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5. CONCLUsION The use of 3D tumor models for the preclinical validation of nanotherapiespromises to be a significant step forward in cancer research. The screeningof new anti-cancer nanocarriers in more predictive models accelerates andupgrades the validation of candidate nanoplatforms, ultimately impactingthe discovery of more effective cancer therapies. These in vitro modelsoffer the possibility of smartly recapitulating crucial biophysical barrierspresent in the native tumor microenvironment, overcoming the issues ofmore simplistic 2D cultures. Up to date, 3D tumor models have beenparticularly relevant for evaluating nanotherapies cell targetingperformance, tumor penetration, internalization and their anti-cancertherapeutic outcome. Synergistically, these in vitro models are also able toserve as high-throughput analysis systems that can greatly reduce the usageof in vivo animal models during validation stages. As we move forward, thescreening of nanotherapeutics in physiologically relevant 3D in vitro modelswill play an increasingly important role in their design and therapeuticperformance analysis before clinical application.AcknowledgementsThis work was developed within the scope of the project CICECO-AveiroInstitute of Materials, UIDB/50011/2020, UIDP/50011/2020 &LA/P/0006/2020, financed by national funds through the FCT/MEC(PIDDAC). This work was also supported by the Programa OperacionalCompetitividade e Internacionaliza%u00e7%u00e3o (POCI), in the component FEDER.This work was also funded by the European Union%u2019s Horizon Europe researchand innovation programme under the scope of project INSPIRE (Grantagreement ID: 101057777). The authors acknowledge the financial supportby the Portuguese Foundation for Science and Technology (FCT) throughDoctoral Grants (DFA/BD/7692/2020, M.V.M., 2023.03472.BDANA, M.T.C.).6. RefeReNCes1. Katrina, A.; Voliani, V. Three-Dimensional Tumor Models : PromotingBreakthroughs in Nanotheranostics Translational Research. Appl Mater Today2020, 19, 100552. https://doi.org/10.1016/j.apmt.2019.100552.2. Shen, S.; Xu, X.; Lin, S.; Zhang, Y.; Liu, H.; Zhang, C.; Mo, R. A NanotherapeuticStrategy to Overcome Chemotherapeutic Resistance of Cancer Stem-likeCells. Nat Nanotechnol 2021, 16 (1), 104%u2013113.https://doi.org/10.1038/s41565-020-00793-0.3. van der Meel, R.; Sulheim, E.; Shi, Y.; Kiessling, F.; Mulder, W. J. M.; Lammers,T. Smart Cancer Nanomedicine. Nat Nanotechnol 2019, 14 (11), 1007%u20131017.https://doi.org/10.1038/s41565-019-0567-y.118Potential of 3D tumor models for nanotherapies pre-clinical screeningVitor M. Gaspar1, Jo%u00e3o F. Mano, et al.