{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T04:00:42Z","timestamp":1786420842948,"version":"build-2736575974"},"reference-count":36,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,5,3]],"date-time":"2021-05-03T00:00:00Z","timestamp":1620000000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In this study, we report the effects of caffeine on angiogenesis in zebrafish embryos both during normal development and after exposure to Fibroblast Growth Factor 2 (FGF2). As markers of angiogenesis, we measured the length and width of intersegmental vessels (ISVs), performed whole-mount in situ hybridization with fli1 and cadh5 vascular markers, and counted the number of interconnecting vessels (ICVs) in sub-intestinal venous plexus (SIVP). In addition, we measured angiogenesis after performing zebrafish yolk membrane (ZFYM) assay with microinjection of fibroblast growth factor 2 (FGF2) and perivitelline tumor xenograft assay with microinjection of tumorigenic FGF2-overexpressing endothelial (FGF2-T-MAE) cells. The results showed that caffeine treatment causes a shortening and thinning of ISVs along with a decreased expression of the vascular marker genes and a decrease in the number of ICVs in the SIVP. Caffeine was also able to block angiogenesis induced by exogenous FGF2 or FGF2-producing cells. Overall, our results are suggestive of the inhibitory effect of caffeine in both direct and indirect angiogenesis.<\/jats:p>","DOI":"10.3390\/ijms22094856","type":"journal-article","created":{"date-parts":[[2021,5,5]],"date-time":"2021-05-05T11:06:01Z","timestamp":1620212761000},"page":"4856","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Caffeine Inhibits Direct and Indirect Angiogenesis in Zebrafish Embryos"],"prefix":"10.3390","volume":"22","author":[{"given":"Ram Manohar","family":"Basnet","sequence":"first","affiliation":[{"name":"Unit of Pharmacology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Zizioli","sequence":"additional","affiliation":[{"name":"Unit of Biotechnology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessia","family":"Musc\u00f2","sequence":"additional","affiliation":[{"name":"Unit of Pharmacology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5176-2839","authenticated-orcid":false,"given":"Dario","family":"Finazzi","sequence":"additional","affiliation":[{"name":"Unit of Biotechnology, DMMT, University of Brescia, 25123 Brescia, Italy"},{"name":"Laboratorio Centrale Analisi Chimico-Cliniche, ASST Spedali Civili, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3294-4121","authenticated-orcid":false,"given":"Sandra","family":"Sigala","sequence":"additional","affiliation":[{"name":"Unit of Pharmacology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Rossini","sequence":"additional","affiliation":[{"name":"Unit of Pharmacology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0985-7356","authenticated-orcid":false,"given":"Chiara","family":"Tobia","sequence":"additional","affiliation":[{"name":"Unit of Experimental Oncology and Immunology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Guerra","sequence":"additional","affiliation":[{"name":"Unit of Experimental Oncology and Immunology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4398-8376","authenticated-orcid":false,"given":"Marco","family":"Presta","sequence":"additional","affiliation":[{"name":"Unit of Experimental Oncology and Immunology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurizio","family":"Memo","sequence":"additional","affiliation":[{"name":"Unit of Pharmacology, DMMT, University of Brescia, 25123 Brescia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.vph.2009.02.002","article-title":"Angiogenesis and its targeting in rheumatoid arthritis","volume":"51","author":"Szekanecz","year":"2009","journal-title":"Vascul. 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