SUSTAINABLE DEVELOPMENT: NEW TOOLS FOR THE SELECTION AND VALIDATION OF MACROMOLECULES FOR MEDICAL APPLICATIONS

A new eco-sustainable biotechnological approach was developed to identify and utilize naturally occurring polymers for the treatment of chronic lung diseases, such as chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF). Among these polymers, lactose-modified hyaluronic acid (Hylach®), designed to bind Galectin-3 (Gal-3), emerged as a promising candidate. Gal-3, a β-galactoside-binding lectin, plays a central role in the pathogenesis of these diseases by promoting inflammation and fibrosis through the synthesis of pro-inflammatory cytokines and extracellular matrix (ECM) molecules. Targeting Gal-3 has therefore been identified as a strategic therapeutic approach to mitigating these pathological processes in COPD and IPF. Through in silico analyses, Hylach derivatives were evaluated for their ability to bind Gal-3. Compounds with 10–40% lactosylation demonstrated the highest binding affinity, optimizing interactions within Gal-3’s carbohydrate-binding domain. These findings guided the selection of specific Hylach derivatives for subsequent experimental studies, minimizing unnecessary synthesis and reducing environmental impact. In vitro studies confirmed the effectiveness of Hylach derivatives, particularly the 30% lactosylated variant. Experiments conducted on 2D cultures of primary human bronchial fibroblasts, as well as lung fibroblasts derived from healthy subjects and IPF patients, revealed that Hylach 30% significantly reduced markers of inflammation, such as reactive oxygen species (ROS), pro-inflammatory cytokines, and matrix metalloproteinases (MMPs). Additionally, it downregulated key ECM molecules, including Collagen I, Collagen III, Elastin, and Fibronectin, in fibroblasts exposed to pro-fibrotic stimuli like TGF-β. Moreover, Hylach derivatives exhibited superior anti-inflammatory effects on inflamed bronchial epithelial cells (16HBE) compared to native HA. To further validate these findings, a three-dimensional (3D) fibroblast culture model was developed to better replicate the tissue microenvironment of chronic lung diseases. Lung fibroblast spheroids, derived from healthy subjects and IPF patients, were used to assess the anti-inflammatory and antifibrotic efficacy of Hylach in a context that better mimics in vivo conditions. These models demonstrated that Hylach 30% consistently reduced the expression of inflammatory and fibrotic molecules, including IL-1β, Gal-3, TNF-α, TGF-β, Collagen I, and Collagen III. The incorporation of both fibroblasts and epithelial cells into a single multicellular spheroidal structure enhanced the model’s complexity and biological relevance. Overall, the findings highlight the anti-inflammatory and antifibrotic potential of Hylach derivatives, particularly the 30% lactosylated variant, in targeting key pathological processes associated with chronic lung diseases. By combining computational and experimental approaches, this study forms the basis for the development of innovative and eco-sustainable therapies for inflammation and fibrosis.