Multi-step femtosecond laser-fabricated membranes for regulated migration of biomolecules and cells

Gopalakrishnan, A.; Denduluri, A. J.; Gallegos, S.; Ramirez, I.; Schneider, S. E.; Cetinkaya, Z.; Kabutz, H.; Hedrick, A.; Jayaram, K.; Neu, C.; Whiting, G. L.

doi:10.64898/2026.05.22.726371

bioRxiv · 10.64898/2026.05.22.726371

Multi-step femtosecond laser-fabricated membranes for regulated migration of biomolecules and cells

Gopalakrishnan, A.·Denduluri, A. J.·Gallegos, S.·Ramirez, I.·Schneider, S. E.·Cetinkaya, Z.·Kabutz, H.·Hedrick, A.·Jayaram, K.·Neu, C.·Whiting, G. L.

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Abstract

Organ-on-chip (OoC) systems enable the recapitulation of key structural and functional characteristics of human tissues within controlled micro-engineered environments. In mechanically active tissues such as musculoskeletal, cardiac, and vascular systems, the incorporation of dynamic physical forces is essential for replicating the biomechanical cues governing cellular morphology and functional responses in-vivo. Without such stimuli, OoC models may fail to capture physiologically relevant tissue behaviors. Porous and semi-permeable membranes are critical components of OoCs, facilitating selective transport of nutrients, gases, and signaling molecules between cellular compartments to support biologically accurate barrier replication. Hence, fabrication strategies that permit precise modulation of membrane permeability are desirable to accommodate for the varying needs in pore size and porosity across organ systems. This study presents a two-stage fabrication process for stretchable, microporous polydimethylsiloxane (PDMS) membranes using femtosecond (fs-) pulse laser drilling. The laser-ablated pores exhibit a characteristic conical morphology, with diameters tapering from the laser entry to exit point. By modulating laser power and number of pulses, 6-15 m exit-end pore diameters were achieved in 50 m thick PDMS films. The membranes demonstrated strong mechanical resilience, with a 5-12% reduction in Youngs modulus after 500 cycles of strain loading. Furthermore, membranes fabricated at lower laser powers exhibited superior retention of elasticity, highlighting the influence of processing parameters on mechanical behavior. Cytocompatibility and permeability assessments confirmed that the membranes supported sustained cell viability and proliferation over at least three days. In size-restricted membrane pore geometries, cellular migration was constrained without any inhibition of biomolecular transport. This selective permeability is critical in multilayer OoC architectures, where a balance between biomolecular diffusion and cellular compartmentalization is necessary to preserve distinct tissue interfaces and functional organization. This work presents fs-laser micro-drilling as a robust and tunable fabrication strategy for producing mechanically resilient, selectively permeable PDMS membranes for physiologically relevant OoC applications.

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Gopalakrishnan, A., Denduluri, A. J., Gallegos, S., Ramirez, I., Schneider, S. E., Cetinkaya, Z., Kabutz, H., Hedrick, A., Jayaram, K., Neu, C., Whiting, G. L.. 2026-05-26. Multi-step femtosecond laser-fabricated membranes for regulated migration of biomolecules and cells. https://doi.org/10.64898/2026.05.22.726371

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