Adhesion and proliferation of fibroblasts on RF plasma‐deposited nanostructured fluorocarbon coatings: Evidence of FAK activation

Rosso, Francesco; Marino, G.; Muscariello, Livio; Cafiero, Gioacchino; Favia, Pietro; D'Aloia, Erica; d’Agostino, Riccardo; Barbarisi, Alfonso

doi:10.1002/jcp.20595

Cited by 26 publications

(21 citation statements)

References 61 publications

(70 reference statements)

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“…ERK 1/2 and FAK activations per sample were calculated using the following formula: phosphoKinase/Total Kinase × 100, where phosphokinase was the densitometer signals intensity of phosph-ERK or phospho-FAK and, Total Kinase was the densitometer signal intensity of total ERK or total FAK as described by Rosso et al [36]. …”

Section: Methodsmentioning

confidence: 99%

Vitronectin Absorbed on Nanoparticles Mediate Cell Viability/Proliferation and Uptake by 3T3 Swiss Albino Mouse Fibroblasts: In Vitro Study

Rosso¹,

Marino²,

Grimaldi³

et al. 2013

BioMed Research International

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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.

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Section: Methodsmentioning

confidence: 99%

Vitronectin Absorbed on Nanoparticles Mediate Cell Viability/Proliferation and Uptake by 3T3 Swiss Albino Mouse Fibroblasts: In Vitro Study

Rosso¹,

Marino²,

Grimaldi³

et al. 2013

BioMed Research International

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“…In our experimental work on cell cultures and tissues we obtained high-resolution ESEM images on natural samples that underwent only a slight fixation (Muscariello et al, 2005;Rosso et al, 2006).…”

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confidence: 99%

Cell surface protein detection with immunogold labelling in ESEM: Optimisation of the method and semi‐quantitative analysis

Muscariello

Rosso

Marino

et al. 2007

Journal Cellular Physiology

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In this work we used a combination of immunogold labelling (IGL) and environmental scanning electron microscopy (ESEM) to detect the presence of a protein on the cell surface. To achieve this purpose we chose as experimental system 3T3 Swiss Albino Mouse Fibroblasts and galectin-3. This protein, whose sub-cellular distribution is still under discussion, is involved in a large number of cell physiological and pathological processes. IGL technique has been utilised by many authors in combination with SEM and TEM to obtain the identification/localisation of receptors and antigens, both in cells and tissues. ESEM represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artefacts and interfere with IGL procedure. The absence of metal coating could yield further advantages for our purpose as the labelling detection is based on the atomic number difference between Nanogold spheres and the biological material. Using the gaseous secondary electron detector (GSED) compositional contrast is easily revealed by the backscattered electrons component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimised to improve the intensity and the specificity of the labelling signal, in order to obtain a semi-quantitative evaluation of the labelling signal.

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“…25,26 The deposition mechanism of such micro/nanostructured coatings was discussed by Milella et al 27 We have previously deposited fluorocarbon thin films by plasma enhanced chemical vapor deposition (PECVD) fed with hexafluoropropylene oxide (C 3 F 6 O). 28 Their peculiar surface characteristics (e.g., roughness, hydrophobicity) have been demonstrated to depend on the experimental conditions, especially on the distance of the substrate from the plasma region in the reactor (glow vs afterglow areas).…”

Section: Introductionmentioning

confidence: 99%

Osteoblast-Like Cell Behavior on Plasma Deposited Micro/Nanopatterned Coatings

et al. 2010

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The behavior of cells in terms of cell-substrate and cell-cell interaction is dramatically affected by topographical characteristics as shape, height, and distance, encountered in their physiological environment. The combination of chemistry and topography of a biomaterial surface influences in turns, important biological responses as inflammatory events at tissue-implant interface, angiogenesis, and differentiation of cells. By disentangling the effect of material chemistry from the topographical one, the possibility of controlling the cell behavior can be provided. In this paper, surfaces with different roughness and morphology were produced by radiofrequency (RF, 13.56 MHz) glow discharges, fed with hexafluoropropylene oxide (C(3)F(6)O), in a single process. Coatings with different micro/nanopatterns and the same uppermost chemical composition were produced by combining two plasma deposition processes, with C(3)F(6)O and tetrafluoroethylene (C(2)F(4)), respectively. The behavior of osteoblast-like cells toward these substrates clearly shows a strict dependence of cell adhesion and proliferation on surface roughness and morphology.

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Adhesion and proliferation of fibroblasts on RF plasma‐deposited nanostructured fluorocarbon coatings: Evidence of FAK activation

Cited by 26 publications

References 61 publications

Vitronectin Absorbed on Nanoparticles Mediate Cell Viability/Proliferation and Uptake by 3T3 Swiss Albino Mouse Fibroblasts: In Vitro Study

Vitronectin Absorbed on Nanoparticles Mediate Cell Viability/Proliferation and Uptake by 3T3 Swiss Albino Mouse Fibroblasts: In Vitro Study

Cell surface protein detection with immunogold labelling in ESEM: Optimisation of the method and semi‐quantitative analysis

Osteoblast-Like Cell Behavior on Plasma Deposited Micro/Nanopatterned Coatings

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