Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro
Chalcones are phenolic compounds produced during the biosynthesis of flavonoids that have numerous biological activities, including anti-inflammatory, antioxidant and anticancer. In this in vitro study, we investigate a newly synthesized chalcone (Chalcone T4) in the context of bone turnover, specif...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-04-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/8/7624 |
| _version_ | 1797605047599628288 |
|---|---|
| author | Iolanda Augusta Fernandes de Matos Natalie Aparecida Rodrigues Fernandes Giovani Cirelli Mariely Araújo de Godoi Letícia Ribeiro de Assis Luis Octávio Regasini Carlos Rossa Junior Morgana Rodrigues Guimarães-Stabili |
| author_facet | Iolanda Augusta Fernandes de Matos Natalie Aparecida Rodrigues Fernandes Giovani Cirelli Mariely Araújo de Godoi Letícia Ribeiro de Assis Luis Octávio Regasini Carlos Rossa Junior Morgana Rodrigues Guimarães-Stabili |
| author_sort | Iolanda Augusta Fernandes de Matos |
| collection | DOAJ |
| description | Chalcones are phenolic compounds produced during the biosynthesis of flavonoids that have numerous biological activities, including anti-inflammatory, antioxidant and anticancer. In this in vitro study, we investigate a newly synthesized chalcone (Chalcone T4) in the context of bone turnover, specifically on the modulation of osteoclast differentiation and activity and osteoblast differentiation. Murine macrophages (RAW 264.7) and pre-osteoblasts (MC3T3-E1) were used as models of osteoclasts and osteoblasts, respectively. Differentiation and activity osteoclasts were induced by RANKL in the presence and absence of non-cytotoxic concentrations of Chalcone T4, added in different periods during osteoclastogenesis. Osteoclast differentiation and activity were assessed by actin ring formation and resorption pit assay, respectively. Expression of osteoclast-specific markers (<i>Nfatc1</i>, <i>Oscar</i>, <i>Acp5</i>, <i>Mmp-9</i> and <i>Ctsk</i>) was determined by RT-qPCR, and the activation status of relevant intracellular signaling pathways (MAPK, AKT and NF-kB) by Western blot. Osteoblast differentiation and activity was induced by osteogenic culture medium in the presence and absence of the same concentrations of Chalcone T4. Outcomes assessed were the formation of mineralization nodules via alizarin red staining and the expression of osteoblast-related genes (<i>Alp</i> e <i>Runx2</i>) by RT-qPCR. Chalcone T4 reduced RANKL-induced osteoclast differentiation and activity, suppressed <i>Oscar</i>, <i>Acp5</i> and <i>Mmp-9</i> expression, and decreased ERK and AKT activation in a dose-dependent manner. <i>Nfact1</i> expression and NF-kB phosphorylation were not modulated by the compound. Mineralized matrix formation and the expression of <i>Alp</i> and <i>Runx2</i> by MC3T3-E1 cells were markedly stimulated by Chalcone T4. Collectively, these results demonstrate that Chalcone T4 inhibits in osteoclast differentiation and activity and stimulates osteogenesis, which indicates a promising therapeutic potential in osteolytic diseases. |
| first_indexed | 2024-03-11T04:55:35Z |
| format | Article |
| id | doaj.art-e833998219a7459e95566ef580465f78 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T04:55:35Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e833998219a7459e95566ef580465f782023-11-17T19:43:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01248762410.3390/ijms24087624Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In VitroIolanda Augusta Fernandes de Matos0Natalie Aparecida Rodrigues Fernandes1Giovani Cirelli2Mariely Araújo de Godoi3Letícia Ribeiro de Assis4Luis Octávio Regasini5Carlos Rossa Junior6Morgana Rodrigues Guimarães-Stabili7Department of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilDepartment of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilDepartment of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilDepartment of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilDepartment of Chemistry and Environmental Sciences, Institute of Biosciences, Humanities and Exact Sciences, São Paulo State University (UNESP), São José do Rio Preto 15054-000, SP, BrazilDepartment of Chemistry and Environmental Sciences, Institute of Biosciences, Humanities and Exact Sciences, São Paulo State University (UNESP), São José do Rio Preto 15054-000, SP, BrazilDepartment of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilDepartment of Diagnosis and Surgery, School of Dentistry of Araraquara, São Paulo State University (UNESP), Araraquara 14801-385, SP, BrazilChalcones are phenolic compounds produced during the biosynthesis of flavonoids that have numerous biological activities, including anti-inflammatory, antioxidant and anticancer. In this in vitro study, we investigate a newly synthesized chalcone (Chalcone T4) in the context of bone turnover, specifically on the modulation of osteoclast differentiation and activity and osteoblast differentiation. Murine macrophages (RAW 264.7) and pre-osteoblasts (MC3T3-E1) were used as models of osteoclasts and osteoblasts, respectively. Differentiation and activity osteoclasts were induced by RANKL in the presence and absence of non-cytotoxic concentrations of Chalcone T4, added in different periods during osteoclastogenesis. Osteoclast differentiation and activity were assessed by actin ring formation and resorption pit assay, respectively. Expression of osteoclast-specific markers (<i>Nfatc1</i>, <i>Oscar</i>, <i>Acp5</i>, <i>Mmp-9</i> and <i>Ctsk</i>) was determined by RT-qPCR, and the activation status of relevant intracellular signaling pathways (MAPK, AKT and NF-kB) by Western blot. Osteoblast differentiation and activity was induced by osteogenic culture medium in the presence and absence of the same concentrations of Chalcone T4. Outcomes assessed were the formation of mineralization nodules via alizarin red staining and the expression of osteoblast-related genes (<i>Alp</i> e <i>Runx2</i>) by RT-qPCR. Chalcone T4 reduced RANKL-induced osteoclast differentiation and activity, suppressed <i>Oscar</i>, <i>Acp5</i> and <i>Mmp-9</i> expression, and decreased ERK and AKT activation in a dose-dependent manner. <i>Nfact1</i> expression and NF-kB phosphorylation were not modulated by the compound. Mineralized matrix formation and the expression of <i>Alp</i> and <i>Runx2</i> by MC3T3-E1 cells were markedly stimulated by Chalcone T4. Collectively, these results demonstrate that Chalcone T4 inhibits in osteoclast differentiation and activity and stimulates osteogenesis, which indicates a promising therapeutic potential in osteolytic diseases.https://www.mdpi.com/1422-0067/24/8/7624chalconeosteoclastogenesisosteogenesisintracellular signaling |
| spellingShingle | Iolanda Augusta Fernandes de Matos Natalie Aparecida Rodrigues Fernandes Giovani Cirelli Mariely Araújo de Godoi Letícia Ribeiro de Assis Luis Octávio Regasini Carlos Rossa Junior Morgana Rodrigues Guimarães-Stabili Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro International Journal of Molecular Sciences chalcone osteoclastogenesis osteogenesis intracellular signaling |
| title | Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro |
| title_full | Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro |
| title_fullStr | Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro |
| title_full_unstemmed | Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro |
| title_short | Chalcone T4 Inhibits RANKL-Induced Osteoclastogenesis and Stimulates Osteogenesis In Vitro |
| title_sort | chalcone t4 inhibits rankl induced osteoclastogenesis and stimulates osteogenesis in vitro |
| topic | chalcone osteoclastogenesis osteogenesis intracellular signaling |
| url | https://www.mdpi.com/1422-0067/24/8/7624 |
| work_keys_str_mv | AT iolandaaugustafernandesdematos chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT natalieaparecidarodriguesfernandes chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT giovanicirelli chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT marielyaraujodegodoi chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT leticiaribeirodeassis chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT luisoctavioregasini chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT carlosrossajunior chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro AT morganarodriguesguimaraesstabili chalconet4inhibitsranklinducedosteoclastogenesisandstimulatesosteogenesisinvitro |