomas Weiss, Daiana Hao, Li Smith, Matthew Jones, Dean Weitzmann, M Neale Alvarez, Jessica |
author_facet | Nesbeth, Paula-Dene Ziegler, Thomas Weiss, Daiana Hao, Li Smith, Matthew Jones, Dean Weitzmann, M Neale Alvarez, Jessica |
author_sort | Nesbeth, Paula-Dene |
collection | PubMed |
description | OBJECTIVES: Untargeted metabolomics analyses by our group have shown that plasma linoleic acid (LA) was inversely associated with bone mineral density Z-score and that bone formation indices were associated with energy-generating metabolic pathways, including fatty acid b-oxidation, in adult cohorts. Here, we examined the effect of increasing LA concentrations on osteoblast precursor cell bioenergetics and osteoblast differentiation to determine whether high LA is detrimental to bone formation. METHODS: We treated MC3T3-E1 pre-osteoblastic cells with 0 µM (control), 1 μM, and 50 μM LA cultured in osteogenic differentiation media supplemented with 50 µM L-ascorbic acid and 2 mM β-glycerophosphate. To assess the effect of LA on early commitment/differentiation, cells were stained for alkaline phosphatase activity and late differentiation using Alizarin Red S staining for mineral deposition, at 7 and 18 days, respectively. To assess cellular bioenergetics, real-time ATP production rates in LA treated (1 or 50 μM) and control MC3T3-E1 cells were measured using an extracellular flux analyzer after 24 hours (normalized for total protein content). Differences in bioenergetic values were determined using one-way ANOVA or Kruskal-Wallis test with Tukey's HSD or Dunn's post hoc tests. RESULTS: While LA had no effect on late differentiation/mineralizing activity of MC3T3 cells, LA dose-dependently decreased commitment/early differentiation. LA also significantly altered the bioenergetic profile of MC3T3-E1 cells by decreasing basal oxygen consumption rate (P < 0.001), as well as mitochondrial and total ATP production rate (P < 0.05 and P < 0.001, respectively). There were no significant changes in glycolytic ATP production rate. CONCLUSIONS: Osteoblast differentiation is a highly bioenergetic process, and this study suggests excess LA may impair ATP production from oxidative phosphorylation. This, in turn, may impede commitment and early differentiation of osteoblasts. Our study supports further clinical and translational investigation into the role of LA and energy metabolism in osteoblast function, as well as bone formation. FUNDING SOURCES: National Institutes of Health. |
format | Online Article Text |
id | pubmed-9193374 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-91933742022-06-14 Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells Nesbeth, Paula-Dene Ziegler, Thomas Weiss, Daiana Hao, Li Smith, Matthew Jones, Dean Weitzmann, M Neale Alvarez, Jessica Curr Dev Nutr Energy and Macronutrient Metabolism OBJECTIVES: Untargeted metabolomics analyses by our group have shown that plasma linoleic acid (LA) was inversely associated with bone mineral density Z-score and that bone formation indices were associated with energy-generating metabolic pathways, including fatty acid b-oxidation, in adult cohorts. Here, we examined the effect of increasing LA concentrations on osteoblast precursor cell bioenergetics and osteoblast differentiation to determine whether high LA is detrimental to bone formation. METHODS: We treated MC3T3-E1 pre-osteoblastic cells with 0 µM (control), 1 μM, and 50 μM LA cultured in osteogenic differentiation media supplemented with 50 µM L-ascorbic acid and 2 mM β-glycerophosphate. To assess the effect of LA on early commitment/differentiation, cells were stained for alkaline phosphatase activity and late differentiation using Alizarin Red S staining for mineral deposition, at 7 and 18 days, respectively. To assess cellular bioenergetics, real-time ATP production rates in LA treated (1 or 50 μM) and control MC3T3-E1 cells were measured using an extracellular flux analyzer after 24 hours (normalized for total protein content). Differences in bioenergetic values were determined using one-way ANOVA or Kruskal-Wallis test with Tukey's HSD or Dunn's post hoc tests. RESULTS: While LA had no effect on late differentiation/mineralizing activity of MC3T3 cells, LA dose-dependently decreased commitment/early differentiation. LA also significantly altered the bioenergetic profile of MC3T3-E1 cells by decreasing basal oxygen consumption rate (P < 0.001), as well as mitochondrial and total ATP production rate (P < 0.05 and P < 0.001, respectively). There were no significant changes in glycolytic ATP production rate. CONCLUSIONS: Osteoblast differentiation is a highly bioenergetic process, and this study suggests excess LA may impair ATP production from oxidative phosphorylation. This, in turn, may impede commitment and early differentiation of osteoblasts. Our study supports further clinical and translational investigation into the role of LA and energy metabolism in osteoblast function, as well as bone formation. FUNDING SOURCES: National Institutes of Health. Oxford University Press 2022-06-14 /pmc/articles/PMC9193374/ http://dx.doi.org/10.1093/cdn/nzac057.018 Text en © The Author 2022. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Energy and Macronutrient Metabolism Nesbeth, Paula-Dene Ziegler, Thomas Weiss, Daiana Hao, Li Smith, Matthew Jones, Dean Weitzmann, M Neale Alvarez, Jessica Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title | Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title_full | Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title_fullStr | Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title_full_unstemmed | Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title_short | Linoleic Acid Reduces Oxidative Phosphorylation and Impairs Early Differentiation of MC3T3-E1 Osteoblast Precursor Cells |
title_sort | linoleic acid reduces oxidative phosphorylation and impairs early differentiation of mc3t3-e1 osteoblast precursor cells |
topic | Energy and Macronutrient Metabolism |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9193374/ http://dx.doi.org/10.1093/cdn/nzac057.018 |
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