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. 2018 Jan 24;8:1515. doi: 10.1038/s41598-018-19894-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Dose- and sex-dependent effects of MP on osteoclast differentiation and activity: (a,c, e) TRAP staining shows that MP increased osteoclastic differentiation in male and female bone marrow-derived preosteoclasts in a dose- and sex-dependent manner. Males showed the formation of large osteoclasts at day (a) 5 that continued to mature at days (c) 7, and (e) 14. At all time-points, male osteoclasts showed a dose-dependent increase in the number of large osteoclasts with increasing MP concentration. Female osteoclasts also showed accelerated osteoclastogenesis in response to increasing MP concentration but were significantly slower than male cells. Day 5 female osteoclast cultures showed TRAP+ cells clusters, with no large osteoclasts. The number of TRAP+ cell clusters increased at low MP doses and some larger osteoclasts were present at high MP doses at day 7. At day 14, female osteoclasts showed the more typical morphology of mature osteoclast with increasing size and number seen in higher MP doses. (b,d,f) Male and female osteoclasts both showed a dose- and sex-dependent increase in activity, with female osteoclast activity significantly less than male at days 5, 7, and 14. (g,h) Osteoclast activity as a function of time showed significant increases in osteoclast activity observed between day 5 and 14 for both male and female osteoclasts. (^@p < 0.05 0 vs 1000 ng/ml, ^@@p < 0.01 0 vs 1000 ng/ml, ^§§P < 0.01, male vs female osteoclast activity, ^ɸp < 0.05 Day 5 vs Day 7).