Supercritical CO2-assisted decellularization: advanced pancreatic tissue platform for diabetes treatment

Introduction: Decellularization is a procedure that aims to remove the cellular and antigenic material of a tissue while preserving its extracellular matrix (ECM) and biological properties. Decellularized ECM offers a natural microenvironment with significant potential for use in cell-based therapies and tissue regeneration [1]. Different protocols have been employed to decellularize tissues and organs. However, many rely on solvents and detergents that compromise the bioactive signals inherent to the native tissue [2]. Thus, it is crucial to establish a method that achieves effective decellularization while maintaining the ECM biochemical integrity. Objectives: In this study, we focus on the pancreatic tissue decellularization to create a native-like matrix suitable for the delivery of functional, insulin-producing β-cells as a therapeutic approach for diabetes treatment [3]. We propose a decellularization protocol based on Supercritical CO₂ (scCO₂) technology as an innovative alternative to solvent-based methods, offering high transfer rates, diffusivity, chemical inertness, and non-toxicity [4]. Conclusions: Both decellularization protocols efficiently removed DNA and SEM confirmed ECM ultrastructure maintenance in both groups. Biochemical analysis revealed that scCO₂ decellularization slightly improves GAG’s preservation, while insoluble collagen is protocol-independent and soluble collagen remains similar in both methods. Proteomic analysis showed that traditional treatment retained more proteoglycans and SLRPs, whereas scCO₂ better preserved some basal membrane and especially elastic fiber proteins. Biocompatibility assays demonstrated that both scaffolds supported cell adhesion, viability, and proliferation. In conclusion, both protocols generate biocompatible scaffolds, with distinct ECM preservation profiles relevant for recellularization, but scCO₂ stands out for being a green technology, with a shorter effective decellularization time and reduced waste.