Ilaria Bononi | Università degli Studi di Ferrara (original) (raw)
Papers by Ilaria Bononi
Antioxidants, 2022
Resveratrol is a powerful antioxidant molecule. In the human diet, its most important source is i... more Resveratrol is a powerful antioxidant molecule. In the human diet, its most important source is in Vitis vinifera grape peel and leaves. Resveratrol exists in two isoforms, cis- and trans. The diastereomeric forms of many drugs have been reported as affecting their activity. The aim of this study was to set up a cellular model to investigate how far resveratrol could counteract cytotoxicity in an oxidant agent. For this purpose, a keratinocyte cell line, which was genetically engineered with jelly fish green fluorescent protein, was treated with the free radical promoter Cumene hydroperoxide. The antioxidant activity of the trans-resveratrol and its diastereomeric mixture was evaluated indirectly in these treated fluorescent-engineered keratinocytes by analyzing the cell number and cell proliferation index. Our results demonstrate that cells, which were pre-incubated with resveratrol, reverted the oxidative damage progression induced by this free radical agent. In conclusion, fluore...
Simian Virus 40, experimentally assayed in vitro in different animal and human cells and in vivo ... more Simian Virus 40, experimentally assayed in vitro in different animal and human cells and in vivo in rodents, was classified as a small DNA tumor virus. In previous studies, many groups identified Simian Virus 40 sequences in healthy individuals and cancer patients using PCR techniques, whereas others failed to detect the viral sequences in human specimens. These conflicting results prompted us to develop a novel indirect ELISA with synthetic pep-tides, mimicking Simian Virus 40 capsid viral protein antigens, named mimotopes. This immunologic assay allowed us to investigate the presence of serum antibodies against Sim-ian Virus 40 and to verify whether Simian Virus 40 is circulating in humans. In this investiga-tion two mimotopes from Simian Virus 40 large T antigen, the viral replication protein and oncoprotein, were employed to analyze for specific reactions to human sera antibodies. This indirect ELISA with synthetic peptides from Simian Virus 40 large T antigen was used to assay ...
Cancers, 2021
Despite improved prognosis for many HPV-positive head and neck squamous cell carcinomas (HNSCCs),... more Despite improved prognosis for many HPV-positive head and neck squamous cell carcinomas (HNSCCs), some cases are still marked by recurrence and metastasis. Our study aimed to identify novel biomarkers for patient stratification. Classical HPV markers: HPV-DNA, p16 and HPV mRNA expression were studied in HNSCC (n = 67) and controls (n = 58) by qPCR. Subsequently, ELISA tests were used for HPV16 L1 antibody and HPV16 E7 oncoprotein detection in serum at diagnosis and follow-up. All markers were correlated to relapse-free survival (RFS) and overall survival (OS). HPV-DNA was found in HNSCCs (29.85%), HPV16-DNA in 95% of cases, HPV16 E7 mRNA was revealed in 93.75%. p16 was overexpressed in 75% of HPV-positive HNSCC compared to negative samples and controls (p < 0.001). Classical markers correlated with improved OS (p < 0.05). Serological studies showed similar proportions of HPV16 L1 antibodies in all HNSCCs (p > 0.05). Serum E7 oncoprotein was present in 30% HPV-positive patie...
La presente invenzione si riferisce all’identificazione di nuovi peptidi sintetici, corrispondent... more La presente invenzione si riferisce all’identificazione di nuovi peptidi sintetici, corrispondenti a specifici epitopi/antigeni della proteina virale capsidica 1, VP1, del virus Polioma BK (BKPyV), per rilevare nel siero e in altri fluidi umani anticorpi contro questo virus, potenzialmente patogeno inclusa l’attività oncogena, impiegando la tecnica immunologica ELISA indiretta
Introduction: in the orthopedic perspective, tissue engineering is focused on the development of ... more Introduction: in the orthopedic perspective, tissue engineering is focused on the development of innovative materials, whose action consists in recruiting bone progenitor cells and in stimulating their proliferation. In this study, we investigated the interaction between human mesenchymal stem cells from adults and two types of biomaterials, which differ for chemical composition and format. To evaluate the effects of porosity on cell adhesion and proliferation comparative analyses were carried out on different forms of two bioceramic materials. Materials and Methods: Bone marrow aspirates from iliac crests were obtained from orthopedic patients who underwent bone marrow harvesting under general anesthesia. Mesenchymal stem cells (hMSCs) were isolated by Ficoll-mediated discontinuous density gradient centrifugation and polystyrene adherence capacity. Cell cultures at the second passage were characterized, by the flow cytometric analysis (FCA) of specific surface antigens. A surface marker-expression pattern, positive (Stro-1, CD29, CD44, CD71, CD73, CD90, CD105) and negative (CD45, CD34, CD235), was employed to characterize the cells obtained. hMSCs were expanded and duplication time evaluated. The AlamarBlue assay was used to evaluate the number of cells attached to the biomaterials and to the control in adhesion and proliferation assay. The quantification of the relative amount of Focal Adhesion Kinase was employed to evaluate the strength of the cell adhesion to the biomaterials after 36 hours from seeding. In order to verify the morphology of the hMSCs attached to bioceramics, stem cells grown on the biomaterials were analyzed by SEM. Moreover, to verify whether the cytoskeletal organization of hMSCs cultured on the analyzed biomaterials was altered, actin fibers were investigated using TRITC conjugated-Phalloidin. Results: the biomaterials assayed in our study provided a favorable environment for hMSCs adhesion and growth, without any evidence of cytotoxic effects. Indeed, hMSCs adhesion and growth assays indicate that shape and porosity influence the cytocompatibility of bioceramics. We may infer that intra- and iter- porosity of biomaterials investigated herein, are determinant factors influencing hMSCs behavior. Besides, the correlation between vitality and a greater amount of Focal Adhesion Kinase at molecular level show how cellular processes may be modulated by structured surface and three dimensional architecture of biomaterials. hMSC SEM analysis showed an elongated spindle-like morphology. The cells adhered to biomaterials with many dendritic projections with some differences in the morphology due to scaffolds structure. Moreover, hMSCs cytoskeletal architecture on biomaterials was indistinguishable from that of hMSCs grown on plastic petri dishes, used as a control. Conclusion: These assays confirm the feasibility to evaluate biomaterials biological features in pre-clinical environment with patients mesenchymal stem cells in order to obtain information about subject specific response. Moreover, the development of new matrices have to consider the possibility to employ bioactive surfaces able to modulate cell adhesion and differentiation processes
DNA sequences of the tumor virus SV40 and the expression of its oncoprotein Tag have been detecte... more DNA sequences of the tumor virus SV40 and the expression of its oncoprotein Tag have been detected in human tumors, such as mesotheliomas, brain tumors, osteosarcomas and lymphomas. Little is known on the SV40 infection of human populations. In our study a new indirect ELISA test has been set employing synthetic peptides corresponding to specific SV40 epitopes of the capsid proteins. Specific antibodies against SV40 have been detected in sera of healthy donors from Casale Monferrato, a town affected by asbestos pollution and with a high prevalence of the mesothelioma. Peripheral blood cells from the same individuals were also analyzed by PCR for the SV40 Tag sequences. Sera and blood specimens harvested in Aviano were also analyzed in the same fashion. Our analyses indicate that 20% of the sera from blood donors carries specific SV40 antibodies, while 38% of the sera from mesothelioma patients are positive for the SV40 antibodies. Moreover, PBMCs from the same donors analyzed by PCR revealed SV40 Tag sequences with a prevalence similar to that detected by the ELISA test. Our serologic data confirm that SV40 is circulating in the human population, whereas the oncologic patients are SV40-positive with prevalence which is two fold compared to the healthy individual
Journal of Hematology & Oncology, 2014
Asbestos is considered the main cause of diseases in workers exposed to this mineral in the workp... more Asbestos is considered the main cause of diseases in workers exposed to this mineral in the workplace as well as an environmental pollutant. The association between asbestos and the onset of different diseases has been reported, but asbestos exposure specific biomarkers are not known. MicroRNAs (miRNAs) are small, single-strand, non-coding RNAs, with potential value as diagnostic, prognostic, and predictive markers in liquid biopsies. Sera collected from workers ex-exposed to asbestos (WEA) fibers were compared with sera from healthy subjects (HS) of similar age, as liquid biopsies. The expression of the circulating miRNA 197-3p was investigated employing two different highly analytical PCR methods, i.e. RT-PCR and dd-PCR. MiR-197-3p levels were tested in sera from WEA compared to HS. MiR-197-3p tested dysregulated in sera from WEA (n=75) compared to HS (n=62). Indeed, miR-197-3p was found to be 2.6 times down-regulated in WEA vs. HS (p=0.0001***). In addition, an inverse correlatio...
International Journal of Molecular Sciences
Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been close... more Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been closely studied in recent years, especially in view of their potential use for treating diseases and damaged tissues and organs. MSCs are capable of self-replication and differentiation into osteoblasts and are considered an important source of cells in tissue engineering for bone regeneration. Several epigenetic factors are believed to play a role in the osteogenic differentiation of MSCs, including microRNAs (miRNAs). MiRNAs are small, single-stranded, non-coding RNAs of approximately 22 nucleotides that are able to regulate cell proliferation, differentiation and apoptosis by binding the 3′ untranslated region (3′-UTR) of target mRNAs, which can be subsequently degraded or translationally silenced. MiRNAs control gene expression in osteogenic differentiation by regulating two crucial signaling cascades in osteogenesis: the transforming growth factor-beta (TGF-β)/bone morphogenic protein (B...
European Journal of Clinical Microbiology & Infectious Diseases
Panton-Valentine leukocidin (PVL) appears to be a virulence factor which, among others, can exace... more Panton-Valentine leukocidin (PVL) appears to be a virulence factor which, among others, can exacerbate the pathogenicity of Staphylococcus aureus infections, especially inducing severe necrotic, deep-seated skin infections, abscesses, and recurrences. These peculiarities have some overlaps with hidradenitis suppurativa (HS). Our main aim was to assess if S. aureus producing PVL could have some role in influencing clinical features and/or course of HS, specifically in the suppuration and recurrence of lesions. This pilot, mono-centric, observational study included all adult subjects affected with HS consecutively referring to our HS clinic over a 3-month period. Clinically evident suppuration and at least 2 weeks wash out from any antibiotic were the main inclusion criteria. Purulent material from HS skin lesions was collected with swabs in order to isolate micro-organisms, with specific regard to S. aureus. Detection of PVL was performed by real-time quantitative PCR (RT-qPCR). We a...
Infectious Agents and Cancer
Background Killian polyp (KP) is a benign lesion that arises from the maxillary sinus. The etiolo... more Background Killian polyp (KP) is a benign lesion that arises from the maxillary sinus. The etiology of KP is unknown. The aim of this study was to investigate the potential involvement of human papilloma- (HPV) and polyoma-viruses (HPyV) infections in the onset of KP. Methods DNA from antral (n = 14) and nasal (n = 14) KP fractions were analyzed for HPV and HPyV sequences, genotypes, viral DNA load and physical status along with expression of viral proteins and p16 cellular protein. Results The oncogenic HPV16 was detected in 3/14 (21.4%) antral KPs, whilst nasal KPs tested HPV-negative (0/14). The mean HPV16 DNA load was 4.65 ± 2.64 copy/104 cell. The whole HPV16 episomal genome was detected in one KP sample, whereas HPV16 DNA integration in two KPs. P16 mRNA level was lower in the KP sample carrying HPV16 episome than in KPs carrying integrated HPV16 and HPV- negative KPs (p
Antioxidants, 2022
Resveratrol is a powerful antioxidant molecule. In the human diet, its most important source is i... more Resveratrol is a powerful antioxidant molecule. In the human diet, its most important source is in Vitis vinifera grape peel and leaves. Resveratrol exists in two isoforms, cis- and trans. The diastereomeric forms of many drugs have been reported as affecting their activity. The aim of this study was to set up a cellular model to investigate how far resveratrol could counteract cytotoxicity in an oxidant agent. For this purpose, a keratinocyte cell line, which was genetically engineered with jelly fish green fluorescent protein, was treated with the free radical promoter Cumene hydroperoxide. The antioxidant activity of the trans-resveratrol and its diastereomeric mixture was evaluated indirectly in these treated fluorescent-engineered keratinocytes by analyzing the cell number and cell proliferation index. Our results demonstrate that cells, which were pre-incubated with resveratrol, reverted the oxidative damage progression induced by this free radical agent. In conclusion, fluore...
Simian Virus 40, experimentally assayed in vitro in different animal and human cells and in vivo ... more Simian Virus 40, experimentally assayed in vitro in different animal and human cells and in vivo in rodents, was classified as a small DNA tumor virus. In previous studies, many groups identified Simian Virus 40 sequences in healthy individuals and cancer patients using PCR techniques, whereas others failed to detect the viral sequences in human specimens. These conflicting results prompted us to develop a novel indirect ELISA with synthetic pep-tides, mimicking Simian Virus 40 capsid viral protein antigens, named mimotopes. This immunologic assay allowed us to investigate the presence of serum antibodies against Sim-ian Virus 40 and to verify whether Simian Virus 40 is circulating in humans. In this investiga-tion two mimotopes from Simian Virus 40 large T antigen, the viral replication protein and oncoprotein, were employed to analyze for specific reactions to human sera antibodies. This indirect ELISA with synthetic peptides from Simian Virus 40 large T antigen was used to assay ...
Cancers, 2021
Despite improved prognosis for many HPV-positive head and neck squamous cell carcinomas (HNSCCs),... more Despite improved prognosis for many HPV-positive head and neck squamous cell carcinomas (HNSCCs), some cases are still marked by recurrence and metastasis. Our study aimed to identify novel biomarkers for patient stratification. Classical HPV markers: HPV-DNA, p16 and HPV mRNA expression were studied in HNSCC (n = 67) and controls (n = 58) by qPCR. Subsequently, ELISA tests were used for HPV16 L1 antibody and HPV16 E7 oncoprotein detection in serum at diagnosis and follow-up. All markers were correlated to relapse-free survival (RFS) and overall survival (OS). HPV-DNA was found in HNSCCs (29.85%), HPV16-DNA in 95% of cases, HPV16 E7 mRNA was revealed in 93.75%. p16 was overexpressed in 75% of HPV-positive HNSCC compared to negative samples and controls (p < 0.001). Classical markers correlated with improved OS (p < 0.05). Serological studies showed similar proportions of HPV16 L1 antibodies in all HNSCCs (p > 0.05). Serum E7 oncoprotein was present in 30% HPV-positive patie...
La presente invenzione si riferisce all’identificazione di nuovi peptidi sintetici, corrispondent... more La presente invenzione si riferisce all’identificazione di nuovi peptidi sintetici, corrispondenti a specifici epitopi/antigeni della proteina virale capsidica 1, VP1, del virus Polioma BK (BKPyV), per rilevare nel siero e in altri fluidi umani anticorpi contro questo virus, potenzialmente patogeno inclusa l’attività oncogena, impiegando la tecnica immunologica ELISA indiretta
Introduction: in the orthopedic perspective, tissue engineering is focused on the development of ... more Introduction: in the orthopedic perspective, tissue engineering is focused on the development of innovative materials, whose action consists in recruiting bone progenitor cells and in stimulating their proliferation. In this study, we investigated the interaction between human mesenchymal stem cells from adults and two types of biomaterials, which differ for chemical composition and format. To evaluate the effects of porosity on cell adhesion and proliferation comparative analyses were carried out on different forms of two bioceramic materials. Materials and Methods: Bone marrow aspirates from iliac crests were obtained from orthopedic patients who underwent bone marrow harvesting under general anesthesia. Mesenchymal stem cells (hMSCs) were isolated by Ficoll-mediated discontinuous density gradient centrifugation and polystyrene adherence capacity. Cell cultures at the second passage were characterized, by the flow cytometric analysis (FCA) of specific surface antigens. A surface marker-expression pattern, positive (Stro-1, CD29, CD44, CD71, CD73, CD90, CD105) and negative (CD45, CD34, CD235), was employed to characterize the cells obtained. hMSCs were expanded and duplication time evaluated. The AlamarBlue assay was used to evaluate the number of cells attached to the biomaterials and to the control in adhesion and proliferation assay. The quantification of the relative amount of Focal Adhesion Kinase was employed to evaluate the strength of the cell adhesion to the biomaterials after 36 hours from seeding. In order to verify the morphology of the hMSCs attached to bioceramics, stem cells grown on the biomaterials were analyzed by SEM. Moreover, to verify whether the cytoskeletal organization of hMSCs cultured on the analyzed biomaterials was altered, actin fibers were investigated using TRITC conjugated-Phalloidin. Results: the biomaterials assayed in our study provided a favorable environment for hMSCs adhesion and growth, without any evidence of cytotoxic effects. Indeed, hMSCs adhesion and growth assays indicate that shape and porosity influence the cytocompatibility of bioceramics. We may infer that intra- and iter- porosity of biomaterials investigated herein, are determinant factors influencing hMSCs behavior. Besides, the correlation between vitality and a greater amount of Focal Adhesion Kinase at molecular level show how cellular processes may be modulated by structured surface and three dimensional architecture of biomaterials. hMSC SEM analysis showed an elongated spindle-like morphology. The cells adhered to biomaterials with many dendritic projections with some differences in the morphology due to scaffolds structure. Moreover, hMSCs cytoskeletal architecture on biomaterials was indistinguishable from that of hMSCs grown on plastic petri dishes, used as a control. Conclusion: These assays confirm the feasibility to evaluate biomaterials biological features in pre-clinical environment with patients mesenchymal stem cells in order to obtain information about subject specific response. Moreover, the development of new matrices have to consider the possibility to employ bioactive surfaces able to modulate cell adhesion and differentiation processes
DNA sequences of the tumor virus SV40 and the expression of its oncoprotein Tag have been detecte... more DNA sequences of the tumor virus SV40 and the expression of its oncoprotein Tag have been detected in human tumors, such as mesotheliomas, brain tumors, osteosarcomas and lymphomas. Little is known on the SV40 infection of human populations. In our study a new indirect ELISA test has been set employing synthetic peptides corresponding to specific SV40 epitopes of the capsid proteins. Specific antibodies against SV40 have been detected in sera of healthy donors from Casale Monferrato, a town affected by asbestos pollution and with a high prevalence of the mesothelioma. Peripheral blood cells from the same individuals were also analyzed by PCR for the SV40 Tag sequences. Sera and blood specimens harvested in Aviano were also analyzed in the same fashion. Our analyses indicate that 20% of the sera from blood donors carries specific SV40 antibodies, while 38% of the sera from mesothelioma patients are positive for the SV40 antibodies. Moreover, PBMCs from the same donors analyzed by PCR revealed SV40 Tag sequences with a prevalence similar to that detected by the ELISA test. Our serologic data confirm that SV40 is circulating in the human population, whereas the oncologic patients are SV40-positive with prevalence which is two fold compared to the healthy individual
Journal of Hematology & Oncology, 2014
Asbestos is considered the main cause of diseases in workers exposed to this mineral in the workp... more Asbestos is considered the main cause of diseases in workers exposed to this mineral in the workplace as well as an environmental pollutant. The association between asbestos and the onset of different diseases has been reported, but asbestos exposure specific biomarkers are not known. MicroRNAs (miRNAs) are small, single-strand, non-coding RNAs, with potential value as diagnostic, prognostic, and predictive markers in liquid biopsies. Sera collected from workers ex-exposed to asbestos (WEA) fibers were compared with sera from healthy subjects (HS) of similar age, as liquid biopsies. The expression of the circulating miRNA 197-3p was investigated employing two different highly analytical PCR methods, i.e. RT-PCR and dd-PCR. MiR-197-3p levels were tested in sera from WEA compared to HS. MiR-197-3p tested dysregulated in sera from WEA (n=75) compared to HS (n=62). Indeed, miR-197-3p was found to be 2.6 times down-regulated in WEA vs. HS (p=0.0001***). In addition, an inverse correlatio...
International Journal of Molecular Sciences
Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been close... more Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been closely studied in recent years, especially in view of their potential use for treating diseases and damaged tissues and organs. MSCs are capable of self-replication and differentiation into osteoblasts and are considered an important source of cells in tissue engineering for bone regeneration. Several epigenetic factors are believed to play a role in the osteogenic differentiation of MSCs, including microRNAs (miRNAs). MiRNAs are small, single-stranded, non-coding RNAs of approximately 22 nucleotides that are able to regulate cell proliferation, differentiation and apoptosis by binding the 3′ untranslated region (3′-UTR) of target mRNAs, which can be subsequently degraded or translationally silenced. MiRNAs control gene expression in osteogenic differentiation by regulating two crucial signaling cascades in osteogenesis: the transforming growth factor-beta (TGF-β)/bone morphogenic protein (B...
European Journal of Clinical Microbiology & Infectious Diseases
Panton-Valentine leukocidin (PVL) appears to be a virulence factor which, among others, can exace... more Panton-Valentine leukocidin (PVL) appears to be a virulence factor which, among others, can exacerbate the pathogenicity of Staphylococcus aureus infections, especially inducing severe necrotic, deep-seated skin infections, abscesses, and recurrences. These peculiarities have some overlaps with hidradenitis suppurativa (HS). Our main aim was to assess if S. aureus producing PVL could have some role in influencing clinical features and/or course of HS, specifically in the suppuration and recurrence of lesions. This pilot, mono-centric, observational study included all adult subjects affected with HS consecutively referring to our HS clinic over a 3-month period. Clinically evident suppuration and at least 2 weeks wash out from any antibiotic were the main inclusion criteria. Purulent material from HS skin lesions was collected with swabs in order to isolate micro-organisms, with specific regard to S. aureus. Detection of PVL was performed by real-time quantitative PCR (RT-qPCR). We a...
Infectious Agents and Cancer
Background Killian polyp (KP) is a benign lesion that arises from the maxillary sinus. The etiolo... more Background Killian polyp (KP) is a benign lesion that arises from the maxillary sinus. The etiology of KP is unknown. The aim of this study was to investigate the potential involvement of human papilloma- (HPV) and polyoma-viruses (HPyV) infections in the onset of KP. Methods DNA from antral (n = 14) and nasal (n = 14) KP fractions were analyzed for HPV and HPyV sequences, genotypes, viral DNA load and physical status along with expression of viral proteins and p16 cellular protein. Results The oncogenic HPV16 was detected in 3/14 (21.4%) antral KPs, whilst nasal KPs tested HPV-negative (0/14). The mean HPV16 DNA load was 4.65 ± 2.64 copy/104 cell. The whole HPV16 episomal genome was detected in one KP sample, whereas HPV16 DNA integration in two KPs. P16 mRNA level was lower in the KP sample carrying HPV16 episome than in KPs carrying integrated HPV16 and HPV- negative KPs (p