Background. L’ipoparatiroidismo postchirurgico rappresenta un’evenienza tutt’altro che rara dopo ... more Background. L’ipoparatiroidismo postchirurgico rappresenta un’evenienza tutt’altro che rara dopo intervento di tiroidectomia totale e/o paratiroidectomia totale. I tentativi di trapiantare tessuto paratiroideo sono iniziati nel 1975 con Wells ed i risultati ancora oggi sono alquanto deludenti. Negli ultimi anni grazie a tecniche di ingegneria tissutale si cerca di costruire paratiroidi artificiali,capaci di secernere paratormone, disponibili per il trapianto in pazienti affetti da ipoparatiroidismo iatrogeno. Pazienti e metodi. I paratireociti sono stati ottenuti da paratiroidi di tre pazienti, uremici cronici in emodialisi, operati per iperparatiroidismo secondario. Le colture cellulari ottenute in RPMI sono state successivamente seminate sugli scaffold di collagene (supporti tridimensionali a lenta biodegradazione). Il collagene rappresenta la componente maggioritaria della matrice extracellulare e quindi costituisce un buon substrato su cui le cellule aderiscono e crescono. I ter...
Fibrous encapsulation is known to occur to many prosthetic implants and is thought to be due to t... more Fibrous encapsulation is known to occur to many prosthetic implants and is thought to be due to the cells not adhering adequately to the surface. For developing new materials able to enhance cellular adhesion by mimicking extracellular matrix components, polyelectrolyte polymers, characterized by tunable surface charges, have been proposed. Here we demonstrate that panoply of cell functions over a two-dimensional substratum is influenced by surface charge. We have at first generated structurally related polyelectrolyte substrata varying in their positive surface charge amount and subsequently evaluated a variety of behaviors of human primary fibroblasts seeded on these polymers. The proportion of adherent, spreading, and proliferating cells was increased significantly on cationic hydrophilic surfaces when compared with the neutral base surface. The extent of cell spreading correlated with cytoskeleton organization as assessed using immunofluorescence techniques. In the key experiment, the presence of cationic charges on cell adhesion-resistant neutral surface increased the synthesis of collagen I and III, the release of their metabolites, and the expression of their mRNA by fibroblasts. Interestingly, the scarce collagen deposits on neutral polymer consisted, for the most part, of collagen I while collagen III was present only in traces probably due to the secretion of metalloproteinase-2 by non-adherent fibroblasts. Taken together, these results show that polyelectrolyte films may promote the attachment of fibroblast cells as well as their normal secretory phenotype. Both effects could be potentially useful in integrating soft connective tissue to the implant, decreasing the chance of its fibrous encapsulation.
The aim of this paper is based on the use of a hyaluronic acid hydrogel of Quercetin tested alone... more The aim of this paper is based on the use of a hyaluronic acid hydrogel of Quercetin tested alone and in combination to an inhibitor of Aurora Kinase type A and B (SNS-314) on human medullary and papillary thyroid cancer cells. Biological investigations were focused on the cellular uptake of the hydrogel, cell viability, antioxidant and cytokines secretion studies. Quercetin delivered from hydrogel show a time and CD44 dependent interaction with both cell lines with significant anti-inflammatory effects. Combination of Quercetin and SNS-314 leads to a synergistic cytotoxic effect on medullary TT and papillary BCPAP cell lines with a significant reduction of the IC50 value. These results, highlights the importance of synergistic effect of the hyaluronic acid hydrogel of Quercetin with SNS-314 in the regulation of human thyroid cancer cell proliferation and emphasize the anti-tumor activity of these molecules. This article is protected by copyright. All rights reserved.
The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the s... more The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the slow release of antibiotic in periodontal pockets. The antibiotics examined were tetracycline and metronidazole. The aim of the study was to evaluate the biocompatibility of the material with periodontal tissue and the efficacy of the 2 prepared systems. Two sites were selected in 2 periodontopathic patients who after non-surgical treatment presented pockets measuring 8 and 7 mm. A sheet of pHEMA containing tetracycline was inserted in one and in the other a sheet containing metronidazole: both were left for 8 days in the chosen pockets. At the start and end of treatment PD and GI clinical indices were measured and the DMDx microbiological test was performed to identify Aa of Pg and Pi. The tissue reaction to pHEMA was evaluated using SEM analysis of two samples collected after 8 days of treatment. The microscopic results showed the optimal biocompatibility of both samples. Differences we...
We study the interaction of 3T3 Swiss albino mouse fibroblasts with polymeric nanoparticles (NPs)... more We study the interaction of 3T3 Swiss albino mouse fibroblasts with polymeric nanoparticles (NPs) and investigate cellular behaviour in terms of viability/cytotoxicity, cell cycle, NPs uptake, MAP kinase (ERK1/2), and focal adhesion kinase (FAK) activation. After incubation of NPs with cell culture media, western blot analysis showed that Vitronectin is retained by NPs, while Fibronectin is not detected. From cytotoxicity studies (MTT and BrdU methods) an LD50 of about 1.5 mg/mL results for NPs. However, NPs in the range 0.01–0.30 mg/mL are able to trigger a statistically significant increase in proliferation and cell cycle progression in dose and time depending manner. Also, biochemical evaluation of ERK1/2 and FAK clearly shows an increasing phosphorylation in a dose and time depending manner. Finally, we found by transmission electron microscopy that NPs are internalised by cells. Competitively blocking VN-integrin receptors with echistatin (1 μg/mL) results in a decrease of viab...
... r to solidify after about 1-2 seconds, completing its solidification reaction after around 60... more ... r to solidify after about 1-2 seconds, completing its solidification reaction after around 60-90 seconds. After this reaction, the glue reaches its maximum mechani-... 7.2.5 TheFuture: Innovative Biotechnology for Hemostasis Control ...
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Immunogold labeling (IGL) technique has been utilized by many authors in combination with scannin... more Immunogold labeling (IGL) technique has been utilized by many authors in combination with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to obtain the identification/localization of receptors and antigens, both in cells and tissues. Environmental scanning electron microscopy (ESEM) represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artifacts and interfere with the IGL procedure. The absence of metal coating could yield further advantages for our purpose as the labeling detection is based on the atomic number difference between nanogold spheres and the biological material. Using the gaseous secondary electron detector, compositional contrast is easily revealed by the backscattered electron component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimized to improve the intensity and the specificity of the labeling signal, in order to obtain a semiquantitative evaluation of the labeling signal.In particular, we used a combination of IGL and ESEM to detect the presence of a protein on the cell surface. To achieve this purpose, we chose as an experimental system 3T3 Swiss albino mouse fibroblasts and galectin-3.
ABSTRACT Nanotechnology is an emerging science that studies how to control matter on an atomic an... more ABSTRACT Nanotechnology is an emerging science that studies how to control matter on an atomic and molecular scale. Generally nanotechnology deals with structures of the size of 100 nanometers or smaller in at least one dimension, and involves developing materials or devices within that size. Nanotechnology ranges from extensions of conventional device physics to completely new approaches based upon molecular self-assembly, from developing new materials with dimensions on the nanoscale to investigating whether we can directly control matter on the atomic
There is a substantial unmet demand for tissues to repair injured, degenerated or congenitally de... more There is a substantial unmet demand for tissues to repair injured, degenerated or congenitally defected tissues. The field of tissue engineering has emerged to fill the void where neither native physiology nor purely artificial implantable materials can sufficiently replace or repair these damaged tissues. While tissues such as bone or skin can effectively repair a small injury given sufficient time,
Background. L’ipoparatiroidismo postchirurgico rappresenta un’evenienza tutt’altro che rara dopo ... more Background. L’ipoparatiroidismo postchirurgico rappresenta un’evenienza tutt’altro che rara dopo intervento di tiroidectomia totale e/o paratiroidectomia totale. I tentativi di trapiantare tessuto paratiroideo sono iniziati nel 1975 con Wells ed i risultati ancora oggi sono alquanto deludenti. Negli ultimi anni grazie a tecniche di ingegneria tissutale si cerca di costruire paratiroidi artificiali,capaci di secernere paratormone, disponibili per il trapianto in pazienti affetti da ipoparatiroidismo iatrogeno. Pazienti e metodi. I paratireociti sono stati ottenuti da paratiroidi di tre pazienti, uremici cronici in emodialisi, operati per iperparatiroidismo secondario. Le colture cellulari ottenute in RPMI sono state successivamente seminate sugli scaffold di collagene (supporti tridimensionali a lenta biodegradazione). Il collagene rappresenta la componente maggioritaria della matrice extracellulare e quindi costituisce un buon substrato su cui le cellule aderiscono e crescono. I ter...
Fibrous encapsulation is known to occur to many prosthetic implants and is thought to be due to t... more Fibrous encapsulation is known to occur to many prosthetic implants and is thought to be due to the cells not adhering adequately to the surface. For developing new materials able to enhance cellular adhesion by mimicking extracellular matrix components, polyelectrolyte polymers, characterized by tunable surface charges, have been proposed. Here we demonstrate that panoply of cell functions over a two-dimensional substratum is influenced by surface charge. We have at first generated structurally related polyelectrolyte substrata varying in their positive surface charge amount and subsequently evaluated a variety of behaviors of human primary fibroblasts seeded on these polymers. The proportion of adherent, spreading, and proliferating cells was increased significantly on cationic hydrophilic surfaces when compared with the neutral base surface. The extent of cell spreading correlated with cytoskeleton organization as assessed using immunofluorescence techniques. In the key experiment, the presence of cationic charges on cell adhesion-resistant neutral surface increased the synthesis of collagen I and III, the release of their metabolites, and the expression of their mRNA by fibroblasts. Interestingly, the scarce collagen deposits on neutral polymer consisted, for the most part, of collagen I while collagen III was present only in traces probably due to the secretion of metalloproteinase-2 by non-adherent fibroblasts. Taken together, these results show that polyelectrolyte films may promote the attachment of fibroblast cells as well as their normal secretory phenotype. Both effects could be potentially useful in integrating soft connective tissue to the implant, decreasing the chance of its fibrous encapsulation.
The aim of this paper is based on the use of a hyaluronic acid hydrogel of Quercetin tested alone... more The aim of this paper is based on the use of a hyaluronic acid hydrogel of Quercetin tested alone and in combination to an inhibitor of Aurora Kinase type A and B (SNS-314) on human medullary and papillary thyroid cancer cells. Biological investigations were focused on the cellular uptake of the hydrogel, cell viability, antioxidant and cytokines secretion studies. Quercetin delivered from hydrogel show a time and CD44 dependent interaction with both cell lines with significant anti-inflammatory effects. Combination of Quercetin and SNS-314 leads to a synergistic cytotoxic effect on medullary TT and papillary BCPAP cell lines with a significant reduction of the IC50 value. These results, highlights the importance of synergistic effect of the hyaluronic acid hydrogel of Quercetin with SNS-314 in the regulation of human thyroid cancer cell proliferation and emphasize the anti-tumor activity of these molecules. This article is protected by copyright. All rights reserved.
The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the s... more The study analyses the possibility of using polyhydroxyethyl methacrylate as a material for the slow release of antibiotic in periodontal pockets. The antibiotics examined were tetracycline and metronidazole. The aim of the study was to evaluate the biocompatibility of the material with periodontal tissue and the efficacy of the 2 prepared systems. Two sites were selected in 2 periodontopathic patients who after non-surgical treatment presented pockets measuring 8 and 7 mm. A sheet of pHEMA containing tetracycline was inserted in one and in the other a sheet containing metronidazole: both were left for 8 days in the chosen pockets. At the start and end of treatment PD and GI clinical indices were measured and the DMDx microbiological test was performed to identify Aa of Pg and Pi. The tissue reaction to pHEMA was evaluated using SEM analysis of two samples collected after 8 days of treatment. The microscopic results showed the optimal biocompatibility of both samples. Differences we...
We study the interaction of 3T3 Swiss albino mouse fibroblasts with polymeric nanoparticles (NPs)... more We study the interaction of 3T3 Swiss albino mouse fibroblasts with polymeric nanoparticles (NPs) and investigate cellular behaviour in terms of viability/cytotoxicity, cell cycle, NPs uptake, MAP kinase (ERK1/2), and focal adhesion kinase (FAK) activation. After incubation of NPs with cell culture media, western blot analysis showed that Vitronectin is retained by NPs, while Fibronectin is not detected. From cytotoxicity studies (MTT and BrdU methods) an LD50 of about 1.5 mg/mL results for NPs. However, NPs in the range 0.01–0.30 mg/mL are able to trigger a statistically significant increase in proliferation and cell cycle progression in dose and time depending manner. Also, biochemical evaluation of ERK1/2 and FAK clearly shows an increasing phosphorylation in a dose and time depending manner. Finally, we found by transmission electron microscopy that NPs are internalised by cells. Competitively blocking VN-integrin receptors with echistatin (1 μg/mL) results in a decrease of viab...
... r to solidify after about 1-2 seconds, completing its solidification reaction after around 60... more ... r to solidify after about 1-2 seconds, completing its solidification reaction after around 60-90 seconds. After this reaction, the glue reaches its maximum mechani-... 7.2.5 TheFuture: Innovative Biotechnology for Hemostasis Control ...
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Immunogold labeling (IGL) technique has been utilized by many authors in combination with scannin... more Immunogold labeling (IGL) technique has been utilized by many authors in combination with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to obtain the identification/localization of receptors and antigens, both in cells and tissues. Environmental scanning electron microscopy (ESEM) represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artifacts and interfere with the IGL procedure. The absence of metal coating could yield further advantages for our purpose as the labeling detection is based on the atomic number difference between nanogold spheres and the biological material. Using the gaseous secondary electron detector, compositional contrast is easily revealed by the backscattered electron component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimized to improve the intensity and the specificity of the labeling signal, in order to obtain a semiquantitative evaluation of the labeling signal.In particular, we used a combination of IGL and ESEM to detect the presence of a protein on the cell surface. To achieve this purpose, we chose as an experimental system 3T3 Swiss albino mouse fibroblasts and galectin-3.
ABSTRACT Nanotechnology is an emerging science that studies how to control matter on an atomic an... more ABSTRACT Nanotechnology is an emerging science that studies how to control matter on an atomic and molecular scale. Generally nanotechnology deals with structures of the size of 100 nanometers or smaller in at least one dimension, and involves developing materials or devices within that size. Nanotechnology ranges from extensions of conventional device physics to completely new approaches based upon molecular self-assembly, from developing new materials with dimensions on the nanoscale to investigating whether we can directly control matter on the atomic
There is a substantial unmet demand for tissues to repair injured, degenerated or congenitally de... more There is a substantial unmet demand for tissues to repair injured, degenerated or congenitally defected tissues. The field of tissue engineering has emerged to fill the void where neither native physiology nor purely artificial implantable materials can sufficiently replace or repair these damaged tissues. While tissues such as bone or skin can effectively repair a small injury given sufficient time,
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