The state of well-being and health of our body is regulated by the fine osmotic and biochemical b... more The state of well-being and health of our body is regulated by the fine osmotic and biochemical balance established between the cells of the different tissues, organs, and systems. Specific districts of the human body are defined, kept in the correct state of functioning, and, therefore, protected from exogenous or endogenous insults of both mechanical, physical, and biological nature by the presence of different barrier systems. In addition to the placental barrier, which even acts as a linker between two different organisms, the mother and the fetus, all human body barriers, including the blood-brain barrier (BBB), blood-retinal barrier, blood-nerve barrier, blood-lymph barrier, and blood-cerebrospinal fluid barrier, operate to maintain the physiological homeostasis within tissues and organs. From a pharmaceutical point of view, the most challenging is undoubtedly the BBB, since its presence notably complicates the treatment of brain disorders. BBB action can impair the delivery of chemical drugs and biopharmaceuticals into the brain, reducing their therapeutic efficacy and/or increasing their unwanted bioaccumulation in the surrounding healthy tissues. Recent nanotechnological innovation provides advanced biomaterials and ad hoc customized engineering and functionalization methods able to assist in brain-targeted drug delivery. In this context, lipid nanocarriers, including both synthetic (liposomes, solid lipid nanoparticles, nanoemulsions, nanostructured lipid carriers, niosomes, proniosomes, and cubosomes) and cell-derived ones (extracellular vesicles and cell membrane-derived nanocarriers), are considered one of the most successful brain delivery systems due to their reasonable biocompatibility and ability to cross the BBB. This review aims to provide a complete and up-to-date point of view on the efficacy of the most varied lipid carriers, whether FDA-approved, involved in clinical trials, or used in in vitro or in vivo studies, for the treatment of inflammatory, cancerous, or infectious brain diseases.
We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeuti... more We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeutic stimuli-responsive cargo and equipping them with antibodies for efficient targeting of lymphoma cells. The post-isolation engineering of the EVs is accomplished by a freeze-thaw method to load therapeuticallyactive zinc oxide nanocrystals (ZnO NCs), obtaining the so-called TrojanNanoHorse (TNH) to recall the biomimetism and cytotoxic potential of this novel nanoconstruct. TNHs are further modified at their surface with anti-CD20 monoclonal antibodies (TNH CD20 ) achieving specific targeting against lymphoid cancer cell line. The in vitro characterization is carried out on CD20+ lymphoid Daudi cell line, CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (B lymphocytes). The TNH shows nanosized structure, high colloidal stability, even over time, and good hemocompatibility. The in vitro characterization shows the high biocompatibility, targeting specificity and cytotoxic capability. Importantly, the selectivity of TNH CD20 demonstrates significantly higher interaction towards the target lymphoid Daudi cell line compared to the CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (lymphocytes). An enhanced cytotoxicity directed against Daudi cancer cells is demonstrated after the TNH CD20 activation with high-energy ultrasound shock-waves (SW). This work demonstrates the efficient re-engineering of EVs, derived from healthy cells, with inorganic nanoparticles and monoclonal antibodies. The obtained hybrid nanoconstructs can be on-demand activated by an external stimulation, here acoustic pressure waves, to exploit a cytotoxic effect conveyed by the ZnO NCs cargo against selected cancer cells.
In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Ram... more In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Raman spectroscopy are combined through a direct imaging approach, to gather structural and chemical information of complex molecular systems such as ion channels in their original plasma membrane. Customized microfabricated sample holder allows to characterize Nav channels embedded in the original plasma membrane extracted from neuronal cells that are derived from healthy human induced pluripotent stem cells. The identification of the channels is accomplished by using two different approaches, one of them widely used in cryo‐EM (the particle analysis method) and the other based on a novel Zernike Polynomial expansion of the images bitmap. This approach allows to carry out a whole series of investigations, one complementary to the other, on the same sample, preserving its state as close as possible to the original membrane configuration.
According to the American Society for Testing and Materials, nanoparticles (NPs) are characterize... more According to the American Society for Testing and Materials, nanoparticles (NPs) are characterized by having two or more dimensions at the nanometer scale and distinctive improved physical/chemical properties compared with their bulk counterpart. Considering their composition, NPs are usually divided into three principal categories: inorganic (metals, metal oxide, ceramic and quantum dots), organic (polymers and lipid-based assembles) and carbon-based NPs . Nano-sized particles exhibit distinctive, chemical and physical properties that denote them as unique nanodevices usable in a wide range of pharmaceutical/therapeutic applications for their capability to affect directly cell vitality or for their drug delivery attitude . They are playing a strategic role in the implementation of new therapeutic solutions against Parkinson's, Alzheimer and infectious diseases, as well as against cancer and many other disorders . Usually, the main challenge is to achieve well-dispersed NPs, even in a colloidal manner, in order to facilitate their use both in in vitro and in vivo tests, in other words, for internalization, vitality, differentiation assays and so on. It is therefore required to oversee their size and shape and hence avoid aggregation and agglomeration in body fluids or in cell culture media, both containing proteins, electrolytes, lipids and other biomolecules. As soon as the NPs come into contact with biological fluids, their surface adsorbs a wide range of biomolecules, forming a protein corona which is assumed to comprise a hard layer and a soft one. The former is closely associated with the NP, while the soft layer consists of loosely bound proteins. Many parameters affect NPs' protein corona composition and subsequently their aggregation state and, ultimately, their therapeutic applicability. The presence of a protein corona on NPs dispersed in a biological fluid is unanimously recognized, as well as the fact that it can hugely affect their size, shape and charge . By controlling and measuring the NPs' colloidal stability and aggregation, researchers can have a clear idea of how these factors affect biodistribution, toxicity and pharmacokinetic parameters. Until now different solutions have been implemented to escape the agglomerates' formation in physiological solutions and media. Nanoparticle dispersion in aqueous media can be described by using the Deryagin, Landau, Verwey and Overbeek (DLVO) theory which takes into account the Van der Waals attractive forces and the electrostatic interactions, allowing assessment of the potential barrier between particles. In this commentary, we will analyze the role of various dispersant enhancers, including chemical, biological and physical approaches. Actually, dispersion stabilizers as a chemical modification of the NP surfaces are routinely applied at the preparation step of the nanomaterials. Such chemical functionalization includes the use of organic coatings, such as various natural and synthetic polymeric layers. Biologically relevant approaches rely on the use of proteins and lipid bilayers, both from synthetic or natural origins, to shield NPs against aggregation in a biologically relevant environment. For physical methods, the application of ultrasound as a pretreatment to improve NP dispersion is applied regularly for deagglomeration of NPs into biological media.
Super-hydrophobic surfaces (SHSs) are bio-inspired, artificial microfabricated interfaces, in whi... more Super-hydrophobic surfaces (SHSs) are bio-inspired, artificial microfabricated interfaces, in which a pattern of cylindrical micropillars is modified to incorporate details at the nanoscale. For those systems, the integration of different scales translates into superior properties, including the ability of manipulating biological solutions. The five Ws, five Ws and one H or the six Ws (6W), are questions, whose answers are considered basic in information-gathering. They constitute a formula for getting the complete story on a subject. According to the principle of the six Ws, a report can only be considered complete if it answers these questions starting with an interrogative word: who, why, what, where, when, how. Each question should have a factual answer. In what follows, SHSs and some of the most promising applications thereof are reviewed following the scheme of the 6W. We will show how these surfaces can be integrated into bio-photonic devices for the identification and detection of a single molecule. We will describe how SHSs and nanoporous silicon matrices can be combined to yield devices with the capability of harvesting small molecules, where the cut-off size can be adequately controlled. We will describe how this concept is utilized for obtaining a direct TEM image of a DNA molecule.
Mesoporous materials are structures characterized by a well-ordered large pore system with unifor... more Mesoporous materials are structures characterized by a well-ordered large pore system with uniform porous dimensions ranging between 2 and 50 nm. Typical samples are zeolite, carbon molecular sieves, porous metal oxides, organic and inorganic porous hybrid and pillared materials, silica clathrate and clathrate hydrates compounds. Improvement in biochemistry and materials science led to the design and implementation of different types of porous materials ranging from rigid to soft two-dimensional (2D) and three-dimensional (3D) skeletons. The present review focuses on the use of three-dimensional printed (3DP) mesoporous scaffolds suitable for a wide range of drug delivery applications, due to their intrinsic high surface area and high pore volume. In the first part, the importance of the porosity of materials employed for drug delivery application was discussed focusing on mesoporous materials. At the end of the introduction, hard and soft templating synthesis for the realization of ordered 2D/3D mesostructured porous materials were described. In the second part, 3DP fabrication techniques, including fused deposition modelling, material jetting as inkjet printing, electron beam melting, selective laser sintering, stereolithography and digital light processing, electrospinning, and two-photon polymerization were described. In the last section, through recent bibliographic research, a wide number of 3D printed mesoporous materials, for in vitro and in vivo drug delivery applications, most of which relate to bone cells and tissues, were presented and summarized in a table in which all the technical and bibliographical details were reported. This review highlights, to a very cross-sectional audience, how the interdisciplinarity of certain branches of knowledge, as those of materials science and nano-microfabrication are, represent a growing valuable aid in the advanced forum for the science and technology of pharmaceutics and biopharmaceutics.
Frontiers in Bioengineering and Biotechnology, Jun 5, 2020
In the last years, different nanotools have been developed to fight cancer cells. They could be a... more In the last years, different nanotools have been developed to fight cancer cells. They could be administered alone, exploiting their intrinsic toxicity, or remotely activated to achieve cell death. In the latter case, ultrasound (US) has been recently proposed to stimulate some nanomaterials because of the US outstanding property of deep tissue penetration and the possibility of focusing. In this study, for the first time, we report on the highly efficient killing capability of amino-propyl functionalized ZnO nanocrystals (ZnO NCs) in synergy with high-energy ultrasound shock waves (SW) for the treatment of cancer cells. The cytotoxicity and internalization of ZnO NCs were evaluated in cervical adenocarcinoma KB cells, as well as the safety of the SW treatment alone. Then, the remarkably high cytotoxic combination of ZnO NCs and SW was demonstrated, comparing the effect of multiple (3 times/day) SW treatments toward a single one, highlighting that multiple treatments are necessary to achieve efficient cell death. At last, preliminary tests to understand the mechanism of the observed synergistic effect were carried out, correlating the nanomaterial surface chemistry to the specific type of stimulus used. The obtained results can thus pave the way for a novel nanomedicine treatment, based on the synergistic effect of nanocrystals combined with highly intense mechanical pressure waves, offering high efficiency, deep and focused tissue penetration, and a reduction of side effects on healthy cells.
Cellular communications take place thanks to a well-connected network of chemicalphysical signals... more Cellular communications take place thanks to a well-connected network of chemicalphysical signals, biomolecules, growth factors, and vesicular messengers that travel inside or between cells. A deep knowledge of the extracellular vesicle (EV) system allows for a better understanding of the whole series of phenomena responsible for cell proliferation and death. To this purpose, here, a thorough immuno-phenotypic characterization of B-cell EV membranes is presented. Furthermore, the cellular membrane of B lymphocytes, Burkitt lymphoma, and human myeloid leukemic cells were characterized through cytofluorimetry assays and fluorescent microscopy analysis. Through cytotoxicity and internalization tests, the tropism of B lymphocyte-derived EVs was investigated toward the parental cell line and two different cancer cell lines. In this study, an innate capability of passive targeting of the native EVs was distinguished from the active targeting capability of monoclonal antibody-engineered EVs, able to selectively drive the vesicles, enhancing their internalization into the target cancer cells. In particular, the specific targeting ability of anti-CD20 engineered EVs towards Daudi cells, highly expressing CD20 marker on their cell membrane, was proved, while almost no internalization events were observed in HL60 cells, since they did not express an appreciable amount of the CD20 marker on their plasma membranes.
Fast diagnosis and more efficient therapies for cancer surely represent one of the huge tasks for... more Fast diagnosis and more efficient therapies for cancer surely represent one of the huge tasks for the worldwide researchers' and clinicians' community. In the last two decades, our understanding of the biology and molecular pathology of cancer mechanisms, coupled with the continuous development of the material science and technological compounds, have successfully improved nanomedicine applications in oncology. This review argues on nanomedicine application of engineered extracellular vesicles (EVs) in oncology. All the most innovative processes of EVs engineering are discussed together with the related degree of applicability for each one of them in cancer nanomedicines.
Conventional technologies for ureteral stent fabrication suffer from major inconveniences such as... more Conventional technologies for ureteral stent fabrication suffer from major inconveniences such as the development of encrustations and bacteria biofilm formation. These drawbacks typically lead to the failure of the device, significant patient discomfort and an additional surgery to remove and replace the stent in the worst cases. This work focuses on the preparation of a new nanocomposite material able to show drug elution properties, biodegradation and eventually potential antibacterial activity. Poly(2-hydroxyethyl methacrylate) or the crosslinked poly(2-hydroxyethyl methacrylate)-co-poly(acrylic acid) hydrogels were prepared by the radical polymerization method and combined with a biodegradable and antibacterial filling agent, i.e., flower-like Zinc Oxide (ZnO) micropowders obtained via the hydrothermal route. The physico-chemical analyses revealed the correct incorporation of ZnO within the hydrogel matrix and its highly mesoporous structure and surface area, ideal for drug incorporation. Two different anti-inflammatory drugs (Ibuprofen and Diclofenac) were loaded within each composite and the release profile was monitored up to two weeks in artificial urine (AU) and even at different pH values in AU to simulate pathological conditions. The addition of mesoporous ZnO micropowders to the hydrogel did not negatively affect the drug loading properties of the hydrogel and it was successfully allowed to mitigate undesirable burst-release effects. Furthermore, the sustained release of the drugs over time was observed at neutral pH, with kinetic constants (k) as low as 0.05 h -1 . By exploiting the pH-tunable swelling properties of the hydrogel, an even more sustained release was achieved in acidic and alkaline conditions especially at short release times, with a further reduction of burst effects (k ≈ 0.01-0.02 h -1 ). The nanocomposite system herein proposed represents a new material formulation for preparing innovative drug eluting stents with intrinsic antibacterial properties.
In the last few years there has been an increasing interest in nanomedicine and in particular in ... more In the last few years there has been an increasing interest in nanomedicine and in particular in developing nanoparticles to fight cancer [1]. Zinc oxide nanocrystals (ZnO NCs), thanks to their peculiar properties, can be employed for cancer diagnosis and therapy. Even if the cytotoxicity mechanism of ZnO NCs is not totally understood yet, it is most probably due to the combination of intracellular Zn cations release and the production of reactive oxygen species: in vitro tests have also shown an increased cytotoxic effect of ZnO on cancer cells with respect to the healthy counterpart [1]. In order to promote the stability of ZnO NCs in physiological environment, increase their biocompatibility and reduce their immunogenic effects, we covered ZnO NCs with a biomimetic phospholipidic bilayer [2] derived from extracellular vesicles, in particular exosomes, obtaining a nanoconstruct called TrojaNanoHorse (TNH). Exosomes are naturally produced by many types of cells as vehicle of interc...
s0010 12.1 Introduction p0010 Zinc oxide (ZnO) is a well-known metal oxide, studied from decades ... more s0010 12.1 Introduction p0010 Zinc oxide (ZnO) is a well-known metal oxide, studied from decades for its semiconducting, piezoelectric, pyroelectric, and optical absorption and emission properties . From the biomedical and biological point of views, it is classified as a "GRAS" (generally recognized as safe) substance by the Food and Drug Administration (FDA) [2]. Actually, it can be easily found in many commercial formulations for healthcare, as in baby-and sun creams preparations [3]. Nevertheless, the materials considered GRAS are above the micrometer range. ZnO, when reduced to the nanoscale, as many other substances, can develop new structural, physicochemical, and optical properties, such as the increase of the surface area-to-volume ratio and a higher chemical reactivity, since a large percentage of atoms in nanosized materials are at the surface and due to quantum effects . Therefore, unique biological and nano-biomedical applications could potentially arise from these new properties, as well as new possible mechanisms of toxicity not present in the bulk counterpart. p0015 The possible interactions between ZnO nanostructures (NSs) and the biological systems are mainly ruled by different morphological parameters such as size, shape, aspect ratio, surface area, surface charge, and chemical reactivity . p0020 Actually, the size of nanoparticles (NPs) is comparable to naturally occurring biological molecules, so their internalization into living cells enables them to affect the cellular behavior and viability.
Generation of artificial vascular grafts as blood vessel substitutes is a primary challenge in bi... more Generation of artificial vascular grafts as blood vessel substitutes is a primary challenge in biomaterial and tissue-engineering research. Ideally, these grafts should be able to recapitulate physiological and mechanical properties of natural vessels and guide the assembly of an endothelial cell lining to ensure hemo-compatibility. In this paper, we advance on this challenging task by designing and fabricating 3D vessel analogues by two-photon laser lithography using a synthetic photoresist. These scaffolds guarantee human endothelial cell adhesion and proliferation, and proper elastic behavior to withstand the pressure exerted by blood flow.
The aim of this study was the design of a 3D scaffold composed of poly(vinyl) alcohol (PVA) for c... more The aim of this study was the design of a 3D scaffold composed of poly(vinyl) alcohol (PVA) for cardiac tissue engineering (CTE) applications.
The state of well-being and health of our body is regulated by the fine osmotic and biochemical b... more The state of well-being and health of our body is regulated by the fine osmotic and biochemical balance established between the cells of the different tissues, organs, and systems. Specific districts of the human body are defined, kept in the correct state of functioning, and, therefore, protected from exogenous or endogenous insults of both mechanical, physical, and biological nature by the presence of different barrier systems. In addition to the placental barrier, which even acts as a linker between two different organisms, the mother and the fetus, all human body barriers, including the blood-brain barrier (BBB), blood-retinal barrier, blood-nerve barrier, blood-lymph barrier, and blood-cerebrospinal fluid barrier, operate to maintain the physiological homeostasis within tissues and organs. From a pharmaceutical point of view, the most challenging is undoubtedly the BBB, since its presence notably complicates the treatment of brain disorders. BBB action can impair the delivery of chemical drugs and biopharmaceuticals into the brain, reducing their therapeutic efficacy and/or increasing their unwanted bioaccumulation in the surrounding healthy tissues. Recent nanotechnological innovation provides advanced biomaterials and ad hoc customized engineering and functionalization methods able to assist in brain-targeted drug delivery. In this context, lipid nanocarriers, including both synthetic (liposomes, solid lipid nanoparticles, nanoemulsions, nanostructured lipid carriers, niosomes, proniosomes, and cubosomes) and cell-derived ones (extracellular vesicles and cell membrane-derived nanocarriers), are considered one of the most successful brain delivery systems due to their reasonable biocompatibility and ability to cross the BBB. This review aims to provide a complete and up-to-date point of view on the efficacy of the most varied lipid carriers, whether FDA-approved, involved in clinical trials, or used in in vitro or in vivo studies, for the treatment of inflammatory, cancerous, or infectious brain diseases.
We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeuti... more We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeutic stimuli-responsive cargo and equipping them with antibodies for efficient targeting of lymphoma cells. The post-isolation engineering of the EVs is accomplished by a freeze-thaw method to load therapeuticallyactive zinc oxide nanocrystals (ZnO NCs), obtaining the so-called TrojanNanoHorse (TNH) to recall the biomimetism and cytotoxic potential of this novel nanoconstruct. TNHs are further modified at their surface with anti-CD20 monoclonal antibodies (TNH CD20 ) achieving specific targeting against lymphoid cancer cell line. The in vitro characterization is carried out on CD20+ lymphoid Daudi cell line, CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (B lymphocytes). The TNH shows nanosized structure, high colloidal stability, even over time, and good hemocompatibility. The in vitro characterization shows the high biocompatibility, targeting specificity and cytotoxic capability. Importantly, the selectivity of TNH CD20 demonstrates significantly higher interaction towards the target lymphoid Daudi cell line compared to the CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (lymphocytes). An enhanced cytotoxicity directed against Daudi cancer cells is demonstrated after the TNH CD20 activation with high-energy ultrasound shock-waves (SW). This work demonstrates the efficient re-engineering of EVs, derived from healthy cells, with inorganic nanoparticles and monoclonal antibodies. The obtained hybrid nanoconstructs can be on-demand activated by an external stimulation, here acoustic pressure waves, to exploit a cytotoxic effect conveyed by the ZnO NCs cargo against selected cancer cells.
In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Ram... more In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Raman spectroscopy are combined through a direct imaging approach, to gather structural and chemical information of complex molecular systems such as ion channels in their original plasma membrane. Customized microfabricated sample holder allows to characterize Nav channels embedded in the original plasma membrane extracted from neuronal cells that are derived from healthy human induced pluripotent stem cells. The identification of the channels is accomplished by using two different approaches, one of them widely used in cryo‐EM (the particle analysis method) and the other based on a novel Zernike Polynomial expansion of the images bitmap. This approach allows to carry out a whole series of investigations, one complementary to the other, on the same sample, preserving its state as close as possible to the original membrane configuration.
According to the American Society for Testing and Materials, nanoparticles (NPs) are characterize... more According to the American Society for Testing and Materials, nanoparticles (NPs) are characterized by having two or more dimensions at the nanometer scale and distinctive improved physical/chemical properties compared with their bulk counterpart. Considering their composition, NPs are usually divided into three principal categories: inorganic (metals, metal oxide, ceramic and quantum dots), organic (polymers and lipid-based assembles) and carbon-based NPs . Nano-sized particles exhibit distinctive, chemical and physical properties that denote them as unique nanodevices usable in a wide range of pharmaceutical/therapeutic applications for their capability to affect directly cell vitality or for their drug delivery attitude . They are playing a strategic role in the implementation of new therapeutic solutions against Parkinson's, Alzheimer and infectious diseases, as well as against cancer and many other disorders . Usually, the main challenge is to achieve well-dispersed NPs, even in a colloidal manner, in order to facilitate their use both in in vitro and in vivo tests, in other words, for internalization, vitality, differentiation assays and so on. It is therefore required to oversee their size and shape and hence avoid aggregation and agglomeration in body fluids or in cell culture media, both containing proteins, electrolytes, lipids and other biomolecules. As soon as the NPs come into contact with biological fluids, their surface adsorbs a wide range of biomolecules, forming a protein corona which is assumed to comprise a hard layer and a soft one. The former is closely associated with the NP, while the soft layer consists of loosely bound proteins. Many parameters affect NPs' protein corona composition and subsequently their aggregation state and, ultimately, their therapeutic applicability. The presence of a protein corona on NPs dispersed in a biological fluid is unanimously recognized, as well as the fact that it can hugely affect their size, shape and charge . By controlling and measuring the NPs' colloidal stability and aggregation, researchers can have a clear idea of how these factors affect biodistribution, toxicity and pharmacokinetic parameters. Until now different solutions have been implemented to escape the agglomerates' formation in physiological solutions and media. Nanoparticle dispersion in aqueous media can be described by using the Deryagin, Landau, Verwey and Overbeek (DLVO) theory which takes into account the Van der Waals attractive forces and the electrostatic interactions, allowing assessment of the potential barrier between particles. In this commentary, we will analyze the role of various dispersant enhancers, including chemical, biological and physical approaches. Actually, dispersion stabilizers as a chemical modification of the NP surfaces are routinely applied at the preparation step of the nanomaterials. Such chemical functionalization includes the use of organic coatings, such as various natural and synthetic polymeric layers. Biologically relevant approaches rely on the use of proteins and lipid bilayers, both from synthetic or natural origins, to shield NPs against aggregation in a biologically relevant environment. For physical methods, the application of ultrasound as a pretreatment to improve NP dispersion is applied regularly for deagglomeration of NPs into biological media.
Super-hydrophobic surfaces (SHSs) are bio-inspired, artificial microfabricated interfaces, in whi... more Super-hydrophobic surfaces (SHSs) are bio-inspired, artificial microfabricated interfaces, in which a pattern of cylindrical micropillars is modified to incorporate details at the nanoscale. For those systems, the integration of different scales translates into superior properties, including the ability of manipulating biological solutions. The five Ws, five Ws and one H or the six Ws (6W), are questions, whose answers are considered basic in information-gathering. They constitute a formula for getting the complete story on a subject. According to the principle of the six Ws, a report can only be considered complete if it answers these questions starting with an interrogative word: who, why, what, where, when, how. Each question should have a factual answer. In what follows, SHSs and some of the most promising applications thereof are reviewed following the scheme of the 6W. We will show how these surfaces can be integrated into bio-photonic devices for the identification and detection of a single molecule. We will describe how SHSs and nanoporous silicon matrices can be combined to yield devices with the capability of harvesting small molecules, where the cut-off size can be adequately controlled. We will describe how this concept is utilized for obtaining a direct TEM image of a DNA molecule.
Mesoporous materials are structures characterized by a well-ordered large pore system with unifor... more Mesoporous materials are structures characterized by a well-ordered large pore system with uniform porous dimensions ranging between 2 and 50 nm. Typical samples are zeolite, carbon molecular sieves, porous metal oxides, organic and inorganic porous hybrid and pillared materials, silica clathrate and clathrate hydrates compounds. Improvement in biochemistry and materials science led to the design and implementation of different types of porous materials ranging from rigid to soft two-dimensional (2D) and three-dimensional (3D) skeletons. The present review focuses on the use of three-dimensional printed (3DP) mesoporous scaffolds suitable for a wide range of drug delivery applications, due to their intrinsic high surface area and high pore volume. In the first part, the importance of the porosity of materials employed for drug delivery application was discussed focusing on mesoporous materials. At the end of the introduction, hard and soft templating synthesis for the realization of ordered 2D/3D mesostructured porous materials were described. In the second part, 3DP fabrication techniques, including fused deposition modelling, material jetting as inkjet printing, electron beam melting, selective laser sintering, stereolithography and digital light processing, electrospinning, and two-photon polymerization were described. In the last section, through recent bibliographic research, a wide number of 3D printed mesoporous materials, for in vitro and in vivo drug delivery applications, most of which relate to bone cells and tissues, were presented and summarized in a table in which all the technical and bibliographical details were reported. This review highlights, to a very cross-sectional audience, how the interdisciplinarity of certain branches of knowledge, as those of materials science and nano-microfabrication are, represent a growing valuable aid in the advanced forum for the science and technology of pharmaceutics and biopharmaceutics.
Frontiers in Bioengineering and Biotechnology, Jun 5, 2020
In the last years, different nanotools have been developed to fight cancer cells. They could be a... more In the last years, different nanotools have been developed to fight cancer cells. They could be administered alone, exploiting their intrinsic toxicity, or remotely activated to achieve cell death. In the latter case, ultrasound (US) has been recently proposed to stimulate some nanomaterials because of the US outstanding property of deep tissue penetration and the possibility of focusing. In this study, for the first time, we report on the highly efficient killing capability of amino-propyl functionalized ZnO nanocrystals (ZnO NCs) in synergy with high-energy ultrasound shock waves (SW) for the treatment of cancer cells. The cytotoxicity and internalization of ZnO NCs were evaluated in cervical adenocarcinoma KB cells, as well as the safety of the SW treatment alone. Then, the remarkably high cytotoxic combination of ZnO NCs and SW was demonstrated, comparing the effect of multiple (3 times/day) SW treatments toward a single one, highlighting that multiple treatments are necessary to achieve efficient cell death. At last, preliminary tests to understand the mechanism of the observed synergistic effect were carried out, correlating the nanomaterial surface chemistry to the specific type of stimulus used. The obtained results can thus pave the way for a novel nanomedicine treatment, based on the synergistic effect of nanocrystals combined with highly intense mechanical pressure waves, offering high efficiency, deep and focused tissue penetration, and a reduction of side effects on healthy cells.
Cellular communications take place thanks to a well-connected network of chemicalphysical signals... more Cellular communications take place thanks to a well-connected network of chemicalphysical signals, biomolecules, growth factors, and vesicular messengers that travel inside or between cells. A deep knowledge of the extracellular vesicle (EV) system allows for a better understanding of the whole series of phenomena responsible for cell proliferation and death. To this purpose, here, a thorough immuno-phenotypic characterization of B-cell EV membranes is presented. Furthermore, the cellular membrane of B lymphocytes, Burkitt lymphoma, and human myeloid leukemic cells were characterized through cytofluorimetry assays and fluorescent microscopy analysis. Through cytotoxicity and internalization tests, the tropism of B lymphocyte-derived EVs was investigated toward the parental cell line and two different cancer cell lines. In this study, an innate capability of passive targeting of the native EVs was distinguished from the active targeting capability of monoclonal antibody-engineered EVs, able to selectively drive the vesicles, enhancing their internalization into the target cancer cells. In particular, the specific targeting ability of anti-CD20 engineered EVs towards Daudi cells, highly expressing CD20 marker on their cell membrane, was proved, while almost no internalization events were observed in HL60 cells, since they did not express an appreciable amount of the CD20 marker on their plasma membranes.
Fast diagnosis and more efficient therapies for cancer surely represent one of the huge tasks for... more Fast diagnosis and more efficient therapies for cancer surely represent one of the huge tasks for the worldwide researchers' and clinicians' community. In the last two decades, our understanding of the biology and molecular pathology of cancer mechanisms, coupled with the continuous development of the material science and technological compounds, have successfully improved nanomedicine applications in oncology. This review argues on nanomedicine application of engineered extracellular vesicles (EVs) in oncology. All the most innovative processes of EVs engineering are discussed together with the related degree of applicability for each one of them in cancer nanomedicines.
Conventional technologies for ureteral stent fabrication suffer from major inconveniences such as... more Conventional technologies for ureteral stent fabrication suffer from major inconveniences such as the development of encrustations and bacteria biofilm formation. These drawbacks typically lead to the failure of the device, significant patient discomfort and an additional surgery to remove and replace the stent in the worst cases. This work focuses on the preparation of a new nanocomposite material able to show drug elution properties, biodegradation and eventually potential antibacterial activity. Poly(2-hydroxyethyl methacrylate) or the crosslinked poly(2-hydroxyethyl methacrylate)-co-poly(acrylic acid) hydrogels were prepared by the radical polymerization method and combined with a biodegradable and antibacterial filling agent, i.e., flower-like Zinc Oxide (ZnO) micropowders obtained via the hydrothermal route. The physico-chemical analyses revealed the correct incorporation of ZnO within the hydrogel matrix and its highly mesoporous structure and surface area, ideal for drug incorporation. Two different anti-inflammatory drugs (Ibuprofen and Diclofenac) were loaded within each composite and the release profile was monitored up to two weeks in artificial urine (AU) and even at different pH values in AU to simulate pathological conditions. The addition of mesoporous ZnO micropowders to the hydrogel did not negatively affect the drug loading properties of the hydrogel and it was successfully allowed to mitigate undesirable burst-release effects. Furthermore, the sustained release of the drugs over time was observed at neutral pH, with kinetic constants (k) as low as 0.05 h -1 . By exploiting the pH-tunable swelling properties of the hydrogel, an even more sustained release was achieved in acidic and alkaline conditions especially at short release times, with a further reduction of burst effects (k ≈ 0.01-0.02 h -1 ). The nanocomposite system herein proposed represents a new material formulation for preparing innovative drug eluting stents with intrinsic antibacterial properties.
In the last few years there has been an increasing interest in nanomedicine and in particular in ... more In the last few years there has been an increasing interest in nanomedicine and in particular in developing nanoparticles to fight cancer [1]. Zinc oxide nanocrystals (ZnO NCs), thanks to their peculiar properties, can be employed for cancer diagnosis and therapy. Even if the cytotoxicity mechanism of ZnO NCs is not totally understood yet, it is most probably due to the combination of intracellular Zn cations release and the production of reactive oxygen species: in vitro tests have also shown an increased cytotoxic effect of ZnO on cancer cells with respect to the healthy counterpart [1]. In order to promote the stability of ZnO NCs in physiological environment, increase their biocompatibility and reduce their immunogenic effects, we covered ZnO NCs with a biomimetic phospholipidic bilayer [2] derived from extracellular vesicles, in particular exosomes, obtaining a nanoconstruct called TrojaNanoHorse (TNH). Exosomes are naturally produced by many types of cells as vehicle of interc...
s0010 12.1 Introduction p0010 Zinc oxide (ZnO) is a well-known metal oxide, studied from decades ... more s0010 12.1 Introduction p0010 Zinc oxide (ZnO) is a well-known metal oxide, studied from decades for its semiconducting, piezoelectric, pyroelectric, and optical absorption and emission properties . From the biomedical and biological point of views, it is classified as a "GRAS" (generally recognized as safe) substance by the Food and Drug Administration (FDA) [2]. Actually, it can be easily found in many commercial formulations for healthcare, as in baby-and sun creams preparations [3]. Nevertheless, the materials considered GRAS are above the micrometer range. ZnO, when reduced to the nanoscale, as many other substances, can develop new structural, physicochemical, and optical properties, such as the increase of the surface area-to-volume ratio and a higher chemical reactivity, since a large percentage of atoms in nanosized materials are at the surface and due to quantum effects . Therefore, unique biological and nano-biomedical applications could potentially arise from these new properties, as well as new possible mechanisms of toxicity not present in the bulk counterpart. p0015 The possible interactions between ZnO nanostructures (NSs) and the biological systems are mainly ruled by different morphological parameters such as size, shape, aspect ratio, surface area, surface charge, and chemical reactivity . p0020 Actually, the size of nanoparticles (NPs) is comparable to naturally occurring biological molecules, so their internalization into living cells enables them to affect the cellular behavior and viability.
Generation of artificial vascular grafts as blood vessel substitutes is a primary challenge in bi... more Generation of artificial vascular grafts as blood vessel substitutes is a primary challenge in biomaterial and tissue-engineering research. Ideally, these grafts should be able to recapitulate physiological and mechanical properties of natural vessels and guide the assembly of an endothelial cell lining to ensure hemo-compatibility. In this paper, we advance on this challenging task by designing and fabricating 3D vessel analogues by two-photon laser lithography using a synthetic photoresist. These scaffolds guarantee human endothelial cell adhesion and proliferation, and proper elastic behavior to withstand the pressure exerted by blood flow.
The aim of this study was the design of a 3D scaffold composed of poly(vinyl) alcohol (PVA) for c... more The aim of this study was the design of a 3D scaffold composed of poly(vinyl) alcohol (PVA) for cardiac tissue engineering (CTE) applications.
Inside Front Cover
On page 402, E. Di Fabrizio and co-workers describe the growth of neuron–ast... more Inside Front Cover
On page 402, E. Di Fabrizio and co-workers describe the growth of neuron–astrocyte co-cultures on nanostructured superhydrophobic substrates. The scanning electron microscopy image on the inside cover shows a flat glial cell monolayer (pink) suspended between adjacent nanostructured pillars. Astrocytes adhere with high affinity to both the top and lateral surface of the pillars and provide an excellent support for cocultured neurons, as shown by a large neuron (blue) spreading upon the glial carpet with its multiple neuritic processes.
Although the detection of methylated cell free DNA represents one of the most promising approache... more Although the detection of methylated cell free DNA represents one of the most promising approaches for relapse risk assessment in cancer patients, the low concentration of cell-free circulating DNA constitutes the biggest obstacle in the development of DNA methylation-based biomarkers from blood. This paper describes a method for the measurement of genomic methylation content directly on circulating tumor cells (CTC), which could be used to deceive the aforementioned problem. Since CTC are disease related blood-based biomarkers, they result essential to monitor tumor's stadiation, therapy, and early relapsing lesions. Within surface's bio-functionalization and cell's isolation procedure standardization, the presented approach reveals a singular ability to detect high 5-methylcytosine CTC-subset content in the whole CTC compound, by choosing folic acid (FA) as transducer molecule. Sensitivity and specificity, calculated for FA functionalized surface (FA-surface), result respectively on about 83% and 60%. FA-surface, allowing the detection and characterization of early metastatic dissemination, provides a unique advance in the comprehension of tumors progression and dissemination confirming the presence of CTC and its association with high risk of relapse. This functionalized surface identifying and quantifying high 5-methylcytosine CTC-subset content into the patient's blood lead significant progress in cancer risk assessment, also providing a novel therapeutic strategy.
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Papers by tania limongi
On page 402, E. Di Fabrizio and co-workers describe the growth of neuron–astrocyte co-cultures on nanostructured superhydrophobic substrates. The scanning electron microscopy image on the inside cover shows a flat glial cell monolayer (pink) suspended between adjacent nanostructured pillars. Astrocytes adhere with high affinity to both the top and lateral surface of the pillars and provide an excellent support for cocultured neurons, as shown by a large neuron (blue) spreading upon the glial carpet with its multiple neuritic processes.