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Structural and biomechanical analysis of femurs from mice treated with diclofenac, miR-15b and miR-365

cris.virtual.author-orcid0000-0002-7182-6905
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cris.virtual.author-orcid0000-0003-0715-0223
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dc.abstract.enPurpose: Nonsteroidal anti-inflammatory drugs (NSAIDs) are used to treat pain, but they have side effects, including the inhibition of bone healing. Diclofenac (DF), a member of the NSAID group, affects bone health adversely. One potential approach to protect bones from the effects of NSAIDs involves the administration of short nucleic acids, such as microRNAs (miRNAs). This study aimed to determine whether two specific miRNAs, miR-15b and miR-365, could mitigate the effects of DF on bone. Methods: We used the C57BL/6J mouse strain and the MC3T3-E1 preosteoblast cell line derived from this mouse strain. Female C57BL/6J mice were treated with DF and miR-15b or miR-365 mimics. After euthanising the mice, we analysed their femurs using micro-computed tomography (μCT) and dynamic mechanical analysis (DMA). In addition, we performed experiments in cultured MC3T3-E1 cells, which were transfected with either miR- 15b or miR-365. We assessed the relative mRNA levels of osteoblast differentiation markers using real-time PCR. Results: Our findings indicated that miR-15b and miR-365 were effective in reversing the detrimental effects of DF on bone mineral density. DF decreased the bone’s storage modulus (E′), while miR-15b and miR-365 ameliorated this effect. In the preosteoblast MC3T3-E1 cells, DF did not significantly regulate marker genes; however, the administration of miR-15b and miR-365 reduced the gene expression of Runx2, Alp, and Satb2. Conclusions: In summary, the impact of DF on the structural and mechanical properties of bone was not mediated by gene regulation in osteoblasts. However, osteoblasts were responsive to the administration of miR-15b and miR-365.
dc.affiliationWydział Medycyny Weterynaryjnej i Nauk o Zwierzętach
dc.affiliation.instituteKatedra Fizjologii, Biochemii i Biostruktury Zwierząt
dc.contributor.authorLehmann, Tomasz P.
dc.contributor.authorTrzaskowska, Aleksandra
dc.contributor.authorPruszyńska-Oszmałek, Ewa
dc.contributor.authorKołodziejski, Paweł
dc.contributor.authorWojtków, Magdalena
dc.contributor.authorPezowicz, Celina
dc.contributor.authorMielcarek, Sławomir
dc.contributor.authorHertel, Agnieszka
dc.contributor.authorJagodziński, Paweł
dc.contributor.authorGłowacki, Maciej
dc.date.access2025-09-24
dc.date.accessioned2025-09-30T10:59:24Z
dc.date.available2025-09-30T10:59:24Z
dc.date.copyright2025-08-26
dc.date.issued2025
dc.description.abstract<jats:sec><jats:title>Purpose</jats:title><jats:p>Nonsteroidal anti-inflammatory drugs (NSAIDs) are used to treat pain, but they have side effects, including the inhibition of bone healing. Diclofenac (DF), a member of the NSAID group, affects bone health adversely. One potential approach to protect bones from the effects of NSAIDs involves the administration of short nucleic acids, such as microRNAs (miRNAs). This study aimed to determine whether two specific miRNAs, miR-15b and miR-365, could mitigate the effects of DF on bone.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>We used the C57BL/6J mouse strain and the MC3T3-E1 pre-osteoblast cell line derived from this mouse strain. Female C57BL/6J mice were treated with DF and miR-15b or miR-365 mimics. After euthanising the mice, we analysed their femurs using micro-computed tomography (μCT) and dynamic mechanical analysis (DMA). In addition, we performed experiments in cultured MC3T3-E1 cells, which were transfected with either miR-15b or miR-365. We assessed the relative mRNA levels of osteoblast differentiation markers using real-time PCR.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Our findings indicated that miR-15b and miR-365 were effective in reversing the detrimental effects of DF on bone mineral density. DF decreased the bone’s storage modulus (E’), while miR-15b and miR-365 ameliorated this effect. In the preosteoblast MC3T3-E1 cells, DF did not significantly regulate marker genes; however, the administration of miR-15b and miR-365 reduced the gene expression of Runx2, Alp, and Satb2.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>In summary, the impact of DF on the structural and mechanical properties of bone was not mediated by gene regulation in osteoblasts. However, osteoblasts were responsive to the administration of miR-15b and miR-365.</jats:p></jats:sec>
dc.description.accesstimeat_publication
dc.description.bibliographyil., bibliogr.
dc.description.financepublication_nocost
dc.description.financecost0,00
dc.description.if0.8
dc.description.number2
dc.description.points100
dc.description.versionfinal_published
dc.description.volume27
dc.identifier.doi10.37190/abb/207864
dc.identifier.eissn2450-6303
dc.identifier.issn1509-409X
dc.identifier.urihttps://sciencerep.up.poznan.pl/handle/item/5047
dc.identifier.weblinkhttps://sciendo.com/pl/article/10.37190/abb/207864
dc.languageen
dc.relation.ispartofActa of Bioengineering and Biomechanics
dc.relation.pages199-209
dc.rightsCC-BY-NC-ND
dc.sciencecloudnosend
dc.share.typeOPEN_JOURNAL
dc.subject.enmiRNA
dc.subject.enfemur
dc.subject.enbone microstructure
dc.subject.enpreosteoblast
dc.subject.endynamic mechanical analysis
dc.titleStructural and biomechanical analysis of femurs from mice treated with diclofenac, miR-15b and miR-365
dc.typeJournalArticle
dspace.entity.typePublication
oaire.citation.issue2
oaire.citation.volume27