A successful application of the 3D printed materials in the biomedical field requires extensive s... more A successful application of the 3D printed materials in the biomedical field requires extensive studies to ensure their biocompatibility at every step of the process. Here, different components suitable for cell applications, including a microfluidic device, were 3D printed using common resins and a deep analysis of their biocompatibility and post printed protocols was conducted.
A 3D culture system based on a photocurable matrix has been developed. The aim is to create a 3D ... more A 3D culture system based on a photocurable matrix has been developed. The aim is to create a 3D printable platform mimicking lung cancer tissue, to study tumor microenvironment evolution, in terms of structural (architecture) and molecular (signalling) components.
3D printed Acrilate PDMS-based devices to design stretchable platforms for cell culture and drug ... more 3D printed Acrilate PDMS-based devices to design stretchable platforms for cell culture and drug testing.
Overview of gene expression analysis. GSEA, gene signature enrichment analysis. Figure S2. DNA se... more Overview of gene expression analysis. GSEA, gene signature enrichment analysis. Figure S2. DNA sequences of single nucleotide polymorphism at codon 762 of PARP1 gene in HT1080, SJSA-1, and SW684 cells. Figure S3. Distribution of trabectedin IC50 as single agent (TR alone) and in combination with veliparib (TR + VEL) or olaparib (TR + OL) among high-PARP1-expressing cells (red triangle) and low-PARP1-expressing cells (blue triangle). Figure S4. Dose- response curve obtained after 72-h treatment with trabectedin (2–0.125nM), olaparib (20–1.25 μM) as single agents and in constant combination. Figure S5. A, western blot analysis of PARylation and PARP1 expression in MES-SA and MES-SA-DX5 leiomyosarcoma cells; B, FISH analysis of PARP1 gene (red) and centromere of chromosome 1 (green) in MESSA and MESSA-DX5. Figure S6. Genomic status as obtained by aCGH analysis of TC-106, 402.91, DMR, SJSA-1, HT1080, SW684: gain (red) and loss (green) of chromosome regions. Figure S7. A, Western blot an...
Concentrations inhibiting 50% of cell viability (IC50) after 72- h treatment with serial dilution... more Concentrations inhibiting 50% of cell viability (IC50) after 72- h treatment with serial dilutions of trabectedin (2–0.125 nM), olaparib (20–1.25 μM), and veliparib (80–5 μM) as single agents (a) or in constant combination (b, c), and their 95% confidence intervals (95% CI). Population doubling time. Combination index ± estimated standard deviation (est st dev) calculated at IC50 by the Chou-Talalay method for trabectedin and olaparib in combination and trabectedin and veliparib in combination in cell lines of different histotypes: leiomyosarcoma (LMS), undifferentiated pleomorphic sarcoma (UPS), myxoid liposarcoma (LMS), dedifferentiated liposarcoma (DLS), fibrosarcoma (FSA), synovial sarcoma (SS), Ewing's sarcoma (ES), and osteosarcoma (OS). Supplier and culture conditions are reported for each cell line. Table S2. Direct correlation between combination index (calculated at IC50) and PARP1 protein intensity expressed as Pearson score (r Pearson). IC50 and 95% confidence interv...
Extracellular vesicles (EVs) are natural carriers produced by many different cell types that have... more Extracellular vesicles (EVs) are natural carriers produced by many different cell types that have a plethora of functions and roles that are still under discovery. This review aims to be a compendium on the current advancement in terms of EV modifications and re-engineering, as well as their potential use in nanomedicine. In particular, the latest advancements on artificial EVs are discussed, with these being the frontier of nanomedicine-based therapeutics. The first part of this review gives an overview of the EVs naturally produced by cells and their extraction methods, focusing on the possibility to use them to carry desired cargo. The main issues for the production of the EV-based carriers are addressed, and several examples of the techniques used to upload the cargo are provided. The second part focuses on the engineered EVs, obtained through surface modification, both using direct and indirect methods, i.e., engineering of the parental cells. Several examples of the current li...
The surface chemistry and charge of zinc oxide nanocrystals influence their behaviour in biologic... more The surface chemistry and charge of zinc oxide nanocrystals influence their behaviour in biological fluids. A novel lipid bilayer assembly is developed to shield ZnO nanocrystals improving their stability and cell internalization.
Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells / Racca... more Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells / Racca, Luisa; Dumontel, Bianca; Miccoli, Beatrice; Canta, Marta; Serpe, Loredana; Canaparo, Roberto; Di Benedetto, Cristiano; Falqui, Andrea; Cauda, Valentina Alice. STAMPA. (2016), pp. 158-158. ((Intervento presentato al convegno Merck Young Chemists Symposium (MYCS 2016) tenutosi a Rimini nel 25-27 novembre 2016. Original Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells
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...
Aim: The effective application of nanoparticles in cancer theranostics is jeopardized by their ag... more Aim: The effective application of nanoparticles in cancer theranostics is jeopardized by their aggregation in biological media, rapid degradation and clearance. The design of biomimetic nanoconstructs with enhanced colloidal stability and non-immunogenicity is therefore essential. We propose naturally stable cell-derived extracellular vesicles to encapsulate zinc oxide (ZnO) nanocrystals as efficacious nanodrugs, to obtain highly biomimetic and stable Trojan nano-horses (TNHs). Materials & methods: Coupling efficiency, biostability, cellular cytotoxicity and internalization were tested. Results: In vitro studies showed a high internalization of TNHs into cancer cells and efficient cytotoxic activity thanks to ZnO intracellular release. Conclusion: TNHs represent an efficient biomimetic platform for future nanotheranostic applications, with biomimetic extracellular vesicle-lipid envelope, facilitated ZnO cellular uptake and potential therapeutic implications.
Enhancing the antitumor activity of the DNA-damaging drugs is an attractive strategy to improve c... more Enhancing the antitumor activity of the DNA-damaging drugs is an attractive strategy to improve current treatment options. Trabectedin is an isoquinoline alkylating agent with a peculiar mechanism of action. It binds to minor groove of DNA inducing single- and double-strand-breaks. These kinds of damage lead to the activation of PARP1, a first-line enzyme in DNA-damage response pathways. We hypothesized that PARP1 targeting could perpetuate trabectedin-induced DNA damage in tumor cells leading finally to cell death. We investigated trabectedin and PARP1 inhibitor synergism in several tumor histotypes both in vitro and in vivo (subcutaneous and orthotopic tumor xenografts in mice). We searched for key determinants of drug synergism by comparative genomic hybridization (aCGH) and gene expression profiling (GEP) and validated their functional role. Trabectedin activated PARP1 enzyme and the combination with PARP1 inhibitors potentiated DNA damage, cell cycle arrest at G2/M checkpoint a...
Malignant Pleural Mesothelioma (MPM) is an aggressive tumor arising from mesothelial cells lining... more Malignant Pleural Mesothelioma (MPM) is an aggressive tumor arising from mesothelial cells lining the pleural cavities characterized by resistance to standard therapies. Most of the molecular steps responsible for pleural transformation remain unclear; however, several growth factor signaling cascades are known to be altered during MPM onset and progression. Transducers of these pathways, such as PIK3CA-mTOR-AKT, MAPK, and ezrin/radixin/moesin (ERM) could therefore be exploited as possible targets for pharmacological intervention. This study aimed to identify ‘druggable’ pathways in MPM and to formulate a targeted approach based on the use of commercially available molecules, such as the multikinase inhibitor sorafenib and the mTOR inhibitor everolimus. We planned a triple approach based on: i) analysis of immunophenotypes and mutational profiles in a cohort of thoracoscopic MPM samples, ii) in vitro pharmacological assays, ii) in vivo therapeutic approaches on MPM xenografts. No mu...
A successful application of the 3D printed materials in the biomedical field requires extensive s... more A successful application of the 3D printed materials in the biomedical field requires extensive studies to ensure their biocompatibility at every step of the process. Here, different components suitable for cell applications, including a microfluidic device, were 3D printed using common resins and a deep analysis of their biocompatibility and post printed protocols was conducted.
A 3D culture system based on a photocurable matrix has been developed. The aim is to create a 3D ... more A 3D culture system based on a photocurable matrix has been developed. The aim is to create a 3D printable platform mimicking lung cancer tissue, to study tumor microenvironment evolution, in terms of structural (architecture) and molecular (signalling) components.
3D printed Acrilate PDMS-based devices to design stretchable platforms for cell culture and drug ... more 3D printed Acrilate PDMS-based devices to design stretchable platforms for cell culture and drug testing.
Overview of gene expression analysis. GSEA, gene signature enrichment analysis. Figure S2. DNA se... more Overview of gene expression analysis. GSEA, gene signature enrichment analysis. Figure S2. DNA sequences of single nucleotide polymorphism at codon 762 of PARP1 gene in HT1080, SJSA-1, and SW684 cells. Figure S3. Distribution of trabectedin IC50 as single agent (TR alone) and in combination with veliparib (TR + VEL) or olaparib (TR + OL) among high-PARP1-expressing cells (red triangle) and low-PARP1-expressing cells (blue triangle). Figure S4. Dose- response curve obtained after 72-h treatment with trabectedin (2–0.125nM), olaparib (20–1.25 μM) as single agents and in constant combination. Figure S5. A, western blot analysis of PARylation and PARP1 expression in MES-SA and MES-SA-DX5 leiomyosarcoma cells; B, FISH analysis of PARP1 gene (red) and centromere of chromosome 1 (green) in MESSA and MESSA-DX5. Figure S6. Genomic status as obtained by aCGH analysis of TC-106, 402.91, DMR, SJSA-1, HT1080, SW684: gain (red) and loss (green) of chromosome regions. Figure S7. A, Western blot an...
Concentrations inhibiting 50% of cell viability (IC50) after 72- h treatment with serial dilution... more Concentrations inhibiting 50% of cell viability (IC50) after 72- h treatment with serial dilutions of trabectedin (2–0.125 nM), olaparib (20–1.25 μM), and veliparib (80–5 μM) as single agents (a) or in constant combination (b, c), and their 95% confidence intervals (95% CI). Population doubling time. Combination index ± estimated standard deviation (est st dev) calculated at IC50 by the Chou-Talalay method for trabectedin and olaparib in combination and trabectedin and veliparib in combination in cell lines of different histotypes: leiomyosarcoma (LMS), undifferentiated pleomorphic sarcoma (UPS), myxoid liposarcoma (LMS), dedifferentiated liposarcoma (DLS), fibrosarcoma (FSA), synovial sarcoma (SS), Ewing's sarcoma (ES), and osteosarcoma (OS). Supplier and culture conditions are reported for each cell line. Table S2. Direct correlation between combination index (calculated at IC50) and PARP1 protein intensity expressed as Pearson score (r Pearson). IC50 and 95% confidence interv...
Extracellular vesicles (EVs) are natural carriers produced by many different cell types that have... more Extracellular vesicles (EVs) are natural carriers produced by many different cell types that have a plethora of functions and roles that are still under discovery. This review aims to be a compendium on the current advancement in terms of EV modifications and re-engineering, as well as their potential use in nanomedicine. In particular, the latest advancements on artificial EVs are discussed, with these being the frontier of nanomedicine-based therapeutics. The first part of this review gives an overview of the EVs naturally produced by cells and their extraction methods, focusing on the possibility to use them to carry desired cargo. The main issues for the production of the EV-based carriers are addressed, and several examples of the techniques used to upload the cargo are provided. The second part focuses on the engineered EVs, obtained through surface modification, both using direct and indirect methods, i.e., engineering of the parental cells. Several examples of the current li...
The surface chemistry and charge of zinc oxide nanocrystals influence their behaviour in biologic... more The surface chemistry and charge of zinc oxide nanocrystals influence their behaviour in biological fluids. A novel lipid bilayer assembly is developed to shield ZnO nanocrystals improving their stability and cell internalization.
Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells / Racca... more Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells / Racca, Luisa; Dumontel, Bianca; Miccoli, Beatrice; Canta, Marta; Serpe, Loredana; Canaparo, Roberto; Di Benedetto, Cristiano; Falqui, Andrea; Cauda, Valentina Alice. STAMPA. (2016), pp. 158-158. ((Intervento presentato al convegno Merck Young Chemists Symposium (MYCS 2016) tenutosi a Rimini nel 25-27 novembre 2016. Original Investigation of cytotoxic effects of different ZnO nanostructures on living cancer cells
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...
Aim: The effective application of nanoparticles in cancer theranostics is jeopardized by their ag... more Aim: The effective application of nanoparticles in cancer theranostics is jeopardized by their aggregation in biological media, rapid degradation and clearance. The design of biomimetic nanoconstructs with enhanced colloidal stability and non-immunogenicity is therefore essential. We propose naturally stable cell-derived extracellular vesicles to encapsulate zinc oxide (ZnO) nanocrystals as efficacious nanodrugs, to obtain highly biomimetic and stable Trojan nano-horses (TNHs). Materials & methods: Coupling efficiency, biostability, cellular cytotoxicity and internalization were tested. Results: In vitro studies showed a high internalization of TNHs into cancer cells and efficient cytotoxic activity thanks to ZnO intracellular release. Conclusion: TNHs represent an efficient biomimetic platform for future nanotheranostic applications, with biomimetic extracellular vesicle-lipid envelope, facilitated ZnO cellular uptake and potential therapeutic implications.
Enhancing the antitumor activity of the DNA-damaging drugs is an attractive strategy to improve c... more Enhancing the antitumor activity of the DNA-damaging drugs is an attractive strategy to improve current treatment options. Trabectedin is an isoquinoline alkylating agent with a peculiar mechanism of action. It binds to minor groove of DNA inducing single- and double-strand-breaks. These kinds of damage lead to the activation of PARP1, a first-line enzyme in DNA-damage response pathways. We hypothesized that PARP1 targeting could perpetuate trabectedin-induced DNA damage in tumor cells leading finally to cell death. We investigated trabectedin and PARP1 inhibitor synergism in several tumor histotypes both in vitro and in vivo (subcutaneous and orthotopic tumor xenografts in mice). We searched for key determinants of drug synergism by comparative genomic hybridization (aCGH) and gene expression profiling (GEP) and validated their functional role. Trabectedin activated PARP1 enzyme and the combination with PARP1 inhibitors potentiated DNA damage, cell cycle arrest at G2/M checkpoint a...
Malignant Pleural Mesothelioma (MPM) is an aggressive tumor arising from mesothelial cells lining... more Malignant Pleural Mesothelioma (MPM) is an aggressive tumor arising from mesothelial cells lining the pleural cavities characterized by resistance to standard therapies. Most of the molecular steps responsible for pleural transformation remain unclear; however, several growth factor signaling cascades are known to be altered during MPM onset and progression. Transducers of these pathways, such as PIK3CA-mTOR-AKT, MAPK, and ezrin/radixin/moesin (ERM) could therefore be exploited as possible targets for pharmacological intervention. This study aimed to identify ‘druggable’ pathways in MPM and to formulate a targeted approach based on the use of commercially available molecules, such as the multikinase inhibitor sorafenib and the mTOR inhibitor everolimus. We planned a triple approach based on: i) analysis of immunophenotypes and mutational profiles in a cohort of thoracoscopic MPM samples, ii) in vitro pharmacological assays, ii) in vivo therapeutic approaches on MPM xenografts. No mu...
Uploads
Papers by Marta Canta