Filiberto Mastrangelo | Università Vita-Salute San Raffaele (original) (raw)

Papers by Filiberto Mastrangelo

Objectives: The dental pulp is composed of cells in contact with the extracellular matrix (ECM), ... more Objectives: The dental pulp is composed of cells in contact with the extracellular matrix (ECM), The main macromolecules in dental pulp ECM are collagenous proteins (especially types I and III collagen), The aim of the work was to evaluate the collagen gene network in the human healthy dental pulp. Methods: Expression of collagen genes was investigated at three different pulp development stages: bud stage (average age 13), during apex maturation (average age 18) and from older subjects (average age 57). A microarray analysis was carried out using high density array containing 21.329 transcripts in replicates, ).The obtained data were statistically analyzed using the Significance Analysis of Microarray system (S.A.M), and Ingenuity Pathways Analysis (I.P.A). RT-PCR analysis was performed to validate microarray results Results: Microarray analysis showed the expression of several collagen genes involved in tooth development at different expression levels: collagen type I (COL1A1), col...

Objectives: In this study was evaluated by immunohistochemical analysis the activity of molecules... more Objectives: In this study was evaluated by immunohistochemical analysis the activity of molecules involved in the process of bone remodeling during the integration period (T1) and at a distance of 10 years (T2) of extraoral autologous bone grafts from two different donor sites: calvaria and iliac crest. Methods: Calvaria and Iliac Crest bone grafts were used to fill bone defects in edentulous sites. Specimens from the donor sites were withdrawn at the moment of grafts insertion (T0), and from regenerated sites four months (T1) and ten years (T2) after grafting. Samples at different experimental times were processed in order to investigate the levels of expression of Bone Sialoprotein (BSP) and pro-apoptotic molecule BAX by immunohystochemical analysis. Moreover was investigated the number of apoptotic cells by the Tunel analysis. Results: In our study, it was possible to evaluate that BSP and BAX were significantly more expressed in samples T1 compared to that observed in samples T0...

Implant Dentistry, 2008

Success in implant dentistry also comes from the implant macrodesign and nanostructure of its sur... more Success in implant dentistry also comes from the implant macrodesign and nanostructure of its surface. Titanium implant surface treatments have been shown to enhance osseointegration, maximize bone healing, and bone-to-implant contact for predictable clinical results. The aim of the study, was to evaluate the geometric macrodesign and the surface nanostructure of a novel dental implant full contact covering (FCC) obtained by electrochemical procedures. FCC implants were analyzed by scanning electronic microscope, profilometer, and x-ray photoelectron spectroscopy and compared with commercial sandblasted and sandblasted, large-grit acid-etched dental implants. Sample analysis allowed to distinguish the different implant macrodesigns, the step and the profile of the coils that cover the fixture, and the surface characteristics. FCC implant showed novel macro-characteristic of crestal module, coils, and apical zone compared with sandblasted and sandblasted and acid-etched dental implants. Moreover, the FCC nanostructure surface showed roughness values statistically higher than the 2 other surfaces, with a more homogeneity in a peaks and valleys arrangement. Finally, the x-ray photoelectron spectroscopy analysis detected differences between the examined surfaces, with the presence of several contaminants according to the different treatment procedures. Research on new macrostructures and nano morphology should result in a better qualitative and quantitative osseointegration response, with a predictability of the clinical results and long-term success of the implants.

Objectives: The aim of this work was to investigate the effects of the peptide VEGF-A165 on the p... more Objectives: The aim of this work was to investigate the effects of the peptide VEGF-A165 on the proliferation and osteogenic differentiation of human Dental Pulp Stem Cells (hDPSCs). Methods: Mesenchymal stem cells were obtained from dental pulp of enclosed thirds molars of young healthy patients. Mesenchymal markers expression was investigated by FACS analysis. Cell differentiation towards osteogenic or adipogenic lineages was evaluated by means of Alizarin Red S or Red Oil O staining, respectively. The expression of receptors (VEGF-1 and -2) and co-receptors (neuropilin-1 and -2) was estimated by RT-PCR. The effects of VEGF-A165 on cell proliferation was evaluated by MTT assay whereas Western Blot analysis was used to measure VEGF-induced changes in ALP and PECAM-1 protein content. Results: Cells, isolated from dental pulp, originated a homogenous population expressing typical mesenchymal markers (CD29, CD90, CD146, CD166 and STRO-1), able to differentiate into osteoblasts and ady...

Objectives: The aim of this study was to evaluate the gene expression of i-NOS in connection with... more Objectives: The aim of this study was to evaluate the gene expression of i-NOS in connection with oxidative stress and the network of antioxidant enzymes (SOD, CAT, GR and GPX) in order to highlight the possible role of Nitric Oxide (NO) in control mechanisms of oxidative stress occurred during tooth development. Methods: Samples were obtained from human third molars pulp at three different stages: bud stage (average age 13), during apex maturation (average age 18) and from older subjects (average age 57). The measurement of gene expression was performed by Microarray analysis. Obtained data were statistically analyzed using the S.A.M system (Significance Analysis of Microarray) and I.P.A. (Ingenuity Pathways analysis). Finally, RT-PCR analysis was performed in order to validate microarray results. Results: Expression of i-NOS gene decreased with progressive tissue maturation. I.P.A software identified networks and functional groups of interesting genes belonging to antioxidant enzy...

The International journal of oral & maxillofacial implants

To evaluate in vivo collagen fiber behavior around two different dental implant necks placed in t... more To evaluate in vivo collagen fiber behavior around two different dental implant necks placed in the mandibular bone of adult pigs. Scanning electron microscopic (SEM) and profilometric analyses were performed on both types of implant necks to evaluate the different surface morphology. Ten dental implants with machined titanium necks and 20 implants with zirconia necks were inserted into the mandibles of five adult pigs. Three months later, the animals were sacrificed; samples from the peri-implant mucosa were obtained and prepared for histologic analysis. Evaluation of collagen fiber orientation in the connective tissue surrounding the implant necks was performed by polarized light microscopy. Inflammation in the peri-implant soft tissues was also measured via the Gingival Index. Postoperative healing was uneventful; all implants, except for one of each type, were osseointegrated after 3 months. SEM and profilometric analyses confirmed that zirconia necks showed Ra, Rq, and Rz value...

Objectives: The aim of this study was to evaluate changes in VEGF and MMPs expression at differen... more Objectives: The aim of this study was to evaluate changes in VEGF and MMPs expression at different times after grafting in extraoral autologous bone grafts from two different donor sites: calvaria and iliac crest. Methods: Calvaria and Iliac Crest bone grafts were used to fill bone defects in edentulous sites, in prediction of implant therapy. Specimens from the donor sites were withdrawn at the moment of grafts insertion (T0), and from regenerated sites six months (T1) and ten years (T2) after grafting. Samples at different experimental times were processed in order to investigate VEGF, MMP2 and MMP9 expression by immunohystochemical analysis. Results: Immunohistochemical analyses showed VEGF, MMP2 and MMP9 significantly increase their expression at T1, while at T2 their expression return to levels comparable to that recorded in specimens from the donor sites (T0). Moreover, in any samples from sites treated with calvaria grafts, all the investigated molecules showed higher levels ...

Objectives: The aim of the study was to evaluate, from a microscopical and morphostructural point... more Objectives: The aim of the study was to evaluate, from a microscopical and morphostructural point of view, the integration of extraoral autologous bone grafts from different donor sites in edentolous area of the jaws. Methods: The donor sites for autologous grafts were Calvaria and Iliac Crest. Samples were withdrawn from the donor sites at the moment of grafts insertion (T0), while samples from regenerated sites were taken four months (T1) and ten years (T2) after grafting. Bone specimens at different experimental times were processed and stained with Hematoxilin-Eosin in order to perform light microscope analysis. Results: Both bone substitutes showed a good integration with the host tissue at light microscope observation. Both samples at T1 showed strong remodeling phenomena, even if in sites regenerated with calvaria grafts a more dynamic aspect could be detected. Moreover, the presence of new blood vessels could be observed in both type of samples. After ten years, both bone gr...

International journal of immunopathology and pharmacology

Bone regeneration procedures allow oral rehabilitation with dental implants also in edentulous ri... more Bone regeneration procedures allow oral rehabilitation with dental implants also in edentulous ridges with severe bone atrophy. The integration of grafted materials with the host tissue can initiate regenerative, inflammatory and apoptotic response. Since molecular mechanisms exist at the basis of such response, the aim of this work is to investigate, by immunohistochemical analyses, the expression of proteins involved in the graft integration process, in parallel to clinical and histological modifications, occurring on sites treated with extraoral autologous bone graft deriving from the parietal region of the calvaria (eAB), intraoral autologous bone graft deriving from mandibular ramus (iAB) and heterologous bone graft from swine (hB) in human patients. In our study, the immunohistochemical expression of BSP, VEGF, eNOS in eAB samples was significantly higher (p < 0.05) compared to values recorded in iAB and hB samples. The inflammatory response, investigated by iNOS expression...

Annals of clinical and laboratory science, 2007

The growth of malignant tumors is determined in large part by the proliferative capacity of the t... more The growth of malignant tumors is determined in large part by the proliferative capacity of the tumor cells. Clinical observations and animal experiments have established that tumor cells elicit immune responses. Histopathologic studies show that many tumors are surrounded by mononuclear cell and mast cell infiltrates. Mast cells are ubiquitous in the body and are critical for allergic reactions. Increasing evidence indicates that mast cells secrete proinflammatory cytokines and are involved in neuro-inflammatory processes and cancer. Mast cells accumulate in the stroma surrounding certain tumors, especially mammary adenocarcinoma, and the molecules they secrete can benefit the tumor. However, mast cells can also increase at the site of tumor growth and participate in tumor rejection. Mast cells may be recruited by tumor-derived chemoattractants and selectively secrete molecules such as growth factors, histamine, heparin, VEGF, and IL-8, as well as proteases that permit the formatio...

Journal of Endodontics, 2009

JOURNAL OF HEALTH SCIENCE, 2008

In the last decade, extracorporeal bone tissue engineering has found more clinical applications d... more In the last decade, extracorporeal bone tissue engineering has found more clinical applications due to the progress and new achievements in the isolation and characterization of stem cells from different sources, as well as, in controlling proliferation and differentiation in vitro. The aim of this study is to evaluate the in vitro behaviour, morphological structure and extracellular matrix synthesis of human dental follicle stem cells (hDFSCs) isolated from human dental bud, when seeded onto a synthetic hydroxyapatite (HA) scaffold (ENGIpore c ). Populations of CD29+, CD90+, CD146+ and CD166+ were sorted by FAC sorter (FACS) analysis and were cultured in osteogenic medium and then, onto the scaffold. These cells were analyzed by optical and electronic microscopy, at week 1 and 6, before and after the differentiation. Light microscopy showed an intense attachment and colonization of the HA scaffold by polygonal-shaped cells. Scanning electron microscopy after six weeks revealed a tri-dimensional organization of the cells and the presence of dense material around the cell clusters. hDFSCs showed participation in protein biosynthesis and demonstrated high proliferation on the synthetic HA scaffold.

Journal of Craniofacial Surgery, 2010

The aim of this work was to investigate the immunohistochemical and histologic patterns occurring... more The aim of this work was to investigate the immunohistochemical and histologic patterns occurring in samples obtained from sites reconstructed with calvarial bone graft, 4 months (T1) and 10 years (T2) after grafting, in comparison to those observed in samples from the calvaria taken from the donor sites at the moment of withdrawal for the grafting (T0).Samples underwent immunohistochemical analysis for bone sialoprotein, matrix metalloproteinases 2 and 9, vascular endothelial growth factor, and Bax expressions; terminal-deoxynucleotidyl-transferase-mediated dUTP nick end-labeling analysis to evaluate the number of apoptotic samples; and morphologic evaluation after hematoxylin and eosin staining.The expression levels of all the investigated molecules, such as the apoptotic cells&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; count, showed a significant (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) increase in T1 samples, that is, 4 months after grafting, although their expression levels recorded after 10 years seemed to be comparable to those in the native bone. Moreover, at observation under the light microscope, T1 samples showed a nonuniform morphology, with mineralized areas surrounded by connective tissues where blood vessels could be distinguished. On the other hand, T0 and T2 samples showed a similar aspect, characterized by uniform areas of mineralized extracellular matrix with cavities containing osteocytes.These data suggest that, after approximately 4 months, it is possible to highlight stronger remodeling phenomena, accompanied by new bone formation and new blood vessel proliferation, which are necessary to the host tissue for graft integration. On the contrary, these phenomena could not be observed in samples obtained after a long period (T2), which reveal a morphologic and an immunohistochemical pattern comparable to that recorded in calvarial native bone (T0).

International Journal of Antimicrobial Agents, 2007

New polymeric composite materials containing a nanohybrid to be used for the controlled release o... more New polymeric composite materials containing a nanohybrid to be used for the controlled release of an antibiotic molecule, chloramphenicol succinate, have been formulated, prepared and characterised. The nanohybrid consists of a layered double hydroxide of Mg-Al hydrotalcitetype, in which the nitrate anions present in the host galleries were replaced with chloramphenicol succinate anions (CFS − ) by a simple ion-exchange reaction. Different amounts of the hybrid material were incorporated in polycaprolactone and processed as films of 0.15 mm thickness. The composite materials were analysed by X-ray diffractometry and thermogravimetry and their mechanical properties were determined. They showed properties even better than those of the pristine polymer. The release process of the antibiotic molecules was found to be very interesting and promising for tuneable drug delivery. It consists of two stages: an initial stage of a very rapid burst, in which a small fraction of drug is released; and a second stage that is much slower, extending for a longer and longer time. This behaviour is profoundly different and much slower than that of a sample in which the antibiotic molecule is directly incorporated into the polymeric matrix. The parameters influencing drug release have been individuated and discussed.

Implant Dentistry, 2013

To evaluate the different behavior of 3-dimensional biomaterial scaffolds-Bovine Bone (BB; Bio-Os... more To evaluate the different behavior of 3-dimensional biomaterial scaffolds-Bovine Bone (BB; Bio-Oss) and Hydroxyapatite (HA; ENGIpore)-during initial bone healing and development. Human dental papilla stem cells (hDPaSCs) were selected with FACsorter cytofluorimetric analysis, cultured with osteogenic medium, and analyzed with Alizarin red stained after differentiation. The obtained osteoblast-like cells (OCs) were cultured with BB and HA. alkaline phosphatase (ALP), OC, MEPE, and runt-related transcription factor 2 (RUNX2) expression markers were investigated performing Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis. After 40 days, samples were analyzed by light and electron microscopy. All the samples showed high in vitro biocompatibility and qualitative differences of OCs adhesion. RT-PCR and Western blot data exhibited similar marker rate, but ALP, OC, MEPE, and RUNX2expression, during initial healing and bone regeneration phase, was higher and faster in human dental papilla onto BB than in HA scaffolds. In biomaterials growth, RUNX2 seems to play an important role as a key regulator in human OCs from dental papilla bone development. Different surface BB scaffold characteristics seem to play a critical role in OCs differentiation showing different time of bone regeneration morphological characteristics as well as higher and faster levels of all observed markers.

Implant Dentistry, 2010

The aim of the study is to investigate human osteoblast-like cell behavior and growth in the pres... more The aim of the study is to investigate human osteoblast-like cell behavior and growth in the presence of 3 different titanium implant surfaces. Human stem cells were first obtained and then sorted by fluorescence-activated cell sorter from mesenchymal stem cell clusters of human dental papilla. The obtained human dental papilla stem cells were induced to differentiate into osteoblast-like cells and were then analyzed by reverse transcriptase polymerase chain reaction and Western blot analyses. The cells proliferated and were cultured onto 3 different titanium discs (sandblasted, sandblasted and large-grit acid-etched, and full contact coverage [FCC]) and analyzed by scanning electron microscope. In all analyses samples, a high cell activity was observed, with typical osteoblast mature morphostructural response on rough surface. The high number of osteoblast-like cells was found on titanium FCC discs. At the same time, scanning electron microscope analysis confirmed the high biocompatibility of this surface. The rapid maturation of the osteoblast-like cells on FCC titanium surface suggests that this structure could play a central role during initial phases of bone healing processes.

BMC Biotechnology, 2009

Background: Stem cells isolated from amniotic fluid are known to be able to differentiate into di... more Background: Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells.

Annals of Clinical & …, 2007

Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal dysplasia characterised by abnorm... more Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal dysplasia characterised by abnormal clavicles, patent sutures and fontanelles, supernumerary teeth, short stature, and a variety of other skeletal changes. The disease gene is CBFA1/RUNX2, which is mapped ...

Platelet-endothelial cell adhesion molecule-1 protein (PECAM-1/CD31) is expressed in numerous phy... more Platelet-endothelial cell adhesion molecule-1 protein (PECAM-1/CD31) is expressed in numerous physiological and pathological processes characterized by an increase of vascular permeability, and in normal and tumour tissues. CD31, member of the immunoglobulin super-family that mediates cell-to-cell adhesion, is a transmembrane glycoprotein, 130-140 kDa, also know as platelet-endothelium cell adhesion molecule (PECAM-1). CD31 is a ligand for CD38 and plays a role in thrombosis and angiogenesis. CD31 is strongly expressed in endothelial cells and weakly expressed in megakaryocytes, platelets, occasional plasma cells, lymphocytes (marginal zone B-cells, peripheral T-cells) and neutrophils. The present study evaluates the angiogenetic processes which are accompanied by an expansion of cystic radicular and keratocystic lesions of the jaw bone. Twelve subjects with maxillary cysts (8 males and 4 females) with an average age of 43 years were selected by the Chieti University Oral Surgery Department. The surgical samples taken were subjected to histological and immunohistochemical analysis. The histological evaluation confirmed the diagnosis of radicular cystisis and keratocystisis. The immunohistochemical analyses were positive for CD31 protein in all the lesions analysed, even though they had different intensities. Using a semiquantive analysis it was possible to highlight, in the radicular cyst samples, an intense expression of the vascular component both in the inflamed area and the adjacent stroma. The lesions with cheratin content showed newly-formed, rather modest, vascularity both in the area showing slight inflammation, where the cellular component is prevalent, and in the adjacent areas showing no sign of inflammation. Therefore, in our observations, angiogenesis could take on a primary role in the development of cystic lesions of the jaw bones. The differences of CD31 expression, in all samples, would advise for a wider monitoring able to evaluate the possible use of such a protein as a diagnostic marker.

International Journal of Cancer, 1997

Studies have shown that microvessel density influences breast-cancer prognosis. Since tumor angio... more Studies have shown that microvessel density influences breast-cancer prognosis. Since tumor angiogenesis is considered to be substantially affected by the excretion of vascular endothelial growth factor (VEGF) from tumor cells, we examined whether VEGF concentration is different in malignant and in non-malignant breast tissue. It was also of interest to discover whether intratumoral VEGF concentration influences disease-free survival (DFS) of breast-cancer patients. Analysis is based on 120 tissue specimens taken from breast fibromas (n 5 23), normal epithelial breast tissue adjacent to fibromas (n 5 8) and invasive breast cancer (n 5 89). VEGF concentration was quantified by using an immunoassay. Microvessel density was determined by immunostaining for factor-VIII-related antigen. Median VEGF concentration is given in pg/mg protein (25%-quantile-75%-quantile) and it was 0 (0-1.8) in normal breast tissue, 9.8 (0.52-43.0) in fibromas and 130.4 (50.8-362.2) in invasive carcinomas. A univariate Cox model revealed that node status, tumor size, estrogen-receptor concentration, histological grading and microvessel density were prognostic factors for disease-free survival in breast cancer. We found a significant correlation between VEGF concentration and microvessel count, but VEGF concentration did not significantly influence diseasefree survival. Although VEGF protein was found at a significantly higher concentration in malignant than in nonmalignant tissue, determination of intratumoral VEGF protein by an enzyme immunoassay was not prognostically relevant in our patient population. Int. J. Cancer 74:455-458, 1997.