Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts
•1,25(OH)2D3 reduces the mechanical loading-induced NO response in primary human osteoblasts•Mechanical loading increases mRNA levels of CYP27B1 in primary human osteoblasts•The conversion rate of 25(OH)D3 to 1,25(OH)2D3 is not affected by mechanical loading in our model•Mechanical loading reduces m...
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| Veröffentlicht in: | The Journal of steroid biochemistry and molecular biology 2016-02, Vol.156, p.32-39 |
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| creator | van der Meijden, K. Bakker, A.D. van Essen, H.W. Heijboer, A.C. Schulten, E.A.J.M. Lips, P. Bravenboer, N. |
| description | •1,25(OH)2D3 reduces the mechanical loading-induced NO response in primary human osteoblasts•Mechanical loading increases mRNA levels of CYP27B1 in primary human osteoblasts•The conversion rate of 25(OH)D3 to 1,25(OH)2D3 is not affected by mechanical loading in our model•Mechanical loading reduces mRNA levels of VDR in primary human osteoblasts
The metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D) is synthesized from its precursor 25-hydroxyvitamin D (25(OH)D) by human osteoblasts leading to stimulation of osteoblast differentiation in an autocrine or paracrine way. Osteoblast differentiation is also stimulated by mechanical loading through activation of various responses in bone cells such as nitric oxide signaling. Whether mechanical loading affects osteoblast differentiation through an enhanced synthesis of 1,25(OH)2D by human osteoblasts is still unknown. We hypothesized that mechanical loading stimulates the synthesis of 1,25(OH)2D from 25(OH)D in primary human osteoblasts. Since the responsiveness of bone to mechanical stimuli can be altered by various endocrine factors, we also investigated whether 1,25(OH)2D or 25(OH)D affect the response of primary human osteoblasts to mechanical loading.
Primary human osteoblasts were pre-incubated in medium with/without 25(OH)D3 (400nM) or 1,25(OH)2D3 (100nM) for 24h and subjected to mechanical loading by pulsatile fluid flow (PFF). The response of osteoblasts to PFF was quantified by measuring nitric oxide, and by PCR analysis. The effect of PFF on the synthesis of 1,25(OH)2D3 was determined by subjecting osteoblasts to PFF followed by 24h post-incubation in medium with/without 25(OH)D3 (400nM).
We showed that 1,25(OH)2D3 reduced the PFF-induced NO response in primary human osteoblasts. 25(OH)D3 did not significantly alter the NO response of primary human osteoblasts to PFF, but 25(OH)D3 increased osteocalcin and RANKL mRNA levels, similar to 1,25(OH)2D3. PFF did not increase 1,25(OH)2D3 amounts in our model, even though PFF did increase CYP27B1 mRNA levels and reduced VDR mRNA levels. CYP24 mRNA levels were not affected by PFF, but were strongly increased by both 25(OH)D3 and 1,25(OH)2D3.
In conclusion, 1,25(OH)2D3 may affect the response of primary human osteoblasts to mechanical stimuli, at least with respect to NO production. Mechanical stimuli may affect local vitamin D metabolism in primary human osteoblasts. Our results suggest that 1,25(OH)2D3 and mechanical loading, both stimuli of the differentiation of |
| doi_str_mv | 10.1016/j.jsbmb.2015.11.014 |
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The metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D) is synthesized from its precursor 25-hydroxyvitamin D (25(OH)D) by human osteoblasts leading to stimulation of osteoblast differentiation in an autocrine or paracrine way. Osteoblast differentiation is also stimulated by mechanical loading through activation of various responses in bone cells such as nitric oxide signaling. Whether mechanical loading affects osteoblast differentiation through an enhanced synthesis of 1,25(OH)2D by human osteoblasts is still unknown. We hypothesized that mechanical loading stimulates the synthesis of 1,25(OH)2D from 25(OH)D in primary human osteoblasts. Since the responsiveness of bone to mechanical stimuli can be altered by various endocrine factors, we also investigated whether 1,25(OH)2D or 25(OH)D affect the response of primary human osteoblasts to mechanical loading.
Primary human osteoblasts were pre-incubated in medium with/without 25(OH)D3 (400nM) or 1,25(OH)2D3 (100nM) for 24h and subjected to mechanical loading by pulsatile fluid flow (PFF). The response of osteoblasts to PFF was quantified by measuring nitric oxide, and by PCR analysis. The effect of PFF on the synthesis of 1,25(OH)2D3 was determined by subjecting osteoblasts to PFF followed by 24h post-incubation in medium with/without 25(OH)D3 (400nM).
We showed that 1,25(OH)2D3 reduced the PFF-induced NO response in primary human osteoblasts. 25(OH)D3 did not significantly alter the NO response of primary human osteoblasts to PFF, but 25(OH)D3 increased osteocalcin and RANKL mRNA levels, similar to 1,25(OH)2D3. PFF did not increase 1,25(OH)2D3 amounts in our model, even though PFF did increase CYP27B1 mRNA levels and reduced VDR mRNA levels. CYP24 mRNA levels were not affected by PFF, but were strongly increased by both 25(OH)D3 and 1,25(OH)2D3.
In conclusion, 1,25(OH)2D3 may affect the response of primary human osteoblasts to mechanical stimuli, at least with respect to NO production. Mechanical stimuli may affect local vitamin D metabolism in primary human osteoblasts. Our results suggest that 1,25(OH)2D3 and mechanical loading, both stimuli of the differentiation of osteoblasts, interact at the cellular level.</description><identifier>ISSN: 0960-0760</identifier><identifier>EISSN: 1879-1220</identifier><identifier>DOI: 10.1016/j.jsbmb.2015.11.014</identifier><identifier>PMID: 26625962</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>1,25-Dihydroxyvitamin D3 ; 25-Hydroxyvitamin D3 ; 25-Hydroxyvitamin D3 1-alpha-Hydroxylase - genetics ; 25-Hydroxyvitamin D3 1-alpha-Hydroxylase - metabolism ; Adult ; Calcitriol - metabolism ; Cells, Cultured ; CYP27B1 ; Female ; Humans ; Male ; Mechanical loading ; Nitric Oxide - metabolism ; Osteoblasts - cytology ; Osteoblasts - metabolism ; Primary human osteoblasts ; Receptors, Calcitriol - metabolism ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Vitamin D - analogs & derivatives ; Vitamin D - metabolism ; Weight-Bearing</subject><ispartof>The Journal of steroid biochemistry and molecular biology, 2016-02, Vol.156, p.32-39</ispartof><rights>2015 Elsevier Ltd</rights><rights>Copyright © 2015 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c274t-e461c15583245f1e39a801686d9fb988284192a044bd53e1e558623a8d1593cb3</citedby><cites>FETCH-LOGICAL-c274t-e461c15583245f1e39a801686d9fb988284192a044bd53e1e558623a8d1593cb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0960076015301400$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26625962$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>van der Meijden, K.</creatorcontrib><creatorcontrib>Bakker, A.D.</creatorcontrib><creatorcontrib>van Essen, H.W.</creatorcontrib><creatorcontrib>Heijboer, A.C.</creatorcontrib><creatorcontrib>Schulten, E.A.J.M.</creatorcontrib><creatorcontrib>Lips, P.</creatorcontrib><creatorcontrib>Bravenboer, N.</creatorcontrib><title>Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts</title><title>The Journal of steroid biochemistry and molecular biology</title><addtitle>J Steroid Biochem Mol Biol</addtitle><description>•1,25(OH)2D3 reduces the mechanical loading-induced NO response in primary human osteoblasts•Mechanical loading increases mRNA levels of CYP27B1 in primary human osteoblasts•The conversion rate of 25(OH)D3 to 1,25(OH)2D3 is not affected by mechanical loading in our model•Mechanical loading reduces mRNA levels of VDR in primary human osteoblasts
The metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D) is synthesized from its precursor 25-hydroxyvitamin D (25(OH)D) by human osteoblasts leading to stimulation of osteoblast differentiation in an autocrine or paracrine way. Osteoblast differentiation is also stimulated by mechanical loading through activation of various responses in bone cells such as nitric oxide signaling. Whether mechanical loading affects osteoblast differentiation through an enhanced synthesis of 1,25(OH)2D by human osteoblasts is still unknown. We hypothesized that mechanical loading stimulates the synthesis of 1,25(OH)2D from 25(OH)D in primary human osteoblasts. Since the responsiveness of bone to mechanical stimuli can be altered by various endocrine factors, we also investigated whether 1,25(OH)2D or 25(OH)D affect the response of primary human osteoblasts to mechanical loading.
Primary human osteoblasts were pre-incubated in medium with/without 25(OH)D3 (400nM) or 1,25(OH)2D3 (100nM) for 24h and subjected to mechanical loading by pulsatile fluid flow (PFF). The response of osteoblasts to PFF was quantified by measuring nitric oxide, and by PCR analysis. The effect of PFF on the synthesis of 1,25(OH)2D3 was determined by subjecting osteoblasts to PFF followed by 24h post-incubation in medium with/without 25(OH)D3 (400nM).
We showed that 1,25(OH)2D3 reduced the PFF-induced NO response in primary human osteoblasts. 25(OH)D3 did not significantly alter the NO response of primary human osteoblasts to PFF, but 25(OH)D3 increased osteocalcin and RANKL mRNA levels, similar to 1,25(OH)2D3. PFF did not increase 1,25(OH)2D3 amounts in our model, even though PFF did increase CYP27B1 mRNA levels and reduced VDR mRNA levels. CYP24 mRNA levels were not affected by PFF, but were strongly increased by both 25(OH)D3 and 1,25(OH)2D3.
In conclusion, 1,25(OH)2D3 may affect the response of primary human osteoblasts to mechanical stimuli, at least with respect to NO production. Mechanical stimuli may affect local vitamin D metabolism in primary human osteoblasts. Our results suggest that 1,25(OH)2D3 and mechanical loading, both stimuli of the differentiation of osteoblasts, interact at the cellular level.</description><subject>1,25-Dihydroxyvitamin D3</subject><subject>25-Hydroxyvitamin D3</subject><subject>25-Hydroxyvitamin D3 1-alpha-Hydroxylase - genetics</subject><subject>25-Hydroxyvitamin D3 1-alpha-Hydroxylase - metabolism</subject><subject>Adult</subject><subject>Calcitriol - metabolism</subject><subject>Cells, Cultured</subject><subject>CYP27B1</subject><subject>Female</subject><subject>Humans</subject><subject>Male</subject><subject>Mechanical loading</subject><subject>Nitric Oxide - metabolism</subject><subject>Osteoblasts - cytology</subject><subject>Osteoblasts - metabolism</subject><subject>Primary human osteoblasts</subject><subject>Receptors, Calcitriol - metabolism</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Vitamin D - analogs & derivatives</subject><subject>Vitamin D - metabolism</subject><subject>Weight-Bearing</subject><issn>0960-0760</issn><issn>1879-1220</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMlOwzAURS0EgjJ8ARLyskgk-NmxEy9YoDIUqagbWFuO80JdZYA4QeLvSWlhyepuzhvuIeQcWAwM1PU6Xoe8zmPOQMYAMYNkj0wgS3UEnLN9MmFasYilih2R4xDWjDEhID0kR1wpLrXiE7J8RreyjXe2olVrC9-8UdsUtF8hDV_NGMEH2pYUrricLueX_I76hr53vrbdF10NtW1oG3ps88qGPpySg9JWAc92eUJeH-5fZvNosXx8mt0uIsfTpI8wUeBAykzwRJaAQtts7JSpQpe5zjKeJaC5ZUmSF1Ig4IgqLmxWgNTC5eKETLd737v2Y8DQm9oHh1VlG2yHYCCVggsmNRtRsUVd14bQYWl23xtgZmPSrM2PSbMxaQDMaHKcutgdGPIai7-ZX3UjcLMFcKz56bEzwXlsHBa-Q9ebovX_HvgGUdKCiw</recordid><startdate>201602</startdate><enddate>201602</enddate><creator>van der Meijden, K.</creator><creator>Bakker, A.D.</creator><creator>van Essen, H.W.</creator><creator>Heijboer, A.C.</creator><creator>Schulten, E.A.J.M.</creator><creator>Lips, P.</creator><creator>Bravenboer, N.</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201602</creationdate><title>Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts</title><author>van der Meijden, K. ; Bakker, A.D. ; van Essen, H.W. ; Heijboer, A.C. ; Schulten, E.A.J.M. ; Lips, P. ; Bravenboer, N.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c274t-e461c15583245f1e39a801686d9fb988284192a044bd53e1e558623a8d1593cb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>1,25-Dihydroxyvitamin D3</topic><topic>25-Hydroxyvitamin D3</topic><topic>25-Hydroxyvitamin D3 1-alpha-Hydroxylase - genetics</topic><topic>25-Hydroxyvitamin D3 1-alpha-Hydroxylase - metabolism</topic><topic>Adult</topic><topic>Calcitriol - metabolism</topic><topic>Cells, Cultured</topic><topic>CYP27B1</topic><topic>Female</topic><topic>Humans</topic><topic>Male</topic><topic>Mechanical loading</topic><topic>Nitric Oxide - metabolism</topic><topic>Osteoblasts - cytology</topic><topic>Osteoblasts - metabolism</topic><topic>Primary human osteoblasts</topic><topic>Receptors, Calcitriol - metabolism</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Vitamin D - analogs & derivatives</topic><topic>Vitamin D - metabolism</topic><topic>Weight-Bearing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>van der Meijden, K.</creatorcontrib><creatorcontrib>Bakker, A.D.</creatorcontrib><creatorcontrib>van Essen, H.W.</creatorcontrib><creatorcontrib>Heijboer, A.C.</creatorcontrib><creatorcontrib>Schulten, E.A.J.M.</creatorcontrib><creatorcontrib>Lips, P.</creatorcontrib><creatorcontrib>Bravenboer, N.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of steroid biochemistry and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>van der Meijden, K.</au><au>Bakker, A.D.</au><au>van Essen, H.W.</au><au>Heijboer, A.C.</au><au>Schulten, E.A.J.M.</au><au>Lips, P.</au><au>Bravenboer, N.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts</atitle><jtitle>The Journal of steroid biochemistry and molecular biology</jtitle><addtitle>J Steroid Biochem Mol Biol</addtitle><date>2016-02</date><risdate>2016</risdate><volume>156</volume><spage>32</spage><epage>39</epage><pages>32-39</pages><issn>0960-0760</issn><eissn>1879-1220</eissn><abstract>•1,25(OH)2D3 reduces the mechanical loading-induced NO response in primary human osteoblasts•Mechanical loading increases mRNA levels of CYP27B1 in primary human osteoblasts•The conversion rate of 25(OH)D3 to 1,25(OH)2D3 is not affected by mechanical loading in our model•Mechanical loading reduces mRNA levels of VDR in primary human osteoblasts
The metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D) is synthesized from its precursor 25-hydroxyvitamin D (25(OH)D) by human osteoblasts leading to stimulation of osteoblast differentiation in an autocrine or paracrine way. Osteoblast differentiation is also stimulated by mechanical loading through activation of various responses in bone cells such as nitric oxide signaling. Whether mechanical loading affects osteoblast differentiation through an enhanced synthesis of 1,25(OH)2D by human osteoblasts is still unknown. We hypothesized that mechanical loading stimulates the synthesis of 1,25(OH)2D from 25(OH)D in primary human osteoblasts. Since the responsiveness of bone to mechanical stimuli can be altered by various endocrine factors, we also investigated whether 1,25(OH)2D or 25(OH)D affect the response of primary human osteoblasts to mechanical loading.
Primary human osteoblasts were pre-incubated in medium with/without 25(OH)D3 (400nM) or 1,25(OH)2D3 (100nM) for 24h and subjected to mechanical loading by pulsatile fluid flow (PFF). The response of osteoblasts to PFF was quantified by measuring nitric oxide, and by PCR analysis. The effect of PFF on the synthesis of 1,25(OH)2D3 was determined by subjecting osteoblasts to PFF followed by 24h post-incubation in medium with/without 25(OH)D3 (400nM).
We showed that 1,25(OH)2D3 reduced the PFF-induced NO response in primary human osteoblasts. 25(OH)D3 did not significantly alter the NO response of primary human osteoblasts to PFF, but 25(OH)D3 increased osteocalcin and RANKL mRNA levels, similar to 1,25(OH)2D3. PFF did not increase 1,25(OH)2D3 amounts in our model, even though PFF did increase CYP27B1 mRNA levels and reduced VDR mRNA levels. CYP24 mRNA levels were not affected by PFF, but were strongly increased by both 25(OH)D3 and 1,25(OH)2D3.
In conclusion, 1,25(OH)2D3 may affect the response of primary human osteoblasts to mechanical stimuli, at least with respect to NO production. Mechanical stimuli may affect local vitamin D metabolism in primary human osteoblasts. Our results suggest that 1,25(OH)2D3 and mechanical loading, both stimuli of the differentiation of osteoblasts, interact at the cellular level.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>26625962</pmid><doi>10.1016/j.jsbmb.2015.11.014</doi><tpages>8</tpages></addata></record> |
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| subjects | 1,25-Dihydroxyvitamin D3 25-Hydroxyvitamin D3 25-Hydroxyvitamin D3 1-alpha-Hydroxylase - genetics 25-Hydroxyvitamin D3 1-alpha-Hydroxylase - metabolism Adult Calcitriol - metabolism Cells, Cultured CYP27B1 Female Humans Male Mechanical loading Nitric Oxide - metabolism Osteoblasts - cytology Osteoblasts - metabolism Primary human osteoblasts Receptors, Calcitriol - metabolism RNA, Messenger - genetics RNA, Messenger - metabolism Vitamin D - analogs & derivatives Vitamin D - metabolism Weight-Bearing |
| title | Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts |
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