A New Approach for On-Chip Production of Biological Microgels Using Photochemical Cross-Linking.

Photochemical cross-linking is a key step for manufacturing microgels in numerous applications, including drug delivery, tissue engineering, material production, and wound healing. Existing photochemical cross-linking techniques in microfluidic devices rely on UV curing, which can cause cell and DNA damage. We address this challenge by developing a microfluidic workflow for producing microgels using visible light-driven photochemical cross-linking of aqueous droplets dispersed in a continuous oil phase. We report a proof-of-concept to construct microgels from the protein Bovine Serum Albumin (BSA) with [Ru(bpy)3]2+ mediated cross-linking. By controlling the capillary number of the continuous and dispersed phases, the volumetric flow rate, and the photochemical reaction time within the microfluidic tubing, we demonstrate the construction of protein microgels with controllable and uniform dimensions. Our technique can, in principle, be applied to a wide range of different proteins with biological and responsive properties. This work therefore bridges the gap between hydrogel manufacturing using visible light and microfluidic microgel templating, facilitating numerous biomedical applications.

In vitro clot model to evaluate fibrin-thrombin effects on fractal dimension of incipient blood clot.

INTRODUCTION: This aim of this study is to investigate the individual effects of varying concentrations of thrombin and fibrinogen on clot microstructure (characterised through the fractal dimension of the incipient clot network, df) and clot formation time (TGP) using a fibrin-thrombin clot model. df and TGP markers are measured using a haemorheological method that has already been investigated for whole blood. METHODS: This is an in vitro study using three thrombin concentrations (0.1, 0.05 and 0.02 NIH/ml) and two fibrinogen concentrations (8 mg/ml and 12 mg/ml) to investigate a fibrin-thrombin clot model. The haemorheological changes were measured at the gel point using df and TGP. RESULTS: Fractal dimension (df) increased with increasing concentrations of thrombin both at 8 mg/ml (1.60±0.024, 1.67±0.022, 1.74±0.079) and 12 mg/ml fibrinogen concentrations (1.63±0.02, 1.87±0.019, 1.95±0.014). On the other hand, TGP decreased for both 8 mg/ml (1089±265, 637±80, 223±22 seconds) and 12 mg/ml fibrinogen concentrations (2008±247, 776±20, 410±20 seconds). In contrast to previous studies investigating whole blood, TGP increased with higher fibrinogen levels. CONCLUSIONS: The findings from this fibrin-thrombin clot model confirmed that df and TGP can detect changes in the incipient clot following manipulation of fibrinogen and thrombin concentration. df increases (indicating stronger clot) with higher concentrations of thrombin and fibrinogen. On the other hand, TGP decreased as expected with higher thrombin level but not with higher fibrinogen concentrations.