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Gesmi MILCOVICH

Ricercatore t.d. art. 24 c. 3 lett. A
Dipartimento Scienze della Vita sede ex Scienze Farmaceutiche Via Campi 103

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Pubblicazioni

2023 - ELASTIN-LIKE RECOMBINAMERS FOR MULTI-MODAL DRUG DELIVERY SYSTEMS [Abstract in Atti di Convegno]
Milcovich, Gesmi; Orbanic, Doriana; Ibáñez-Fonseca, Arturo; Contessotto, Paolo; Flora, Tatjana; Marsico, Grazia; Spelat, Renza; Ferro, Federico; Amenitsch, Heinz; Capaldo, Pietro; Carlos Rodriguez-Cabello, José; Pandit, Abhay
abstract

2023 - WETTABILITY AND HYDRODYNAMICS KEY HALLMARKS ON DRUGS’ DISSOLUTION RATE [Abstract in Atti di Convegno]
Biasin, Alice; Pribac, Federico; Milcovich, Gesmi; Franceschinis, Erica; Hasa, Dritan; Voinovich, Dario; Grassi, Gabriele; Grassi, Mario; Abrami, Michela
abstract

2022 - Designing of polymeric gels mimicking the normal and fibrotic liver tissues: Effect of viscoelasticity on cells adhesion and survival [Relazione in Atti di Convegno]
Fornasier, Claudia; Farra, Rossella; Milcovich, Gesmi; Biasin, Alice; Hai Truong, Nhung; Grassi, Gabriele; Grassi, Mario; Abrami, Michela
abstract

Chronic hepatitis B/C virus, alcohol abuse, and non-alcoholic fatty liver disease are the major causes of liver fibrosis (LF), that leads to the progressive impairment of liver function (cirrhosis), often resulting in hepatocellular carcinoma. Despite the etiology, a key element in LF generation is represented by hepatic stellate cells (HSCs) that, from a quiescent condition, differentiate into proliferative and migratory myofibroblasts (cell activation) which secrete Extra Cellular Matrix (ECM) making the tissue increasingly stiffer. Interestingly, HCS activation is triggered by stiff environments, typical of fibrotic liver tissues (shear modulus G > (5 - 6) kPa for human beings). To study the activation process and novel drugs to down-modulate HSC activation, we developed proper substrates which can effectively mimic the viscoelastic properties of healthy and pathological liver tissue. Therefore, we designed alginates-based gels resembling the healthy and the pathological liver tissues from a viscoelastic point of view. For this purpose, hydrogels characterized by different alginate concentrations were produced to indicatively match healthy and pathological stiffness (crosslinking by CaCl2). Then, to improve cells adhesion and survival on the gel surface, the water phase was substituted by a proper culture medium. Furthermore, additional studies were carried out in order to add ECM-like substrates to the alginate solution, like collagen or fibronectin or matrigel, to promote cells-gel interactions. Each gels was characterized by stress and frequency sweep tests to get the linear viscoelastic range, the relaxation spectrum (generalized Maxwell model fitting to frequency sweep data) and the shear modulus G (stiffness). Finally, the optimal gel systems effectively mimicking healthy or pathological liver tissues were successfully considered for cell seeding and followed their adhesion and survival up to three days

2022 - Drug delivery and in vivo absorption [Capitolo/Saggio]
Abrami, Michela; Grassi, Lucia; Farra, Rossella; Dapas, Barbara; di Vittorio, Rosario; Grassi, Gabriele; Milcovich, Gesmi; Grassi, Mario
abstract

It is probably not well known that 4000 years B.C. Sumerians were able to prepare many medicaments and illness was considered a sort of divine punishment while healing was the consequent purification, a viewpoint that have heavily affected the western world up to the modern age. The scientific darkness permeating Europe after the end of the Roman Empire favoured the survival of this concept while the golden age of the Arab Science (9th -13th centuries) and Renaissance represented the seeds of a paramount change. However, it is only at the end of the 19th century that, under a rigorous experimental Galilean approach, the real origins of many diseases were discovered. As a matter of fact, the modern age of drug delivery started after the 2nd world war and the first example of mathematical modelling in this field dates back to 1961. The clear affirmation of mathematical modelling in the biopharmaceutical field took place in the last twenty years of the 20th century thanks to valuable researchers such as Peppas and Langer. The third millennium opened with new important challenges such as the simultaneous modelling of drug release, Adsorption, Distribution Metabolism and Elimination (ADME), processes that rule drug fate in vivo. Thus, the aim of this chapter is to describe and to comment the most recent advances in the mathematical modelling of drug release and ADME processes in the light of Paracelsus’ belief of the human body as a chemical plant.

2022 - Effect of chest physiotherapy on cystic fibrosis sputum nanostructure: an experimental and theoretical approach [Articolo su rivista]
Abrami, Michela; Maschio, Massimo; Conese, Massimo; Confalonieri, Marco; Salton, Francesco; Gerin, Fabio; Dapas, Barbara; Farra, Rossella; Adrover, Alessandra; Milcovich, Gesmi; Fornasier, Claudia; Biasin, Alice; Grassi, Mario; Grassi, Gabriele
abstract

Cystic fibrosis (CF) is a disease characterized by the production of viscous mucoid secretions in multiple organs, particularly the airways. The pathological increase of proteins, mucin and biological polymers determines their arrangement into a three-dimensional polymeric network, affecting the whole mucus and impairing the muco-ciliary clearance which promotes inflammation and bacterial infection. Thus, to improve the efficacy of the drugs usually applied in CF therapy (e.g., mucolytics, anti-inflammatory and antibiotics), an in-depth understanding of the mucus nanostructure is of utmost importance. Drug diffusivity inside a gel-like system depends on the ratio between the diffusing drug molecule radius and the mesh size of the network. Based on our previous findings, we propose the combined use of rheology and low field NMR to study the mesh size distribution of the sputum from CF patients. Specifically, we herein explore the effects of chest physiotherapy on CF sputum characteristic as evaluated by rheology, low field NMR and the drug penetration through the mucus via mathematical simulation. These data show that chest physiotherapy has beneficial effects on patients, as it favourably modifies sputum and enhances drug penetration through the respiratory mucus.

2022 - Effect of cystic fibrosis sputum rheology on lungs drug delivery by inhalation [Relazione in Atti di Convegno]
Abrami, Michela; Maschio, Massimo; Conese, Massimo; Confalonieri, Marco; Salton, Francesco; Gerin, Fabio; Dapas, Barbara; Farra, Rossella; Adrover, Alessandra; Milcovich, Gesmi; Fornasier, Claudia; Biasin, Alice; Grassi, Gabriele; Grassi, Mario
abstract

Cystic Fibrosis (CF), one of the most common lethal genetic diseases in people of Caucasian origin, causes a severe increase of mucus viscosity (due to water loss) in many organs, mainly the airways. As a consequence, increased viscosity impairs muco-ciliary clearance determining sputum (pathological mucus) stagnation, thus promoting bacterial lung infections. In the case of drugs administered by inhalation (usually, mucolytics, anti-inflammatory and antibiotics), the structure of the Airway Surface Liquid (ASL) lining the walls of the bronchial tree requires an in-depth understanding in order to improve the efficacy of the drugs. Indeed, ASL is composed by a thin inner water-like serous fluid named periciliary layer, where cells cilia beat, and by a thicker outer non-Newtonian viscoelastic fluid representing the Mucus Layer (ML). In particular, ML, is a heterogeneous medium made up of mainly physical cross-linked polymeric regions, embedded in a less viscous water-like fluid. Although drug transport inside ASL can depend on different factors, drug diffusion is one of the most important. Thus, the estimation of the drug diffusion coefficient (D) is a key factor: at this purpose the Lustig-Peppas model for not swelling systems represents an effective tool. Indeed, this model refers to both the network mesh size and the volume fraction of the solid (polymeric) network permeating the ML. Interestingly, based on the Flory, Chui and Schurz theories, it is possible to express D directly as a function of the ML shear modulus (G). Thus, assuming that ASL rheological properties are similar to those of sputum, it is possible evaluating D and the kinetics of drug penetration inside ASL. Therefore, this work provided to measure CF sputum G by fitting the generalized Maxwell model to frequency sweep data, to evaluate D and to simulate drug diffusion through the ASL layer by means of Fick's equation

2022 - Reduction of Cardiac Fibrosis by Interference with YAP-Dependent Transactivation [Articolo su rivista]
Garoffolo, G.; Casaburo, M.; Amadeo, F.; Salvi, M.; Bernava, G.; Piacentini, L.; Chimenti, I.; Zaccagnini, G.; Milcovich, G.; Zuccolo, E.; Agrifoglio, M.; Ragazzini, S.; Baasansuren, O.; Cozzolino, C.; Chiesa, M.; Ferrari, S.; Carbonaro, D.; Santoro, R.; Manzoni, M.; Casalis, L.; Raucci, A.; Molinari, F.; Menicanti, L.; Pagano, F.; Ohashi, T.; Martelli, F.; Massai, D.; Colombo, G. I.; Messina, E.; Morbiducci, U.; Pesce, M.
abstract

Background: Conversion of cardiac stromal cells into myofibroblasts is typically associated with hypoxia conditions, metabolic insults, and/or inflammation, all of which are predisposing factors to cardiac fibrosis and heart failure. We hypothesized that this conversion could be also mediated by response of these cells to mechanical cues through activation of the Hippo transcriptional pathway. The objective of the present study was to assess the role of cellular/nuclear straining forces acting in myofibroblast differentiation of cardiac stromal cells under the control of YAP (yes-associated protein) transcription factor and to validate this finding using a pharmacological agent that interferes with the interactions of the YAP/TAZ (transcriptional coactivator with PDZ-binding motif) complex with their cognate transcription factors TEADs (TEA domain transcription factors), under high-strain and profibrotic stimulation. Methods: We employed high content imaging, 2-dimensional/3-dimensional culture, atomic force microscopy mapping, and molecular methods to prove the role of cell/nuclear straining in YAP-dependent fibrotic programming in a mouse model of ischemia-dependent cardiac fibrosis and in human-derived primitive cardiac stromal cells. We also tested treatment of cells with Verteporfin, a drug known to prevent the association of the YAP/TAZ complex with their cognate transcription factors TEADs. Results: Our experiments suggested that pharmacologically targeting the YAP-dependent pathway overrides the profibrotic activation of cardiac stromal cells by mechanical cues in vitro, and that this occurs even in the presence of profibrotic signaling mediated by TGF-β1 (transforming growth factor beta-1). In vivo administration of Verteporfin in mice with permanent cardiac ischemia reduced significantly fibrosis and morphometric remodeling but did not improve cardiac performance. Conclusions: Our study indicates that preventing molecular translation of mechanical cues in cardiac stromal cells reduces the impact of cardiac maladaptive remodeling with a positive effect on fibrosis.

2021 - Drug delivery and mathematical modeling: an historical perspective [Relazione in Atti di Convegno]
Grassi, Mario; Abrami, Michela; Grassi, Lucia; Farra, Rossella; Dapas, Barbara; Di Vittorio, Rosario; Milcovich, Gesmi; Grassi, Gabriele
abstract

It is probably not well known that 4000 years BC Sumerians were able to prepare many medicaments and that the same were used to do Assyrians, Babylonians and Egyptians 3000 years BC. Interestingly, the idea that illness is a sort of divine punishment and healing is the consequent purication, was born in that time and this view heavily aected the western world up to the modern age. If Hippocrates and Galenus, the two most famous witnesses of the Greek and Roman medicinal world, started detaching from this viewpoint, the scientic darkness permeating Europe after the end of the Roman Empire rearmed this way of conceiving illness. Only in the 9th -13th centuries, thanks to the golden age of the Arab Science, something changed and, undoubtedly, represented the basis for the big cultural and scientic revolution represented by Renaissance. Interestingly, in the rst half of the 16th century, Paracelsus, with a very modern vision, conceived human body as a chemical laboratory. However, it is only at the end of the 19th century that, under a rigorous experimental Galilean approach, the real origins of many diseases were discovered. Consequently, this can be considered as the beginning of the modern pharmaceutics whose task is to optimize and improve the clinical eects of drugs. Since then, drug delivery has developed and after the 2nd world war it entered in its modern age with the realization of the rst controlled release system (1952). Notably, the rst example of mathematical modelling trying to simulate drug release from a delivery system appeared 9 years later (1961) and the clear armation of mathematical modelling in the biopharmaceutical eld took place in the last twenty years of the 20th century thanks to valuable researchers such as Peppas and Langer. The third millennium opens with new important challenges for delivery systems designing and mathematical modelling such as the overcoming of biological barriers. This means stopping focusing only on the mathematical modelling of drug release but also considering adsorption, distribution and elimination processes that rule drug fate in vivo. The theoretical bridge joining these two aspects of mathematical modelling could be the ancient, but evergreen, mass balance stating that the administered dose does not disappears but it spreads among tissues before complete elimination/metabolization. In so doing, the idea of Paracelsus about human body has been denitely accepted

2020 - Modulation of Efficient Diiodo-BODIPY in vitro Phototoxicity to Cancer Cells by Carbon Nano-Onions [Articolo su rivista]
Bartelmess, J.; Milcovich, G.; Maffeis, V.; D'Amora, M.; Bertozzi, S. M.; Giordani, S.
abstract

Photodynamic therapy is currently one of the most promising approaches for targeted cancer treatment. It is based on responses of vital physiological signals, namely, reactive oxygen species (ROS), which are associated with diseased condition development, such as tumors. This study presents the synthesis, incorporation, and application of a diiodo-BODIPY–based photosensitizer, based on a non-covalent functionalization of carbon nano-onions (CNOs). In vitro assays demonstrate that HeLa cells internalize the diiodo-BODIPY molecules and their CNO nanohybrids. Upon cell internalization and light exposure, the pyrene–diiodo-BODIPY molecules induce an increase of the ROS level of HeLa cells, resulting in remarkable photomediated cytotoxicity and apoptosis. Conversely, when HeLa cells internalize the diiodo-BODIPY/CNO nanohybrids, no significant cytotoxicity or ROS basal level increase can be detected. These results define a first step toward the understanding of carbon nanomaterials that function as molecular shuttles for photodynamic therapeutics, boosting the modulation of the photosensitizer.

2020 - Soft nanoonions: A dynamic overview onto catanionic vesicles temperature-driven transition [Articolo su rivista]
Milcovich, G.; Antunes, F. E.; Grassi, M.; Asaro, F.
abstract

Catanionic vesicles are emerging interesting structures for bioapplications. They selfgenerate by a pairing of oppositely charged ionic surfactants that assemble into hollow structures. Specifically, the anionic-cationic surfactant pair assumes a double-tailed zwitterionic behavior. In this work, the multilamellar-to-unilamellar thermal transition of several mixed aqueous systems, with a slight excess of the anionic one, were investigated. Interestingly, it was found that the anionic counterion underwent a dissociation as a consequence of a temperature increase, leading to the mentioned thermal transition. The present work proposed the spectroscopic techniques, specifically multinuclear NMR and PGSTE (pulsed gradient stimulated echo), as a key tool to study such systems, with high accuracy and effectiveness, while requiring a small amount of the sample. The results presented herein evidence encouraging perspectives, forecasting the application of the studied vesicular nanoreservoirs, for e.g., drug delivery.

2019 - Antibacterial drug release from a biphasic gel system: Mathematical modelling [Articolo su rivista]
Abrami, M.; Golob, S.; Pontelli, F.; Chiarappa, G.; Grassi, G.; Perissutti, B.; Voinovich, D.; Halib, N.; Murena, L.; Milcovich, G.; Grassi, M.
abstract

Bacterial infections represent an important drawback in the orthopaedic field, as they can develop either immediately after surgery procedures or after some years. Specifically, in case of implants, they are alleged to be troublesome as their elimination often compels a surgical removal of the infected implant. A possible solution strategy could involve a local coating of the implant by an antibacterial system, which requires to be easily applicable, biocompatible and able to provide the desired release kinetics for the selected antibacterial drug. Thus, this work focusses on a biphasic system made up by a thermo-reversible gel matrix (Poloxamer 407/water system) hosting a dispersed phase (PLGA micro-particles), containing a model antibacterial drug (vancomycin hydrochloride). In order to understand the key parameters ruling the performance of this delivery system, we developed a mathematical model able to discriminate the drug diffusion inside micro-particles and within the gel phase, eventually providing to predict the drug release kinetics. The model reliability was confirmed by fitting to experimental data, proposing as a powerful theoretical approach to design and optimize such in situ delivery systems.

2019 - Biomechanical cues towards cancer tissues characterization [Poster]
Milcovich, G; Saccomano, M; Ulloa Severino, L; Alves, F; Casalis, L.
abstract

2019 - Geometry and strain sensing dictate YAP-dependent mechanical signaling in human cardiac stromal cells [Poster]
Pesce, M; Amadeo, F; Salvi, M; Milcovich, G; Gianoli, S; Parisse, P; Pagani, F; Angelini, F; Messina, E; Agrifoglio, M; Casalis, L; Molinari, F; Chimenti, I; Morbiducci, U.
abstract

2019 - Gold nanoparticles/electrochemically expanded graphite composite: A bifunctional platform toward glucose sensing and SERS applications [Articolo su rivista]
Xiang, X.; Feng, S.; Chen, J.; Feng, J.; Hou, Y.; Ruan, Y.; Weng, X.; Milcovich, G.
abstract

An integrated nanogold/expanded graphite based sensor was fabricated by a former electrochemical etching of the pencil lead electrode (PLE) and a later in-situ deposition of gold nanoparticles (AuNPs). The electrochemical pretreatment of PLE (EPLE) created a 3D graphene-like surface, enhanced the electrode surface area and facilitated the electron transfer ability within 5 min without any hazardous chemicals added. The obtained AuNPs/EPLE sensor had an excellent electrochemical response to glucose with a wide linear concentration range, from 0.05 to 38 mM and 38 to 60 mM, and a low detection limit of 5 μM (S/N = 3). Furthermore, the AuNPs/EPLE sensor was successfully employed to detect the glucose in human serum samples, and the results agreed well with those measured in hospital. Finally, the sensor exhibited efficient and reproducible surface-enhanced Raman scattering activities for the probe molecules.

2019 - Mathematical modelling of antibacterial release from a biphasic gel system [Relazione in Atti di Convegno]
Grassi, Mario; Abrami, Michela; Golob, Samuel; Pontelli, Fabio; Chiarappa, Gianluca; Perissutti, Beatrice; Voinovich, Dario; Halib, Nadia; Murena, Luigi; Milcovich, Gesmi; Grassi, Gabriele
abstract

Bacterial infections represent an important problem in the orthopaedic field as they can develop either immediately after the surgical intervention or after some years [1]. In particular, they can be very problematic in the case of implants as, often, their elimination requires the surgical removal of the infected implant. Accordingly, a possible solution strategy is to act locally by coating the implant by an antibacterial system that has to be easily applicable, biocompatible (it must not hinder implant osseointegration) and able to provide the desired release kinetics of the selected antibacterial drug. In this frame, this paper focuses the attention on a biphasic polymeric system made up by a thermos-reversible hydrogel, constituted by Poloxamer 407, hosting a dispersed phase represented by polylactic-co-glycolic acid 50:50 (PLGA) micro-particles containing the antibacterial drug (vancomycin hydrochloride). While below room temperature, the Poloxamer 407/water system behaves as a solution and it is easily spreadable on the implant surface, upon temperature rise to the physiological value, the Poloxamer 407/water solution undergoes gelation. Basically, gelation ensures that the PLGA micro-particles remain in situ, between the implant surface and the growing bone. On the contrary, the controlled drug delivery is due to vancomycin hydrochloride release from PLGA micro-particles, acting as the reservoir phase. The primary aim of this paper is to develop a mathematical model able to properly describe the in vitro vancomycin hydrochloride release from the biphasic system.

2019 - Mathematical modelling of antibacterial release from a biphasic gel system [Poster]
Grassi, M; Abrami, M; Golob, S; Pontelli, F; Chiarappa, G; Perissutti, B; Voinovich, D; Halib, N; Murena, L; Milcovich, G; Grassi, G.
abstract

2019 - Nitrogen and sulfur co-doped carbon nanodots toward bovine hemoglobin: A fluorescence quenching mechanism investigation [Articolo su rivista]
Wang, J.; Xiang, X.; Milcovich, G.; Chen, J.; Chen, C.; Feng, J.; Hudson, S. P.; Weng, X.; Ruan, Y.
abstract

A deep understanding of the molecular interactions of carbon nanodots with biomacromolecules is essential for wider applications of carbon nanodots both in vitro and in vivo. Herein, nitrogen and sulfur co-doped carbon dots (N,S-CDs) with a quantum yield of 16% were synthesized by a 1-step hydrothermal method. The N,S-CDs exhibited a good dispersion, with a graphite-like structure, along with the fluorescence lifetime of approximately 7.50 ns. Findings showed that the fluorescence of the N,S-CDs was effectively quenched by bovine hemoglobin as a result of the static fluorescence quenching. The mentioned quenching mechanism was investigated by the Stern-Volmer equation, temperature-dependent quenching, and fluorescence lifetime measurements. The binding constants, number of binding sites, and the binding average distance between the energy donor N,S-CDs and acceptor bovine hemoglobin were calculated as well. These findings will provide for valuable insights on the future bioapplications of N,S-CDs.

2018 - Co-reductive fabrication of carbon nanodots with high quantum yield for bioimaging of bacteria [Articolo su rivista]
Wang, J.; Liu, X.; Milcovich, G.; Chen, T. -Y.; Durack, E.; Mallen, S.; Ruan, Y.; Weng, X.; Hudson, S. P.
abstract

A simple and straightforward synthetic approach for carbon nanodots (C-dots) is proposed. The strategy is based on a one-step hydrothermal chemical reduction with thiourea and urea, leading to high quantum yield C-dots. The obtained C-dots are welldispersed with a uniform size and a graphite-like structure. A synergistic reduction mechanism was investigated using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The findings show that using both thiourea and urea during the one-pot synthesis enhances the luminescence of the generated C-dots. Moreover, the prepared C-dots have a high distribution of functional groups on their surface. In this work, C-dots proved to be a suitable nanomaterial for imaging of bacteria and exhibit potential for application in bioimaging thanks to their low cytotoxicity.

2018 - Stabilization of unilamellar catanionic vesicles induced by β-cyclodextrins: A strategy for a tunable drug delivery depot [Articolo su rivista]
Milcovich, Gesmi; Antunes, Filipe E.; Grassi, Mario; Asaro, Fioretta
abstract

The limited stability of catanionic vesicles has discouraged their wide use for encapsulation and controlled release of active substances. Their structure can easily break down to form lamellar phases, micelles or rearrange into multilamellar vesicles, as a consequence of small changes in their composition. However, despite the limited stability, catanionic vesicles possess an attractive architecture, which is able to efficiently encapsulate both hydrophobic and hydrophilic molecules. Therefore, improving the stability of the vesicles, as well as the control on unilamellar structures, are prerequisites for their wider application range. This study focuses on the impact of β-cyclodextrins for the stabilization of SDS/CTAB catanionic vesicles. Molar ratio and sample preparation procedures have been investigated to evaluate the temperature stability of catanionic vesicles. Diffusion and spectroscopic techniques evidenced that when β-cyclodextrins are added, unilamellar structures are stabilized above the multilamellar-unilamellar vesicles critical temperature. The results evidence encouraging perspectives for the use of vesicular nanoreservoirs for drug depot applications.

2017 - Modulating carbohydrate-based hydrogels as viscoelastic lubricant substitute for articular cartilages [Articolo su rivista]
Milcovich, Gesmi; Antunes, Filipe E.; Farra, Rossella; Grassi, Gabriele; Grassi, Mario; Asaro, Fioretta
abstract

Viscosupplementation is a therapeutic approach for osteoarthritis treatment, where the synovial fluid,the natural lubricant of the joints, is replaced by viscoelastic solutions with rheological properties compa-rable or better than the starting material. This study presents the development of an innovative platformfor viscosupplementation, based on the optimization of polysaccharide-based colloidal hydrogel, aimingto reduce on-site enzyme degradation and enhance the possibility of hyaluronic acid substitution withalternative biomaterials. Catanionic vesicles are proposed as physical crosslinker that can guarantee theformation of a ‘soft’, tunable network, offering a dual-therapeutic approach: on the mechanical reliefperspective, as well as on the drug/gene delivery strategy. This research focuses on the fabrication andoptimization of colloidal networks, driven by the synergistic interaction among catanionic vesicles andcationic modified cellulose polymers. This study tests the hypothesis that cellulose-like polymers can bearranged into functional matrix, mimicking the mechanical properties of healthy synovial fluids.

2017 - Recent advances in smart biotechnology: Hydrogels and nanocarriers for tailored bioactive molecules depot [Articolo su rivista]
Milcovich, G.; Lettieri, S.; Antunes, F. E.; Medronho, B.; Fonseca, A. C.; Coelho, J. F. J.; Marizza, P.; Perrone, F.; Farra, R.; Dapas, B.; Grassi, G.; Grassi, M.; Giordani, S.
abstract

Over the past ten years, the global biopharmaceutical market has remarkably grown, with ten over the top twenty worldwide high performance medical treatment sales being biologics. Thus, biotech R&D (research and development) sector is becoming a key leading branch, with expanding revenues. Biotechnology offers considerable advantages compared to traditional therapeutic approaches, such as reducing side effects, specific treatments, higher patient compliance and therefore more effective treatments leading to lower healthcare costs. Within this sector, smart nanotechnology and colloidal self-assembling systems represent pivotal tools able to modulate the delivery of therapeutics. A comprehensive understanding of the processes involved in the selfassembly of the colloidal structures discussed therein is essential for the development of relevant biomedical applications. In this review we report the most promising and best performing platforms for specific classes of bioactive molecules and related target, spanning from siRNAs, gene/plasmids, proteins/growth factors, small synthetic therapeutics and bioimaging probes.

2017 - Synthetic/ECM-inspired hybrid platform for hollow microcarriers with ROS-triggered nanoporation hallmarks [Articolo su rivista]
Milcovich, G.; Contessotto, P.; Marsico, G.; Ismail, S.; Pandit, A.
abstract

Reactive oxygen species (ROS) are key pathological signals expressed in inflammatory diseases such as cancer, ischemic conditions and atherosclerosis. An ideal drug delivery system should not only be responsive to these signals but also should not elicit an unfavourable host response. This study presents an innovative platform for drug delivery where a natural/synthetic composite system composed of collagen type I and a synthesized polythioether, ensures a dual stimuli-responsive behaviour. Collagen type I is an extracellular matrix constituent protein, responsive to matrix metalloproteinases (MMP) cleavage per se. Polythioethers are stable synthetic polymers characterized by the presence of sulphur, which undergoes a ROS-responsive swelling switch. A polythioether was synthesised, functionalized and tested for cytotoxicity. Optimal conditions to fabricate a composite natural/synthetic hollow sphere construct were optimised by a template-based method. Collagen-polythioether hollow spheres were fabricated, revealing uniform size and ROS-triggered nanoporation features. Cellular metabolic activity of H9C2 cardiomyoblasts remained unaffected upon exposure to the spheres. Our natural/synthetic hollow microspheres exhibit the potential for use as a pathological stimuli-responsive reservoir system for applications in inflammatory diseases.

2017 - The pH of the human nail plate [Capitolo/Saggio]
Murdan, S.; Milcovich, G.; Goriparthi, G. S.
abstract

In this chapter, measurements of the nail plate pH are reported. Measurements were conducted in vivo in 37 volunteers with healthy finger- and toenails, using a skin pH meter. The pH of unwashed and washed fin- gernails and the big toenails was measured, and the influence of washing, anatomical site (fin- gers/toes), side (left/right), finger digit (digits 1–5), and gender were determined. The pH of the nail plate surface was around 5. There was no significant difference between the sides, i.e., the right or left hand/foot, among the ten fin- gernails and between the two great toenails. However, toenails had a significantly higher pH than fingernails. Washing the nails caused an immediate, but transient, increase in pH, which was not sustained with time, and pH returned to prewashing levels within 20 min. In males, washing did not significantly influ- ence finger- or toenail plate pH. In females, however, washed fingernails had a signifi- cantly higher pH than unwashed ones, while there was no difference in the pH of the toe- nails. The pH of the nail plate interior, mea- sured after tape stripping, was found to be slightly lower than that at its surface.

2016 - Design of a protease and ROS responsive biomaterial approach to target drug delivery in inflamed tissue microenvironment [Articolo su rivista]
Milcovich, G; Contessotto, P; Pandit, A
abstract

2016 - Design of a protease and ROS responsive biomaterial approach to target drug delivery in inflamed tissue microenvironment [Poster]
Milcovich, G; Contessotto, P; Pandit, A.
abstract

2016 - Scaffold and scaffold-free self-assembled systems in regenerative medicine [Articolo su rivista]
Thomas, D.; Gaspar, D.; Sorushanova, A.; Milcovich, G.; Spanoudes, K.; Mullen, A. M.; O'Brien, T.; Pandit, A.; Zeugolis, D. I.
abstract

Self-assembly in tissue engineering refers to the spontaneous chemical or biological association of components to form a distinct functional construct, reminiscent of native tissue. Such self-assembled systems have been widely used to develop platforms for the delivery of therapeutic and/or bioactive molecules and various cell populations. Tissue morphology and functional characteristics have been recapitulated in several self-assembled constructs, designed to incorporate stimuli responsiveness and controlled architecture through spatial confinement or field manipulation. In parallel, owing to substantial functional properties, scaffold-free cell-assembled devices have aided in the development of functional neotissues for various clinical targets. Herein, we discuss recent advancements and future aspirations in scaffold and scaffold-free self-assembled devices for regenerative medicine purposes.

2016 - Thermo-responsive hydrogels from cellulose-based polyelectrolytes and catanionic vesicles for biomedical application [Articolo su rivista]
Milcovich, Gesmi; Antunes, Filipe; Golob, Samuel; Farra, Rossella; Grassi, Mario; Voinovich, Dario; Grassi, Gabriele; Asaro, Fioretta
abstract

In this study, negatively charged catanionic vesicles/hydrophobically modified hydroxyethylcellulose polymers thermo-responsive hydrogels have been fabricated. Vesicular aggregates were found to act as multifunctional junctions for networking of modified-cellulose water solutions. The contributions of the electrostatic and hydrophobic interactions were evaluated by changing either vesicles composition or the polymer hydrophobic substitution. Thermal-induced size and lamellarity of hydrogelenclosed vesicles were detected, with further polygonal shape changes induced by cellulose-based polymer addition. The thermal transition was also found to tune hydrogel mechanical behaviour. The network formation was further assessed through molecular insights, which allow to determine the arrangement of the polymer chains on the vesicles’ surface. The examined systems exhibited interesting thermo-responsive characteristics. Thus, vesicularly crosslinked hydrogels herein presented can offer a wide variety of applications, i.e. in biomedical field, as multi-drug delivery systems, thanks to their ability to provide for different environments to guest molecules, comprising bulk water, vesicles’ interior and bilayers, sites on polymeric chains.

2016 - Towards an elastin-based hydrogel platform for the repair of myocardial infarction [Poster]
Contessotto, P; Milcovich, G; Spelat, R; Marsico, G; Chaturvedi, A; Orbanic, D; Flora, T; Da Costa, M; Rackauskas, M; Amenitsch, H; Mccartin, M; Rodriguez-Cabello, Jc; Joshi, L; Dockery, P; Kilcoyne, M; Pandit, A.
abstract

2016 - Towards an elastin-based hydrogel platform for the repair of myocardial infarction [Articolo su rivista]
Contessotto, P; Milcovich, G; Spelat, R; Marsico, G; Chaturvedi, A; Orbanic, D; Flora, T; Da Costa, M; Rackauskas, M; Amenitsch, H; Mccartin, M; Rodriguez-Cabello, Jc; Joshi, L; Dockery, P; Kilcoyne, M; Pandit, A
abstract

2015 - Fabrication of ROS and Protease Responsive Drug Delivery Systems [Poster]
Milcovich, G; Contessotto, P; Marsico, G; Pandit, A.
abstract

2014 - Fabrication of a multi-modal delivery system using a elastin-like recombinamer [Poster]
Milcovich, G; Contessotto, P; Marsico, G; Orbanic, D; Flora, T; Rodriguez-Cabello, Jc; Pandit, A.
abstract

2014 - Physical characterization of alginate-Pluronic F127 gel for endoluminal NABDs delivery [Articolo su rivista]
Abrami, M.; D'Agostino, I.; Milcovich, G.; Fiorentino, S.; Farra, R.; Asaro, F.; Lapasin, R.; Grassi, G.; Grassi, M.
abstract

Here we focus the attention on the physical characteristics of a highly biocompatible hydrogel made up of crosslinked alginate and Pluronic F127 (PF127). This is a composite polymeric blend we propose for artery endoluminal delivery of an emerging class of molecules named nucleic acid based drugs (NABDs). The physical characterization of our composite gel, i.e. mesh size distribution and PF127–alginate mutual organization after crosslinking, can significantly determine the NABDs release kinetics. Thus, to explore these aspects, different technical approaches, i.e. rheology, low/high field NMR and TEM, were used. While rheology provided information at the macroscopic and nano-level, the other three approaches gave details at the nano-level. We observe that Pluronic micelles, organizing in cubic ordered domains, generate, upon alginate crosslinking, the formation of meshes (z150 nm) larger than those occurring in a Pluronic-free alginate network (z25 nm). Nevertheless, smaller alginate meshes are still on and can just host un-structured Pluronic micelles and water. Accordingly, the gel structure is quite inhomogeneous, where big meshes (filled by crystalline Pluronic) co-exist with smaller meshes (hosting water and un-structured PF127 micelles). While big meshes offer a considerable hindering action on a diffusing solute, smaller ones represent a sort of free space where solute diffusion is faster. The presence of big and small meshes indicates that drug release may follow a double kinetics characterized by a fast and slow release. Notably, this behavior is considered appropriate for endoluminal drug release to the arterial wall.

2013 - Guar gum/borax hydrogel: Rheological, low field NMR and release characterizations [Articolo su rivista]
Coviello, T.; Matricardi, P.; Alhaique, F.; Farra, R.; Tesei, G.; Fiorentino, S.; Asaro, F.; Milcovich, G.; Grassi, M.
abstract

Guar gum (GG) and Guar gum borax (GGb) hydrogels are studied by means of rheology, Low Field NMR and model drug release tests. These three approaches are used to estimate the mesh size (ξ) of the polymeric network. A comparison with similar Scleroglucan systems is carried out. In the case of GGb, the rheological and Low Field NMR estimations of ξ lead to comparable results, while the drug release approach seems to underestimate ξ. Such discrepancy is attributed to the viscous effect of some polymeric chains that, although bound to the network to one end, can freely fluctuate among meshes. The viscous drag exerted by these chains slows down drug diffusion through the polymeric network. A proof for this hypothesis is given by the case of Scleroglucan gel, where the viscous contribution is not so significant and a good agreement between the rheological and release test approaches was found.

2013 - Hydrogels from biocompatible polyelectrolytes and catanionic vesicles [Poster]
Asaro, F; Milcovich, G; Antunes, F E
abstract

2013 - Structure and dynamics of polymer-surfactant hydrogels for bioactive molecule delivery [Poster]
Milcovich, G; Antunes, Fe; Asaro, F; Grassi, M.
abstract

2013 - Surfactant polymer and Silica nanoparticles [Poster]
Asaro, F; Milcovich, G
abstract

2012 - Caratterizzazione reologico/NMR di gel composti da alginato-pluronico per il rilascio di farmaci basati sul DNA [Relazione in Atti di Convegno]
D’Agostino, I.; Milcovich, G.; Asaro, Fioretta; Fiorentino, S. M.; Lapasin, Romano; Grassi, Gabriele; Grassi, Mario
abstract

2012 - Caratterizzazione reologico/NMR di gel composti da alginato-pluronico per il rilascio di farmaci basati sul DNA [Poster]
D’Agostino, I; Milcovich, G; Asaro, F; Fiorentino, S; Lapasin, R; Grassi, G; Grassi, M.
abstract

2012 - Effect of β-cyclodextrin on multi-to uni-lamellar thermal transition of catanionic vesicles [Poster]
Milcovich, G; Asaro, F.
abstract

2012 - Insights into catanionic vesicles thermal transition by NMR spectroscopy [Capitolo/Saggio]
Milcovich, G.; Asaro, F.
abstract

Oppositely charged ionic surfactants can self-assemble into hollow structures, called catanionic vesicles, where the anionic-cationic surfactant pair assumes a double-tailed zwitterionic attitude. In the present work, multilamellar-to-unilamellar thermal transition of a mixed aqueous system of sodium dodecyl sulphate (SDS) and cetyl trimethyl ammonium bromide (CTAB), with a slight excess of the anionic one, has been investigated by 1H, 2H, 14N NMR spectra and 23Na transverse relaxation measurements. It has been inferred that an increase of the temperature enhances the SDS counterion dissociation, which can be considered as one of the driving forces of the mentioned transition. Moreover, interesting 23Na T2 changes with temperature have been detected for unilamellar aggregates.

2012 - Microstructure of Pluronic F127 Alginate Composite Hydrogel Probed by NMR and Rheology [Poster]
D’Agostino, I; Milcovich, G; Asaro, F; Fiorentino, S; Lapasin, R; Grassi, G; Grassi, M.
abstract

2011 - An assessment of the human nail plate pH [Articolo su rivista]
Murdan, S.; Milcovich, G.; Goriparthi, G. S.
abstract

Purpose of Study: To measure the pH of the surface of healthy nail plates. Procedures: The surface pH of human fingernails and big toenails was measured in vivo using a skin pH meter. The influence of washing, anatomical site (fingers/toes), side (left/right), digit (digits 1-5) and gender was determined. The pH of the nail interior was also measured. Results: The pH of the nail plate surface was around 5, with toenails having a significantly higher pH than fingernails. Immediately after hand washing, the nail surface pH increased significantly, from pH 5.1 ± 0.4 to 5.3 ± 0.5. However, this was not sustained with time, and the pH returned to prewashing levels within 20 min. Gender had an influence on unwashed, but not washed, nail plate pH. The pH of the nail plate interior was lower than that of its surface. Conclusions: It was possible to measure nail plate pH, and baseline values are provided. Copyright © 2011 S. Karger AG, Basel.

2011 - Insights into catanionic vesicles thermal transition by NMR Spectroscopy [Poster]
Milcovich, G; Asaro, F.
abstract

2011 - Insights into catanionic vesicles thermal transition by NMR Spectroscopy [Poster]
Milcovich, G; Asaro, F.
abstract

2010 - An in vivo determination of the human nail plate pH [Poster]
Murdan, S; Milcovich, G; Goriparthi, G.
abstract