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Bio-Design and Manufacturing  2022 Vol.5 No.4 P.660-673

http://doi.org/10.1007/s42242-022-00202-6


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


Author(s):  Hui L. Ma, Wanlu Li, Mian Wang, Laudemir C. Varanda, Janice R. Perussi, Y. Shrike Zhang & Emanuel Carrilho

Affiliation(s):  Instituto de Química de São Carlos, Universidade de São Paulo, USP, Av. Trabalhador São-carlense, São Carlos, SP 13566-590, Brazil; more

Corresponding email(s):   yszhang@research.bwh.harvard.edu, emanuel@iqsc.usp.br

Key Words:  Malignant melanoma, 3D cell culture, Hypericin, Microemulsion, Photodynamic therapy


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Hui L. Ma, Wanlu Li, Mian Wang, Laudemir C. Varanda, Janice R. Perussi, Y. Shrike Zhang & Emanuel Carrilho. In vitro 3D malignant melanoma model for the evaluation of hypericin-loaded oil-in-water microemulsion in photodynamic therapy[J]. Journal of Zhejiang University Science D, 2022, 5(4): 660-673.

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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.

巴西圣保罗大学Carrilho与哈佛医学院Y. Shrike Zhang等 | 体外3D黑色素瘤模型用于光动力治疗的评估

本研究论文聚焦体外3D黑色素瘤模型用于光动力治疗的评估。体外3D黑色素瘤模型能够更为准确地重现体内肿瘤的生理变化,因而成为药物筛选和疾病模拟的有效工具。本文基于甲基丙烯酰明胶(GelMA)这一广泛应用于3D 细胞培养的组织工程支架制造的水凝胶,建立了负载金丝桃素(Hy) 的微乳液(ME),以构建3D黑色素瘤体外模型,同时评估Hy-ME在光动力治疗黑色素瘤中的效率。该研究使用的ME通过自发乳化法生产,从而提高了Hy在肿瘤部位的生物利用度。进一步通过光交联Hy-ME、6% GelMA和B16F10 、Balb/c 3T3细胞,建立了体外3D黑色素瘤模型。结果显示,含有Hy-ME的水凝胶比包含游离Hy的水凝胶表现出更高的光毒性和细胞积累,表明开发的Hy-ME微乳液体系可以作为潜在的Hy载体。此外,3D培养的细胞比2D培养的细胞显示更低的光动力治疗敏感性。综上,该研究揭示了基于GelMA 水凝胶的由模具构建的体外疾病模型,具有用户友好且成本低廉的优势,为探究皮肤癌和其他皮肤相关疾病的纳米载体递送效率提供了有效平台。

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