Herein, we demonstrate the translation of Two-Photon Grayscale Lithography (2GL®), as well as Aligned 2-Photon Lithography (A2PL®), to biomedical applications. Specifically, we will present a novel workflow of aligned two-photon polymerization (2PP) microfabrication for 3D cell assays and perfusion inside microfluidic devices. For completeness, we also reveal how 2GL® can be applied to artificial intelligence (AI) generated topographies for enhanced and scalable 2.5D cell culturing. The versatility offered by both aligned and 2GL® printing holds great promise for various applications in biotechnology, tissue engineering, and microfluidics, creating new opportunities for innovation within established biomedical and pharmaceutical industries.
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