Periodic Reporting for period 1 - PACGEL (Plasma-ACtivated hydroGEL: new frontiers solutions in cardiac regenerative medicine)
Periodo di rendicontazione: 2023-04-01 al 2025-03-31
Cardiovascular diseases are the leading cause of mortality, accounting for 45 % of global deaths in Europe. However, cardiac regenerative medicine solutions are currently missing (except for heart transplantation). Human heart diseases models are thus needed for the development and preclinical validation of new effective therapies.
In this context, PACGEL (Plasma-ACtivated hydrogGEL: new frontiers solutions in cardiac regenerative medicine) project aims to provide new strategies in cardiac regenerative medicine by developing injectable hydrogels with cardioprotective function and biomimetic in vitro 3D hydrogel-based models reproducing different oxidative stages of fibrotic cardiac scar tissues.
PACGEL will accelerate the fundamental knowledge in the development of new promising strategies for cardiac regenerative medicine, including new treatments and new preclinical models of human pathological cardiac tissue for their screening. Furthermore, models derived from PACGEL will reduce the costs and time for new therapy development, and the number of needed experimental animals in agreement of 3Rs (Reduction, Replacement, Refinement) principle.
In this context, PACGEL (Plasma-ACtivated hydrogGEL: new frontiers solutions in cardiac regenerative medicine) project aims to provide new strategies in cardiac regenerative medicine by developing injectable hydrogels with cardioprotective function and biomimetic in vitro 3D hydrogel-based models reproducing different oxidative stages of fibrotic cardiac scar tissues.
PACGEL will accelerate the fundamental knowledge in the development of new promising strategies for cardiac regenerative medicine, including new treatments and new preclinical models of human pathological cardiac tissue for their screening. Furthermore, models derived from PACGEL will reduce the costs and time for new therapy development, and the number of needed experimental animals in agreement of 3Rs (Reduction, Replacement, Refinement) principle.
In PACGEL project, atmospheric pressure plasma jet device was employed as a tool to generate unique mixtures of reactive oxygen species (ROS) in media, with the aim to mimic the in vivo-like heart microenvironment, under mild to severe conditions of oxidative stress. First, the effect of various ROS doses on human and rat cardiac cells (fibroblasts and cardiomyocytes) were studied in vitro with the aim to find out the maximum ROS dose above which oxidative stresses were induced in cells. Then, a proper ROS dose (below the above mentioned limit) able to promote cardioprotective effects on cardiomyocytes was selected and added into polymer solutions, to prepare injectable hydrogels for perspective use as a new therapeutic approach. ROS generation and stability versus time were studied using different polymer solutions. The physicochemical characterization of polymers after plasma treatment suggested the potential degradation of the polymer backbone with increasing ROS dose, and depending on polymer chemistry. A careful selection of polymer chemistry and ROS dose is thus needed leading to hydrogels with enhanced stiffness and the ability for a controlled ROS release.
For the first time the biological responses of several ROS concentration were studied on various human and rat cells (fibroblasts and cardiomyocytes) highlighting ROS and cell-dependent responses. Results derived from PACGEL depicted the relevance of ROS dose generated from atmospheric pressure plasma jet device versus conventional ROS mixtures (e.g. hydrogen peroxide as usually employed to reproduce oxidative stress conditions). Experimental results evidenced the importance to control the atmospheric pressure plasma jet for the generation of needed ROS concentration without significantly affecting the physicochemical properties of the hydrogels. Thanks to PACGEL experimental results, polymer chemistry could be linked to ROS chemistry in view to design optimal hydrogel systems for both injectable therapeutical approaches and 3D in vitro modelling of tissues.
PACGEL project allowed to study the effects of ROS from a biological and physicochemical point of view. Findings pave the way towards future research for the understanding of the advanced biological response (e.g. gene expression) of relevant human cardiac cells in 2D and 3D cultures. The controlled ROS release from plasma-activated hydrogels and derived biological response observed in PACGEL open promising opportunities for future in vivo validation.
Experimental data and results derived from PACGEL project are the subject of publications currently under preparation.
PACGEL project allowed to study the effects of ROS from a biological and physicochemical point of view. Findings pave the way towards future research for the understanding of the advanced biological response (e.g. gene expression) of relevant human cardiac cells in 2D and 3D cultures. The controlled ROS release from plasma-activated hydrogels and derived biological response observed in PACGEL open promising opportunities for future in vivo validation.
Experimental data and results derived from PACGEL project are the subject of publications currently under preparation.