{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T17:26:18Z","timestamp":1775064378219,"version":"3.50.1"},"reference-count":42,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2019,7,24]],"date-time":"2019-07-24T00:00:00Z","timestamp":1563926400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Innovation of Spain","award":["MAT2014-59134R"],"award-info":[{"award-number":["MAT2014-59134R"]}]},{"name":"Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT\/MEC)","award":["M-ERA-NET\/0004\/2015 (PAIRED) and UID\/AGR\/04033\/2019 (CITAB)"],"award-info":[{"award-number":["M-ERA-NET\/0004\/2015 (PAIRED) and UID\/AGR\/04033\/2019 (CITAB)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Oleanolic (OA) and ursolic (UA) acids are recognized triterpenoids with anti-cancer properties, showing cell-specific activity that can be enhanced when loaded into polymeric nanoparticles. The cytotoxic activity of OA and UA was assessed by Alamar Blue assay in three different cell lines, i.e., HepG2 (Human hepatoma cell line), Caco-2 (Human epithelial colorectal adenocarcinoma cell line) and Y-79 (Human retinoblastoma cell line). The natural and synthetic mixtures of these compounds were tested as free and loaded in polymeric nanoparticles in a concentration range from 2 to 32 \u00b5mol\/L. The highest tested concentrations of the free triterpene mixtures produced statistically significant cell viability reduction in HepG2 and Caco-2 cells, compared to the control (untreated cells). When loaded in the developed PLGA nanoparticles, no differences were recorded for the tested concentrations in the same cell lines. However, in the Y-79 cell line, a decrease on cell viability was observed when testing the lowest concentration of both free triterpene mixtures, and after their loading into PLGA nanoparticles.<\/jats:p>","DOI":"10.3390\/pharmaceutics11080362","type":"journal-article","created":{"date-parts":[[2019,7,24]],"date-time":"2019-07-24T10:48:19Z","timestamp":1563965299000},"page":"362","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":77,"title":["In Vitro Cytotoxicity of Oleanolic\/Ursolic Acids-Loaded in PLGA Nanoparticles in Different Cell Lines"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7524-9914","authenticated-orcid":false,"given":"Am\u00e9lia M.","family":"Silva","sequence":"first","affiliation":[{"name":"Centre for Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Tr\u00e1s-os Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Department of Biology and Environment, UTAD, Quinta de Prados, 5001-801 Vila Real, Portugal"}]},{"given":"Helen L.","family":"Alvarado","sequence":"additional","affiliation":[{"name":"Department of Biology and Environment, UTAD, Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Department of Pharmacy and Pharmaceutical Technology and Physical Chemistry, Faculty of Pharmacy, University of Barcelona, Ave. Joan XXIII s\/n, 08028 Barcelona, Spain"}]},{"given":"Guadalupe","family":"Abrego","sequence":"additional","affiliation":[{"name":"Department of Biology and Environment, UTAD, Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Department of Chemical and Instrumental Analysis, Faculty of Chemistry and Pharmacy, University of El Salvador, Final 25 Ave. Norte, 3026 San Salvador, El Salvador"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1564-3661","authenticated-orcid":false,"given":"Carlos","family":"Martins-Gomes","sequence":"additional","affiliation":[{"name":"Centre for Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Tr\u00e1s-os Montes e Alto Douro (UTAD), Quinta de Prados, 5001-801 Vila Real, Portugal"},{"name":"Department of Biology and Environment, UTAD, Quinta de Prados, 5001-801 Vila Real, Portugal"}]},{"given":"Maria L.","family":"Gardu\u00f1o-Ramirez","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Qu\u00edmicas, Universidad Aut\u00f3noma del Estado de Morelos, Av. Universidad No. 1001, Col Chamilpa, 62209 Cuernavaca, Mexico"}]},{"given":"Mar\u00eda L.","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Pharmaceutical Technology and Physical Chemistry, Faculty of Pharmacy, University of Barcelona, Ave. Joan XXIII s\/n, 08028 Barcelona, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4989-4821","authenticated-orcid":false,"given":"Ana C.","family":"Calpena","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Pharmaceutical Technology and Physical Chemistry, Faculty of Pharmacy, University of Barcelona, Ave. Joan XXIII s\/n, 08028 Barcelona, Spain"}]},{"given":"Eliana B.","family":"Souto","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra (FFUC), P\u00f3lo das Ci\u00eancias da Sa\u00fade, 3000-548 Coimbra, Portugal"},{"name":"CEB\u2014Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2019,7,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1155\/2015\/620472","article-title":"Antimicrobial activity of oleanolic and ursolic acids: An update","volume":"2015","author":"Jesus","year":"2015","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3139","DOI":"10.2217\/nnm-2016-0251","article-title":"Potential use of nanocarriers with pentacyclic triterpenes in cancer treatments","volume":"11","author":"Valdes","year":"2016","journal-title":"Nanomedicine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"20614","DOI":"10.3390\/molecules201119721","article-title":"Ursolic Acid\u2014A Pentacyclic Triterpenoid with a Wide Spectrum of Pharmacological Activities","volume":"20","year":"2015","journal-title":"Molecules"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.jff.2018.02.012","article-title":"Chemical characterization and bioactive properties of decoctions and hydroethanolic extracts of Thymus carnosus Boiss","volume":"43","year":"2018","journal-title":"J. Funct. Foods"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3617","DOI":"10.1039\/C8FO00456K","article-title":"Thymus pulegioides L. as a rich source of antioxidant, anti-proliferative and neuroprotective phenolic compounds","volume":"9","author":"Taghouti","year":"2018","journal-title":"Food Funct."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1016\/j.bmcl.2015.12.064","article-title":"Targeting cancer cells with oleanolic and ursolic acid derived hydroxamates","volume":"26","author":"Wiemann","year":"2016","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_7","first-page":"1","article-title":"Synthesis, structural studies, and cytotoxic evaluation of novel ursolic acid hybrids with capabilities to arrest breast cancer cells in mitosis","volume":"19","author":"Pattnaik","year":"2016","journal-title":"J. Asian Nat. Prod. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejmech.2016.04.051","article-title":"Urea derivates of ursolic, oleanolic and maslinic acid induce apoptosis and are selective cytotoxic for several human tumor cell lines","volume":"119","author":"Sommerwerk","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1055\/s-0029-1186102","article-title":"Pentacyclic triterpenes of the lupane, oleanane and ursane group as tools in cancer therapy","volume":"75","author":"Laszczyk","year":"2009","journal-title":"Planta Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2714","DOI":"10.1080\/13880209.2016.1180538","article-title":"Anti-proliferative and antibacterial in vitro evaluation of the polyurethane nanostructures incorporating pentacyclic triterpenes","volume":"54","author":"Oprean","year":"2016","journal-title":"Pharm. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1016\/j.phymed.2015.05.004","article-title":"Antiproliferative activity of rosamultic acid is associated with induction of apoptosis, cell cycle arrest, inhibition of cell migration and caspase activation in human gastric cancer (SGC-7901) cells","volume":"22","author":"Sui","year":"2015","journal-title":"Phytomedicine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.jep.2010.07.014","article-title":"Cytotoxicity and bioavailability studies on a decoction of Oldenlandia diffusa and its fractions separated by HPLC","volume":"131","author":"Ganbold","year":"2010","journal-title":"J. Ethnopharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1002\/ijc.24433","article-title":"Triterpenes augment the inhibitory effects of anticancer drugs on growth of human esophageal carcinoma cells in vitro and suppress experimental metastasis in vivo","volume":"125","author":"Yamai","year":"2009","journal-title":"Int. J. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.ijpharm.2013.11.025","article-title":"Design of cationic lipid nanoparticles for ocular delivery: Development, characterization and cytotoxicity","volume":"461","author":"Fangueiro","year":"2014","journal-title":"Int. J. Pharm."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"565","DOI":"10.3892\/ol.2011.275","article-title":"Induction of apoptosis in the SW620 colon carcinoma cell line by triterpene-enriched extracts from Ganoderma lucidum through activation of caspase-3","volume":"2","author":"Ji","year":"2011","journal-title":"Oncol. Lett."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1007\/s10156-012-0387-6","article-title":"The use of the microplate alamar blue assay (MABA) to assess the susceptibility of Mycobacterium lepraemurium to anti-leprosy and other drugs","volume":"18","year":"2012","journal-title":"J. Infect. Chemother."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.ejmech.2014.04.084","article-title":"Solid lipid nanoparticles for hydrophilic biotech drugs: Optimization and cell viability studies (Caco-2 & HEPG-2 cell lines)","volume":"81","author":"Severino","year":"2014","journal-title":"Eur. J. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.jff.2019.01.010","article-title":"Thymus carnosus extracts induce anti-proliferative activity in Caco-2 cells through mechanisms that involve cell cycle arrest and apoptosis","volume":"54","year":"2019","journal-title":"J. Funct. Foods"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejpb.2014.02.005","article-title":"Nanotoxicology applied to solid lipid nanoparticles and nanostructured lipid carriers-A systematic review of in vitro data","volume":"87","author":"Doktorovova","year":"2014","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_20","unstructured":"Grumezescu, A.M. (2016). Advances in Nanobiomaterials for Oncology Nanomedicine. Applications of NanoBioMaterials, Elsevier."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1290","DOI":"10.2174\/1567201813666160713121122","article-title":"Nanoparticles for Cancer Targeting: Current and Future Directions","volume":"13","author":"Swain","year":"2016","journal-title":"Curr. Drug Deliv."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/j.ijpharm.2018.06.064","article-title":"Mediterranean essential oils as precious matrix components and active ingredients of lipid nanoparticles","volume":"548","author":"Carbone","year":"2018","journal-title":"Int. J. Pharm."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Carbone, C. (2019). Clotrimazole-Loaded Mediterranean Essential Oils NLC: A Synergic Treatment of Candida Skin Infections. Pharmaceutics, 11.","DOI":"10.3390\/pharmaceutics11050231"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1016\/j.biotechadv.2012.08.005","article-title":"Nanoencapsulation of polyphenols for protective effect against colon-rectal cancer","volume":"31","author":"Santos","year":"2013","journal-title":"Biotechnol. Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.nano.2015.01.004","article-title":"Design and optimization of oleanolic\/ursolic acid-loaded nanoplatforms for ocular anti-inflammatory applications","volume":"11","author":"Alvarado","year":"2015","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.jchromb.2015.01.009","article-title":"Development and validation of a high-performance liquid chromatography method for the quantification of ursolic\/oleanic acids mixture isolated from Plumeria obtusa","volume":"983\u2013984","author":"Alvarado","year":"2015","journal-title":"J. Chromatogr. B"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.colsurfb.2015.03.062","article-title":"Nanoemulsions for dermal controlled release of oleanolic and ursolic acids: In vitro, ex vivo and in vivo characterization","volume":"130","author":"Alvarado","year":"2015","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_28","unstructured":"Hern\u00e1ndez Ch\u00e1vez, L.I. (2004). Estudio Qu\u00edmico y Evaluaci\u00f3n Farmacol\u00f3gica de la Actividad Antidiab\u00e9tica de las hojas de Plumeria Obtusa, Especie Empleada en la Medicina Tradicional Mexicana Para Los S\u00edntomas de la Diabetes Mellitus. [Ph.D. Thesis, Universidad de Colima]."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0378-5173(89)90281-0","article-title":"Nanocapsule formation by interfacial polymer deposition following solvent displacement","volume":"55","author":"Fessi","year":"1989","journal-title":"Int. J. Pharm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.ijpharm.2013.12.022","article-title":"In vitro evaluation of permeation, toxicity and effect of praziquantel-loaded solid lipid nanoparticles against Schistosoma mansoni as a strategy to improve efficacy of the schistosomiasis treatment","volume":"463","author":"Souza","year":"2014","journal-title":"Int. J. Pharm."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.tiv.2004.03.012","article-title":"Comparison of alamar blue and MTT assays for high through-put screening","volume":"18","author":"Hamid","year":"2004","journal-title":"Toxicol. In Vitro"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/S0022-1759(98)00028-3","article-title":"A novel one-step, highly sensitive fluorometric assay to evaluate cell-mediated cytotoxicity","volume":"213","author":"Nociari","year":"1998","journal-title":"J. Immunol. Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1093\/jnci\/53.2.347","article-title":"Characteristics of an established cell line of retinoblastoma","volume":"53","author":"Reid","year":"1974","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1016\/S1470-2045(14)70376-6","article-title":"Evidence-based care for trilateral retinoblastoma","volume":"15","author":"Dimaras","year":"2014","journal-title":"Lancet Oncol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1436","DOI":"10.1016\/S0140-6736(11)61137-9","article-title":"Retinoblastoma","volume":"379","author":"Dimaras","year":"2012","journal-title":"Lancet"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1007\/s00418-014-1285-z","article-title":"Re-characterization of established human retinoblastoma cell lines","volume":"143","author":"Busch","year":"2015","journal-title":"Histochem. Cell Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1097\/CMR.0000000000000137","article-title":"Ursolic acid and resveratrol synergize with chloroquine to reduce melanoma cell viability","volume":"25","author":"Junco","year":"2015","journal-title":"Melanoma Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.lfs.2016.01.017","article-title":"Ursolic acid (UA): A metabolite with promising therapeutic potential","volume":"146","author":"Kashyap","year":"2016","journal-title":"Life Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"493","DOI":"10.3748\/wjg.v8.i3.493","article-title":"Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15","volume":"8","author":"Li","year":"2002","journal-title":"World J. Gastroenterol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2652","DOI":"10.1016\/j.ejmech.2011.03.050","article-title":"In vitro and in vivo anticancer activity evaluation of ursolic acid derivatives","volume":"46","author":"Shao","year":"2011","journal-title":"Eur. J. Med. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1016\/j.tiv.2009.12.008","article-title":"Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines","volume":"24","author":"Yan","year":"2010","journal-title":"Toxicol. In Vitro"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/S0921-4526(02)00552-5","article-title":"The first observation of the boson peak from water vapour deposited amorphous ice","volume":"316\u2013317","author":"Li","year":"2002","journal-title":"Phys. B Condens. Matter"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/11\/8\/362\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:09:05Z","timestamp":1760188145000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/11\/8\/362"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,7,24]]},"references-count":42,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2019,8]]}},"alternative-id":["pharmaceutics11080362"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics11080362","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,7,24]]}}}