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Herein, the synthesis, characterization, and cellular uptake on folate receptor (FR)-positive breast cancer cells of nanosystems based on CNCs and a chitosan (CS) derivative are investigated. The physical adsorption of the CS derivative, containing a targeting ligand (folic acid, FA) and an imaging agent (fluorescein isothiocyanate, FITC), on the surface of the CNCs was studied as an eco-friendly methodology to functionalize CNCs. The fluorescent CNCs\/FA-CS-FITC nanosystems with a rod-like morphology showed good stability in simulated physiological and non-physiological conditions and non-cytotoxicity towards MDA-MB-231 breast cancer cells. These functionalized CNCs presented a concentration-dependent cellular internalization with a 5-fold increase in the fluorescence intensity for the nanosystem with the higher FA content. Furthermore, the exometabolic profile of the MDA-MB-231 cells exposed to the CNCs\/FA-CS-FITC nanosystems disclosed a moderate impact on the cells\u2019 metabolic activity, limited to decreased choline uptake and increased acetate release, which implies an anti-proliferative effect. The overall results demonstrate that the CNCs\/FA-CS-FITC nanosystems, prepared by an eco-friendly approach, have a high affinity towards FR-positive cancer cells and thus might be applied as nanocarriers with imaging properties for active targeted therapy.<\/jats:p>","DOI":"10.3390\/nano11082057","type":"journal-article","created":{"date-parts":[[2021,8,12]],"date-time":"2021-08-12T10:54:41Z","timestamp":1628765681000},"page":"2057","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Cellulose Nanocrystals\/Chitosan-Based Nanosystems: Synthesis, Characterization, and Cellular Uptake on Breast Cancer Cells"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5600-2396","authenticated-orcid":false,"given":"Ricardo J. B.","family":"Pinto","sequence":"first","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"given":"Nicole S.","family":"Lameirinhas","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0028-6910","authenticated-orcid":false,"given":"Gabriela","family":"Guedes","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7377-8532","authenticated-orcid":false,"given":"Gustavo H.","family":"Rodrigues da Silva","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"given":"P\u00e1r\u00e1stu","family":"Oskoei","sequence":"additional","affiliation":[{"name":"Department of Biology, CESAM\u2014Centre for Environmental and Marine Studies, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9220-3440","authenticated-orcid":false,"given":"Stefan","family":"Spirk","sequence":"additional","affiliation":[{"name":"Institute of Bioproducts and Paper Technology, Graz University of Technology, Inffeldgasse 23, 8010 Graz, Austria"}]},{"given":"Helena","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Department of Biology, CESAM\u2014Centre for Environmental and Marine Studies, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4289-9256","authenticated-orcid":false,"given":"Iola F.","family":"Duarte","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9212-2704","authenticated-orcid":false,"given":"Carla","family":"Vilela","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"given":"Carmen S. R.","family":"Freire","sequence":"additional","affiliation":[{"name":"Department of Chemistry, CICECO\u2014Aveiro Institute of Materials, University of Aveiro, 3810-193 Aveiro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Vilela, C., Pinto, R.J.B., Pinto, S., Marques, P.A.A.P., Silvestre, A.J.D., and Freire, C.S.R. (2018). Polysaccharide Based Hybrid Materials: Metals and Metal Oxides, Graphene and Carbon Nanotubes, Springer. [1st ed.].","DOI":"10.1007\/978-3-030-00347-0"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3358","DOI":"10.1002\/anie.200460587","article-title":"Cellulose: Fascinating biopolymer and sustainable raw material","volume":"44","author":"Klemm","year":"2005","journal-title":"Angew. Chem. Int. 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