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Three-dimensional culture models: emerging platforms for screening the antitumoral efficacy of nanomedicines

    Larissa Bueno Tofani

    School of Pharmaceutical Science of Ribeirao Preto, University of Sao Paulo (USP), Ribeirao Preto, Sao Paulo, 14040-903, Brazil

    ,
    Marcela Tavares Luiz

    *Author for correspondence: Tel.: +55 163 301 6998;

    E-mail Address: marcela.tavares@unesp.br

    School of Pharmaceutical Science of Sao Paulo State University (UNESP), Araraquara, Sao Paulo, 14800-903, Brazil

    ,
    Jessyca Aparecida Paes Dutra

    School of Pharmaceutical Science of Sao Paulo State University (UNESP), Araraquara, Sao Paulo, 14800-903, Brazil

    ,
    Juliana Palma Abriata

    School of Pharmaceutical Science of Ribeirao Preto, University of Sao Paulo (USP), Ribeirao Preto, Sao Paulo, 14040-903, Brazil

    &
    Marlus Chorilli

    **Author for correspondence: Tel.: +55 163 301 6998;

    E-mail Address: marlus.chorilli@unesp.br

    School of Pharmaceutical Science of Sao Paulo State University (UNESP), Araraquara, Sao Paulo, 14800-903, Brazil

    Published Online:https://doi.org/10.2217/nnm-2022-0205

    Nanomedicines have been investigated for delivering drugs to tumors due to their ability to accumulate in the tumor tissues. 2D in vitro cell culture has been used to investigate the antitumoral potential of nanomedicines. However, a 2D model cannot adequately mimic the in vivo tissue conditions because of the lack of cell–cell interaction, a gradient of nutrients and the expression of genes. To overcome this limitation, 3D cell culture models have emerged as promising platforms that better replicate the complexity of native tumors. For this purpose, different techniques can be used to produce 3D models, including scaffold-free, scaffold-based and microfluidic-based models. This review addresses the principles, advantages and limitations of these culture methods for evaluating the antitumoral efficacy of nanomedicines.

    Tweetable abstract

    3D cell culture has emerged to replicate more effectively the complexity of native tumors than the 2D model traditionally used. Different 3D cell culture approaches have been developed and studied, with promising results in the nanotechnology of cancer.

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

    References

    • 1. Sung H, Ferlay J, Siegel RL et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA. Cancer J. Clin. 71(3), 209–249 (2021).
    • 2. Zhong S, Jeong JH, Chen Z et al. Targeting tumor microenvironment by small-molecule inhibitors. Transl. Oncol. 13(1), 57–69 (2020).
    • 3. Yeini E, Ofek P, Albeck N et al. Targeting glioblastoma: advances in drug delivery and novel therapeutic approaches. Adv. Ther. 4, 2000124 (2020).
    • 4. Wu J. The enhanced permeability and retention (EPR) effect: the significance of the concept and methods to enhance its application. J. Pers. Med. 11(8), 771–778 (2021).
    • 5. Francia V, Montizaan D, Salvati A. Interactions at the cell membrane and pathways of internalization of nano-sized materials for nanomedicine. Beilstein J. Nanotechnol. 11, 338–353 (2020).
    • 6. Luiz MT, Delello Di Filippo L, Tofani LB et al. Highlights in targeted nanoparticles as a delivery strategy for glioma treatment. Int. J. Pharm. 604, 120758 (2021).
    • 7. Ratemi E, Sultana Shaik A, Al Faraj A, Halwani R. Alternative approaches for the treatment of airway diseases: focus on nanoparticle medicine. Clin. Exp. Allergy 46(8), 1033–1042 (2016).
    • 8. Bharti C, Gulati N, Nagaich U, Pal A. Mesoporous silica nanoparticles in target drug delivery system: a review. Int. J. Pharm. Investig. 5(3), 124 (2015).
    • 9. Vega-Vásquez P, Mosier NS, Irudayaraj J. Nanoscale drug delivery systems: from medicine to agriculture. Front. Bioeng. Biotechnol. 8, 1–16 (2020).
    • 10. Salehipour M, Rezaei S, Mosafer J et al. Recent advances in polymer-coated iron oxide nanoparticles as magnetic resonance imaging contrast agents. J. Nanoparticle Res. 23(2), 1–35 (2021).
    • 11. Bonilla AM, Gonzalez PH. Hybrid polymeric-magnetic nanoparticles in cancer treatments. Curr. Pharm. Des. 23(35), 5392–5402 (2017).
    • 12. Milligan JJ, Saha S. A nanoparticle's journey to the tumor: strategies to overcome first-pass metabolism and their limitations. Cancers (Basel). 14(7), 1741–1757 (2022).
    • 13. Roma-Rodrigues C, Rivas-García L, Baptista PV, Fernandes AR. Gene therapy in cancer treatment: why go nano? Pharmaceutics 12(3), 1–35 (2020).
    • 14. Bayón-Cordero L, Alkorta I, Arana L. Application of solid lipid nanoparticles to improve the efficiency of anticancer drugs. Nanomaterials 9(3), 474–494 (2019).
    • 15. Pammolli F, Magazzini L, Riccaboni M. The productivity crisis in pharmaceutical R&D. Nat. Rev. Drug Discov. 10(6), 428–438 (2011).
    • 16. Amiri-Kordestani L, Pazdur R. Oncology approvals in 2020: a year of firsts in the midst of a pandemic. Nat. Rev. Clin. Oncol. 18(3), 129–130 (2021).
    • 17. Basavaraj S, Betageri GV. Can formulation and drug delivery reduce attrition during drug discovery and development – review of feasibility, benefits and challenges. Acta Pharm. Sin. B 4(1), 3–17 (2014).
    • 18. Krüger A, Gonçalves Maltarollo V, Wrenger C, Kronenberger T. ADME profiling in drug discovery and a new path paved on silica. In: Drug Discovery and Development - New Advances. IntechOpen, London, UK (2020).
    • 19. Fontoura JC, Viezzer C, dos Santos FG et al. Comparison of 2D and 3D cell culture models for cell growth, gene expression and drug resistance. Mater. Sci. Eng. C 107, 110264 (2020).
    • 20. Junod SW. FDA and clinical drug trials: a short history, Food and Drug Administration, United States (2013). www.fda.gov
    • 21. Shoemaker RH. The NCI60 human tumour cell line anticancer drug screen. Nat. Rev. Cancer 6(10), 813–823 (2006).
    • 22. Barretina J, Caponigro G, Stransky N et al. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483(7391), 603–607 (2012).
    • 23. Ghandi M, Huang FW, Jané-Valbuena J et al. Next-generation characterization of the cancer cell line encyclopedia. Nature 569(7757), 503–508 (2019).
    • 24. Rustamov V, Keller F, Klicks J, Hafner M, Rudolf R. Bone sialoprotein shows enhanced expression in early, high-proliferation stages of three-dimensional spheroid cell cultures of breast cancer cell line MDA-MB-231. Front. Oncol. 9 (2019).
    • 25. Fischbach C, Chen R, Matsumoto T et al. Engineering tumors with 3D scaffolds. Nat. Methods 4(10), 855–860 (2007).
    • 26. Pampaloni F, Reynaud EG, Stelzer EHK. The third dimension bridges the gap between cell culture and live tissue. Nat. Rev. Mol. Cell Biol. 8(10), 839–845 (2007).
    • 27. Duval K, Grover H, Han L-H et al. Modeling physiological events in 2D vs. 3D cell culture. Physiology 32(4), 266–277 (2017). • Provides an overview of traditional 2D and 3D cell culture models, as well as the challenges, differences, implications, advantages and limitations of each model.
    • 28. Imamura Y, Mukohara T, Shimono Y et al. Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer. Oncol. Rep. 33(4), 1837–1843 (2015).
    • 29. Jensen C, Teng Y. Is it time to start transitioning from 2D to 3D cell culture? Front. Mol. Biosci. 7 (2020).
    • 30. Yakavets I, Francois A, Benoit A et al. Advanced co-culture 3D breast cancer model for investigation of fibrosis induced by external stimuli: optimization study. Sci. Rep. 10(1), 21273 (2020).
    • 31. Carvalho S, Silveira MJ, Domingues M et al. Multicellular quadruple colorectal cancer spheroids as an in vitro tool for antiangiogenic potential evaluation of nanoparticles. Adv. Ther. 2200282, 1–11 (2023).
    • 32. Melissaridou S, Wiechec E, Magan M et al. The effect of 2D and 3D cell cultures on treatment response, EMT profile and stem cell features in head and neck cancer. Cancer Cell Int. 19(1), 1–10 (2019).
    • 33. Goswami S, Sahai E, Wyckoff JB et al. Erratum: macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop. Cancer Res. 65(15), 7031 (2005).
    • 34. Xin X, Yang H, Zhang F, Yang ST. 3D cell coculture tumor model: a promising approach for future cancer drug discovery. Process Biochem. 78, 148–160 (2019).
    • 35. Lis R, Touboul C, Mirshahi P et al. Tumor associated mesenchymal stem cells protects ovarian cancer cells from hyperthermia through CXCL12. Int. J. Cancer 128(3), 715–725 (2011).
    • 36. Huang B, Gao J. Application of 3D cultured multicellular spheroid tumor models in tumor-targeted drug delivery system research. J. Control. Rel. 270, 246–259 (2018).
    • 37. Cheng J, Li L, Liu Y et al. Interleukin-1α induces immunosuppression by mesenchymal stem cells promoting the growth of prostate cancer cells. Mol. Med. Rep. 6(5), 955–960 (2012).
    • 38. Costa EC, Moreira AF, de Melo-Diogo D et al. 3D tumor spheroids: an overview on the tools and techniques used for their analysis. Biotechnol. Adv. 34(8), 1427–1441 (2016).
    • 39. Gunti S, Hoke ATK, Vu KP, London NR. Organoid and spheroid tumor models: techniques and applications. Cancers (Basel) 13(4), 1–18 (2021).
    • 40. Alzeeb G, Metges JP, Corcos L, Le Jossic-Corcos C. Three-dimensional culture systems in gastric cancer research. Cancers (Basel) 12(10), 1–20 (2020).
    • 41. Souza AG, Silva IBB, Campos-Fernandez E et al. Comparative assay of 2D and 3D cell culture models: proliferation, gene expression and anticancer drug response. Curr. Pharm. Des. 24(15), 1689–1694 (2018). •• A comparative study between 2D and 3D cell culture to determine the most suitable model for preclinical in vitro drug testing. Evaluates the proliferation, genetic expression and chemoresistance of prostate tumor cell lines, PC-3, LNCaP and DU145 after treatment with antineoplastic drugs (paclitaxel and docetaxel), demonstrating the similar behavior of the 3D cell culture models tested with in vivo systems and their promising use as a more reliable tool for drug development.
    • 42. Luiz MT, Viegas JSR, Abriata JP et al. Docetaxel-loaded folate-modified TPGS-transfersomes for glioblastoma multiforme treatment. Mater. Sci. Eng. C 124, 112033 (2021).
    • 43. Chen X, Yuan M, Zhang Q et al. Synergistic combination of doxorubicin and paclitaxel delivered by blood brain barrier and glioma cells dual targeting liposomes for chemotherapy of brain glioma. Curr. Pharm. Biotechnol. 17(7), 636–650 (2016).
    • 44. Arora D, Bhunia BK, Janani G, Mandal BB. Bioactive three-dimensional silk composite in vitro tumoroid model for high throughput screening of anticancer drugs. J. Colloid Interface Sci. 589, 438–452 (2021).
    • 45. Ferreira LP, Gaspar VM, Monteiro MV et al. Screening of dual chemo-photothermal cellular nanotherapies in organotypic breast cancer 3D spheroids. J. Control. Rel. 331, 85–102 (2021).
    • 46. Miranda MA, Silva LB, Carvalho IPS et al. Targeted uptake of folic acid-functionalized polymeric nanoparticles loading glycoalkaloidic extract in vitro and in vivo assays. Colloids Surfaces B Biointerfaces 192, 111106 (2020).
    • 47. Oner E, Kotmakci M, Baird AM et al. Development of EphA2 siRNA-loaded lipid nanoparticles and combination with a small‐molecule histone demethylase inhibitor in prostate cancer cells and tumor spheroids. J. Nanobiotechnol. 19, 71 (2021).
    • 48. Amaral RLF, Miranda M, Marcato PD, Swiech K. Comparative analysis of 3D bladder tumor spheroids obtained by forced floating and hanging drop methods for drug screening. Front. Physiol. 22, 8 (2017).
    • 49. Ware MJ, Colbert K, Keshishian V et al. Generation of homogenous three-dimensional pancreatic cancer cell spheroids using an improved hanging drop technique. Tissue Eng. C Methods 22(4), 312–321 (2016).
    • 50. Badea MA, Balas M, Dinischiotu A. Biological properties and development of hypoxia in a breast cancer 3D model generated by hanging drop technique. Cell Biochem. Biophys. 80(1), 63–73 (2021).
    • 51. Cavo M, Delle Cave D, D'Amone E et al. A synergic approach to enhance long-term culture and manipulation of MiaPaCa-2 pancreatic cancer spheroids. Sci. Rep. 10(1), 1–11 (2020).
    • 52. Yakavets I, Yankovsky I, Millard M et al. The alteration of temoporfin distribution in multicellular tumor spheroids by β-cyclodextrins. Int. J. Pharm. 529(1–2), 568–575 (2017).
    • 53. Santos JM, Camões SP, Filipe E et al. Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing. Stem Cell Res. Ther. 6(1), 1–19 (2015).
    • 54. Türker E, Demirçak N, Arslan-Yildiz A. Scaffold-free three-dimensional cell culturing using magnetic levitation. Biomater. Sci. 6(7), 1745–1753 (2018).
    • 55. Rijal G, Bathula C, Li W. Application of synthetic polymeric scaffolds in breast cancer 3D tissue cultures and animal tumor models. Int. J. Biomater. 2017, 1-9 (2017).
    • 56. Miranda MA, Marcato PD, Mondal A et al. Cytotoxic and chemosensitizing effects of glycoalkaloidic extract on 2D and 3D models using RT4 and patient derived xenografts bladder cancer cells. Mater. Sci. Eng. C 119, 111460 (2021).
    • 57. Jia W, Jiang X, Liu W et al. Effects of three-dimensional collagen scaffolds on the expression profiles and biological functions of glioma cells. Int. J. Oncol. 52(6), 1787–1800 (2018).
    • 58. Herreros-Pomares A, Zhou X, Calabuig-Fariñas S et al. 3D printing novel in vitro cancer cell culture model systems for lung cancer stem cell study. Mater. Sci. Eng. C. 122, 111914 (2021).
    • 59. Khot MI, Perry SL, Maisey T et al. Inhibiting ABCG2 could potentially enhance the efficacy of hypericin-mediated photodynamic therapy in spheroidal cell models of colorectal cancer. Photodiagnosis Photodyn. Ther. 23, 221–229 (2018).
    • 60. Berger Fridman I, Ugolini GS, Vandelinder V et al. High throughput microfluidic system with multiple oxygen levels for the study of hypoxia in tumor spheroids. Biofabrication 13, 0350 (2021).
    • 61. Liu W, Liu D, Hu R et al. An integrated microfluidic 3D tumor system for parallel and high-throughput chemotherapy evaluation. Analyst 145(20), 6447–6455 (2020).
    • 62. Lazzari G, Couvreur P, Mura S. Multicellular tumor spheroids: a relevant 3D model for the: in vitro preclinical investigation of polymer nanomedicines. Polym. Chem. 8(34), 4947–4969 (2017).
    • 63. Kamatar A, Gunay G, Acar H. Natural and synthetic biomaterials for engineering multicellular tumor spheroids. Polymers (Basel) 12(11), 1–23 (2020).
    • 64. Wang A, Madden LA, Paunov VN. Advanced biomedical applications based on emerging 3D cell culturing platforms. J. Mater. Chem. B 8(46), 10487–10501 (2020).
    • 65. Pinto B, Henriques AC, Silva PMA, Bousbaa H. Three-dimensional spheroids as in vitro preclinical models for cancer research. Pharmaceutics 12, 1186 (2020). • A comprehensive overview of 3D tumor systems and how they can be obtained, focusing in their applicability in cancer field as tool to evaluation of therapeutic nanomedicine. Provides important features about the 3D culture cancer cells can better replicate the in vivo tumor environment and how these cultures are emerging as preclinical anticancer screening and accurate low-cost cancer models for preclinical drug screening.
    • 66. Shen H, Cai S, Wu C et al. Recent advances in three-dimensional multicellular spheroid culture and future development. Micromachines 12(1), 1–21 (2021).
    • 67. Mohammad-Hadi L, MacRobert AJ, Loizidou M, Yaghini E. Photodynamic therapy in 3D cancer models and the utilisation of nanodelivery systems. Nanoscale 10(4), 1570–1581 (2018). •• The review presents recent advances of use 3D model as tool to evaluate photodynamic therapy and nanodelivery systems as cancer therapy.
    • 68. Saydé T, Hamoui O El, Alies B et al. Biomaterials for three-dimensional cell culture: from applications in oncology to nanotechnology. Nanomaterials 11(2), 1–29 (2021).
    • 69. Tomás-Bort E, Kieler M, Sharma S et al. 3D approaches to model the tumor microenvironment of pancreatic cancer. Theranostics 10(11), 5074–5089 (2020).
    • 70. Van Zundert I, Fortuni B, Rocha S. From 2D to 3D cancer cell models – the enigmas of drug delivery research. Nanomaterials 10(11), 1–30 (2020).
    • 71. Pitingolo G, Nizard P, Riaud A, Taly V. Beyond the on/off chip trade-off: a reversibly sealed microfluidic platform for 3D tumor microtissue analysis. Sensors Actuators B Chem. 274, 393–401 (2018).
    • 72. Shang M, Soon RH, Lim CT et al. Microfluidic modelling of the tumor microenvironment for anti-cancer drug development. Lab Chip 19(3), 369–386 (2019).
    • 73. Feiner-Gracia N, Glinkowska Mares A, Buzhor M et al. Real-time ratiometric imaging of micelles assembly state in a microfluidic cancer-on-a-chip. ACS Appl. Bio Mater. 4(1), 669–681 (2021).
    • 74. Agarwal P, Wang H, Sun M et al. Microfluidics enabled bottom-up engineering of 3D vascularized tumor for drug discovery. ACS Nano 11(7), 6691–6702 (2017).
    • 75. Bidan N, Lores S, Vanhecke A et al. Before in vivo studies: in vitro screening of sphingomyelin nanosystems using a relevant 3D multicellular pancreatic tumor spheroid model. Int. J. Pharm. 617, 121577 (2022).
    • 76. Heinrich MA, Mostafa AMRH, Morton JP et al. Translating complexity and heterogeneity of pancreatic tumor: 3D in vitro to in vivo models. Adv. Drug Deliv. Rev. 174, 265–293 (2021).
    • 77. Vaidya B, Parvathaneni V, Kulkarni NS et al. Cyclodextrin modified erlotinib loaded PLGA nanoparticles for improved therapeutic efficacy against non-small-cell lung cancer. Int. J. Biol. Macromol. 122, 338–347 (2019).
    • 78. Yu Q, Tang X, Zhao W et al. Mild hyperthermia promotes immune checkpoint blockade-based immunotherapy against metastatic pancreatic cancer using size-adjustable nanoparticles. Acta Biomater. 133, 244–256 (2021).
    • 79. Abdolahinia ED, Nadri S, Rahbarghazi R et al. Enhanced penetration and cytotoxicity of metformin and collagenase conjugated gold nanoparticles in breast cancer spheroids. Life Sci. 231, 116545 (2019).
    • 80. Dhanwal V, Katoch A, Singh A et al. Self-assembled organic nanoparticles of benzimidazole analogue exhibit enhanced uptake in 3D tumor spheroids and oxidative stress induced cytotoxicity in breast cancer. Mater. Sci. Eng. C 97, 467–478 (2019).
    • 81. Rozhina E, Batasheva S, Gomzikova M et al. Multicellular spheroids formation: the synergistic effects of halloysite nanoclay and cationic magnetic nanoparticles. Colloids Surfaces A Physicochem. Eng. Asp. 565, 16–24 (2019).
    • 82. Stocke NA, Sethi P, Jyoti A et al. Toxicity evaluation of magnetic hyperthermia induced by remote actuation of magnetic nanoparticles in 3D micrometastasic tumor tissue analogs for triple negative breast cancer. Biomaterials 120, 115–125 (2017).
    • 83. Evans JC, Malhotra M, Sweeney K et al. Folate-targeted amphiphilic cyclodextrin nanoparticles incorporating a fusogenic peptide deliver therapeutic siRNA and inhibit the invasive capacity of 3D prostate cancer tumours. Int. J. Pharm. 532(1), 511–518 (2017).
    • 84. Fitzgerald KA, Guo J, Raftery RM et al. Nanoparticle-mediated siRNA delivery assessed in a 3D co-culture model simulating prostate cancer bone metastasis. Int. J. Pharm. 511(2), 1058–1069 (2016).
    • 85. Elberskirch L, Knoll T, Königsmark R et al. Microfluidic 3D intestine tumor spheroid model for efficient in vitro investigation of nanoparticular formulations. J. Food Compos. Anal. 63, 103307 (2019). • The authors generated a combination of static and microfluidic chip 3D culture to mimic the 3D gastrointestinal tract to evaluate and characterize the muco-adhering and muco-permeating properties of nanoparticles with surface modification muco-adhesive proved by Carbopol® or the muco-permeating Pluronic® F127.