Cell biology

The research activities of this area at the cross-roads of materials science, nanotechnology and cell biology, primarily focusing on how newly-discovered materials and interfacial processes can be developed and used for specific applications. In particular, our research emphasizes cytomechanic studies correlating cytomechanical profiles to the underlying molecular and cellular mechanisms to isolate and study extracellular vesicles, with materials engineering promoting and driving cell adhesion and differentiation.

Cellular homeostasis and cytomechanics

The surface of living cells is strictly related to many cellular processes such as adhesion, signalling, transport, energy transformation, tumour metastasis etc. Cellular functions are mediated by a plethora of specific biomolecules including cell-to-cell adhesion proteins and those that constitute the highly complex and dynamic architecture of the cytoskeleton which, in turn, connects structurally and functionally the intracellular environments with the extracellular matrix via other adhesion molecules.

Our research activities focus on the biomolecular and cytomechanical cellular behaviour of primary fibroblast cultures related to juvenile hereditary Parkinson’s disease and of epithelial tumour cell lines. Nanomechanical properties of single cells obtained by advanced microscopic techniques allow to correlate morpho-mechanical cellular state changes upon interaction with pharmacological agents and the underlying biological \ biochemical processes.

From Left to right: Cover image of Ref. 4, selected publications; CLSM micrographs of organization of F-actin (stained with phalloidin, in red) and microtubules (stained with a-Tub, in green) filaments in fibroblasts cells (nuclei stained with DAPI, in blue) (Ref.3);  Laser scanning confocal microscopy (left) and Scanning Force Microscopy image (right) of MCF-7 cells grown on Fn modified Petri dish. Cells were stained with TRITC-labelled phalloidin to reveal F-actin and with Hoechst to detect nuclei. When grown on Fn, the cells exhibited short actin-rich extensions around their perimeters, as indicated by arrows, as well as cytoplasmic extensions at the cell edges.
From Left to right: Cover image of Ref. 4, selected publications; CLSM micrographs of organization of F-actin (stained with phalloidin, in red) and microtubules (stained with a-Tub, in green) filaments in fibroblasts cells (nuclei stained with DAPI, in blue) (Ref.3); Laser scanning confocal microscopy (left) and Scanning Force Microscopy image (right) of MCF-7 cells grown on Fn modified Petri dish. Cells were stained with TRITC-labelled phalloidin to reveal F-actin and with Hoechst to detect nuclei. When grown on Fn, the cells exhibited short actin-rich extensions around their perimeters, as indicated by arrows, as well as cytoplasmic extensions at the cell edges.

Analysis of Extracellular vesicles 

A recently developed research activity concerns the EVs isolation from fibroblasts of parkin-mutant patients and the characterization of their biochemical differences compared to EV derived from control samples. In particular, EVs are isolated from culture media by an optimized differential centrifugation protocol and their size and morphology determined by flow cytometry, electron and atomic force microscopy. We are also performing a mass spectrometry analysis to investigate their lipidomic profiles.

Materials for cell engineering

Cell engineering is a very promising research field which aims to induce specific biological processes such as proliferation and differentiation, cell-to-cell interaction, biomolecular production and extracellular matrix (ECM) formation, at the cell/material interface. Specifically cell migration is a phenomenon that is involved in different physiological processes such as morphogenesis, wound healing and tumour invasion. Biochemical or biophysical stimuli such as chemotaxis, galvanotaxis extracellular matrix compliance/stiffness and topology can influence cell migration in terms of speed, direction and persistence. The control of the cell environment by multiple physicochemical cues has therefore emerged as a key factor to enable functionality, modulate response, and affect cell behaviour.

In particular our recent studies have shown that cells are able to recognize the mechanical properties of a substratum over which they move and that these properties direct the motion through a phenomenon called durotaxis. Thus, mechanical interactions between a cell and its underlying substratum play a crucial role in modulating cell motility. We are investigating combinations of external stimuli ranging from chemical to mechanical and electrical cues in the cellular microenvironment. Moreover we are carrying out cell tracking analysis to deepen our understanding of the mechanisms underlying cell motility.

. Mechanical variables influencing cell migration. Cells movement is random on substrate with a homogeneously distributed elasticity. However on a surface with an asymmetric elasticity, cells move by durotaxis towards regions with high elasticity. Representative tissue-stiffness and Young's modulus, E. Representative examples of durotactic regulation of cell behaviour on double sheeted PDMS substrates exhibiting a mechanical gradient due to the micropattern underlying a thin membrane. (On the lower right) Immunofluorescent staining showing the response of human fibroblasts to micro-patterned PDMS double sheet substrates with mechanical gradient.

Plasma processing to control cell adhesion

The nature of the interface between cells and materials can stimulate a repulsive (or adhesive) response that can causes the cells to separate (adhere) to the desired material. This effect dramatically depends on surface properties of the material. Plasma surface modification is applicable whenever the surface of the device has to be bioactive or bioinert by changing chemical/topographical features of a material surface without affecting its bulk properties. As an example, the irreversible, undesired adhesion of biomolecules and cells (i.e. ‘biofouling’) can be controlled by a plasma assisted deposition of Polyethylene oxide (PEO)-like coatings. On the other hand, surfaces containing polar groups (i.e NH2, COOH, OH etc. …) or deposition of coatings in which biological molecules are dispersed in an organic matrix can be produced by plasma in order to impart bioactivity and biocompatibility to the surface. Finally, surfaces with micro- and nanostructured coatings can dramatically improve cell/material interactions due to topographical cues.

Sketch of the potentialities of plasma processing of surfaces in order to produce micro-nanostructured surfaces (left); unfouling surfaces (top), biomimetic/bioactive surfaces in which a coating embedding active biomolecules are deposited (right) and functional surfaces containing chemical groups like amino, carbonyl, alcoholic and carboxylic ones (bottom)
Sketch of the potentialities of plasma processing of surfaces in order to produce micro-nanostructured surfaces (left); unfouling surfaces (top), biomimetic/bioactive surfaces in which a coating embedding active biomolecules are deposited (right) and functional surfaces containing chemical groups like amino, carbonyl, alcoholic and carboxylic ones (bottom)

Plasma processing of cells and biological liquids 

Cold atmospheric pressure plasmas are emerging as an exciting development for therapeutics. These plasmas are very efficient sources of highly reactive oxygen and nitrogen species (RONS), UV radiation, electromagnetic fields and charged particles. Experiments show that cold atmospheric plasmas allow efficient, contact-free and painless disinfection, without damaging healthy tissue. In healthcare, new horizons are being opened for wound healing, tissue regeneration, cancer therapy, and treatment of chronic wounds assisted by plasma technology.

Our facilities have been designed to allow us to study the response of different type of cells to different plasma doses and to correlate the chemical composition of plasma treated cell culture media with cell behaviour.

Negative effect of plasma dose (different treatment times) on Saos2 tumor cell lines cytoskeleton.
Negative effect of plasma dose (different treatment times) on Saos2 tumor cell lines cytoskeleton.

Facilities & Labs

NanoFab Lab @ Lecce

Lab di Caratterizzazione @ Lecce

Bio Lab @ Lecce

S.Li.M. Lab @ Roma

Bio Lab @ URT Bari

Chemical-Structural Characterization Lab@ URT Bari

Wet chemistry Lab@ URT Bari

Plasma Technologies Lab@ URT Bari

People

Franco_CalabiFranco

Calabi

CNR Senior Research

Antonio_GaballoAntonio

Gaballo

CNR Research

Viso_donnaPaola

Priore

CNR PostDoc

Ilaria_PalamaIlaria E.

Palamà

CNR Research

Barbara_CorteseBarbara

Cortese

CNR Research

Eloisa_SardellaEloisa

Sardella

CNR Researcher

Stefano_LeporattiStefano

Leporatti

CNR Research

Pietro-FaviaPietro

Favia

Associate Professor

Roberto_GristinaRoberto

Gristina

CNR Researcher

fabio_palumbor150Fabio

Palumbo

CNR Researcher

Publications

  1. Lippolis, R. A. Siciliano, C. Pacelli, A. Ferretta, M. F. Mazzeo, S. Scacco, F. Papa, A. Gaballo, C. Dell’Aquila, M. De Mari, S. Papa , T. Cocco, Altered protein expression  pattern in skin fibroblasts from parkin-mutant early-onset Parkinson’s disease patients. Biochimica Biophysica Acta, 1852, 1960-1970 (2015). ISSN: 0925-4439; doi: 10.1016/j.bbadis.2015.06.015
  2. Vergara, P. Simeone, F. Julien, M. Trerotola, A. Giudetti, L. Capobianco, A. Tinelli, C. Bellomo,
I. Fournie, A. Gaballo, S. Alberti, M. Salzet, M. Maffia, Translating epithelial mesenchymal transition markers into the clinic: Novel insights from proteomics EuPA Open Proteomics, 10, 31-41, (2016) ISSN: 2212-9685; doi:10.1016/j.euprot.2016.01.003
  3. Vergara, M. Ferraro, M.F. Cascione, L.L. del Mercato, S. Leporatti, A. Ferretta, P. Tanzarella, C. Pacelli, A. Santino, M. Maffia, T. Cocco, R. Rinaldi, A. Gaballo, Cytoskeletal Alterations and Biomechanical Properties of parkin-Mutant Human Primary Fibroblasts. Cell Biochemistry Biophysics, 71, 1395-1404, (2015). ISSN:1085-9195; doi: 10.1007/s12013-014-0362-1
  4. Ferretta, A. Gaballo, P. Tanzarella, C. Piccoli, N. Capitanio, B. Nico, T. Annese, M. Di Paola, C. Dell’aquila, M. De Mari, E. Ferranini, V. Bonifati, C. Pacelli, T. Cocco, Effect of resveratrol on mitochondrial function: implications in parkin-associated familiar Parkinson’s disease Biochimica Biophysica Acta, 1842, 902-915 (2014). ISSN: 0925-4439; doi: 10.1016/j.bbadis.2014.02.010
  5. Vergara, P. Simeone, D. Latorre, M.F. Cascione, S. Leporatti, M. Trerotola, A.M. Giudetti, L. Capobianco, P. Lunetti, A. Rizzello, R. Rinaldi, S. Alberti, M. Maffia. Proteomics analysis of E-Cadherin knockdown in epithelial breast cancer cells J. Biotechnology Special Issue EuroBiotech 22, 3-11 (2015). ISSN: 0168-1656; doi:10.1016/j.jbiotec.2014.10.034.
  6. E Palamà, S. D’Amone, B. Cortese, Chapter 22: Mechanical guidance of cell migration, Nanomaterials and Regenerative Medicine (Y. Lin and T. Gong, ed.) IAPCOBP Publishing, (2016) (invited chapter). ISBN: 978-953-56942-3-6. DOI: 10.5599/obp.9.0
  7. Pagani, R. C. Paolicelli, E. Murana, B. Cortese, S. Di Angelantonio, E. Zurolo, E.Guiducci, T. A. Ferreira, S. Garofalo, M. Catalano, G. D’Alessandro, A. Porzia, G.Peruzzi, F. Mainiero, C. Limatola, C.T. Gross, D. Ragozzino, Defective microglial development in the hippocampus of CX3CR1 deficient mice Front. Neurosci. 9,111. (2015) ISSN: 1662-5102; doi: 10.3389/fncel.2015.00111. ISSN: 1662-453X.
  8. Cortese, I.E Palamà, S. D’Amone, G. Gigli. Influence of electrotaxis on cell behaviourIntegr.Biol6,817 – 830, (2014). ISSN: 1520-6602; doi: 10.1039/C4IB00142G Integrative Biology 2014 HOT Articles.
  9. Sardella, E.R. Fisher, J.C. Shearer, M.G. Trulli, R. Gristina, P. Favia. N2/H2O plasma assisted functionalization of Poly(epsilon-caprolactone) porous scaffolds: acid/basic character versus cell behavior. Plasma processes and Polymers 12-8, 786-798 (2015). ISSN: 1612-8850; doi: 10.1002/ppap.201400201
  10. Yang, G. Camporeale, E. Sardella, G. Dilecce, J.S. Wu, F. Palumbo, P. Favia; Deposition of Hydroxyl Functionalized Films by means of water Aereosol assisted Atmospheric pressure plasma Plasma processes and polymers 11-11, 1102-1111 (2014) ISSN: 1612-8850; doi: 10.1002/ppap.201400066
  11. Trizio, E. Sardella, E. Francioso, G. Dilecce, V. Rizzi, P. Cosma, M. Schmidt, M. Hansch, T. von Woedtke, P. Favia, R. Gristina; Investigation of air-DBD effects on biological liquids for in vitro studies on eukaryotic cells, Clinical Plasma Medicine 3-2, 62-71 (2015). ISSN: 22128166; doi: 1016/j.cpme.2015.09.003.

Other selected publications

  1. C. Pacelli, D. De Rasmo, A. Signorile, I. Grattagliano, G. di Tullio, A. D’Orazio, B.Nico, G. P. Comi, D. Ronchi, E. Ferranini, D. Pirolo, P. Seibel, S. Schubert, A. Gaballo, G. Villani, T. Cocco Mitochondrial defect and PGC-1α dysfunction in parkin-associated familial Parkinson’s disease. Biochimica Biophysica Acta, 1812,1041-1053, (2011) ISSN: 0925-4439; doi: 10.1016/j.bbadis.2010.12.022
  2. N. Denora, V. Laquintana, A. Lopalco, R. M. Iacobazzi, A. Lopedota, A. Cutrignelli, G. Iacobellis, C. Annese, M. F. Cascione, S. Leporatti, M. Franco In vitro targeting and imaging the translocator protein TSPO 18-kDa through G(4)-PAMAM-FITC labeled dendrimers J. Contr. 172, 1111-1125 (2013). ISSN: 0168-3659; doi: 10.1016/j.jconrel.2013.09.024
  3. D. Vergara, P. Simeone, D. Toraldo, P. del Boccio, V. Vergaro, S. Leporatti, D. Pieragostino, A. Tinelli, S. De Domenico, S. Alberti, A. Urbani, M. Salzet, A. Santino, and M. Maffia Resveratrol downregulates Akt/GSK and ERK signalling pathways in OVCAR-3 ovarian cancer cells Molecular BioSystems 8, 1078-1087 (2012). ISSN: 1742-206X; doi: 10.1039/c2mb05486h
  4. Leporatti, D. Vergara, A. Zacheo, V. Vergaro, G. Maruccio, R. Cingolani, R. Rinaldi Cytomechanical and topological investigation of MCF-7 cells by scanning force microscopyNanotechnology 20 055103 (2009). (Paper of the month, Cover Page Issue) ISSN: 0957-4484; doi: 10.1088/0957-4484/20/5/055103
  5. I.E. Palamà, S. D’Amone, A.M.L. Coluccia, G. Gigli. Micropatterned polyelectrolyte multilayer films promote alignment and myogenic differentiation of C2C12 cells in standard growth medium. Biotechnology & Bioengineering, 110, 586-596, (2013). ISSN: 1097-0290; doi: 10.1002/bit.24626.
  6. B. Cortese,  M.O. Riehle, S. D’Amone, G. Gigli, Influence of Variable Substrate geometry on Wettability and Cellular Responses, Journal of Colloid and Interface Science 394, 582–589, (2013). ISSN: 0021-9797, doi: 10.1016/j.jcis.2012.11.051.
  7. I.E. Palamà, S. D’Amone, A.M.L. Coluccia, M. Biasiucci, G. Gigli Cell self-patterning on uniform PDMS-surface with controlled mechanical cues Integrative Biology, 4, 228-236, (2012). doi: 10.1039/c2ib00116k; ISSN 1757-9708; (Highlight: the paper is the 5th position of the “top ten most accessed articles” of Integrative Biology web site in the month of December 2011)
  8. B. Cortese, G. Gigli, M. Riehle, Mechanical Gradient Cues for Guided Cell Motility and Control of Cell Behaviour on uniform substrates, Adv. Funct. Mater., 19, 2961-2968, (2009). ISSN: 1616-3028; doi: 10.1002/adfm.200900918;
  9. B. Cortese, C. Piliego, I. Viola, S. D’Amone, R. Cingolani, G. Gigli, Engineering transfer of micro and nanometer scale features by surface energy modification, Langmuir, 25, 7025–7031, (2009). ISSN 0743-7463; doi: 10.1021/la900248j.
  10. G. Da ponte, E. Sardella, F. Fanelli, R. d’Agostino, R. Gristina, P. Favia; Plasma deposition of PEO-like coatings wirth aereosol assisted dielectric barrier discharges PLASMA PROCESSES AND POLYMERS 9-11 1176-1183 (2012). ISSN: 16128850; doi: 10.1002/ppap.201100201

Patents

Process for the production by plasma of nanometric thickness coatings allowing controlled release of silver ions of other elements, or of molecules of biomedical interest, from solid products, and products thus coated R. D’agostino, P. Favia, F. Fracassi, E. Sardella, C. Costagliola, A. Mangone. Patent WO2013021409-A1: E. Sardella, P. Favia et al. WO2013021409 (2013)

Abstract: Process for the production by plasmochemical deposition of a film having a nanometric thickness, optionally multilayered, permitting carrying out in a controlled, uniform and long lasting way, release of substances of interest in a surrounding medium containing liquids, from a substrate including the substance to be released as micro/nano particles, or from a layer deposited on the substrate including the substance to be released as micro/nano particles, or from a layer of the substance to be released deposited on the substrate, or from a substrate that is the substance to be released optionally in the form of particles. The substances to be released can be metals, compounds having anti-bacterial properties, biologically active molecules such as drugs, hormones, vegetable extracts, peptides, lipids, protides and glucides. The layer with the substance to be released, be it organic or inorganic, is obtained by plasmochemical deposition optionally having a structure similar to polyethylene oxide (PEO) or polyethylene glycol (PEG), called PEO-like polymers, constituted, in a variable percentage da ethylene oxide units (-CH2CH2O-, EO); barrier film is obtained by depositing by plasma at least one organic or inorganic layer, optionally with a PEO-like structure, wherein chemical composition, degree of crosslinking and thickness are adjustable by the plasmo chemical deposition process parameters, and allow to adjust the release of the active substance according to specific needs. The structures on which the above said films can be deposited are: medical-surgical devices, common handworks, structures known as scaffolds, and the above defined substances to be released themselves. The invention also relates to medical-surgical devices, common handworks and scaffolds coated by a substrate and barrier layer, as well as to biologically active substances coated by at least one barrier layer.

Project

  1. My First AIRC Grant – Role of Electro/Mechanical cues in the control and guidance of Glioma Progression (MFAG)  2015 n. 16803 (2015-2018)
  2. SITEMA – Sviluppo di nuove metodologie e strumenti Innovativi per la diagnoSi ed il trattamento Terapeutico di tumori Epiteliali uMAni. Regione Puglia Bando “Aiuti a Sostegno Cluster Tecnologici Regionali” (2015-2017)
  3. RINOVATIS – Rigenerazione di tessuti nervosi ed osteocartilaginei mediante innovativi approcci di Tissue Engineering, PON MIUR PON02_00563_3448479, (2013-2015)

Awards

  1. Yang, G. Camporeale, E. Sardella, G. Dilecce, F. Palumbo, P. Favia, J.-S. Wu; One-step Atmospheric Pressure Plasma Synthesized Polyethylene Embedded with Tunable Amount of Lysozyme; Best Oral paper at 8th Asia-Pacific International Symposium on the Basics and Applications of Plasma Technology APSPT8 20th-22nd December, 2013, Hsinchu, Taiwan performed by the first author
  2. Camporeale, Y.W. Yang, E. Sardella, G. Dilecce, F. Palumbo, J.S. Wu, P. Favia; New protein carrier systems deposited by atmospheric pressure glow discharge fed with water-ethylene mixture Young Scientist Lecture Competition for the first author at 8th Int. Conf. on Surface, Caotings and Nano-Structured Materials; (NANOSMAT) 22-25 September 2013, Granada, Spain
  3. Trizio, R. Gristina, E. Sardella, E. Francioso, G. Dilecce, M. Schmidt, T. von Woedtke, P. Favia; Effects of air DBD on eukaryotic cells and biological liquids, Best Poster award at the 22nd International Symposium on Plasma Chemistry July 5-10, 2015 Antwerp, Belgium

Latest News

I° meeting TecnoMed Puglia

Lecce, 05 dicembre 2019 - Aula Rita Levi Montalcini - CNR NANOTEC Lecce

Si terrà domani, giovedì 05 dicembre, con inizio alle ore 14.00 presso l'aula Rita Levi Montalcini del Cnr Nanotec, il "I° meeting TecnoMed Puglia: Tecnopolo per la medicina di precisione". Il meeting mira a fare il punto sulle attività programmate, sullo stato di avanzamento e sugli highlights.

Puoi scaricare la locandina da qui

Jam session Nanotec... note di scienza su scala nanometrica

Lecce, 27 settembre 2019 - ex monastero degli Olivetani "CAR-T: l'alba di una nuova era"  con: Attilio Guarini (IRCCS Istituto Tumori “Giovanni Paolo II” di Bari)  introduce e modera: Marco Ferrazzoli (Ufficio Stampa CNR Roma) a cura di: Gabriella Zammillo 

Le CAR-T (Chimeric Antigens Receptor Cells-T) sono cellule modificate in laboratorio a partire dai linfociti T. Rappresentano una nuova strategia di cura che sfrutta il sistema immunitario per combattere alcuni tipi di tumore come linfomi aggressivi a grandi cellule e leucemie linfoblastiche acute a cellule B. Il prof Attilio Guarini, ematologo all’Istituto tumori Giovanni Paolo II di Bari, le definisce la “vis sanatrix naturae della antica medicina salernitana”, trattandosi del potenziamento dell’attività citotossica dei linfociti del paziente opportunamente ingegnerizzati per riconoscere e contrastare alcuni tipi di cellule tumorali.

 

Le CAR-T possono quindi essere definite un “farmaco vivente” proprio perché prodotto a partire dalle cellule dello stesso paziente aprendo così ad un nuovo mondo, considerato che i farmaci convenzionali sono prodotti da sostanze chimiche o, in alternativa, sono anticorpi prodotti in laboratorio dai biologi. Un trattamento estremamente complesso e costoso, non sempre applicabile, ma laddove possibile, dai risultati incoraggianti per le aspettative di vita. Lo sviluppo di nuove tecnologie per la produzione di CAR-T è parte integrante delle attività di ricerca condotte dal TecnoMed Puglia, il TecnoPolo per la Medicina di Precisione, coordinato da Giuseppe Gigli direttore del Cnr Nanotec di Lecce, e che nel suo nucleo fondatore vede anche l’IRCCS Istituto Tumori “Giovanni Paolo II” di Bari, il Centro di malattie neurodegenerative e dell’invecchiamento cerebrale dell’Università di Bari con sede presso l’Ospedale " G. Panico" di Tricase e la Regione Puglia.

 

L'evento apre la nuova stagione della rassegna divulgativa "Jam session Nanotec: note di scienza su scala nanometrica", un progetto Cnr Nanotec di Gabriella Zammillo, realizzato in collaborazione con Liberrima.

A condurre e moderare la serata, Marco Ferrazzoli, capo ufficio stampa dal CNR. Puoi scaricare la locandina da qui

Notte dei Ricercatori

Lecce, 27 settembre 2019

 

ex monastero degli Olivetani, ore 18:00 - 24:00

 
 Ritorna puntuale la Notte dei Ricercatori, l’evento più atteso dai tanti appassionati di scienza, ghiotti di conoscenza senza distinzione di età. E sempre più densa di contenuti è la partecipazione del @CnrNanotec che, per l’edizione 2019,  ha reso ancora più appetibile il calendario degli appuntamenti programmati all’interno del progetto europeo #ERN-Apulia  coordinato da Unisalento, tracciando un ideale tour tra gli intriganti campi del sapere che si dipana attraverso narrazioni, illustrazioni, laboratori hand-on, dibattiti, giochi per grandi e piccini e rappresentazioni teatrali. Clicca qui per il programma completo delle attività di Nanotec.   Per l'evento completo apri il link: www.laricercaviendinotte.it