On-Chip Synthesis of Hyaluronic Acid-Based Nanoparticles for Selective Inhibition of CD44+ Human Mesenchymal Stem Cell Proliferation
Enrica Chiesa,
Federica Riva,
Rossella Dorati,
Antonietta Greco,
Stefania Ricci,
Silvia Pisani,
Maddalena Patrini,
Tiziana Modena,
Bice Conti,
Ida Genta
Affiliations
Enrica Chiesa
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
Federica Riva
Department of Public Health, Experimental and Forensic Medicine, Histology and Embryology Unit, University of Pavia, Via Forlanini 10, 27100 Pavia, Italy
Rossella Dorati
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
Antonietta Greco
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
Stefania Ricci
Department of Public Health, Experimental and Forensic Medicine, Histology and Embryology Unit, University of Pavia, Via Forlanini 10, 27100 Pavia, Italy
Silvia Pisani
Immunology and Transplantation Laboratory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. Matteo, 27100 Pavia, Italy
Maddalena Patrini
Department of Physics, University of Pavia, Via Bassi 6, 27100 Pavia, Italy
Tiziana Modena
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
Bice Conti
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
Ida Genta
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy
In this study, an innovative microfluidics-based method was developed for one-step synthesis of hyaluronic acid (HA)-based nanoparticles (NPs), by exploiting polyelectrolytic interactions between HA and chitosan (CS), in order to improve reliability, reproducibility and possible scale-up of the NPs preparation. The on-chip synthesis, using a staggered herringbone micromixer, allowed to produce HA/CS NPs with tailored-made size and suitable for both parenteral (117.50 ± 4.51 nm) and loco-regional (349.15 ± 38.09 nm) administration, mainly composed by HA (more than 85% wt) with high negative surface charge (< −20 mV). HA/CS NPs were successfully loaded with a challenging water-insoluble molecule, Everolimus (EVE), an FDA- and EMA-approved anticancer drug able to lead to cell cycle arrest, reduced angiogenesis and promotion of apoptosis. HA/CS NPs resulted to be massively internalized in CD44+ human mesenchymal stem cells via CD44 receptor-mediated endocytosis. HA/CS NPs selectiveness towards CD44 was highlighted by blocking CD44 receptor by anti-CD44 primary antibody and by comparison to CS-based NPs cellular uptake. Eventually, high effectiveness in inhibiting cell proliferation was demonstrated on-chip synthetized EVE loaded HA/CS NPs by tracking in vitro DNA synthesis.
