Periodic Reporting for period 2 - PEPSA-MATE (Nanopeptides and Nanosaccharides for Advanced and Sustainable Materials)
Periodo di rendicontazione: 2023-03-01 al 2025-08-31
The purpose of PEPSA-MATE is to create a multidisciplinary team – including early career researchers – to derive novel products from polysaccharides (complex carbohydrates) and peptides nanoparticles using experimental and computer-based theoretical design approaches and innovative ultrasonic fabrication technologies. Being eco-friendly alternatives to conventional fossil fuel-derived materials, the engineered nanoparticles are used as drug-delivery systems and biodegradable plastic matrix.
PEPSA-MATE outcomes expand knowledge base and research capability in Chemical biological Sciences and Technology by providing much-needed understanding and knowledge required to engineer new materials based on naturally sourced nanomaterials.
PEPSA-MATE involves key participation of junior researchers providing strong support and training for the development of their careers.
PEPSA-MATE develops intersectoral collaborations between a SME and academic partners to boost entrepreneurship creativity towards sustainable products and processes.
PEPSA-MATE brings together interdisciplinary and complementary expertise in the field of nanomaterials, biopolymers, polysaccharides, cell biology, acoustics, and computational modelling.
PEPSA-MATE outcomes expand knowledge base and research capability in Chemical biological Sciences and Technology by providing much-needed understanding and knowledge required to engineer new materials based on naturally sourced nanomaterials.
PEPSA-MATE involves key participation of junior researchers providing strong support and training for the development of their careers.
PEPSA-MATE develops intersectoral collaborations between a SME and academic partners to boost entrepreneurship creativity towards sustainable products and processes.
PEPSA-MATE brings together interdisciplinary and complementary expertise in the field of nanomaterials, biopolymers, polysaccharides, cell biology, acoustics, and computational modelling.
Briefly, during the second reporting period, we have engineered a library of modified glycogen nanoparticles. We engineered glycogen nanoparticles with tuneable structural and functional properties for the fabrication of advanced materials
First, we developed a simple and cost-effective method to reduce the size of glycogen nanoparticles in a controlled manner, achieving diameters ranging from 80 nm to 15 nm. These nanoparticles were subsequently functionalized with amine and carboxyl groups, as well as Protein A and Protein G, to enable antibody conjugation and the loading of RNA molecules, peptides, and small hydrophobic drugs. To investigate the interactions between glycogen and nucleic acids, molecular dynamics (MD) simulations were performed. These simulations characterized the binding between 20-nucleotide segments of luciferase-coding mRNA and amine-modified glycogen branches under both physiological (pH 7.4) and endosomal (pH 5) conditions. Next, we utilized biodegradable phytoglycogen nanoparticles (PG NPs)—highly branched glucose-based polysaccharides—as scaffolds for coupling adhesive dopamine motifs (for use as biodegradable underwater adhesives) and gold nanoparticles (for potential biosensing applications). The extent of chemical modification of PG NPs was assessed by NMR spectroscopy. The size distribution and colloidal stability of PG NPs in aqueous suspension were evaluated using dynamic light scattering (DLS), particle tracking, and chromatography. The morphology of the modified PG NPs was examined by electron microscopy (EM), atomic force microscopy (AFM), and super-resolution microscopy, while surface charge was determined via electrophoretic mobility measurements. Therapeutic peptides with anticancer and antimicrobial activity were synthesized using solid-phase peptide synthesis protocols and characterized by fluorescence spectroscopy and circular dichroism (CD). These positively charged peptides were either chemically conjugated to glycogen nanoparticles or encapsulated within negatively charged glycogen nanoparticles via electrostatic interactions. In parallel, porous microparticles based on hyaluronic acid microsponges (MSPs) were fabricated and loaded with therapeutic peptides for oral delivery. The stability of the microsponges and the retention of the peptide payload were evaluated in biological media mimicking the gastrointestinal tract, using fluorescence spectroscopy. Finally, glycogen nanoparticles were loaded with hydrophobic drugs, including GSA-10 and resveratrol, which are molecules under investigation for the potential treatment of multiple sclerosis and HIV, respectively. The interactions and bioactivities of these drug-loaded glycogen nanoparticles were assessed in T cells, cancer cells, and oligodendrocytes, focusing on cellular uptake kinetics and intracellular trafficking, using confocal microscopy and flow cytometry.
First, we developed a simple and cost-effective method to reduce the size of glycogen nanoparticles in a controlled manner, achieving diameters ranging from 80 nm to 15 nm. These nanoparticles were subsequently functionalized with amine and carboxyl groups, as well as Protein A and Protein G, to enable antibody conjugation and the loading of RNA molecules, peptides, and small hydrophobic drugs. To investigate the interactions between glycogen and nucleic acids, molecular dynamics (MD) simulations were performed. These simulations characterized the binding between 20-nucleotide segments of luciferase-coding mRNA and amine-modified glycogen branches under both physiological (pH 7.4) and endosomal (pH 5) conditions. Next, we utilized biodegradable phytoglycogen nanoparticles (PG NPs)—highly branched glucose-based polysaccharides—as scaffolds for coupling adhesive dopamine motifs (for use as biodegradable underwater adhesives) and gold nanoparticles (for potential biosensing applications). The extent of chemical modification of PG NPs was assessed by NMR spectroscopy. The size distribution and colloidal stability of PG NPs in aqueous suspension were evaluated using dynamic light scattering (DLS), particle tracking, and chromatography. The morphology of the modified PG NPs was examined by electron microscopy (EM), atomic force microscopy (AFM), and super-resolution microscopy, while surface charge was determined via electrophoretic mobility measurements. Therapeutic peptides with anticancer and antimicrobial activity were synthesized using solid-phase peptide synthesis protocols and characterized by fluorescence spectroscopy and circular dichroism (CD). These positively charged peptides were either chemically conjugated to glycogen nanoparticles or encapsulated within negatively charged glycogen nanoparticles via electrostatic interactions. In parallel, porous microparticles based on hyaluronic acid microsponges (MSPs) were fabricated and loaded with therapeutic peptides for oral delivery. The stability of the microsponges and the retention of the peptide payload were evaluated in biological media mimicking the gastrointestinal tract, using fluorescence spectroscopy. Finally, glycogen nanoparticles were loaded with hydrophobic drugs, including GSA-10 and resveratrol, which are molecules under investigation for the potential treatment of multiple sclerosis and HIV, respectively. The interactions and bioactivities of these drug-loaded glycogen nanoparticles were assessed in T cells, cancer cells, and oligodendrocytes, focusing on cellular uptake kinetics and intracellular trafficking, using confocal microscopy and flow cytometry.
Innovations in functional nanomaterials have contributed significantly to the transformation of many industries, including manufacturing, healthcare, cosmetics, textiles, and electronics. The first-generation of nanoparticle is primarily based on non-degradable organic and inorganic materials. However, the potential impact of synthetic and non-degradable NPs on human health and ecosystems (e.g. microplastic pollution) is raising significant social and environmental concerns because of their potential long-term cytotoxicity and accumulation. Hence, sustainable, green, non-toxic, and degradable alternatives to first-generation man-made synthetic nanomaterials are urgently needed. Such nanomaterials need to be cost effective, simple, reproducible, and scalable, and can be exploited as building blocks for engineering biodegradable, non-toxic functional materials. PEPSA-MATE pursues a sustainable-oriented approach in turning nanoparticles naturally sourced on a large-scale from sweet corn, and custom-designed peptides into products using innovative technologies.
IMPACT: During the secondments ESR involved in PEPSA-MATE developed communication and independent thinking skills and professional maturity. The partners produced 27 joint OA publications of high scientific value and 1 international patent application. The partners applied for new joint research grants (Horizon Europe RIA/IA, MSCA SE) and have organized two international conference on Peptide Materials 2023, 26-29 October, Sorrento Italy and the international symposium on Macromolecular Bioscience and Biomaterials, Frascati on 6-9 July 2025 for promoting the results of the ongoing research activities. PEPSA-MATE partners engaged numerous companies, stakeholders, and users to communicate the research outputs and identify possible opportunities to transfer the developed technologies. The coordinator set up a webpage to communicate and disseminate the results generated in the 2nd RP. The partners have implemented the outreach plan to target multiple audiences and provided interactive learning experiences to engage hundreds of high school students by using virtual reality headsets for an immersive explorative experience of the human body and nanomedicines.
IMPACT: During the secondments ESR involved in PEPSA-MATE developed communication and independent thinking skills and professional maturity. The partners produced 27 joint OA publications of high scientific value and 1 international patent application. The partners applied for new joint research grants (Horizon Europe RIA/IA, MSCA SE) and have organized two international conference on Peptide Materials 2023, 26-29 October, Sorrento Italy and the international symposium on Macromolecular Bioscience and Biomaterials, Frascati on 6-9 July 2025 for promoting the results of the ongoing research activities. PEPSA-MATE partners engaged numerous companies, stakeholders, and users to communicate the research outputs and identify possible opportunities to transfer the developed technologies. The coordinator set up a webpage to communicate and disseminate the results generated in the 2nd RP. The partners have implemented the outreach plan to target multiple audiences and provided interactive learning experiences to engage hundreds of high school students by using virtual reality headsets for an immersive explorative experience of the human body and nanomedicines.