Massimo Fresta | University "Magna Graecia" of Catanzaro (original) (raw)

Papers by Massimo Fresta

Biomedicines

The incidence of cancer is increasing dramatically, affecting all ages of the population and reac... more The incidence of cancer is increasing dramatically, affecting all ages of the population and reaching an ever higher worldwide mortality rate. The lack of therapies’ efficacy is due to several factors such as a delay in diagnosis, tumor regrowth after surgical resection and the occurrence of multidrug resistance (MDR). Tumor-associated immune cells and the tumor microenvironment (TME) deeply affect the tumor’s progression, leading to several physicochemical changes compared to physiological conditions. In this scenario, macrophages play a crucial role, participating both in tumor suppression or progression based on the polarization of onco-suppressive M1 or pro-oncogenic M2 phenotypes. Moreover, much evidence supports the pivotal role of macrophage-derived extracellular vesicles (EVs) as mediators in TME, because of their ability to shuttle the cell–cell and organ–cell communications, by delivering nucleic acids and proteins. EVs are lipid-based nanosystems with a broad size range d...

Materials Science and Engineering: C, 2021

In the last few decades, several natural and synthetic polymers have been used as starting materi... more In the last few decades, several natural and synthetic polymers have been used as starting material for the development of innovative polymeric nanoparticles able to encapsulate biologically active substances and to modulate their biopharmaceutical features and/or therapeutic efficacy. This investigation focused on the comparison of the physico-chemical properties of nanosystems made up of two of the most successfully used biodegradable biomaterials, namely poly(lactic-co-glycolic acid) (PLGA) and zein, belonging to the synthetic and natural family of polymers, respectively. Rutin, a polyphenolic bioflavonoid characterized by peculiar antioxidant properties, was chosen as the model drug to be encapsulated in the polymeric systems. The results demonstrated a greater ability of zein-based nanosystems to effectively retain the active compound with respect to the PLGA particles. The integration of rutin in the protein matrix favored a controlled drug leakage, and was influenced by the surfactant used to stabilize the formulation. Moreover, rutin-loaded zein nanoparticles showed significant in vitro antioxidant activity, evidencing a synergistic action between the intrinsic antioxidant activity of the protein and the pharmacological properties of the active compound. Finally, the intracellular localization of the zein nanosystems was demonstrated through confocal laser scanning microscopy.

International Journal of Pharmaceutics, 2002

Our research on topical application of lidocaine-loaded non-ionic surfactant vesicles (NSVs) was ... more Our research on topical application of lidocaine-loaded non-ionic surfactant vesicles (NSVs) was prompted by the great interest on new delivery systems for local anaesthetics. This study is focused on a novel formulation of NSVs entrapping lidocaine in the form of a free base (LID) and a hydrochloride (LIDHCl). NSVs were prepared from polyoxyethylene sorbitan monolaurate (Tween20™) and cholesterol. The effect of vesicle composition and environmental pH condition (8.6-5.5) on drug encapsulation efficiency (e.e.) was investigated. Experimental strategies involved: freeze-fracture, microscopy technique, dynamic light scattering, permeation through Silastic™ and mouse abdominal skin, in vitro release kinetics of vesicle-entrapped drugs, fluorescence quenching analyses. Diffusion experiments showed that the flux of charged lidocaine through Silastic™ membrane was possible only after the vesicle encapsulation. Permeation through mouse abdominal skin of LIDHCl loaded vesicles showed a higher flux and a shorter lag time with respect to classical liposome formulations, while LID permeation rate was quite similar for NSV and liposome formulations. Vesicles were also prepared in the presence of dicetylphosphate (DCP) and N-cetylpyridinium chloride (CP) to obtain negatively and positively charged vesicles respectively, but in this case the e.e. of the drug was negligible. The possible reason of the remarkable lower e.e. observed with charged vesicles was investigated by means of fluorescence quenching experiments.

Gels

For many years, corneal transplantation has been the first-choice treatment for irreversible dama... more For many years, corneal transplantation has been the first-choice treatment for irreversible damage affecting the anterior part of the eye. However, the low number of cornea donors and cases of graft rejection highlighted the need to replace donor corneas with new biomaterials. Tissue engineering plays a fundamental role in achieving this goal through challenging research into a construct that must reflect all the properties of the cornea that are essential to ensure correct vision. In this review, the anatomy and physiology of the cornea are described to point out the main roles of the corneal layers to be compensated and all the requirements expected from the material to be manufactured. Then, a deep investigation of alginate as a suitable alternative to donor tissue was conducted. Thanks to its adaptability, transparency and low immunogenicity, alginate has emerged as a promising candidate for the realization of bioengineered materials for corneal regeneration. Chemical modificat...

Pharmaceutical Research, 2014

Purpose To synthesize a new polymeric prodrug based on α,βpoly(N-2-hydroxyethyl)(2-aminoethylcarb... more Purpose To synthesize a new polymeric prodrug based on α,βpoly(N-2-hydroxyethyl)(2-aminoethylcarbamate)-d,l-aspartamide copolymer bearing amine groups in the side chain (PHEA-EDA), covalently linked to the anticancer drug doxorubicin and to test its potential application in anticancer therapy. Methods The drug was previously derivatized with a biocompatible and hydrophilic linker, leading to a doxorubicin derivative highly reactive with amino groups of PHEA-EDA. The PHEA-EDA-DOXO prodrug was characterized in terms of chemical stability. The pharmacokinetics, biodistribution and cytotoxicity of the product was investigated in vitro and in vivo on human breast cancer MCF-7 and T47D cell lines and NOD-SCID mice bearing a MCF-7 human breast carcinoma xenograft. Data collected were compared to those obtained using free doxorubicin. Results The final polymeric product is water soluble and easily hydrolysable in vivo, due to the presence of ester and amide bonds along the spacer between the drug and the polymeric backbone. In vitro tests showed a retarded cytotoxic effect on tumor cells, whereas a significant improvement of the in vivo antitumor activity of PHEA-EDA-DOXO and a survival advantage of the treated NOD-SCID mice was evidenced, compared to that of free doxorubicin. Conclusions The features of the PHEA-EDA-DOXO provide a potential protection of the drug from the plasmatic enzymatic degradation and clearance, an improvement of the blood pharmacokinetic parameters and a suitable body biodistribution. The data collected support the promising rationale of the proposed macromolecular prodrug PHEA-EDA-DOXO for further potential development and application in the treatment of solid cancer diseases. KEY WORDS antitumor activity. biodistribution. doxorubicin. PHEA-EDA. polymeric prodrug Chiara Di Meo and Felisa Cilurzo contributed equally.

Antioxidants

N-Acylethanolamine acid amidase (NAAA) is an N-terminal cysteine hydrolase that preferentially ca... more N-Acylethanolamine acid amidase (NAAA) is an N-terminal cysteine hydrolase that preferentially catalyzes the hydrolysis of endogenous lipid mediators such as palmitoylethanolamide, which has been shown to exhibit neuroprotective and antinociceptive properties by engaging peroxisome proliferator-activated receptor-α. A few potent NAAA inhibitors have been developed, including α-acylamino-β-lactone derivatives, which are very strong and effective, but they have limited chemical and plasmatic stability, compromising their use as systemic agents. In the present study, as an example of a molecule belonging to the chemical class of N-(2-oxo-3-oxetanyl)amide NAAA inhibitors, URB866 was entrapped in poly(lactic-co-glycolic acid) nanoparticles in order to increase its physical stability. The data show a monomodal pattern and a significant time- and temperature-dependent stability of the molecule-loaded nanoparticles, which also demonstrated a greater ability to effectively retain the compoun...

Pharmaceutics

The regeneration of cardiac tissue is a multidisciplinary research field aiming to improve the he... more The regeneration of cardiac tissue is a multidisciplinary research field aiming to improve the health condition of the post-heart attack patient. Indeed, myocardial tissue has a poor ability to self-regenerate after severe damage. The scientific efforts focused on the research of a biomaterial able to adapt to heart tissue, thus guaranteeing the in situ release of active substances or growth promoters. Many types of hydrogels were proposed for this purpose, showing several limitations. The aim of this study was to suggest a new usage for glyceryl monooleate-based lyotropic liquid crystals (LLCs) as a biocompatible and inert material for a myocardial application. The main advantages of LLCs are mainly related to their easy in situ injection as lamellar phase and their instant in situ transition in the cubic phase. In vivo studies proved the biocompatibility and the inertia of LLCs after their application on the myocardial tissue of mice. In detail, the cardiac activity was monitored ...

Frontiers in Pharmacology, 2021

Advances in nanotechnology have favored the development of novel colloidal formulations able to m... more Advances in nanotechnology have favored the development of novel colloidal formulations able to modulate the pharmacological and biopharmaceutical properties of drugs. The peculiar physico-chemical and technological properties of nanomaterial-based therapeutics have allowed for several successful applications in the treatment of cancer. The size, shape, charge and patterning of nanoscale therapeutic molecules are parameters that need to be investigated and modulated in order to promote and optimize cell and tissue interaction. In this review, the use of polymeric nanoparticles as drug delivery systems of anticancer compounds, their physico-chemical properties and their ability to be efficiently localized in specific tumor tissues have been described. The nanoencapsulation of antitumor active compounds in polymeric systems is a promising approach to improve the efficacy of various tumor treatments.

European Food Research and Technology, 2019

This investigation is focused on the granulometric and rheological analysis of two kinds of wheat... more This investigation is focused on the granulometric and rheological analysis of two kinds of wheat flour, i.e. type 0 and type 00. The granulometric analysis was carried out using a laser diffraction particle size analyzer. The particle size distribution of the type 0 and 00 flours, expressed as the mass median diameter (D (v,0.5)), was 80.922 µm and 71.686 µm, respectively. The rheological characterization of physical mixtures of wheat flour and water was carried out by means of small deformation dynamic oscillatory measurements. The rheological measurements evidenced a different behavior of the two samples of flour when they were mixed with water. Namely, the progressive addition of water to the type 0 wheat flour favored a decrease of G′ and G″ values, emphasizing that the dough becomes less full-bodied, while its structure remains unmodified; in fact, the phase-angle was not influenced by the water content. On the contrary, the type 00 wheat flour mixed with increased water content provided a dough characterized by lower elastic and viscous moduli, even though the behavior of the phase angle progressively varied. The different results may be due to the different particle size distribution of the flour and the different percentage of gluten present in each. Thanks to this parallel study of the particle size distribution of flours and the rheological behavior of the derived doughs, new food preparation procedures and new research on adulteration in the bread production can be designed.

Biomedicines

Bergamot essential oil (BEO) and Ammonium glycyrrhizinate (AG), naturally derived compounds, have... more Bergamot essential oil (BEO) and Ammonium glycyrrhizinate (AG), naturally derived compounds, have remarkable anti-inflammatory properties, thus making them suitable candidates for the treatment of skin disorders. Despite this, their inadequate physicochemical properties strongly compromise their topical application. Ultradeformable nanocarriers containing both BEO and AG were used to allow their passage through the skin, thus maximizing their therapeutic activity. Physicochemical characterization studies were performed using Zetasizer Nano ZS and Turbiscan Lab®. The dialysis method was used to investigate the release profile of the active compounds. In vivo studies were performed on human healthy volunteers through the X-Rite spectrophotometer. The nanosystems showed suitable features for topical cutaneous administration in terms of mean size, surface charge, size distribution, and long-term stability/storability. The co-delivery of BEO and AG in the deformable systems improved both...

Journal of Medicinal Chemistry, 2003

The structure-activity relationship studies on 2-quinolinecarboxamide peripheral benzodiazepine r... more The structure-activity relationship studies on 2-quinolinecarboxamide peripheral benzodiazepine receptor (PBR) ligands have been refined with the aim of using these ligands as carriers of radionuclides and boron atoms. Some new ligands show enhanced affinity and steroidogenic activity with respect to reference compound 1 and are interesting candidates for radiolabeling and PET studies. Moreover, carborane derivative 3q, representing the first example of PBR ligand bearing a carborane cage, can be useful to explore an alternative mechanism in BNCT.

Current Bioactive Compounds, 2007

The use of natural compounds has recently been reconsidered in modern clinical practice. A furthe... more The use of natural compounds has recently been reconsidered in modern clinical practice. A further advancement of these compounds is represented by the possibility of using innovative drug delivery systems that can improve the biopharmaceutical features of the delivered compounds. This review examines the recent developments in the field of delivery of natural drugs belonging to several therapeutic classes. In particular, the examined therapeutic classes are: antitumoral drugs, i.e. paclitaxel, doxorubicin, tea catechins and hypericin; anti-infective drugs, i.e. Melaleuca alternifolia and Artemisia arborescens L.; and antinflammatory/antioxidant drugs, i.e. cannabinoids and glycyrrhizinates used for topical application. In this review we highlight the utility of a suitable drug delivery system to improve the biopharmaceutical aspects of these drugs. The examined carriers are: vesicular carriers (liposomes, Ethosomes ® , ultradeformable liposomes), nano-and microparticles, innovative gels, microemulsions.

Bioorganic & Medicinal Chemistry, 2002

The interaction between ofloxacin, as a model drug of the fluoroquinolone class, and biomembranes... more The interaction between ofloxacin, as a model drug of the fluoroquinolone class, and biomembranes was examined as the possible initial step in a transmembrane diffusion process. Dipalmitoylphosphatidylcholine was used for the preparation of biomembrane models. The influence of environmental conditions and protonation on molecular physicochemical behavior, and hence on the membrane interaction, was investigated by differential scanning calorimetry (DSC). This technique has been shown to be very effective in the interpretation of interactions of drug microspeciations with biomembranes. These findings suggest that the interaction occurred owing to ionic and hydrophobic forces showing how the passage through the membrane is mainly favored in the pH interval 6-7.4. It was demonstrated that a pH gradient through model membranes may be responsible for a poorly homogeneous distribution of ofloxacin (or other related fluoroquinolones), which justifies the in vivo accumulation properties of this drug. DSC experiments, which are in agreement with computational data, also showed that the complexing capability of ofloxacin with regard to Mg(++) or Ca(++) may govern the drug entrance into bacterial cells before the DNA Girase inhibition and could ensure the formation of hydrophobic and more fluid phospholipid domains on the surface of the model membrane. These regions are more permeable with regard to various solutes, as well as ofloxacin, allowing a so-called 'self-promoted entrance pathway'. The combination of experimental methodologies with computational data allowed a further rationalization of the results and opened new perspectives into the mechanism of action of ofloxacin, namely its interaction with lipid bilayers and drug-divalent cation complex formation, which might be extended to the entire fluoroquinolone class. Ofloxacin accumulation within Escherichia coli ATCC 25922 was measured as a function of time. Also in this example, the environmental conditions influenced ofloxacin penetration and accumulation. The in vitro experiments, reported here, show that a suitable balance of hydrophilic and hydrophobic fluoroquinolone properties needs to occur for there to be increased drug permeation.

Digestive and Liver Disease, 2017

Current Drug Targets, 2015

The ability of some surfactants to self-assemble in a water/oil bi-phase environment thus forming... more The ability of some surfactants to self-assemble in a water/oil bi-phase environment thus forming supramolecular structure leading to the formation of w/o/w multiple emulsions was investigated. The w/o/w multiple emulsions obtained by self-assembling (one-step preparation method) were compared with those prepared following the traditional two-step procedure. Methyl-nicotinate was used as a hydrophilic model drug. The formation of the multiple emulsion structure was evidenced by optical microscopy, which showed a mean size of the inner oil droplets of 6 μm and 10 μm for one-step and two-step multiple emulsions, respectively. The in vitrobiopharmaceutical features of the various w/o/w multiple emulsion formulations were evaluated by means of viscosimetry studies, drug release and in vitro percutaneous permeation experiments through human stratum corneum and viable epidermis membranes. The self-assembled multiple emulsions allowed a more gradual percutaneous permeation (a zero-order permeation rate) than the two-step ones. The in vivotopical carrier properties of the two different multiple emulsions were evaluated on healthy human volunteers by using the spectrophotometry of reflectance, an in vivonon invasive method. These multiple emulsion systems were also compared with conventional emulsion formulations. Our findings demonstrated that the multiple emulsions obtained by self-assembling were able to provide a more sustained drug delivery into the skin and hence a longer therapeutic action than two-step multiple emulsions and conventional emulsion formulations. Finally, our findings showed that the supramolecular micro-assembly of multiple emulsions was able to influence not only the biopharmaceutical characteristics but also the potential in vivotherapeutic response.

Journal of Controlled Release, 2005

The aim of this work was the evaluation of various ethosomal suspensions made up of water, phosph... more The aim of this work was the evaluation of various ethosomal suspensions made up of water, phospholipids and ethanol at various concentrations for their potential application in dermal administration of ammonium glycyrrhizinate, a useful drug for the treatment of various inflammatory-based skin diseases. Physicochemical characterization of ethosomes was carried out by photon correlation spectroscopy and freeze fracture electron microscopy. The percutaneous permeation of ammonium glycyrrhizinate/ethosomes was evaluated in vitro through human stratum corneum and epidermis membranes by using Franz's cells and compared with the permeation profiles of drug solutions either in water or in a water-ethanol mixture. Reflectance spectrophotometry was used as a non-invasive technique to evaluate the carrier toxicity, the drug permeation and the anti-inflammatory activity of ammonium glycyrrhizinate in a model of skin erythema in vivo on human volunteers. Ethosomal suspensions had mean sizes ranging from 350 nm to 100 nm as a function of ethanol and lecithin quantities, i.e., high amounts of ethanol and a low lecithin concentration provided ethosome suspensions with a mean size of approximately 100 nm and a narrow size distribution. In vitro and in vivo experiments were carried out by using an ethosome formulation made up of ethanol 45% (v/v) and lecithin 2% (w/v). The ethosome suspension showed a very good skin tolerability in human volunteers, also when applied for a long period (48 h). Ethosomes elicited an increase of the in vitro percutaneous permeation of both methylnicotinate and ammonium glycyrrhizinate. Ethosomes were able to significantly enhance the anti-inflammatory activity of ammonium glycyrrhizinate compared to the ethanolic or aqueous solutions of this drug. Some in vivo experiments also showed the ability of ethosome to ensure a skin accumulation and a sustained release of the ammonium glycyrrhizinate.

Journal of Colloid and Interface Science, 2000

The influence exerted by the antimicrobial and antimicotic nonapeptide Leucinostatin A (Leu A) on... more The influence exerted by the antimicrobial and antimicotic nonapeptide Leucinostatin A (Leu A) on a biological membrane model made up of dipalmitoylphosphatidylcholine (DPPC) was investigated. Drug-membrane interactions, studied by means of differential scanning calorimetry and Fourier-transform infrared spectroscopy, depend on the behavior of the molecule which positions its hydrophilic part toward the bilayer phospholipid polar heads, while it inserts its hydrophobic portion into the membrane phospholipid acyl chain moiety. Calorimetric experiments showed that the peptide undergoes self-aggregation within the bilayer structure when present at molar fractions higher than 0.03. Peptide-membrane interactions as a function of time were analyzed as well. The latter demonstrated that Leu A inserts rapidly into the outer bilayers of DPPC membranes. 45 Ca 2+ uptake by DPPC vesicles gave a reason for the ionophoric activity of Leu A against both mouse thymocytes and artificial membranes, which seems to be correlated to the self-assembling property of the peptide within the bilayers.

European Journal of Pharmaceutics and Biopharmaceutics, 2012

Topical application of anticancer drugs for the treatment of malignancies represents a new challe... more Topical application of anticancer drugs for the treatment of malignancies represents a new challenge in dermatology, potentially being an alternative therapeutic approach for the efficacious treatment of nonmelanoma skin cancer, that is, actinic keratoses, and malignant lesions of the skin caused by ultraviolet radiation. Anti-proliferative and antimitotic drugs, including many of the taxanes, are currently under investigation for the treatment of cutaneous malignant transformation of actinic keratoses, particularly the squamous cell carcinoma. Paclitaxel-loaded ethosomes Ò are proposed as topical drug delivery systems for the treatment of this pathology due to their suitable physicochemical characteristics and enhanced skin penetration ability for deep dermal delivery. Our in vitro data show that the skin application of paclitaxel-loaded ethosomes Ò improved the permeation of paclitaxel in a stratum corneum-epidermis membrane model and increased its anti-proliferative activity in a squamous cell carcinoma model as compared to the free drug. The results obtained encouraged the use of the paclitaxel-loaded ethosomes Ò as the formulation for the potential treatment of squamous cell carcinoma, a malignant transformation of actinic keratoses.

Pharmaceutics, 2020

Mesoporous silicon microparticles (MSMPs) can incorporate drug-carrying nanoparticles (NPs) into ... more Mesoporous silicon microparticles (MSMPs) can incorporate drug-carrying nanoparticles (NPs) into their pores. An NP-loaded MSMP is a multistage vector (MSV) that forms a Matryoshka-like structure that protects the therapeutic cargo from degradation and prevents its dilution in the circulation during delivery to tumor cells. We developed an MSV constituted by 1 µm discoidal MSMPs embedded with PEGylated liposomes containing oxaliplatin (oxa) which is a therapeutic agent for colorectal cancer (CRC). To obtain extra-small liposomes able to fit the 60 nm pores of MSMP, we tested several liposomal formulations, and identified two optimal compositions, with a prevalence of the rigid lipid 1,2-distearoyl-sn-glycero-3-phosphocholine and of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. To improve the MSV assembly, we optimized the liposome-loading inside the MSMP and achieved a five-fold increase of the payload using an innovative lyophilization appro...

Biomedicines

The incidence of cancer is increasing dramatically, affecting all ages of the population and reac... more The incidence of cancer is increasing dramatically, affecting all ages of the population and reaching an ever higher worldwide mortality rate. The lack of therapies’ efficacy is due to several factors such as a delay in diagnosis, tumor regrowth after surgical resection and the occurrence of multidrug resistance (MDR). Tumor-associated immune cells and the tumor microenvironment (TME) deeply affect the tumor’s progression, leading to several physicochemical changes compared to physiological conditions. In this scenario, macrophages play a crucial role, participating both in tumor suppression or progression based on the polarization of onco-suppressive M1 or pro-oncogenic M2 phenotypes. Moreover, much evidence supports the pivotal role of macrophage-derived extracellular vesicles (EVs) as mediators in TME, because of their ability to shuttle the cell–cell and organ–cell communications, by delivering nucleic acids and proteins. EVs are lipid-based nanosystems with a broad size range d...

Materials Science and Engineering: C, 2021

In the last few decades, several natural and synthetic polymers have been used as starting materi... more In the last few decades, several natural and synthetic polymers have been used as starting material for the development of innovative polymeric nanoparticles able to encapsulate biologically active substances and to modulate their biopharmaceutical features and/or therapeutic efficacy. This investigation focused on the comparison of the physico-chemical properties of nanosystems made up of two of the most successfully used biodegradable biomaterials, namely poly(lactic-co-glycolic acid) (PLGA) and zein, belonging to the synthetic and natural family of polymers, respectively. Rutin, a polyphenolic bioflavonoid characterized by peculiar antioxidant properties, was chosen as the model drug to be encapsulated in the polymeric systems. The results demonstrated a greater ability of zein-based nanosystems to effectively retain the active compound with respect to the PLGA particles. The integration of rutin in the protein matrix favored a controlled drug leakage, and was influenced by the surfactant used to stabilize the formulation. Moreover, rutin-loaded zein nanoparticles showed significant in vitro antioxidant activity, evidencing a synergistic action between the intrinsic antioxidant activity of the protein and the pharmacological properties of the active compound. Finally, the intracellular localization of the zein nanosystems was demonstrated through confocal laser scanning microscopy.

International Journal of Pharmaceutics, 2002

Our research on topical application of lidocaine-loaded non-ionic surfactant vesicles (NSVs) was ... more Our research on topical application of lidocaine-loaded non-ionic surfactant vesicles (NSVs) was prompted by the great interest on new delivery systems for local anaesthetics. This study is focused on a novel formulation of NSVs entrapping lidocaine in the form of a free base (LID) and a hydrochloride (LIDHCl). NSVs were prepared from polyoxyethylene sorbitan monolaurate (Tween20™) and cholesterol. The effect of vesicle composition and environmental pH condition (8.6-5.5) on drug encapsulation efficiency (e.e.) was investigated. Experimental strategies involved: freeze-fracture, microscopy technique, dynamic light scattering, permeation through Silastic™ and mouse abdominal skin, in vitro release kinetics of vesicle-entrapped drugs, fluorescence quenching analyses. Diffusion experiments showed that the flux of charged lidocaine through Silastic™ membrane was possible only after the vesicle encapsulation. Permeation through mouse abdominal skin of LIDHCl loaded vesicles showed a higher flux and a shorter lag time with respect to classical liposome formulations, while LID permeation rate was quite similar for NSV and liposome formulations. Vesicles were also prepared in the presence of dicetylphosphate (DCP) and N-cetylpyridinium chloride (CP) to obtain negatively and positively charged vesicles respectively, but in this case the e.e. of the drug was negligible. The possible reason of the remarkable lower e.e. observed with charged vesicles was investigated by means of fluorescence quenching experiments.

Gels

For many years, corneal transplantation has been the first-choice treatment for irreversible dama... more For many years, corneal transplantation has been the first-choice treatment for irreversible damage affecting the anterior part of the eye. However, the low number of cornea donors and cases of graft rejection highlighted the need to replace donor corneas with new biomaterials. Tissue engineering plays a fundamental role in achieving this goal through challenging research into a construct that must reflect all the properties of the cornea that are essential to ensure correct vision. In this review, the anatomy and physiology of the cornea are described to point out the main roles of the corneal layers to be compensated and all the requirements expected from the material to be manufactured. Then, a deep investigation of alginate as a suitable alternative to donor tissue was conducted. Thanks to its adaptability, transparency and low immunogenicity, alginate has emerged as a promising candidate for the realization of bioengineered materials for corneal regeneration. Chemical modificat...

Pharmaceutical Research, 2014

Purpose To synthesize a new polymeric prodrug based on α,βpoly(N-2-hydroxyethyl)(2-aminoethylcarb... more Purpose To synthesize a new polymeric prodrug based on α,βpoly(N-2-hydroxyethyl)(2-aminoethylcarbamate)-d,l-aspartamide copolymer bearing amine groups in the side chain (PHEA-EDA), covalently linked to the anticancer drug doxorubicin and to test its potential application in anticancer therapy. Methods The drug was previously derivatized with a biocompatible and hydrophilic linker, leading to a doxorubicin derivative highly reactive with amino groups of PHEA-EDA. The PHEA-EDA-DOXO prodrug was characterized in terms of chemical stability. The pharmacokinetics, biodistribution and cytotoxicity of the product was investigated in vitro and in vivo on human breast cancer MCF-7 and T47D cell lines and NOD-SCID mice bearing a MCF-7 human breast carcinoma xenograft. Data collected were compared to those obtained using free doxorubicin. Results The final polymeric product is water soluble and easily hydrolysable in vivo, due to the presence of ester and amide bonds along the spacer between the drug and the polymeric backbone. In vitro tests showed a retarded cytotoxic effect on tumor cells, whereas a significant improvement of the in vivo antitumor activity of PHEA-EDA-DOXO and a survival advantage of the treated NOD-SCID mice was evidenced, compared to that of free doxorubicin. Conclusions The features of the PHEA-EDA-DOXO provide a potential protection of the drug from the plasmatic enzymatic degradation and clearance, an improvement of the blood pharmacokinetic parameters and a suitable body biodistribution. The data collected support the promising rationale of the proposed macromolecular prodrug PHEA-EDA-DOXO for further potential development and application in the treatment of solid cancer diseases. KEY WORDS antitumor activity. biodistribution. doxorubicin. PHEA-EDA. polymeric prodrug Chiara Di Meo and Felisa Cilurzo contributed equally.

Antioxidants

N-Acylethanolamine acid amidase (NAAA) is an N-terminal cysteine hydrolase that preferentially ca... more N-Acylethanolamine acid amidase (NAAA) is an N-terminal cysteine hydrolase that preferentially catalyzes the hydrolysis of endogenous lipid mediators such as palmitoylethanolamide, which has been shown to exhibit neuroprotective and antinociceptive properties by engaging peroxisome proliferator-activated receptor-α. A few potent NAAA inhibitors have been developed, including α-acylamino-β-lactone derivatives, which are very strong and effective, but they have limited chemical and plasmatic stability, compromising their use as systemic agents. In the present study, as an example of a molecule belonging to the chemical class of N-(2-oxo-3-oxetanyl)amide NAAA inhibitors, URB866 was entrapped in poly(lactic-co-glycolic acid) nanoparticles in order to increase its physical stability. The data show a monomodal pattern and a significant time- and temperature-dependent stability of the molecule-loaded nanoparticles, which also demonstrated a greater ability to effectively retain the compoun...

Pharmaceutics

The regeneration of cardiac tissue is a multidisciplinary research field aiming to improve the he... more The regeneration of cardiac tissue is a multidisciplinary research field aiming to improve the health condition of the post-heart attack patient. Indeed, myocardial tissue has a poor ability to self-regenerate after severe damage. The scientific efforts focused on the research of a biomaterial able to adapt to heart tissue, thus guaranteeing the in situ release of active substances or growth promoters. Many types of hydrogels were proposed for this purpose, showing several limitations. The aim of this study was to suggest a new usage for glyceryl monooleate-based lyotropic liquid crystals (LLCs) as a biocompatible and inert material for a myocardial application. The main advantages of LLCs are mainly related to their easy in situ injection as lamellar phase and their instant in situ transition in the cubic phase. In vivo studies proved the biocompatibility and the inertia of LLCs after their application on the myocardial tissue of mice. In detail, the cardiac activity was monitored ...

Frontiers in Pharmacology, 2021

Advances in nanotechnology have favored the development of novel colloidal formulations able to m... more Advances in nanotechnology have favored the development of novel colloidal formulations able to modulate the pharmacological and biopharmaceutical properties of drugs. The peculiar physico-chemical and technological properties of nanomaterial-based therapeutics have allowed for several successful applications in the treatment of cancer. The size, shape, charge and patterning of nanoscale therapeutic molecules are parameters that need to be investigated and modulated in order to promote and optimize cell and tissue interaction. In this review, the use of polymeric nanoparticles as drug delivery systems of anticancer compounds, their physico-chemical properties and their ability to be efficiently localized in specific tumor tissues have been described. The nanoencapsulation of antitumor active compounds in polymeric systems is a promising approach to improve the efficacy of various tumor treatments.

European Food Research and Technology, 2019

This investigation is focused on the granulometric and rheological analysis of two kinds of wheat... more This investigation is focused on the granulometric and rheological analysis of two kinds of wheat flour, i.e. type 0 and type 00. The granulometric analysis was carried out using a laser diffraction particle size analyzer. The particle size distribution of the type 0 and 00 flours, expressed as the mass median diameter (D (v,0.5)), was 80.922 µm and 71.686 µm, respectively. The rheological characterization of physical mixtures of wheat flour and water was carried out by means of small deformation dynamic oscillatory measurements. The rheological measurements evidenced a different behavior of the two samples of flour when they were mixed with water. Namely, the progressive addition of water to the type 0 wheat flour favored a decrease of G′ and G″ values, emphasizing that the dough becomes less full-bodied, while its structure remains unmodified; in fact, the phase-angle was not influenced by the water content. On the contrary, the type 00 wheat flour mixed with increased water content provided a dough characterized by lower elastic and viscous moduli, even though the behavior of the phase angle progressively varied. The different results may be due to the different particle size distribution of the flour and the different percentage of gluten present in each. Thanks to this parallel study of the particle size distribution of flours and the rheological behavior of the derived doughs, new food preparation procedures and new research on adulteration in the bread production can be designed.

Biomedicines

Bergamot essential oil (BEO) and Ammonium glycyrrhizinate (AG), naturally derived compounds, have... more Bergamot essential oil (BEO) and Ammonium glycyrrhizinate (AG), naturally derived compounds, have remarkable anti-inflammatory properties, thus making them suitable candidates for the treatment of skin disorders. Despite this, their inadequate physicochemical properties strongly compromise their topical application. Ultradeformable nanocarriers containing both BEO and AG were used to allow their passage through the skin, thus maximizing their therapeutic activity. Physicochemical characterization studies were performed using Zetasizer Nano ZS and Turbiscan Lab®. The dialysis method was used to investigate the release profile of the active compounds. In vivo studies were performed on human healthy volunteers through the X-Rite spectrophotometer. The nanosystems showed suitable features for topical cutaneous administration in terms of mean size, surface charge, size distribution, and long-term stability/storability. The co-delivery of BEO and AG in the deformable systems improved both...

Journal of Medicinal Chemistry, 2003

The structure-activity relationship studies on 2-quinolinecarboxamide peripheral benzodiazepine r... more The structure-activity relationship studies on 2-quinolinecarboxamide peripheral benzodiazepine receptor (PBR) ligands have been refined with the aim of using these ligands as carriers of radionuclides and boron atoms. Some new ligands show enhanced affinity and steroidogenic activity with respect to reference compound 1 and are interesting candidates for radiolabeling and PET studies. Moreover, carborane derivative 3q, representing the first example of PBR ligand bearing a carborane cage, can be useful to explore an alternative mechanism in BNCT.

Current Bioactive Compounds, 2007

The use of natural compounds has recently been reconsidered in modern clinical practice. A furthe... more The use of natural compounds has recently been reconsidered in modern clinical practice. A further advancement of these compounds is represented by the possibility of using innovative drug delivery systems that can improve the biopharmaceutical features of the delivered compounds. This review examines the recent developments in the field of delivery of natural drugs belonging to several therapeutic classes. In particular, the examined therapeutic classes are: antitumoral drugs, i.e. paclitaxel, doxorubicin, tea catechins and hypericin; anti-infective drugs, i.e. Melaleuca alternifolia and Artemisia arborescens L.; and antinflammatory/antioxidant drugs, i.e. cannabinoids and glycyrrhizinates used for topical application. In this review we highlight the utility of a suitable drug delivery system to improve the biopharmaceutical aspects of these drugs. The examined carriers are: vesicular carriers (liposomes, Ethosomes ® , ultradeformable liposomes), nano-and microparticles, innovative gels, microemulsions.

Bioorganic & Medicinal Chemistry, 2002

The interaction between ofloxacin, as a model drug of the fluoroquinolone class, and biomembranes... more The interaction between ofloxacin, as a model drug of the fluoroquinolone class, and biomembranes was examined as the possible initial step in a transmembrane diffusion process. Dipalmitoylphosphatidylcholine was used for the preparation of biomembrane models. The influence of environmental conditions and protonation on molecular physicochemical behavior, and hence on the membrane interaction, was investigated by differential scanning calorimetry (DSC). This technique has been shown to be very effective in the interpretation of interactions of drug microspeciations with biomembranes. These findings suggest that the interaction occurred owing to ionic and hydrophobic forces showing how the passage through the membrane is mainly favored in the pH interval 6-7.4. It was demonstrated that a pH gradient through model membranes may be responsible for a poorly homogeneous distribution of ofloxacin (or other related fluoroquinolones), which justifies the in vivo accumulation properties of this drug. DSC experiments, which are in agreement with computational data, also showed that the complexing capability of ofloxacin with regard to Mg(++) or Ca(++) may govern the drug entrance into bacterial cells before the DNA Girase inhibition and could ensure the formation of hydrophobic and more fluid phospholipid domains on the surface of the model membrane. These regions are more permeable with regard to various solutes, as well as ofloxacin, allowing a so-called 'self-promoted entrance pathway'. The combination of experimental methodologies with computational data allowed a further rationalization of the results and opened new perspectives into the mechanism of action of ofloxacin, namely its interaction with lipid bilayers and drug-divalent cation complex formation, which might be extended to the entire fluoroquinolone class. Ofloxacin accumulation within Escherichia coli ATCC 25922 was measured as a function of time. Also in this example, the environmental conditions influenced ofloxacin penetration and accumulation. The in vitro experiments, reported here, show that a suitable balance of hydrophilic and hydrophobic fluoroquinolone properties needs to occur for there to be increased drug permeation.

Digestive and Liver Disease, 2017

Current Drug Targets, 2015

The ability of some surfactants to self-assemble in a water/oil bi-phase environment thus forming... more The ability of some surfactants to self-assemble in a water/oil bi-phase environment thus forming supramolecular structure leading to the formation of w/o/w multiple emulsions was investigated. The w/o/w multiple emulsions obtained by self-assembling (one-step preparation method) were compared with those prepared following the traditional two-step procedure. Methyl-nicotinate was used as a hydrophilic model drug. The formation of the multiple emulsion structure was evidenced by optical microscopy, which showed a mean size of the inner oil droplets of 6 μm and 10 μm for one-step and two-step multiple emulsions, respectively. The in vitrobiopharmaceutical features of the various w/o/w multiple emulsion formulations were evaluated by means of viscosimetry studies, drug release and in vitro percutaneous permeation experiments through human stratum corneum and viable epidermis membranes. The self-assembled multiple emulsions allowed a more gradual percutaneous permeation (a zero-order permeation rate) than the two-step ones. The in vivotopical carrier properties of the two different multiple emulsions were evaluated on healthy human volunteers by using the spectrophotometry of reflectance, an in vivonon invasive method. These multiple emulsion systems were also compared with conventional emulsion formulations. Our findings demonstrated that the multiple emulsions obtained by self-assembling were able to provide a more sustained drug delivery into the skin and hence a longer therapeutic action than two-step multiple emulsions and conventional emulsion formulations. Finally, our findings showed that the supramolecular micro-assembly of multiple emulsions was able to influence not only the biopharmaceutical characteristics but also the potential in vivotherapeutic response.

Journal of Controlled Release, 2005

The aim of this work was the evaluation of various ethosomal suspensions made up of water, phosph... more The aim of this work was the evaluation of various ethosomal suspensions made up of water, phospholipids and ethanol at various concentrations for their potential application in dermal administration of ammonium glycyrrhizinate, a useful drug for the treatment of various inflammatory-based skin diseases. Physicochemical characterization of ethosomes was carried out by photon correlation spectroscopy and freeze fracture electron microscopy. The percutaneous permeation of ammonium glycyrrhizinate/ethosomes was evaluated in vitro through human stratum corneum and epidermis membranes by using Franz's cells and compared with the permeation profiles of drug solutions either in water or in a water-ethanol mixture. Reflectance spectrophotometry was used as a non-invasive technique to evaluate the carrier toxicity, the drug permeation and the anti-inflammatory activity of ammonium glycyrrhizinate in a model of skin erythema in vivo on human volunteers. Ethosomal suspensions had mean sizes ranging from 350 nm to 100 nm as a function of ethanol and lecithin quantities, i.e., high amounts of ethanol and a low lecithin concentration provided ethosome suspensions with a mean size of approximately 100 nm and a narrow size distribution. In vitro and in vivo experiments were carried out by using an ethosome formulation made up of ethanol 45% (v/v) and lecithin 2% (w/v). The ethosome suspension showed a very good skin tolerability in human volunteers, also when applied for a long period (48 h). Ethosomes elicited an increase of the in vitro percutaneous permeation of both methylnicotinate and ammonium glycyrrhizinate. Ethosomes were able to significantly enhance the anti-inflammatory activity of ammonium glycyrrhizinate compared to the ethanolic or aqueous solutions of this drug. Some in vivo experiments also showed the ability of ethosome to ensure a skin accumulation and a sustained release of the ammonium glycyrrhizinate.

Journal of Colloid and Interface Science, 2000

The influence exerted by the antimicrobial and antimicotic nonapeptide Leucinostatin A (Leu A) on... more The influence exerted by the antimicrobial and antimicotic nonapeptide Leucinostatin A (Leu A) on a biological membrane model made up of dipalmitoylphosphatidylcholine (DPPC) was investigated. Drug-membrane interactions, studied by means of differential scanning calorimetry and Fourier-transform infrared spectroscopy, depend on the behavior of the molecule which positions its hydrophilic part toward the bilayer phospholipid polar heads, while it inserts its hydrophobic portion into the membrane phospholipid acyl chain moiety. Calorimetric experiments showed that the peptide undergoes self-aggregation within the bilayer structure when present at molar fractions higher than 0.03. Peptide-membrane interactions as a function of time were analyzed as well. The latter demonstrated that Leu A inserts rapidly into the outer bilayers of DPPC membranes. 45 Ca 2+ uptake by DPPC vesicles gave a reason for the ionophoric activity of Leu A against both mouse thymocytes and artificial membranes, which seems to be correlated to the self-assembling property of the peptide within the bilayers.

European Journal of Pharmaceutics and Biopharmaceutics, 2012

Topical application of anticancer drugs for the treatment of malignancies represents a new challe... more Topical application of anticancer drugs for the treatment of malignancies represents a new challenge in dermatology, potentially being an alternative therapeutic approach for the efficacious treatment of nonmelanoma skin cancer, that is, actinic keratoses, and malignant lesions of the skin caused by ultraviolet radiation. Anti-proliferative and antimitotic drugs, including many of the taxanes, are currently under investigation for the treatment of cutaneous malignant transformation of actinic keratoses, particularly the squamous cell carcinoma. Paclitaxel-loaded ethosomes Ò are proposed as topical drug delivery systems for the treatment of this pathology due to their suitable physicochemical characteristics and enhanced skin penetration ability for deep dermal delivery. Our in vitro data show that the skin application of paclitaxel-loaded ethosomes Ò improved the permeation of paclitaxel in a stratum corneum-epidermis membrane model and increased its anti-proliferative activity in a squamous cell carcinoma model as compared to the free drug. The results obtained encouraged the use of the paclitaxel-loaded ethosomes Ò as the formulation for the potential treatment of squamous cell carcinoma, a malignant transformation of actinic keratoses.

Pharmaceutics, 2020

Mesoporous silicon microparticles (MSMPs) can incorporate drug-carrying nanoparticles (NPs) into ... more Mesoporous silicon microparticles (MSMPs) can incorporate drug-carrying nanoparticles (NPs) into their pores. An NP-loaded MSMP is a multistage vector (MSV) that forms a Matryoshka-like structure that protects the therapeutic cargo from degradation and prevents its dilution in the circulation during delivery to tumor cells. We developed an MSV constituted by 1 µm discoidal MSMPs embedded with PEGylated liposomes containing oxaliplatin (oxa) which is a therapeutic agent for colorectal cancer (CRC). To obtain extra-small liposomes able to fit the 60 nm pores of MSMP, we tested several liposomal formulations, and identified two optimal compositions, with a prevalence of the rigid lipid 1,2-distearoyl-sn-glycero-3-phosphocholine and of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. To improve the MSV assembly, we optimized the liposome-loading inside the MSMP and achieved a five-fold increase of the payload using an innovative lyophilization appro...