Silva-Junior Duilio Benicio E, Ribeiro Ney Robson Bezerra, Junior David Martins Nunes, Tardivo Lucas Fernando Presa, Peixoto Rodrigo Chaves, Teixeira Lucas Novaes, Hammock Bruce D, Clemente-Napimoga Juliana Trindade, Napimoga Marcelo Henrique, Abdalla Henrique Ballassini
Faculdade São Leopoldo Mandic, Campinas, Brazil.
Department of Entomology and nematology and UCD Comprehensive Cancer Center, University of California, Davis, CA, United States of America.
Biochim Biophys Acta Mol Cell Biol Lipids. 2025 Oct;1870(7):159669. doi: 10.1016/j.bbalip.2025.159669. Epub 2025 Jul 31.
EpFAs are crucial mediators in resolving inflammation and regulating various biological processes. However, their activity is constrained by the rapid metabolism mediated by the soluble epoxide hydrolase (sEH), which converts EpFAs into inactive or even pro-inflammatory diols. Nevertheless, the specific effects of soluble epoxide hydrolase inhibition (sEHI) and EpFAs on osteogenic cell metabolism remain unclear. Cultures of the human immortalized osteoblast-like cell line (SAOS-2) were treated with varying concentrations (0.1-10 μM) of the potent sEHI TPPU or EpFAs (epoxyeicosatrienoic acids [EETs], epoxydocosapentaenoic acids [EDPs], and epoxyeicosatetraenoic acids [EEQs], derived from arachidonic acid [ARA], eicosapentaenoic acid [EPA], and docosahexaenoic acid [DHA], respectively). Cellular metabolic activity and proliferation were evaluated. Osteogenic potential was assessed through alkaline phosphatase activity, mineral nodule formation, and the expression of osteogenic markers, including Runx-2, Osx, Col1, Bsp, Opg, Ocn, Opn, and sEH. Treatment with TPPU and EpFAs enhanced cellular metabolic activity during the first 48 h without affecting proliferation. Alkaline phosphatase activity and mineral nodule formation assays revealed that TPPU significantly stimulated osteogenic differentiation, while EpFAs, particularly EETs, EEQs, and EDPs, promoted osteogenesis predominantly at later stages. Furthermore, TPPU modulated the expression of key osteogenic markers, enhancing differentiation. Notably, EDPs were found to disrupt the synergistic effects between sEHI and EpFAs during the mineralization process. These findings suggest that sEHI enhances mineralization and may facilitate tissue regeneration in vitro. The differential effects of EpFAs and their interplay with sEHI provide insights into potential therapeutic strategies for bone tissue engineering and regeneration.
环氧脂肪酸（EpFAs）是解决炎症和调节各种生物过程的关键介质。然而，它们的活性受到可溶性环氧化物水解酶（sEH）介导的快速代谢的限制，sEH会将EpFAs转化为无活性甚至促炎的二醇。尽管如此，可溶性环氧化物水解酶抑制（sEHI）和EpFAs对成骨细胞代谢的具体影响仍不清楚。用人永生化成骨细胞样细胞系（SAOS-2）培养物，分别用不同浓度（0.1-10 μM）的强效sEHI TPPU或EpFAs（分别衍生自花生四烯酸[ARA]、二十碳五烯酸[EPA]和二十二碳六烯酸[DHA]的环氧二十碳三烯酸[EETs]、环氧二十二碳五烯酸[EDPs]和环氧二十碳四烯酸[EEQs]）处理。评估细胞代谢活性和增殖情况。通过碱性磷酸酶活性、矿化结节形成以及成骨标志物（包括Runx-2、Osx、Col1、Bsp、Opg、Ocn、Opn和sEH）的表达来评估成骨潜力。用TPPU和EpFAs处理在最初48小时内增强了细胞代谢活性，但不影响增殖。碱性磷酸酶活性和矿化结节形成试验表明，TPPU显著刺激成骨分化，而EpFAs，特别是EETs、EEQs和EDPs，主要在后期促进成骨。此外，TPPU调节关键成骨标志物的表达，增强分化。值得注意的是，发现EDPs在矿化过程中破坏了sEHI和EpFAs之间的协同作用。这些发现表明，sEHI增强矿化作用，并可能在体外促进组织再生。EpFAs的不同作用及其与sEHI的相互作用为骨组织工程和再生的潜在治疗策略提供了见解。
