Engineering Gelation Kinetics in Living Silk Hydrogels by Differential Dynamic Microscopy Microrheology and Machine Learning

Abstract

Microbes embedded in hydrogels comprise one form of living material. Discovering formulations that balance potentially competing mechanical and biological properties in living hydrogels--for example gel time of the hydrogel formulation and viability of the embedded organisms--can be challenging. In this work, a pipeline is developed to automate characterization of the gel time of hydrogel formulations. Using this pipeline, living materials comprised of enzymatically crosslinked silk and embedded E. coli--formulated from within a 4D parameter space--are engineered to gel within a pre-selected timeframe. Gelation time is estimated using a novel adaptation of microrheology analysis using differential dynamic microscopy (DDM). In order to expedite the discovery of gelation regime boundaries, Bayesian machine learning models are deployed with optimal decision-making under uncertainty. The rate of learning is observed to vary between AI-assisted planning and human planning, with the fastest rate occurring during AI-assisted planning following a round of human planning. For a subset of formulations gelling within a targeted timeframe of 5-15 minutes, fluorophore production within the embedded cells is substantially similar across treatments, evidencing that gel time can be tuned independent of other material properties--at least over a finite range--while maintaining biological activity.