title: Vitronectin absorbed on nanoparticles mediate cell viability/proliferation and uptake by 3T3 Swiss albino mouse fibroblasts: in vitro study.
abstract: We study the interaction of 3T3 Swiss albino mouse fibroblasts with polymeric nanoparticles (NPs) and investigate cellular behaviour in terms of viability/cytotoxicity, cell cycle, NPs uptake, MAP kinase (ERK1/2), and focal adhesion kinase (FAK) activation. After incubation of NPs with cell culture media, western blot analysis showed that Vitronectin is retained by NPs, while Fibronectin is not detected. From cytotoxicity studies (MTT and BrdU methods) an LD50 of about 1.5 mg/mL results for NPs. However, NPs in the range 0.01-0.30 mg/mL are able to trigger a statistically significant increase in proliferation and cell cycle progression in dose and time depending manner. Also, biochemical evaluation of ERK1/2 and FAK clearly shows an increasing phosphorylation in a dose and time depending manner. Finally, we found by transmission electron microscopy that NPs are internalised by cells. Competitively blocking VN-integrin receptors with echistatin (1 μg/mL) results in a decrease of viability/proliferation, cell cycle progression, cellular uptake, and FAK/ERK activation showing the involvement of Vitronectin receptors in signal transduction. In conclusion, our results show that cell surface NPs interactions are mediated by absorbed plasma proteins (i.e., Vitronectin) that represent an external stimuli, switched to the nucleus by FAK enzyme, which in turn modulate fibroblasts viability/proliferation.
authors.affiliation: Laboratory of Applied Biotechnology, Department of Anaesthesiological, Surgical and Emergency Sciences, Second University of Napoli, Via Costantinopoli 16, 80138 Napoli, Italy. francesco.rosso@unina2.it
authors.forename: F
authors.initials: F
authors.lastname: Rosso
authors.forename: G
authors.initials: G
authors.lastname: Marino
authors.forename: A
authors.initials: A
authors.lastname: Grimaldi
authors.forename: G
authors.initials: G
authors.lastname: Cafiero
authors.forename: E
authors.initials: E
authors.lastname: Chiellini
authors.forename: F
authors.initials: F
authors.lastname: Chiellini
authors.forename: M
authors.initials: M
authors.lastname: Barbarisi
authors.forename: A
authors.initials: A
authors.lastname: Barbarisi
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D010766:Phosphorylation; D015398:Signal Transduction; D041701:Swiss 3T3 Cells; D019096:Vitronectin / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D005353:Fibronectins; D019096:Vitronectin; D051416:Focal Adhesion Protein-Tyrosine Kinases