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In vitro 3D malignant melanoma model for the evaluation of hypericin-loaded oil-in-water microemulsion in photodynamic therapy

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Abstract

Advances in biomimetic three-dimensional (3D) melanoma models have brought new prospects of drug screening and disease modeling, since their physiological relevancy for recapitulating in vivo tumor architectures is more accurate than traditional two-dimensional (2D) cell culture. Gelatin methacryloyl (GelMA) is widely used as a tissue-engineered scaffold hydrogel for 3D cell culture. In the present study, an in vitro 3D malignant melanoma model based on GelMA was fabricated to evaluate the efficiency of hypericin (Hy)-loaded microemulsion (ME) in photodynamic therapy against melanoma. The ME was produced by the spontaneous emulsification method to enhance the bioavailability of Hy at tumor sites. Hy-loaded MEs were applied to a 3D malignant melanoma model made using 6% GelMA and the co-culture of B16F10 and Balb/c 3T3 cells, followed by crosslinking using violet light (403 nm). The observation revealed excellent cell viability and the presence of F-actin cytoskeleton network. Hy-loaded MEs exhibited higher phototoxicity and cell accumulation (about threefold) than free Hy, and the cells cultured in the 3D system displayed lower susceptibility (about 2.5-fold) than those in 2D culture. These findings indicate that the developed MEs are potential delivery carriers for Hy; furthermore, GelMA hydrogel-based modeling in polydimethylsiloxane (PDMS) molds is a user-friendly and cost-effective in vitro platform to investigate drug penetration and provide a basis for evaluating nanocarrier efficiency for skin cancer and other skin-related diseases.

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Acknowledgements

This work was supported in part by the FAPESP-INCTBio (Process 2014/50867-3) and FAPESP-CEPOF (2013/07276-1). The authors gratefully acknowledge Dr. A.O. Ribeiro from Universidade Federal do ABC, Santo André-SP, Brazil, for kindly providing the Hypericin and CAPES fellowship—Finance Code 001 to HLM; YSZ acknowledges support by the Brigham Research Institute; LCV thanks FAPESP for financial support (Process 2013/01284-2).

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Authors and Affiliations

  1. Instituto de Química de São Carlos, Universidade de São Paulo, USP, Av. Trabalhador São-carlense, São Carlos, SP, 13566-590, Brazil

    Hui L. Ma, Laudemir C. Varanda, Janice R. Perussi & Emanuel Carrilho

  2. Instituto Nacional de Ciência e Tecnologia de Bioanalítica, INCTBio, Campinas, SP, 13083-970, Brazil

    Hui L. Ma & Emanuel Carrilho

  3. Division of Engineering in Medicine, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA, 02139, USA

    Hui L. Ma, Wanlu Li, Mian Wang & Y. Shrike Zhang

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  1. Hui L. Ma
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  2. Wanlu Li
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  4. Laudemir C. Varanda
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Contributions

HLM: conceptualization, methodology, visualization, investigation, writing—original draft preparation. WL: conceptualization, methodology. MW: methodology. LCV: methodology, writing—review & editing. JRP: resources. YSZ: conceptualization, methodology, writing—review & editing, supervision. EC: conceptualization, writing—review & editing, project administration, resources, supervision.

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Correspondence to Y. Shrike Zhang or Emanuel Carrilho.

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Ma, H.L., Li, W., Wang, M. et al. In vitro 3D malignant melanoma model for the evaluation of hypericin-loaded oil-in-water microemulsion in photodynamic therapy. Bio-des. Manuf. 5, 660–673 (2022). https://doi.org/10.1007/s42242-022-00202-6

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