Publications by authors named "Massimiliano Papi"

82 Publications

Inhibiting the Growth of 3D Brain Cancer Models with Bio-Coronated Liposomal Temozolomide.

Pharmaceutics 2021 Mar 12;13(3). Epub 2021 Mar 12.

Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 291, 00161 Rome, Italy.

Nanoparticles (NPs) have emerged as an effective means to deliver anticancer drugs into the brain. Among various forms of NPs, liposomal temozolomide (TMZ) is the drug-of-choice for the treatment and management of brain tumours, but its therapeutic benefit is suboptimal. Although many possible reasons may account for the compromised therapeutic efficacy, the inefficient tumour penetration of liposomal TMZ can be a vital obstacle. Recently, the protein corona, i.e., the layer of plasma proteins that surround NPs after exposure to human plasma, has emerged as an endogenous trigger that mostly controls their anticancer efficacy. Exposition of particular biomolecules from the corona referred to as protein corona fingerprints (PCFs) may facilitate interactions with specific receptors of target cells, thus, promoting efficient internalization. In this work, we have synthesized a set of four TMZ-encapsulating nanomedicines made of four cationic liposome (CL) formulations with systematic changes in lipid composition and physical-chemical properties. We have demonstrated that precoating liposomal TMZ with a protein corona made of human plasma proteins can increase drug penetration in a 3D brain cancer model derived from U87 human glioblastoma multiforme cell line leading to marked inhibition of tumour growth. On the other side, by fine-tuning corona composition we have also provided experimental evidence of a non-unique effect of the corona on the tumour growth for all the complexes investigated, thus, clarifying that certain PCFs (i.e., APO-B and APO-E) enable favoured interactions with specific receptors of brain cancer cells. Reported results open new perspectives into the development of corona-coated liposomal drugs with enhanced tumour penetration and antitumour efficacy.
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http://dx.doi.org/10.3390/pharmaceutics13030378DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999290PMC
March 2021

Visco-Hyperelastic Characterization of the Equine Immature Zona Pellucida.

Materials (Basel) 2021 Mar 5;14(5). Epub 2021 Mar 5.

Dipartimento di Meccanica, Matematica e Management, Politecnico di Bari, 70125 Bari, Italy.

This article presents a very detailed study on the mechanical characterization of a highly nonlinear material, the immature equine zona pellucida (ZP) membrane. The ZP is modeled as a visco-hyperelastic soft matter. The Arruda-Boyce constitutive equation and the two-term Prony series are identified as the most suitable models for describing the hyperelastic and viscous components, respectively, of the ZP's mechanical response. Material properties are identified via inverse analysis based on nonlinear optimization which fits nanoindentation curves recorded at different rates. The suitability of the proposed approach is fully demonstrated by the very good agreement between AFM data and numerically reconstructed force-indentation curves. A critical comparison of mechanical behavior of two immature ZP membranes (i.e., equine and porcine ZPs) is also carried out considering the information on the structure of these materials available from electron microscopy investigations documented in the literature.
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http://dx.doi.org/10.3390/ma14051223DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961360PMC
March 2021

PE_PGRS3 ensures provision of the vital phospholipids cardiolipin and phosphatidylinositols by promoting the interaction between and host cells.

Virulence 2021 Dec;12(1):868-884

Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario "A. Gemelli", Rome, Italy.

PE_PGRS proteins of () constitute a large family of complex modular proteins whose role is still unclear. Among those, we have previously shown, using the heterologous expression in , that PE_PGRS3 containing a unique arginine-rich C-terminal domain, promotes adhesion to host cells. In this study, we investigate the role of PE_PGRS3 and its C-terminal domain directly in using functional deletion mutants. The results obtained here show that PE_PGRS3 is localized on the mycobacterial cell wall and its arginine-rich C-terminal region protrudes from the mycobacterial membrane and mediates entry into epithelial cells. Most importantly, this positively charged helical domain specifically binds phosphorylated phosphatidylinositols and cardiolipin, whereas it is unable to bind other phospholipids. Interestingly, administration of cardiolipin and phosphatidylinositol but no other phospholipids was able to turn-off expression of 3 activated by phosphate starvation conditions. These findings suggest that PE_PGRS3 has the key role to serve as a bridge between mycobacteria and host cells by interacting with specific host phospholipids and extracting them from host cells, for their direct integration or as a source of phosphate, during phases of TB pathogenesis when is short of phosphate supply.
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http://dx.doi.org/10.1080/21505594.2021.1897247DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8007152PMC
December 2021

Face masks and nanotechnology: Keep the blue side up.

Nano Today 2021 Apr 13;37:101077. Epub 2021 Jan 13.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Rome, Italy.

Coronavirus Disease 2019 (COVID-19) is one of the biggest challenges of the 21st century. While researchers are working on vaccine development and elucidating the mechanism of action and evolution of the harmful SARS-CoV-2, the current most important public health measure, second only to social distancing, is the obligatory wearing of facial protection. The Centers for Disease Control and Prevention recommended in April 2020 that the public wear face coverings in areas with high rates of transmission based on epidemiological evidence on the strong relationship between mask wearing and pandemic control. This protection against SARS-CoV-2 and other airborne pathogens, boost the design and production of innovative solutions by industry stakeholders. Nanoparticles, nanofibers, and other pioneering technologies based on nanomaterials have been introduced in mask production chains to improve performance and confer antiviral properties. During an emergency like COVID-19, these products directly available to the public should be carefully analyzed in terms of efficacy and possible long-term effects on the wearers' skin and lungs as well as on the environment. This opinion paper provides a wealth of information on the role of nanotechnologies in improving the performance of facial masks and on possible future consequences caused by a poorly regulated use of nanotechnology in textiles.
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http://dx.doi.org/10.1016/j.nantod.2021.101077DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7833187PMC
April 2021

Graphene-Based Materials: Biological and Biomedical Applications.

Int J Mol Sci 2021 Jan 12;22(2). Epub 2021 Jan 12.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, 00168 Rome, Italy.

This editorial aims to summarize the eleven scientific papers published in the Special Issue "Graphene-Based Materials: Biological and Biomedical Applications" [...].
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http://dx.doi.org/10.3390/ijms22020672DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7826772PMC
January 2021

Fourier Transform Infrared Spectroscopy as a useful tool for the automated classification of cancer cell-derived exosomes obtained under different culture conditions.

Anal Chim Acta 2020 Dec 9;1140:219-227. Epub 2020 Oct 9.

Dipartimento di Neuroscienze, Sezione di Fisica, Università Cattolica Del Sacro Cuore, Roma, Italy; Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy. Electronic address:

Exosomes possess great potential as cancer biomarkers in personalized medicine due to their easy accessibility and capability of representing their parental cells. To boost the translational process of exosomes in diagnostics, the development of novel and effective strategies for their label-free and automated characterization is highly desirable. In this context, Fourier Transform Infrared Spectroscopy (FTIR) has great potential as it provides direct access to specific biomolecular bands that give compositional information on exosomes in terms of their protein, lipid and genetic content. Here, we used FTIR spectroscopy in the mid-Infrared (mid-IR) range to study exosomes released from human colorectal adenocarcinoma HT-29 cancer cells cultured in different media. To this purpose, cells were studied in well-fed condition of growth, with 10% of exosome-depleted FBS (EVd-FBS), and under serum starvation with 0.5% EVd-FBS. Our data show the presence of statistically significant differences in the shape of the Amide I and II bands in the two conditions. Based on these differences, we showed the possibility to automatically classify cancer cell-derived exosomes using Principal Component Analysis combined with Linear Discriminant Analysis (PCA-LDA); we tested the effectiveness of the classifier with a cross-validation approach, obtaining very high accuracy, precision, and recall. Aside from classification purposes, our FTIR data provide hints on the underlying cellular mechanisms responsible for the compositional differences in exosomes, suggesting a possible role of starvation-induced autophagy.
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http://dx.doi.org/10.1016/j.aca.2020.09.037DOI Listing
December 2020

Celecoxib Exerts Neuroprotective Effects in β-Amyloid-Treated SH-SY5Y Cells Through the Regulation of Heme Oxygenase-1: Novel Insights for an Old Drug.

Front Cell Dev Biol 2020 29;8:561179. Epub 2020 Sep 29.

Department of Healthcare Surveillance and Bioethics, Section of Pharmacology, Università Cattolica del Sacro Cuore, Rome, Italy.

The formation and aggregation of amyloid-β-peptide (Aβ) into soluble and insoluble species represent the pathological hallmarks of Alzheimer's disease (AD). Over the last few years, however, soluble Aβ (sAβ) prevailed over fibrillar Aβ (fAβ) as determinant of neurotoxicity. One of the main therapeutic strategies for challenging neurodegeneration is to fight against neuroinflammation and prevent free radical-induced damage: in this light, the heme oxygenase/biliverdin reductase (HO/BVR) system is considered a promising drug target. The aim of this work was to investigate whether or not celecoxib (CXB), a selective inhibitor of the pro-inflammatory cyclooxygenase-2, modulates the HO/BVR system and prevents lipid peroxidation in SH-SY5Y neuroblastoma cells. Both sAβ (6.25-50 nM) and fAβ (1.25-50 nM) dose-dependently over-expressed inducible HO (HO-1) after 24 h of incubation, reaching statistical significance at 25 and 6.25 nM, respectively. Interestingly, CXB (1-10 μM, for 1 h) further enhanced Aβ-induced HO-1 expression through the nuclear translocation of the transcriptional factor Nrf2. Furthermore, 10 μM CXB counteracted the Aβ-induced ROS production with a mechanism fully dependent on HO-1 up-regulation; nevertheless, 10 μM CXB significantly counteracted only 25 nM sAβ-induced lipid peroxidation damage in SH-SY5Y neurons by modulating HO-1. Both carbon monoxide (CORM-2, 50 nM) and bilirubin (50 nM) significantly prevented ROS production in Aβ-treated neurons and favored both the slowdown of the growth rate of Aβ oligomers and the decrease in oligomer/fibril final size. In conclusion, these results suggest a novel mechanism through which CXB is neuroprotective in subjects with early AD or mild cognitive impairment.
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http://dx.doi.org/10.3389/fcell.2020.561179DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7550645PMC
September 2020

Searching for the Mechanical Fingerprint of Pre-diabetes in T1DM: A Case Report Study.

Front Bioeng Biotechnol 2020 29;8:569978. Epub 2020 Sep 29.

Dipartimento di Neuroscienze, Sezione di Fisica, Università Cattolica del Sacro Cuore, Rome, Italy.

We report the case of a 38 year-old Caucasian man enrolled in a study aimed at investigating the physical properties of red blood cells (RBCs) using advanced microscopy techniques, including Atomic Force Microscopy (AFM). At the time of his first enrolment in the study, he had normal Fasting Plasma Glucose (FPG) values, a BMI of 24.1, and no other symptoms of diabetes, including fatigue, high triglycerides, low HDL cholesterol, and altered inflammatory and corpuscular RBC indices. The subject reported no family history of diabetes, obesity, and cardiovascular diseases. Despite his apparently healthy conditions, the biomechanics of his RBCs was altered, showing increased values of stiffness and viscosity. More than 1 year after the mechanical measurements, the subject was admitted to the Operational Unit of Diabetology of the Policlinico Gemelli Hospital with high blood glucose and glycosylated hemoglobin (HbA1c) levels and diagnosed with type 1 diabetes (T1DM). Here, we show these data, and we discuss the hypothesis that RBC mechanical properties could be sensitive to changes occurring during the pre-diabetic phase of T1DM.
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http://dx.doi.org/10.3389/fbioe.2020.569978DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7552738PMC
September 2020

Antimicrobial and Antibiofilm Properties of Graphene Oxide on .

Antibiotics (Basel) 2020 Oct 13;9(10). Epub 2020 Oct 13.

Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, 20123 Rome, Italy.

The aim of this study was to evaluate the antibacterial properties of graphene oxide (GO) against in vitro conditions and when used to coat dentin surface to prevent adhesion. The ATCC strain of 29212 has been used to perform a viability test. The pellet was suspended in ultrapure water, NaCl, PBS buffer, CaCl and MgCl, Luria-Bertani broth solutions. The viability was evaluated by the colony forming unit counting method. Atomic force microscopy images and the measure of surface zeta potential variation were analyzed. Dentin discs were covered with a film of GO ( = 15) or were not treated ( = 15). Bacterial suspension was added to each sample of dentine discs and microbial counts were calculated. Statistically significant differences between two groups were assessed by a two-tailed unpaired -test. Bacteria cell morphology was investigated with scanning electron microscopy. The highest growth inhibition was obtained in ddHO and CaCl solution while, in PBS and NaCl, GO had poor antibacterial efficacy with a growth enhancing effect in the latter. GO on dentin discs demonstrated high antibacterial activity. GO film has demonstrated acceptable adhesion properties to root dentin and a role in the inhibition of bacterial film proliferation and biofilm formation.
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http://dx.doi.org/10.3390/antibiotics9100692DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602102PMC
October 2020

Enhanced Chemotherapy for Glioblastoma Multiforme Mediated by Functionalized Graphene Quantum Dots.

Materials (Basel) 2020 Sep 17;13(18). Epub 2020 Sep 17.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, 00185 Roma, Italy.

Glioblastoma is the most aggressive and lethal brain cancer. Current treatments involve surgical resection, radiotherapy and chemotherapy. However, the life expectancy of patients with this disease remains short and chemotherapy leads to severe adverse effects. Furthermore, the presence of the blood-brain barrier (BBB) makes it difficult for drugs to effectively reach the brain. A promising strategy lies in the use of graphene quantum dots (GQDs), which are light-responsive graphene nanoparticles that have shown the capability of crossing the BBB. Here we investigate the effect of GQDs on U87 human glioblastoma cells and primary cortical neurons. Non-functionalized GQDs (NF-GQDs) demonstrated high biocompatibility, while dimethylformamide-functionalized GQDs (DMF-GQDs) showed a toxic effect on both cell lines. The combination of GQDs and the chemotherapeutic agent doxorubicin (Dox) was tested. GQDs exerted a synergistic increase in the efficacy of chemotherapy treatment, specifically on U87 cells. The mechanism underlying this synergy was investigated, and it was found that GQDs can alter membrane permeability in a manner dependent on the surface chemistry, facilitating the uptake of Dox inside U87 cells, but not on cortical neurons. Therefore, experimental evidence indicates that GQDs could be used in a combined therapy against brain cancer, strongly increasing the efficacy of chemotherapy and, at the same time, reducing its dose requirement along with its side effects, thereby improving the life quality of patients.
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http://dx.doi.org/10.3390/ma13184139DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7560355PMC
September 2020

Graphene Quantum Dots' Surface Chemistry Modulates the Sensitivity of Glioblastoma Cells to Chemotherapeutics.

Int J Mol Sci 2020 Aug 31;21(17). Epub 2020 Aug 31.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, 00168 Rome, Italy.

Recent evidence has shown that graphene quantum dots (GQDs) are capable of crossing the blood-brain barrier, the barrier that reduces cancer therapy efficacy. Here, we tested three alternative GQDs' surface chemistries on two neural lineages (glioblastoma cells and mouse cortical neurons). We showed that surface chemistry modulates GQDs' biocompatibility. When used in combination with the chemotherapeutic drug doxorubicin, GDQs exerted a synergistic effect on tumor cells, but not on neurons. This appears to be mediated by the modification of membrane permeability induced by the surface of GQDs. Our findings highlight that GQDs can be adopted as a suitable delivery and therapeutic strategy for the treatment of glioblastoma, by both directly destabilizing the cell membrane and indirectly increasing the efficacy of chemotherapeutic drugs.
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http://dx.doi.org/10.3390/ijms21176301DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503375PMC
August 2020

Expression of Pinopodes in the Endometrium from Recurrent Pregnancy Loss Women. Role of Thrombomodulin and Ezrin.

J Clin Med 2020 Aug 13;9(8). Epub 2020 Aug 13.

Dipartimento di Scienze della Salute della Donna, del Bambino e di Sanità Pubblica, Fondazione Policlinico Universitario A. Gemelli, Istituto di Ricovero e Cura a Carattere Scientifico (I.R.C.C.S.), 00168 Roma, Italy.

Background: Pinopode expression has been suggested as a marker of endometrial receptivity.

Methods: We set up an experimental study comparing endometrial tissue from recurrent pregnancy loss (RPL, = 30) and fertile control (CTR, = 20) women in terms of pinopode expression/morphology; expression of thrombomodulin (TM) and ezrin; cytoskeletal organization. Endometrial samples were collected during implantation window and evaluated by scanning electron microscopy, western blot, and immunofluorescence.

Results: We found that RPL endometrial tissue showed: (i) increased pinopodes density (* < 0.05); (ii) a reduced diameter of pinopodes (* < 0.05); (iii) a decreased TM and ezrin expression ( < 0.05). Additionally, confocal images showed a significantly reduced expression of phosphorylated ()-ezrin, confirming the results obtained through immunoblot analysis. Immunofluorescence staining showed that in CTR samples, junctions between cells are intact and clearly visible, whereas actin filaments appear completely lost in RPL endometrial samples; this suggests that, due to the impaired expression and activity of TM and ezrin, actin does not bind to plasma membrane in order to orchestrate the cytoskeletal actin filaments.

Conclusions: Our findings suggest that an impaired expression of TM and expression/activation of ezrin may affect the connection between the TM and actin cytoskeleton, impairing the organization of cytoskeleton and, eventually, the adequate pinopode development.
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http://dx.doi.org/10.3390/jcm9082634DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7464296PMC
August 2020

Noncanonical type 2B von Willebrand disease associated with mutations in the VWF D'D3 and D4 domains.

Blood Adv 2020 07;4(14):3405-3415

Dipartimento di Medicina e Chirurgia Traslazionale, Facoltà di Medicina e Chirurgia Agostino Gemelli, Università Cattolica S. Cuore, Rome, Italy.

We observed a 55-year-old Italian man who presented with mucosal and cutaneous bleeding. Results of his blood analysis showed low levels of von Willebrand factor (VWF) antigen and VWF activity (both VWF ristocetin cofactor and VWF collagen binding), mild thrombocytopenia, increased ristocetin-induced platelet aggregation, and a deficiency of high-molecular-weight multimers, all typical phenotypic hallmarks of type 2B von Willebrand disease (VWD). The analysis of the VWF gene sequence revealed heterozygous in cis mutations: (1) c.2771G>A and (2) c.6532G>T substitutions in the exons 21 and 37, respectively. The first mutation causes the substitution of an Arg residue with a Gln at position 924, in the D'D3 domain. The second mutation causes an Ala to Ser substitution at position 2178 in the D4 domain. The patient's daughter did not present the same fatherly mutations but showed only the heterozygous polymorphic c.3379C>T mutation in exon 25 of the VWF gene causing the p.P1127S substitution, inherited from her mother. The in vitro expression of the heterozygous in cis VWF mutant rVWFWT/rVWF924Q-2178S confirmed and recapitulated the ex vivo VWF findings. Molecular modeling showed that these in cis mutations stabilize a partially stretched and open conformation of the VWF monomer. Transmission electron microscopy and atomic force microscopy showed in the heterozygous recombinant form rVWFWT/rVWF924Q-2178S a stretched conformation, forming strings even under static conditions. Thus, the heterozygous in cis mutations 924Q/2178S promote conformational transitions in the VWF molecule, causing a type 2B-like VWD phenotype, despite the absence of typical mutations in the A1 domain of VWF.
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http://dx.doi.org/10.1182/bloodadvances.2020002334DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7391138PMC
July 2020

Graphene Oxide-Linezolid Combination as Potential New Anti-Tuberculosis Treatment.

Nanomaterials (Basel) 2020 Jul 22;10(8). Epub 2020 Jul 22.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, 00168 Roma, Italy.

Global pandemic management represents a serious issue for health systems. In some cases, repurposing of existing medications might help find compounds that have the unexpected potential to combat microorganisms. In the same way, changing cell-drug interaction by nanotechnology could represent an innovative strategy to fight infectious diseases. Tuberculosis (TB) remains one of the most alarming worldwide infectious diseases and there is an urgent need for new drugs and treatments, particularly for the emergence and spread of drug-resistant () strains. New nanotechnologies based on carbon nanomaterials are now being considered to improve anti-TB treatments, and graphene oxide (GO) showed interesting properties as an anti-TB drug. GO, which preferentially accumulates in the lungs and is degraded by macrophagic peroxidases, can trap and in a dose-dependent manner, reducing the entry of bacilli into macrophages. In this paper, combinations of isoniazid (INH), amikacin (AMK) and linezolid (LZD) and GO anti-mycobacterial properties were evaluated against H37Rv by using a checkerboard assay or an in vitro infection model. Different GO effects have been observed when incubated with INH, AMK or LZD. Whereas the INH and AMK anti-mycobacterial activities were blocked by GO co-administration, the LZD bactericidal effect increased in combination with GO. GO-LZD significantly reduced extracellular mycobacteria during infection and was able to kill internalized bacilli. GO-LZD co-administration is potentially a new promising anti-TB treatment at the forefront in fighting emerging antibiotic-resistant strains where LZD administration is suggested. This innovative pharmacological approach may lead to reduced treatment periods and decreased adverse effects. More importantly, we demonstrate how nanomaterials-drugs combinations can represent a possible strategy to quickly design drugs for pandemics treatment.
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http://dx.doi.org/10.3390/nano10081431DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466666PMC
July 2020

Circulating miRNAs in Small Extracellular Vesicles Secreted by a Human Melanoma Xenograft in Mouse Brains.

Cancers (Basel) 2020 Jun 19;12(6). Epub 2020 Jun 19.

Institute for Biomedical Technologies (ITB), CNR, 20090 Segrate, Italy.

The identification of liquid biomarkers remains a major challenge to improve the diagnosis of melanoma patients with brain metastases. Circulating miRNAs packaged into tumor-secreted small extracellular vesicles (sEVs) contribute to tumor progression. To investigate the release of tumor-secreted miRNAs by brain metastasis, we developed a xenograft model where human metastatic melanoma cells were injected intracranially in nude mice. The comprehensive profiles of both free miRNAs and those packaged in sEVs secreted by the melanoma cells in the plasma demonstrated that most (80%) of the sEV-associated miRNAs were also present in serum EVs from a cohort of metastatic melanomas, included in a publicly available dataset. Remarkably, among them, we found three miRNAs (miR-224-5p, miR-130a-3p and miR-21-5p) in sEVs showing a trend of upregulation during melanoma progression. Our model is proven to be valuable for identifying miRNAs in EVs that are unequivocally secreted by melanoma cells in the brain and could be associated to disease progression.
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http://dx.doi.org/10.3390/cancers12061635DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352810PMC
June 2020

Graphene Oxide Nano-Concentrators Selectively Modulate RNA Trapping According to Metal Cations in Solution.

Front Bioeng Biotechnol 2020 25;8:421. Epub 2020 May 25.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Rome, Italy.

With recent advances in nanotechnology, graphene nanomaterials are being translated to applications in the fields of biosensing, medicine, and diagnostics, with unprecedented power. Graphene is a carbon allotrope derived from graphite exfoliation made of an extremely thin honeycomb of sp2 hybridized carbons. In comparison with the bulk materials, graphene and its water-soluble derivative graphene oxide have a smaller size suitable for diagnostic platform miniaturization as well as high surface area and consequently loading of a large number of biological probes. In this work, we propose a nanotechnological method for concentrating total RNA solution and/or enriching small RNA molecules. To this aim, we exploited the unique trapping effects of GO nanoflakes in the presence of divalent cations (i.e., calcium and magnesium) that make it flocculate and precipitate, forming complex meshes that are positively charged. Here, we demonstrated that GO traps can concentrate nucleic acids in the presence of divalent cations and that small RNAs can be selectively released from GO-magnesium traps. GO nano-concentrators will allow better analytical performance with samples available in small amounts and will increase the sensitivity of sequencing platforms by short RNA selection.
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http://dx.doi.org/10.3389/fbioe.2020.00421DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261913PMC
May 2020

Graphene-based scaffolds for tissue engineering and photothermal therapy.

Nanomedicine (Lond) 2020 06 8;15(14):1411-1417. Epub 2020 Jun 8.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Roma, Italy.

Researchers have been creating 3D replicas of damaged tissues for millions of patients by using innovative biomaterials. While these scaffolds have regenerative properties, it would be beneficial if they could be utilized for local therapies, such as for cancer treatment. This report discusses the main advances in graphene scaffold design for near-infrared (NIR) photothermal therapy (PTT). NIR-PTT is a promising alternative for cancer cell killing, mediated by an increase of temperature due to NIR light-absorbers delivered to the tumor proximity. Graphene is a bidimensional material largely exploited in nanomedicine for its unique properties, such as high growth factor loading, which induces cell differentiation and its capacity to absorb NIR light. Here we cover aspects of future research in multifunctional graphene implants for cancer therapy and tissue regeneration.
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http://dx.doi.org/10.2217/nnm-2020-0050DOI Listing
June 2020

A time-dependent study of nano-mechanical and ultrastructural properties of internal limiting membrane under ocriplasmin treatment.

J Mech Behav Biomed Mater 2020 10 18;110:103853. Epub 2020 May 18.

Istituto di Oftalmologia, Università Cattolica del Sacro Cuore, Roma, Italy.

Vitreomacular traction (VMT) syndrome has only been surgically treated for a long time. Recently, enzymatic vitreolysis with ocriplasmin has emerged as a possible option to release VMT and, in some cases, close full thickness macular holes (FTMHs). Despite its clinical relevance, gathering information about the ocriplasmin-induced alterations of the Inner Limiting Membrane (ILM) of the retina in a clinical study is a complex task, mainly because of the inter-individual variability among patients. To obtain more insights into the mechanism underlying the drug action, we studied in-vitro the mechanical and morphological changes of the ILM using Atomic Force Microscopy (AFM). To this aim, we measured the ILM average Young's modulus (YM), hysteresis (H) and adhesion work (A) over time under ocriplasmin treatment. Our data unveil a time-dependent increase in the membrane YM of 19% of its initial value, along with changes in its adhesive and dissipative behavior. Such modifications well correlate with the morphological alterations detected in the AFM imaging mode. Taken all together, the results here presented provide more insights into the mechanism underlying the ocriplasmin action in-vivo, suggesting that it is only able to alter the top-most layer of the vitreal side of the membrane, not compromising the inner ILM structure.
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http://dx.doi.org/10.1016/j.jmbbm.2020.103853DOI Listing
October 2020

Unravelling the Potential of Graphene Quantum Dots in Biomedicine and Neuroscience.

Int J Mol Sci 2020 May 25;21(10). Epub 2020 May 25.

Department of Neuroscience, Università Cattolica del Sacro Cuore, 00168 Roma, Italy.

Quantum dots (QDs) are semiconducting nanoparticles that have been gaining ground in various applications, including the biomedical field, thanks to their unique optical properties. Recently, graphene quantum dots (GQDs) have earned attention in biomedicine and nanomedicine, thanks to their higher biocompatibility and low cytotoxicity compared to other QDs. GQDs share the optical properties of QD and have proven ability to cross the blood-brain barrier (BBB). For this reason, GQDs are now being employed to deepen our knowledge in neuroscience diagnostics and therapeutics. Their size and surface chemistry that ease the loading of chemotherapeutic drugs, makes them ideal drug delivery systems through the bloodstream, across the BBB, up to the brain. GQDs-based neuroimaging techniques and theranostic applications, such as photothermal and photodynamic therapy alone or in combination with chemotherapy, have been designed. In this review, optical properties and biocompatibility of GQDs will be described. Then, the ability of GQDs to overtake the BBB and reach the brain will be discussed. At last, applications of GQDs in bioimaging, photophysical therapies and drug delivery to the central nervous system will be considered, unraveling their potential in the neuroscientific field.
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http://dx.doi.org/10.3390/ijms21103712DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7279214PMC
May 2020

3D Graphene Scaffolds for Skeletal Muscle Regeneration: Future Perspectives.

Front Bioeng Biotechnol 2020 5;8:383. Epub 2020 May 5.

Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Rome, Italy.

Although skeletal muscle can regenerate after injury, in chronic damages or in traumatic injuries its endogenous self-regeneration is impaired. Consequently, tissue engineering approaches are promising tools for improving skeletal muscle cells proliferation and engraftment. In the last decade, graphene and its derivates are being explored as novel biomaterials for scaffolds production for skeletal muscle repair. This review describes 3D graphene-based materials that are currently used to generate complex structures able not only to guide cell alignment and fusion but also to stimulate muscle contraction thanks to their electrical conductivity. Graphene is an allotrope of carbon that has indeed unique mechanical, electrical and surface properties and has been functionalized to interact with a wide range of synthetic and natural polymers resembling native musculoskeletal tissue. More importantly, graphene can stimulate stem cell differentiation and has been studied for cardiac, neuronal, bone, skin, adipose, and cartilage tissue regeneration. Here we recapitulate recent findings on 3D scaffolds for skeletal muscle repairing and give some hints for future research in multifunctional graphene implants.
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http://dx.doi.org/10.3389/fbioe.2020.00383DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7214535PMC
May 2020

Biocompatibility assessment of sub-5 nm silica-coated superparamagnetic iron oxide nanoparticles in human stem cells and in mice for potential application in nanomedicine.

Nanoscale 2020 Jan;12(3):1759-1778

Institute of Translational Pharmacology (IFT), Department of Biomedical Sciences, National Research Council (CNR), via del Fosso del Cavaliere 100, 00133 Rome, Italy.

Ultrasmall superparamagnetic iron oxide nanoparticles with a size <5 nm are emerging nanomaterials for their excellent biocompatibility, chemical stability, and tunable surface modifications. The applications explored include dual-modal or multi-modal imaging, drug delivery, theranostics and, more recently, magnetic resonance angiography. Good biocompatibility and biosafety are regarded as the preliminary requirements for their biomedical applications and further exploration in this field is still required. We previously synthesized and characterized ultrafine (average core size of 3 nm) silica-coated superparamagnetic iron oxide fluorescent nanoparticles, named sub-5 SIO-Fl, uniform in size, shape, chemical properties and composition. The cellular uptake and in vitro biocompatibility of the as-synthesized nanoparticles were demonstrated in a human colon cancer cellular model. Here, we investigated the biocompatibility of sub-5 SIO-Fl nanoparticles in human Amniotic Mesenchymal Stromal/Stem Cells (hAMSCs). Kinetic analysis of cellular uptake showed a quick nanoparticle internalization in the first hour, increasing over time and after long exposure (48 h), the uptake rate gradually slowed down. We demonstrated that after internalization, sub-5 SIO-Fl nanoparticles neither affect hAMSC growth, viability, morphology, cytoskeletal organization, cell cycle progression, immunophenotype, and the expression of pro-angiogenic and immunoregulatory paracrine factors nor the osteogenic and myogenic differentiation markers. Furthermore, sub-5 SIO-Fl nanoparticles were intravenously injected into mice to investigate the in vivo biodistribution and toxicity profile for a time period of 7 weeks. Our findings showed an immediate transient accumulation of nanoparticles in the kidney, followed by the liver and lungs, where iron contents increased over a 7-week period. Histopathology, hematology, serum pro-inflammatory response, body weight and mortality studies demonstrated a short- and long-term biocompatibility and biosafety profile with no apparent acute and chronic toxicity caused by these nanoparticles in mice. Overall, these results suggest the feasibility of using sub-5 SIO-Fl nanoparticles as a promising agent for stem cell magnetic targeting as well as for diagnostic and therapeutic applications in oncology.
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http://dx.doi.org/10.1039/c9nr09683cDOI Listing
January 2020

Carbon nanomaterials: a new way against tuberculosis.

Expert Rev Med Devices 2019 Oct 30;16(10):863-875. Epub 2019 Sep 30.

Fondazione Policlinico Universitario A. Gemelli, IRCCS , Roma , Italy.

: Tuberculosis (TB) remains one of the most alarming worldwide infectious diseases primarily in low-income countries, where the infection shows a higher and unvaried prevalence. In the last years, the emergence and spread of () strains resistant to first-line anti-TB drugs are the cause of major concern and prompted the implementation of new treatments, including the development of new drugs and the repurposing of old ones. : In this review, we discuss solutions against TB based on nanomaterials (NMTs), alone or combined with current anti-TB drugs. We will summarize drug delivery platforms tested in or models and their activity against mycobacteria. We will describe how the new nanotechnologies based on carbon nanomaterials, like carbon nanotubes and graphene oxide are now facing the panorama of the medical fight against TB. : We foresee that in the next decade carbon nanomaterials will be at the forefront in fighting emerging antibiotic-resistant strains by shortening treatment periods, reducing adverse effects and mitigating antibiotic use. However, toxicity and biodegradation studies should be done prior to the clinical translation of carbon nanomaterials.
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http://dx.doi.org/10.1080/17434440.2019.1671820DOI Listing
October 2019

Dynamic structural determinants underlie the neurotoxicity of the N-terminal tau 26-44 peptide in Alzheimer's disease and other human tauopathies.

Int J Biol Macromol 2019 Dec 27;141:278-289. Epub 2019 Aug 27.

European Brain Research Institute (EBRI), Viale Regina Elena 295, 00161 Rome, Italy; Institute of Translational Pharmacology (IFT)-CNR, Via Fosso del Cavaliere 100, 00133 Rome, Italy. Electronic address:

The intrinsically disordered tau protein plays a pivotal role in the pathogenesis of Alzheimer's disease (AD) and other human tauopathies. Abnormal post-translational modifications of tau, such as truncation, are causally involved in the onset/development of these neurodegenerative diseases. In this context, the AD-relevant N-terminal fragment mapping between 26 and 44 amino acids of protein (tau26-44) is interesting, being endowed with potent neurotoxic effects in vitro and in vivo. However, the understanding of the mechanism(s) of tau26-44 toxicity is a challenging task because, similarly to the full-length tau, it does not have a unique 3D structure but exists as dynamic ensemble of conformations. Here we use Atomic Force Spectroscopy, Small Angle X-ray Scattering and Molecular Dynamics simulation to gather structural and functional information on the tau26-44. We highlight the presence, the type and the location of its temporary secondary structures and we unveil the occurrence of relevant transient tertiary conformations that could contribute to tau26-44 toxicity. Data are compared with those obtained on the biologically-inactive, reverse-sequence (tau44-26 peptide) which has the same mass, charge, aminoacidic composition as well as the same overall unfolded character of tau26-44.
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http://dx.doi.org/10.1016/j.ijbiomac.2019.08.220DOI Listing
December 2019

Nanomechanical mapping helps explain differences in outcomes of eye microsurgery: A comparative study of macular pathologies.

PLoS One 2019 7;14(8):e0220571. Epub 2019 Aug 7.

Fondazione Policlinico A. Gemelli IRCCS, Roma, Italia.

Many ocular diseases are associated with an alteration of the mechanical and the material properties of the eye. These mechanically-related diseases include macular hole and pucker, two ocular conditions due to the presence of abnormal physical tractions acting on the retina. A complete relief of these tractions can be obtained through a challenging microsurgical procedure, which requires the mechanical peeling of the internal limiting membrane of the retina (ILM). In this paper, we provide the first comparative study of the nanoscale morphological and mechanical properties of the ILM in macular hole and macular pucker. Our nanoscale elastic measurements unveil a different bio-mechanical response of the ILM in the two pathologies, which correlates well to significant differences occurring during microsurgery. The results here presented pave the way to the development of novel dedicated microsurgical protocols based on the material ILM properties in macular hole or pucker. Moreover, they contribute to clarify why, despite a common aetiology, a patient might develop one disease or the other, an issue which is still debated in literature.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0220571PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6685617PMC
March 2020

Converting the personalized biomolecular corona of graphene oxide nanoflakes into a high-throughput diagnostic test for early cancer detection.

Nanoscale 2019 Aug;11(32):15339-15346

Istituto di Fisica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, Rome, Italy.

Advances in nanotechnology are introducing the exciting possibility of cancer identification at early stages via analysis of the personalized biomolecular corona (BC), i.e. the dynamic "halo" of proteins that adsorbs onto NPs following exposure to patients' plasma. In this study, we develop a blood test for early cancer detection based on the characterization of the BC that forms around Graphene Oxide (GO) nanoflakes. Among its elective properties, GO binds low amounts of albumin, the most abundant protein in the blood and one of the most enriched proteins in the BC of many nanomaterials. This unique property of GO allows strong adsorption of poorly concentrated plasma proteins without abundant protein depletion. In our study, GO nanometric flakes have been used to analyze BCs from 50 subjects, half of them diagnosed with pancreatic cancer and half of them being healthy volunteers. Pancreatic cancer was chosen as the model of a high mortality disease with poor survival rates due to its delayed diagnosis. The receiver operating characteristic (ROC) curve analysis was applied to measure the diagnostic accuracy of the BC-based test. We obtained an area under the curve (AUC) of 0.96 and the test discriminated cancer patients from healthy subjects with a sensitivity of 92%. Finally, a double-blind validation was made using a second test dataset (10 healthy subjects + 10 pancreatic cancer patients) and it confirmed the results obtained on the first training dataset. Being highly accurate, fast, inexpensive and easy to perform, we believe that the BC-enabled blood test has the potential to become a turning point in early detection of cancer and other diseases.
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http://dx.doi.org/10.1039/c9nr01413fDOI Listing
August 2019

A novel method for post-mortem interval estimation based on tissue nano-mechanics.

Int J Legal Med 2019 Jul 27;133(4):1133-1139. Epub 2019 Mar 27.

Institute of Physics, Fondazione Policlinico Universitario A. Gemelli IRCCS, Università Cattolica del Sacro Cuore, Rome, Italy.

Forensic estimation of post-mortem interval relies on different methods, most of which, however, have practical limitations or provide insufficient results, still lacking a gold standard method. In order to better understand the phenomenon of rigor mortis and its applicability to the post-mortem interval estimation, we decided to use atomic force microscopy, a tool often employed to measure mechanical properties of adherent cells. Thus, we surgically removed skeletal muscle samples of three forensic cases from 0 to 120 h post-mortem and quantitatively evaluate two parameters: the Young's modulus (E), which gives information about the sample stiffness, and the hysteresis (H), which estimates the contribution of viscous forces. Despite being a preliminary study, the obtained results show that the temporal behavior of E well correlates with the expected evolution of rigor mortis between 0 and 48 h post-mortem, and then monotonically decreases over time. Unfortunately, it is strongly affected by inter-individual variability. However, we found that H provides measurable data along a time-dependent curve back to the starting point, and these data measured on different subjects collapse onto a single master curve, getting rid of the inter-individual variability. Although a larger sampling should be performed to improve the result reliability, this finding is strongly suggestive that the evaluation of rigor mortis should involve the measure of the nanoscale dissipative behavior of muscular tissues.
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http://dx.doi.org/10.1007/s00414-019-02034-zDOI Listing
July 2019

Biocompatible N-acetyl cysteine reduces graphene oxide and persists at the surface as a green radical scavenger.

Chem Commun (Camb) 2019 Apr;55(29):4186-4189

Istituto di Fisica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168, Rome, Italy.

We demonstrate that N-acetyl cysteine (NAC) reduces graphene oxide (GO) at room temperature. This represents a new green method to produce reduced GO (rGO). NAC adheres to the rGO surface as demonstrated by several spectroscopy techniques and avoids GO-mediated oxidation of glutathione. This method offers new opportunities for the production of green biocompatible rGO and NAC-based therapies.
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http://dx.doi.org/10.1039/c9cc00429gDOI Listing
April 2019

Microfluidic manufacturing of surface-functionalized graphene oxide nanoflakes for gene delivery.

Nanoscale 2019 Feb;11(6):2733-2741

Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 291, 00161, Rome, Italy.

Graphene oxide (GO) is a single-atomic-layered material made of a sheet of oxidized carbon atoms arranged in a honeycomb structure. Thanks to the notable physical and chemical properties of GO, GO-based nanomaterials have applications in many fields of research, including gene delivery. It has been reported that pristine GO can absorb single-stranded DNA and RNA through π-π stacking, which cannot be used as a gene carrier because it is hard to load double-stranded DNA (dsDNA). To tackle this issue, this work was aimed at developing a hybrid nanoparticle (NP) system made of GO coated with cationic lipids (hereafter referred to as GOCL) with suitable physical-chemical properties for gene delivery applications. To this end, nanosized GO flakes (nGO) were coated with the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) by microfluidic mixing. Comprehensive characterization of GOCL NPs was performed by a combination of dynamic light scattering (DLS), micro-electrophoresis and atom force microscopy (AFM). Our results show that GOCL NPs exhibit adequate size (<150 nm) and surface charge (ξ = +15 mV) for gene delivery purposes. Complexes made of GOCL NPs and plasmid DNA (pDNA) were used to transfect human cervical cancer cells (HeLa) and human embryonic kidney (HEK-293) cells. Pristine nGO and DOTAP cationic liposomes were used as a reference. GOCL NPs exhibited a similar TE but a much higher cell viability compared with DOTAP cationic liposomes. Confocal fluorescence microscopy provided a reasonable explanation for the superior performance of GOCL/DNA complexes showing that they are much more numerous, regular in size and homogeneously distributed than DOTAP/DNA complexes, thus splitting their gene payload over the entire cell population. Because of the imperative demand for efficient and safe nanocarriers, this study will contribute to the development of novel surface-functionalized GO-based hybrid gene vectors.
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http://dx.doi.org/10.1039/c8nr09245aDOI Listing
February 2019

A protein chimera self-assembling unit for drug delivery.

Biotechnol Prog 2019 03 27;35(2):e2769. Epub 2018 Dec 27.

Istituto di Biochimica e Biochimica Clinica, Università Cattolica del Sacro Cuore, Rome, Italy.

In the modern view of selective drug delivery of bioactive molecules, the attention is moving onto the setup of the perfect carrier more than in the optimization of the active compound. In this respect, virus-like particles constitute bioinspired nanodevices with the intrinsic ability to transport a large class of molecules, ranging from smart drugs to small interfering RNAs. In this work, we demonstrate the efficacy of a novel construct obtained by fusing a self-assembling protein from the human Rotavirus A, VP6, with the Small Ubiquitin Modifier domain, which maintains the ability to form nanoparticles and nanotubes and is able to be used as a drug carrier, even without specific targeting epitopes. The high expression and purification yield, combined with low toxicity of the empty particles, clearly indicate a good candidate for future studies of selective drug delivery. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2769, 2019.
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http://dx.doi.org/10.1002/btpr.2769DOI Listing
March 2019

Graphene oxide coatings prevent Candida albicans biofilm formation with a controlled release of curcumin-loaded nanocomposites.

Nanomedicine (Lond) 2018 11 15;13(22):2867-2879. Epub 2018 Nov 15.

Fondazione Policlinico A. Gemelli IRCCS - Università Cattolica Sacro Cuore. Istituto di Fisica, Largo Francesco Vito 1, 00168, Rome, Italy.

Aim: Fabrication of graphene oxide (GO)-based medical devices coatings that limit adhesion of Candida albicans, a main issue of healthcare-associated infections.

Methods: The GO composites noncovalently functionalized with curcumin (CU), a hydrophobic molecule with active antimicrobial action, polyethylene glycol (PEG) that hinders the absorption of biomolecules or a combination of CU and PEG (GO-CU-PEG) were drop-casted on surfaces and antifungal efficacy was assessed.

Results: We demonstrate that GO-CU-PEG coatings can reduce fungal adhesion, proliferation and biofilm formation. Furthermore, in an aqueous environment, surfaces release curcumin-PEG nanocomposites that have a minimum inhibitory concentration of 9.25 μg/ml against C. albicans.

Conclusion: Prevention of early cell adhesion and creation of a proximal environment unfavorable for growth make these GO-supported biomaterials attractive for innovative medical device manufacturing.
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http://dx.doi.org/10.2217/nnm-2018-0183DOI Listing
November 2018