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"Endothelial Progenitor Cells (EPCs), a minor subpopulation of the mononuclear cell fraction in peripheral blood, play a critical role in cancer development as they contribute to angiogenesis-mediated pathological neovascularization. In response to tumor cytokines, including VEGF, EPCs mobilize from the bone marrow into the peripheral circulation and move to the tumor bed where they incorporate into sprouting neovessels. In the present study, we evaluated the effects of everolimus (Afinitor, Novartis), a rapamycin analogue, alone or in combination with chloroquine, a 4-alkylamino substituted quinoline family member, one of the autophagy inhibitors, on EPCs biological functions. We found that either everolimus or chloroquine induce growth inhibition on EPCs in a dose-dependent manner after 72 h from the beginning of incubation. The combined administration of the two drugs to EPC was synergistic in inducing growth inhibition; in details, the maximal pharmacological synergism between everolimus and chloroquine in inducing growth inhibition on EPCs cells was recorded when chloroquine was administered 24 h before everolimus. Moreover, we have studied the mechanisms of cell death induced by the two agents alone or in combination on EPCs and we have found that the synergistic effect of combination on EPC growth inhibition was paralleled by increased apoptosis induction and reduced autophagy. These effects occurred together with biochemical features that are typical of reduced autophagic death such as increased co-immunoprecipitation between Beclin 1 and Bcl-2. Chloroquine antagonized the inhibition of the activity of Akt→4EBP1 axis mediated by everolimus and at the same time it blocked the feed-back activation of Erk-1\/2 induced by RAD in EPCs. These data suggest a new strategy in order to block angiogenesis in tumours in which this process plays a key role in both the sustainment and spreading of cancer cells"

The aim of the present study was to perform an immunophenotypic analysis of human gingival fibroblast cells and its eventual modulation by Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF).Gingival fibroblasts were derived from gingival biopsy of 15 healthy subjects. The presence of fibroblast cells in culture and the absence of epithelial cells was performed with fluorescence microscopy using vimentin and cytokeratin markers, respectively. Molecular expression of gingival fibroblast cell membrane was carried out with monoclonal antibodies by flow cytometry analysis. Human recombinant GM-CSF at the concentration of 200 ng/ml was used for the in vitro stimulation of gingival fibroblasts. Statistical analysis was performed using the Student "t"-test.Human gingival fibroblasts express a wide surface molecular panel including mainly CD59, CD99, CD9, CD95, CD55, CD63, CD26, CD117, CD71 and CD86. The GM-CSF seems to regulate the CD49B expression positively and the CD40 and CD103 expression negatively.Results show that GM-CSF is able to modulate the in vitro expression of some membrane molecules of gingival fibroblasts and therefore it may regulate, in vivo, peculiar specific biological functions of gingival tissue.

Background: The aim of this cross-sectional study was to (i) determine the prevalence and distribution of developmental grooves in a young population and (ii) to evaluate the local periodontal conditions. Methods: Two hundred and fifty-one students with a mean age of 22.9 ± 4.7, attending the School of Dentistry and Dental Hygiene of Vita-Salute San Raffaele University (Milan, Italy) were included. The subjects underwent a clinical evaluation by two calibrated examiners. The following clinical parameters were recorded for each site presenting a radicular groove and for each corresponding site on an adjacent tooth used as control: probing pocket depth, plaque index, bleeding on probing, recession depth. Results: The prevalence of radicular grooves at patient and tooth level was 15.9% and 5%, respectively. When compared to control sites, the number of teeth with a radicular groove that presented plaque and bleeding on probing was higher. The logistic regression analysis showed that the presence of radicular grooves was significantly associated with the presence of plaque (OR, 6.14, p < 0.001) and of bleeding on probing (OR, 2.91, p = 0.01). Conclusions: The presence of radicular grooves increases the possibility of developing gingival inflammation by acting as a plaque retentive factor.

The aim of this cross-sectional study is to (i) determine the prevalence, extent, severity, and distribution of gingival recessions and patient perception in a young population and (ii) to identify potential risk indicators. Two hundred fifty-one students with a mean age of 22.9 ± 4.7, attending the School of Dentistry and Dental Hygiene of Vita-Salute San Raffaele University (Milan, Italy) were included. The subjects had undergone a clinical evaluation, by two calibrated examiner, and a questionnaire. Demographic and clinical data were collected to evaluate association of these factors with gingival recessions. The prevalence of gingival recessions at patient and tooth level was 39% and 5.2%, respectively. The only factor associated with the presence of GR was age. On the other hand, age and smoking were associated with the extent, whereas BOP, NCCLs and KT were associated with the severity. Out of 98 subjects presenting at least one GR, 63 (64%) were conscious of the presence of the GR. NCCLs were also strongly associated with the perception of the recession by the patient. There is a low prevalence of buccal gingival recessions in this sample of Italian students. More than 50% of the sample was aware of the problem. Almost all patients presenting symptomatology or aesthetic concern requested appropriate therapy. The findings highlight the low relevance of gingival recessions in daily practice and the importance of controlling potential risk indicators in young populations.

Aim Reports regarding prevalence of peri-implant diseases show widely varying prevalence rates, which can be explained partially by variable diagnostic criteria adopted. Furthermore, several different factors have been associated with peri-implant diseases. Hence, the aim of this cross-sectional study is to (a) determine the prevalence, extent and severity of peri-implant diseases in patients enrolled in a university dental clinic and (b) to evaluate the association between peri-implantitis and patient/implant-related factors. Material and Methods A total of 237 subjects from the Dental Department of Vita-Salute San Raffaele University (Milan, Italy) with 831 implants with more than 1 year of follow-up after loading were clinically evaluated. Implants showing bleeding on probing (BOP), with or without suppuration, and/or probing pocket depth (PPD) >= 4 mm, were radiographically analysed. Demographic and clinical data were collected to evaluate by multilevel regression analysis association with peri-implantitis. Results The prevalence of peri-implant mucositis and peri-implantitis was 38.8% and 35%, respectively. Patients with a FMBS > 25%, having >= 4 implants as well as implants with plaque, PPD >= 4 mm or less than 1 mm of keratinized mucosa presented higher odds ratios for peri-implantitis. Conclusions Peri-implant diseases are frequent conditions affecting >70% of the patients. Several patient/implant-related factors may influence the risk for peri-implantitis.

The aim of the present case series was to evaluate the outcomes of the modified coronally advanced tunnel technique (MCAT) using the width of keratinized tissue (KTW) as an indicator to apply the connective tissue graft (CTG) specifically. Seven patients requiring treatment for the presence of multiple gingival RT1 recession defects in the maxilla were enrolled in the study. A total of 36 recessions were treated with MCAT, and the CTG was applied in 16 sites presenting < 2 mm of KTW at baseline. The mean root coverage from baseline to 1 year postsurgery was 90% for the sites treated with MCAT alone and 93.7% for those treated with MCAT+CTG. The increase of KTW was higher in the sites treated with CTG than in the sites treated without it. Within the limitations of the present case series, it can be concluded that the proposed surgical technique is extremely effective in gaining root coverage and reducing the amount of connective tissue harvested from the donor site.

Endoglin (CD105, an accessory component of the TGF-beta receptor complex) expression and distribution on different human tumour cells and its role in cellular proliferation were evaluated. We examined: 1) sixteen human carcinoma cell lines, 2) eight human sarcoma cell lines, 3) five miscellaneous tumour cell lines. HECV (endothelial cells) were employed as a positive control for endoglin expression. Normal Human Dermal Fibroblasts (NHDF) and 293 cells (epithelial kidney cells) were used as normal controls for connective and epithelial tissues, respectively. The results showed that CD105 was poorly expressed in the majority of human carcinoma cells (10/16), whereas it was highly expressed in most human sarcoma cells (7/8), and differently expressed by miscellaneous tumour cell lines. These data reflect endoglin expression by the normal counterparts of tumour cell lines, i.e. NHDF and 293 cells. However, CD105 levels in sarcoma cell lines, even though consistently lower than in NHDF, were significantly higher than those observed in carcinoma cells. Interestingly, CD105 presented a strong expression in the cytoplasm of MDA-MB-453 (breast carcinoma), NPA (papillary thyroid carcinoma), COLO-853 (melanoma) and SaOS-2 (osteosarcoma), but was weakly expressed on their cell membrane. This differential expression in the cytoplasm and on the membrane of some tumour cells, suggests a complex mechanism of translocation for this protein. The analysis of clonal growth in soft agar of some cell lines, characterized by high CD105 expression, showed an increased colony formation potential that was antagonized by the addition of anti-CD105 blocking mAb. The results indicated that endoglin is differentially expressed in human carcinoma and sarcoma cells and its overexpression modulates the proliferative rate of human solid tumour cells. Moreover, these data suggest that CD105 is involved in the regulation of TGF-beta effects in human solid malignancies, and therefore it could play an important role in tumour diagnosis and treatment.

The Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) is a hematopoietic growth factor that regulates the in vitro and in vivo proliferation and differentiation of hematopoietic cells through the interaction with a specific heterodimeric receptor complex (GM-CSFR), consisting of an alpha and a beta chain with molecular weights of 80 and 120 KDa, respectively. We have studied the expression of the GM-CSFR (alpha chain) on the surface of the human osteosarcoma cell line SaOS-2 and the in vitro effects of different concentrations (10, 100, and 200 ng/ml) of GM-CSF on GM-CSFR expression and the biological activity of SaOS-2 cells. Our data show that SaOS-2 cells express GM-CSFR and that GM-CSF can down-regulate the expression of its own receptor on these cells. Furthermore, to evaluate the biological effects of GM-CSF on SaOS-2 cells, we have investigated cell proliferation and differentiation of these cells treated with different doses of the growth factor through: (1) a morphological analysis of typical osteoblast differentiation markers such as osteopontin and BSP-II; (2) measurement of alkaline phosphatase (ALP) activity; (3) production of bone ECM components (collagen I, fibronectin, tenascin, and laminin); (4) production of interleukin-6 (IL-6) and osteocalcin in the culture medium. The results show that the in vitro treatment of SaOS-2 cells with recombinant human GM-CSF causes a decreased cell proliferation and an increased production of osteopontin, BSP-II, ALP, IL-6, and most but not all ECM components. These findings suggest that GM-CSF can regulate proliferation and differentiation of osteoblast-like SaOS-2 cells and could also play an unexpected role in the maturation of bone tissue.