High-throughput mechanical nuclear envelope rupture and the intracellular dynamics of massive wound repair

The dynamics of nuclear envelope rupture and wound repair are critical biological processes that play essential roles in cell homeostasis. Previous studies on nuclear envelope repair dynamics have mainly focused on small ruptures induced by low-throughput tools such as laser ablation and atomic force microscopy. Recent findings also suggest that nuclear envelope rupture can aid the gene delivery process. We present a device that deterministically porates the cell membrane and nuclear envelope in high throughput, with applications in single-cell studies of wound repair dynamics and in statistical assessments of cell poration. The device consists of sharp nanostructures in arrays of microchannels, enabling precise and localized disruption at both the plasma membrane and nuclear envelope while preserving cell viability. The distribution of endosomal sorting complexes required for transport (ESCRT) proteins after mechanical disruption at both the plasma membrane and nuclear envelope highlights the cell recovery strategy for severe wounds. Interestingly, during this extreme rupture, resources seem to be allocated more towards nuclear envelope repair. The nanostructured microfluidics also facilitate rapid protein expression following naked plasmid DNA transfection, a feat that is only possible through nuclear envelope rupture. High-throughput mechanoporation on membrane systems provides a new perspective on studying complex wound repair dynamics and enables a means to deliver cargo to intracellular and intranuclear domains. This technology opens promising avenues for further research in the field of intracellular delivery and wound healing mechanisms.