Ms. Stefana Orobeti Profile

Stefana Orobeti

at Academia Romana

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Proceedings Article | 12 March 2024 Presentation + Paper

Polymeric scaffolds fabricated by two photon polymerization for 3D cancer cell invasion assay

Alexandra Bran, Florin Jipa, Stefana Orobeti, Emanuel Axente, Livia Sima, Felix Sima, Koji Sugioka

Proceedings Volume 12873, 128730B (2024) https://doi.org/10.1117/12.3001418

KEYWORDS: Cancer, Polymers, Melanoma, Two photon polymerization, Collagen, In vitro testing, 3D modeling, Ultrafast phenomena

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Three dimensionally (3D) engineered scaffolds are a viable alternative to investigate cells in physiologically relevant configurations. Two photon polymerization (TPP) is a 3D maskless laser direct writing technology that employs a focused femtosecond (fs) laser beam to produce a localized chemical reaction with high precision that ultimately leads to polymerization of a photosensitive material inside the focal volume. TPP has the capability of creating synthetic polymer constructs with 3D complex architectures and high resolution far beyond the diffraction limit. TPP was demonstrated to fulfill technical requirements necessary for fabricating personalized 3D scaffolds for tissue engineering and regenerative medicine applications. Herein, we propose the use of polymeric scaffolds fabricated by TPP for cancer research, specifically as model structures for cancer cell invasion assessment in 3D environments. In particular, the aim is to evaluate cancer cell interaction with confined spaces developed in a woodpile-like polymeric scaffold with pore dimensions less than 1 μm in microchannel cross-section. TPP of negative photoresist SU-8 was conducted using a 3D Lithography platform produced by Nanoscribe GmbH. Scaffolds with uniform networks of pores with sizes down to 0.66 μm were successfully produced, which were then used for melanoma cancer cell invasion assays. The scaffolds demonstrated potential for use in testing the invasion potential of melanoma cancer cells in comparison to normal melanocytes. Time-lapse microscopy observations were carried out to assess the optimal intervals for cell analysis in interaction with scaffolds. Preliminary in vitro tests suggested that melanoma-melanocytes co-culture may exhibit a more invasive potential in narrower spaces as compared to normal melanocytes alone, while an inhibitory effect on melanocyte invasion may be attributed to melanoma cells present in co-culture.

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