Estratégia Universal de Mecano-ajuste de Hidrogeis: Goma Gelana Gamma-Irradiada para Revolucionar Modelos In Vitro ao Mimetizar Tecidos Vivos de forma Prolongada e Estável.

The major goal of the proposed research is to solve a long-standing dilemma in the field of biomaterials and 3D biological constructs – the fact that extracellular matrix components, as those that exist in the human body and properly interact with living cells, lack the 3D structure capacities to be implemented at the laboratory level with the confidence and stability required to model health and disease processes. Being able to solve that is prone to disrupt the in vitro testing pipelines across academic and industrial vectors, and enable the comprehensive analysis of longer-term, complex biological processes that so far are only possible with the use of animals. To do this, the proposed research will bridge far beyond the current state of the art in 3 major objectives: First, by exploring for the first time ever the gramma-induced breakage of gellan gum polymer molecular weight in the context of living cell encapsulation and biomedical modelling (Fig. 1, Fig. 2). Second, by using that to create a mechanotuner hydrogel component which plans to be universal, e.g. to virtually function with any hydrogel composition or material that needs to be studied by different scientists, industries, and applications. This is a very ambitious goal which we are in the position to propose as we have factual preliminary data (supplementary file) from our lab, which demonstrates feasibility within our initial explorations of the proposed methodologies (Fig. 1 and 3). Third, we aim to demonstrate why the capacity to stabilize and modulate the mechanics of 3D constructs made of soft hydrogels is so powerful, focusing on 3 major applications: Application 1) To sustain and regulate stem cell differentiation and 3D tissue formation in collagen hydrogels, which are soft and extremely challenging to manipulate in 3D constructs due to unwanted contraction (Fig 3). We plan to demonstrate how GGG can prevent that and allow for the on-demand stiffening of collagen, and how that can then enhance stem cell differentiation towards osteogenic lineage, effectively converting fat into bone. Application 2) To revolutionize 3D cancer cell models by tuning cancer invasiveness and add enhanced physiological relevance by mechanically tuning constructs to precisely match different tissue states (healthy, pre-malignant and tumoral), showing how those critically alter the behavior and aggressiveness of the cancer cells (Figure 4). This will demonstrate how important it is to adjust those characteristics to obtain in vitro responses that mimic what happens in the human body; Application 3) To create new avenues in biofabrication and additive manufacturing by inventing a totally new bioink framework where stability and absence of unwanted contraction may lead to highly functional printed structures that can finally realize the long-standing promise of organ regeneration and precision medicine. To do this, the proposed research plan and team integrates a broad range of disciplines, from materials science, physics, and chemistry, to cell and stem cell biology; from bio- and tissue-engineering, to cancer microenvironment biology, as well as biofabrication. All these expertise’s will be applied to the ambitious goal of creating a new biomaterial and hydrogel product, and to showcase its potential to revolutionize soft tissue 3D models across virtually unlimited biomedical applications.