Nuclear fractal dimension in oral squamous cell carcinoma: a novel method for the evaluation of grading, staging, and survival.

Fractal dimension (FD) in tissue specimens from patients with oral squamous cell carcinoma (OSCC) was evaluated. FD values in different stages of OSCC, and the correlations with clinicopathological variables and patient survival were investigated. Histological sections from OSCC and control non-neoplastic mucosa specimens were stained with hematoxylin-eosin for pathological analysis and with Feulgen for nuclear evaluation. FD in OSCC groups vs. controls revealed statistically significant differences (P < 0.001). In addition, a progressive increase of FD from stage I and II lesions and stage III and IV lesions was observed, with statistically significant differences (P = 0.003). Moreover, different degrees of tumor differentiation showed a significant difference in the average nuclear FD values (P = 0.001). A relationship between FD and patients' survival was also detected with lower FD values associated to longer survival time and higher FD values with shorter survival time (P = 0.034). These data showed that FD significantly increased during OSCC progression. Thus, FD could represent a novel prognostic tool for OSCC, as FD values significantly correlated with patient survival. Fractal geometry could give insights into tumor morphology and could become an useful tool for analyzing irregular tumor growth patterns.

Blood vessels are concentrated within the implant surface concavities: a histologic study in rabbit tibia.

Angiogenesis plays a key role in bone formation and maintenance. Bone formation has been reported to initiate in the concavities rather than the convexities in a hydroxyapatite substratum and the implant threads of dental implants. The aim of the present study was to evaluate the number of the blood vessels inside the concavities and around the convexities of the threads of implants in a rabbit tibia model. A total of 32 thread-shaped implants blasted with apatitic calcium phosphate (TCP/HA blend) (Resorbable Blast Texturing, RBT) (Maestro, BioHorizons(®), Birmingham, AL, USA) were inserted in 8 rabbits. Each rabbit received 4 implants, 2 in the right and 2 in left tibia. Implants were retrieved after 1, 2, 4, and 8 weeks and treated to obtain thin ground sections. Statistically significant differences were found in the number of vessels that had formed in the concavities rather than the convexities of the implants after 1 (p = 0.000), and 2 weeks (p = 0.000), whilst no significant differences after 4 (p = 0.546) and 8 weeks (p = 0.275) were detected. The present results supported the hypothesis that blood vessel formation was stimulated by the presence of the concavities, which may provide a suitable environment in which mechanical forces, concentrations and gradients of chemotactic molecules and blood clot retention may all drive vascular and bone cell migration.

Histological and immunohistochemical evaluation of the peri-implant soft tissues around machined and acid-etched titanium healing abutments: a prospective randomised study.

A close spatial correlation has been described between the roughness of intraoral materials and the rate of bacterial colonisation. The aim of the present study in man was to conduct a comparative immunohistochemical evaluation of the inflammatory infiltrate, microvessel density, the nitric oxide synthases 1 and 3 and the vascular endothelial growth factor expression, the proliferative activity, and the B and T lymphocyte and histiocyte positivity in the peri-implant soft tissues around machined and acid-etched titanium healing caps. Ten patients participated in this study. The patients were enrolled consecutively. All patients received dental implants left to heal in a non-submerged mode. Healing caps were inserted in all implants. Half of the implants were supplied randomly with machined caps of titanium (control), while the other half were provided randomly with acid-etched titanium caps (test). After a 6-month healing period, a gingival biopsy was performed with a circular scalpel around the healing caps of both groups. The inflammatory infiltrate was mostly present in test specimens. Their extension was much larger than that of the control samples. A higher number of T and B lymphocytes were observed in test specimens. Higher values of microvessel density and a higher expression of vascular endothelial growth factor intensity were observed in the test samples. Furthermore, the Ki-67, NOS1 and NOS3 expression was significantly higher in the test specimens. All these results showed that the tissues around test healing caps underwent a higher rate of restorative processes, most probably correlated to the higher inflammation processes observed in these tissues.

Immunohistochemical expression of vascular endothelial growth factor (VEGF) in different types of odontogenic cysts.

The objective of the present study was to evaluate vascular endothelial growth factor (VEGF) expression in different types of odontogenic cysts. A total of 25 parakeratotic odontogenic keratocysts (POKCs), 16 orthokeratotic odontogenic keratocysts (OOKCs), and 28 follicular cysts (FCs) were evaluated semiquantitatively for immunohistochemical analysis of VEGF in epithelial cells, endothelial cells of blood vessels, inflammatory cells and focally stromal cells. A significant different expression of VEGF in all cell components was found in keratocysts compared to FCs. The POKCs (80%) and OOKCs (68%) showed more than 50% VEGF positive epithelial cells, whereas the majority of FCs (71%) were either negative in the epithelium or showed less than 10% positive cells. Similarly, the POKCs (88%) and OOKCs (68%) showed more than 50% positive endothelial cells, whereas the FCs (75%) were either negative or showed less than 10% VEGF positive endothelial cells. The highest percentage of cases with score 2 positivity in the stromal cells was observed in POKCs (68%); OOKCs showed a score 2 positivity in 44%, score 1 in 31% and score 0 in 25%, whereas 68% of FCs showed a score 0, 25% a score 1 and only 7% of cases showed a score 2. No statistically significant differences were observed between POKCs and OOKCs in VEGF expression in the epithelial and endothelial cells, whereas the positivity score in stromal cells was significantly higher in POKCs compared to OOKCs. The present results can support the hypothesis that angiogenesis is an active mechanism in the invasive growth of the OKC.

Immunohistochemical analysis of inflammatory infiltrate in aggressive and chronic periodontitis: a comparative study.

This immunohistochemical study evaluated the inflammatory infiltrate with its cluster differentiation markers (CD 4, CD 8, CD 20, and CD 68) in aggressive and chronic periodontitis gingival tissues in order to identify the specific cell distribution. Twenty-seven human gingival biopsies were obtained and analyzed. Fourteen patients were suffering from chronic periodontitis and six from aggressive periodontitis; seven patients with healthy gingiva were included as the control group. The specimens were immunohistochemically stained for anti-CD 4 (T helper cells), anti-CD 8 (T cytotoxic/suppressor), anti CD-20 (B plasma cells) and anti CD-68 (macrophages). Chronic periodontitis samples were mainly dominated by CD 4 and CD 8+ cells. On the contrary, in aggressive periodontitis patients all four cell types (CD 4, CD 8, CD 20 and CD 68 + cells, respectively) were remarkably increased. CD 20+ cells were significantly (p < 0.05) more prevalent in aggressive versus chronic periodontitis. The control samples expressed lower CD 4, CD 8, CD 20 and CD 68+ cells confirming a none inflammatory state. The present study demonstrates prevalence for CD 20+ cells in aggressive periodontitis lesions. However, further studies need to be performed to confirm and identify a clear pattern of inflammatory cells and hereafter the mechanisms sustaining the disease.

Sinus lift with autologous bone alone or in addition to equine bone: an immunohistochemical study in man.

PURPOSE: The aim of the present study was to perform a comparative histological and immunohistochemical evaluation of microvessel density, vascular endothelial growth factor, and nitric oxide synthase (NOS) expression in sinus augmentation using autologous bone alone or in combination with equine bone. MATERIALS AND METHODS: Sixteen patients underwent sinus augmentation with autologous bone and a 50:50 mixture of autologous and equine bone. Control cores were harvested from preexisting nonaugmented bone under the sinus floor. The specimens were processed for immunohistochemistry. RESULTS: The greatest values of microvessel density were found in the sites grafted with autologous bone with significant differences between control versus autologous group (P < 0.01) and control versus autologous + equine group (P < 0.01). The higher and lower intensities of vascular endothelial growth factor and NOS3 expression were prevalent in the sites grafted with autologous bone with significant differences with the controls (P < 0.05). No significant differences (P > 0.05) were found in the NOS1 expression among the groups. CONCLUSION: The results obtained showed that the mixture of autologous and equine bone was biocompatible, and its use was associated with new blood vessels ingrowth during healing, which has been found to be extremely important for bone formation.

Detection of Helicobacter pylori in saliva and esophagus.

The route of Helicobacter pylori transmission remains unclear and the currently suggested route is person-to-person transfer by faecal-oral and oral-oral mode. The aim of this study was to verify the presence of H. pylori in esophagus and saliva of humans. Saliva samples, mucosal biopsies from esophagus, gastric antrum and fundus were collected from 19 patients with positive Urea Breath Test (UBT). Gastric biopsies were used for H. pylori colture and antimicrobial susceptibility tests whereas saliva samples were collected to detect H. pylori with a Nested-PCR targeting 16S rRNA gene as well as esophagus biopsies which were also investigated with immunohistochemical staining. Helicobacter pylori was isolated in 18 patients both in gastric antrum and fundus. The molecular analysis, confirmed by comparative sequences evaluation, gave positive results in all saliva and esophageal samples whereas the immunohistochemistry revealed the presence of H. pylori in 15.8% (3/19) of the esophagus samples. Our data suggest that saliva and esophagus may be considered reservoirs for H. pylori in humans and emphasize the need to use more susceptible techniques for H. pylori detection, in particular in over-crowded sites. Identification of the transmission route of H. pylori is crucial in developing an effective plan of surveillance by finding new means of disease management.

Loss of expression of TGF-beta1, TbetaRI, and TbetaRII correlates with differentiation in human oral squamous cell carcinomas.

Despite advances in biological and molecular characteristics, the prognosis of oral squamous cell carcinomas is still very unfavourable and is based on the classical clinicopathological parameters. However, tumors with similar clinicopathological characteristics may differ dramatically in their clinical outcome. Thus, the identification of novel prognostic factors is necessary to improve prognostic and therapeutic approaches. Transforming growth factor-beta1 (TGF-beta1) is a potent growth inhibitor of epithelial cell proliferation, thus, inactivation of TGF-beta1 signalling may play a role in cancer. The expression levels of TGF-beta1 and its type I and type II receptors (TbetaRI and TbetaRII) were assessed by immunohistochemical and Western blot analyses in 22 oral squamous cell carcinoma lesions, in their normal adjacent mucosa and in the squamous carcinoma cell lines FaDu and CAL27. Immunohistochemistry on 22 oral carcinomas and case-matched normal oral mucosae demonstrated that TGF-beta1, TbetaRI, and TbetaRII were intensively and homogeneously expressed in all normal epithelia. In contrast, TGF-beta1 and its receptors were significantly reduced in poorly (G3) differentiated tumors as compared to moderately (G2) and well differentiated (G1) lesions (p=2.8 x 10(-3), p=1.3 x 10(-3), p=2.8 x 10(-3) and p=1.3 x 10(-3), respectively). The progressive reduction of the expression levels was confirmed by Western blotting. The oral squamous carcinoma cell lines Cal27 and FaDu demonstrated a reduced and a lack of TbetaRI expression, respectively. A significant decrease of TbetaRII expression, as compared to Cal27 cells, was shown in FaDu cells. Thus, the decreased expression of TbetaRII combined with the absence of TbetaRI could account for the resistance of FaDu cells to the growth-inhibiting effect of TGF-beta1. TGF-beta1 and TGF-beta1 receptor expression significantly decreased as tumors became less differentiated and thus more aggressive, suggesting a functional role of these molecules in oral tumor progression.

P16 expression in odontogenic tumors.

AIMS AND BACKGROUND: The aim of the study was to examine the immunohistochemical expression of a cell-cycle-related factor (p16) in order to elucidate its role in the growth and diffusion of odontogenic tumors. STUDY DESIGN: Thirty-six odontogenic tumors were divided into two groups according to their clinical behavior: group A and group B composed of tumors at low and high risk of recurrences, respectively. The ANOVA test was used to detect differences between the two groups. RESULTS: p16 was expressed in both groups, but with different localization. A statistically significant difference was found in p16 expression of peripheral cells, with an increase in the expression in group B compared to group A (P < 0.05). In addition, there was no significant difference in p16 positive expression of the central cells of odontogenic tumors, which was high in both groups. CONCLUSIONS: The present data show a correlation between p16 expression and the biological behavior of odontogenic tumors.

Expression of transforming growth factor beta1 in ameloblastomas.

The aim of the present study was to compare the expression of transforming growth factor (TGF) beta1 in ameloblastomas (AMs) with different risk of recurrence by immunohistochemistry. A total of 29 cases of AMs were evaluated. The tumors were divided into 2 groups: group A (10 cases) composed of unicystic and peripheral AMs, associated with a low risk of recurrence, and group B (19 cases) composed of solid AMs, associated with a high risk of recurrence. Statistical evaluation showed significant differences between groups A and B lesions, with higher values of positivity for TGF-beta1 in stromal cells in group B tumors (P = 0.0308). No statistically significant differences of immunoreactivity were found in vessels and odontogenic epithelium between the 2 groups. The increased TGF-beta1 expression in tumors with a high risk of recurrence could be explained with the fact that although TGF-beta acts as a potent tumor suppressor in the early stages of tumor progression, later it seems to enhance the invasive phenotype of the tumor.

KAI-1 protein expression in odontogenic cysts.

The KAI-1 tumor suppressor gene is widely distributed in normal tissues and its down-regulation may be correlated with the invasive phenotype and metastases in several different epithelial tumors. The aim of the present study was an evaluation of KAI-1 expression in radicular cysts (RC), follicular cysts (FC), orthokeratinized keratocysts (OOKC), and parakeratinized keratocysts (POKC). Eighty-five odontogenic cysts, 28 RC, 22 FC, and 35 OKC (16 OOKC, 19 POKC) were selected. All the POKC were negative and only four of 16 of the OOKC were positive for KAI-1. On the contrary, all RC and FC cases were positive and immunoreactivity for KAI-1 was detected throughout all the layers of the cyst epithelium. The lack of KAI-1 expression in POKC could help to explain the differences in the clinical and pathologic behavior of OKC and, according to what has been reported for epithelial tumors, could be related to the increased aggressive behavior and invasiveness of OKC.

Comparative evaluation of the peri-implant bone tissue mineral density around unloaded titanium dental implants.

OBJECTIVE: The mechanical properties of bone are greatly influenced by the percentages of organic and mineral constituents. Nevertheless, the information about the mineral content on a microscopic scale in peri-implant bone is scarce. The aim of this work was to analyze the bone mineral density of peri-implant bone under different techniques. DESIGN: Five unloaded titanium dental implants with a micro-structured surface (three XiVE plus and two Frialit 2, DENTSPLY-Friadent, Mannheim, Germany) were retrieved from the mandible of five patients after a 6-month period. scanning electron microscopy with backscattered electron signal (BSE), light microscopy (LM) with a double staining technique, fluorescence microscopy and confocal laser microscopy were used for measuring microscopic mineral content variations in peri-implant bone. Histomorphometry and image intensity (grey level) were evaluated using a software package for image analysis. RESULTS: The low mineral density index (LMDI) for LM was of 29.2+/-3.1 (mean+/-S.D.), while the high mineral density index (HMDI) was of 88.2+/-3.6 (mean+/-S.D.). The one-way ANOVA analysis showed a significant difference (P<0.001) among the groups. The pairwise Holm-Sidak test identified the differences among HMDI indexes for both LM and SEM values and also for cross-evaluation of the LMDI and HMDI values. The comparison between LMDI indexes for both SEM and LM did not show any significance. The fluorescence microscopy analysis showed clearly the difference between old (high mineralized) and new (low mineralized) bone tissue near the implant surface. Under confocal laser microscopy the same sections showed the area of bone modelling closest to implant surface. CONCLUSION: In this study it was found that bone around unloaded implants showed a low mineral density index under all the investigation methods used. It was also found that the conventional LM technique with the double staining method was able to intensely stain the bone area with a low mineral content.

Inflammatory infiltrate, microvessel density, nitric oxide synthase expression, vascular endothelial growth factor expression, and proliferative activity in peri-implant soft tissues around titanium and zirconium oxide healing caps.

BACKGROUND: The aim of the present study in humans was to conduct a comparative immunohistochemical evaluation of vascular endothelial growth factor (VEGF) and nitric oxide synthase (NOS) expression, inflammatory infiltrate, proliferative activity expression, and microvessel density (MVD) in peri-implant soft tissues of titanium and zirconium oxide healing caps. METHODS: Five patients, three men and two women (aged 30 to 66 years; mean: 49 years), participated in this study. All patients received dental implants that were 3.8 mm in diameter and 11 mm in length. All implants were left to heal in a non-submerged (single-stage) mode. Healing caps (3.8 mm in diameter and 3.0 mm in height) were inserted in all implants. Half of the implants were supplied with standard, prefabricated caps of commercially pure titanium, whereas the other half were provided with test zirconium oxide caps. After a 6-month healing period, a gingival biopsy was performed with a circular scalpel (5.5 mm in diameter) around the healing caps of both groups, without unscrewing or removing the healing caps. The dimensions of the gingival biopsies were 1.7 mm (5.5-3.8 mm) in thickness and 3 mm in height. RESULTS: Statistically significant differences were found in the microvessel density between titanium and zirconium oxide healing caps and group II (P < or = 0.0001). Statistically significant differences were likewise found in the low and high intensities of NOS1, NOS3, and VEGF (P < or = 0.0001). In conclusion, the high intensity of NOS1, NOS3, and VEGF were mostly expressed in the titanium group, whereas the low intensity of NOS1, NOS3, and VEGF were mostly expressed in the zirconium oxide group. CONCLUSIONS: In our specimens, the inflammatory infiltrate was mostly present in the titanium specimens. Their extension was much larger than that of the zirconium oxide specimens. Higher values of MVD were observed in the titanium specimens (29.1 versus 15.8). In addition, a higher expression of VEGF intensity was observed in the peri-implant tissues of titanium healing caps, whereas predominantly lower expressions of VEGF intensity were noted around the zirconium oxide healing caps. The Ki-67 expression was higher in the titanium specimens. All these data revealed that the tissues around titanium healing caps underwent a higher rate of inflammation-associated processes, most probably correlated to the higher inflammation processes observed in these tissues. A higher intensity expression of NOS1 and NOS3 was recorded in the tissues around titanium, whereas, on the contrary, a lower intensity of expression was found in the tissues around zirconium oxide specimens. These latter data indicate that the higher expression of these two mediators could be correlated to the higher amount of bacteria present around the titanium samples.

Screw vs cement-implant-retained restorations: an experimental study in the beagle. Part 2. Immunohistochemical evaluation of the peri-implant tissues.

Crestal bone loss has been reported to occur around dental implants. Even if the causes of this bone loss are not completely understood, the presence of a microgap between implant and abutment with a possible contamination of the internal portion of the implants has been suggested. The aim of this study was to see if there were differences in the vascular endothelial growth factor (VEGF) expression, microvessel density (MVD), proliferative activity (MIB-1), and inflammatory infiltrate in the soft tissues around implants with screwed and cemented abutments. Sandblasted and acid-etched implants were inserted in the mandibles of 6 Beagle dogs. Ten 3.5- x 10-mm root-form implants were inserted in each mandible. A total of 60 implants (30 with screwed abutments and 30 with cemented abutments) were used. After 12 months, all the bridges were removed and all abutments were checked for mobility. A total of 8 loosened screws (27%) were found in the screwed abutments, whereas no loosening was observed in cemented abutments. A gingival biopsy was performed in 8 implants with cemented abutments, in 8 implants with screwed abutments, and in 8 implants with unscrewed abutments. No statistically significant differences were found in the inflammatory infiltrate and in the MIB-1 among the different groups. No statistically significant difference was found in the MVD between screwed and cemented abutments (P = .2111), whereas there was a statistically significant difference in MVD between screwed and unscrewed abutments (P = .0277) and between cemented and unscrewed abutments (P = .0431). A low intensity of VEGF was prevalent in screwed and in cemented abutments, whereas a high intensity of VEGF was prevalent in unscrewed abutments. These facts could be explained by the effects induced, in the abutments that underwent a screw loosening, by the presence of bacteria inside the hollow portion of the implants or by enhanced reparative processes.

Suppression of the functionally coupled cyclooxygenase-2/prostaglandin E synthase as a basis of simvastatin-dependent plaque stabilization in humans.

BACKGROUND: The clinical benefits of statins are attributed to changes in plaque composition that lead to reduced metalloproteinase (MMP) activity and plaque stabilization. However, the molecular mechanism of this effect is unclear. Recently, we demonstrated enhanced expression of isoforms of inducible cyclooxygenase (COX) and PGE synthase (COX-2/mPGES) in human symptomatic plaque and provided evidence that this is associated with MMP-induced plaque rupture. The aim of this study was to characterize the effect of simvastatin on inflammatory infiltration and the expression of COX-2/mPGES and MMPs in human carotid plaques. METHODS AND RESULTS: Seventy patients with symptomatic carotid artery stenosis were randomized to the American Heart Association Step 1 diet plus simvastatin (40 mg/d) or the American Heart Association Step 1 diet alone for 4 months before endarterectomy. Plaques were subjected to analysis of COX-1, COX-2, mPGES, MMP-2 and MMP-9, lipid and oxidized LDL (oxLDL) content, and collagen content by immunocytochemistry, Western blot, and reverse transcription-polymerase chain reaction, whereas zymography was used to detect MMP activity. Immunocytochemistry was also used to identify CD68+ macrophages, CD3+ T-lymphocytes, smooth muscle cells (SMCs), and HLA-DR+ inflammatory cells. Plaques from the simvastatin group had fewer (P<0.0001) macrophages, T-lymphocytes, and HLA-DR+ cells; less (P<0.0001) immunoreactivity for COX-2/mPGES and MMPs; reduced (P<0.0001) gelatinolytic activity; increased (P<0.0001) collagen content; and reduced (P<0.0001) lipid and oxLDL content. Interestingly, COX-2/mPGES inhibition by simvastatin was completely reversed by mevalonate in vitro. CONCLUSIONS: This study demonstrates that simvastatin decreases inflammation and inhibits COX-2/mPGES expression in plaque macrophages, and this effect in turn may contribute to plaque stabilization by inhibition of MMP-induced plaque rupture.

Oxygen and life span: chronic hypoxia as a model for studying HIF-1alpha, VEGF and NOS during aging.

To test if oxygen sensitive mechanisms are affected by hypoxia, we studied hypoxia inducible factor-1alpha (HIF-1alpha), vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) expression by immunohistochemical analysis in young and old rat carotid bodies (CBs) using hypoxia as a model for modulating aging. Four groups of male age-matched Wistar rats (3 and 24 months) were used. Two groups were kept in room air, and two groups were kept under chronic intermittent hypoxia for 12 days. In aged carotid body and in hypoxia the increased expression of HIF-1alpha, VEGF, iNOS is less evident as compared to the young one. Electron microscopy sections showed a reduced mitochondrial number and area in the aged CBs and during hypoxia. Less responsiveness to hypoxia could be evidenced in the aged rats as compared to the young rats, suggesting an age dependency of the oxygen sensitive mechanisms.

Vascular endothelial growth factor (VEGF) expression in healthy and inflamed human dental pulps.

Vascular endothelial growth factor (VEGF) is a glycoprotein that has the capability to increase vascular proliferation and permeability. VEGF has been found to be expressed in several different types of tumors, and it may contribute to the progression of malignant tumors. Immunostaining for VEGF and factor VIII was performed in normal healthy pulps and in irreversible pulpitis. In both cases the vessels were always positive for VEGF. Our immunohistochemical data show that the expression of VEGF was strongly positive in the inflammatory infiltrate in irreversible pulpitis. VEGF expression in the stromal cells in healthy pulps ranged from 20 to 100% (with a mean of 68.82), and in irreversible pulpitis ranged from 0 to 100% (with a mean of 62.35%); this difference was statistically significant (p = 0.05). This down-regulation in the stromal cells in irreversible pulpitis could be due to the presence, in a low compliance system such as the dental pulp, of inflammatory infiltrate. VEGF is probably a factor implicated in the etiology and progression of pulpitis. The microvessel density in healthy pulps was 90.00 +/- 27.5, while, in irreversible pulpitis, it was 56.68 +/- 21.15. This difference was statistically significant (p = 0.001). The decrease in microvessel density in irreversible pulpitis could be related to failing vascular function and blood flow decrease.

Osteocyte density in the peri-implant bone of implants retrieved after different time periods (4 weeks to 27 years).

Bone tissue is characterized by a constant turnover in response to mechanical stimuli, and osteocytes play an essential role in bone mechanical adaptation. However, little to no information has been published regarding osteocyte density as a function of implantation time in vivo. The aim of this retrospective histological study was to evaluate the osteocyte density of the peri-implant bone in implants retrieved because of different reasons in a time period from 4 weeks to 27 years. A total of 18 samples were included in the present study. Specimens were divided into 3 groups depending on the loading history of the implants: loading between 4 weeks and 7 months (group 1); loading between 1 and 5 years (group 2); loading between 14 and 27 years (group 3). All the samples were histologically evaluated and osteocyte density was obtained using the ratio of the number of osteocytes to the bone-area (mm(2) ). The osteocyte density values significantly increased in the Group 2 (1-5 years) compared with Group 1 (4 weeks-7 months), and significantly decreased in the Group 3 (14-27 years) compared to Group 2. No significant differences were detected between Group 1 and Group 3. The decrease in osteocyte density observed in samples that were in vivo for long periods of time under loading is possibly because of the fact that once the bone structure is well aligned and biomechanically competent, a lower number of osteocytes are necessary to keep the tissue homeostasis under loading.

Matrix metalloproteinases 2, 3, 8, 9, and 13 in the peri-implant soft tissues around titanium and zirconium oxide healing caps.

PURPOSE: This human study sought to compare, from an immunohistochemical point of view, matrix metalloproteinases (MMPs) 2, 3, 8, 9, and 13 in the soft tissues around titanium and zirconium oxide healing caps. MATERIALS AND METHODS: Five patients participated in this study. All patients received 3.8 × 11-mm dental implants, which were left to heal in a nonsubmerged (single-stage) mode. Healing caps (3.8 mm in diameter and 3.0 mm in height) were inserted in all implants. Half of the implants were randomly supplied with standard, prefabricated caps of commercially pure titanium (control), while the other half were randomly provided with zirconium oxide caps (test). After a 6-month healing period, gingival biopsy specimens were obtained with a circular scalpel around the healing caps of both groups, without unscrewing or removing the healing caps, and the samples underwent immunohistochemical processing for MMPs 2, 3, 8, 9, and 13. RESULTS: Statistically significant differences were found in the values of MMP-8 in the cells of the inflammatory infiltrate, with higher values for the titanium samples. Statistically significantly higher values were found, also in the titanium samples, for MMP-9 in the endothelial cells of the blood vessels. No statistically significant differences were found for any other MMPs. CONCLUSIONS: The present results showed that the soft tissues around titanium healing caps underwent a higher rate of restorative processes, most probably correlated to the MMP levels observed in the tissues.

Hemostasis control in endodontic surgery: a comparative study of calcium sulfate versus gauzes and versus ferric sulfate.

INTRODUCTION: Calcium sulfate (CaS) is a simple, biocompatible material with a long history of safe use in different fields of medicine. CaS is a rapidly resorbing material that leaves behind a calcium phosphate lattice, which promotes bone regeneration and hemostasis. The aim of this study was a clinical evaluation of the hemostatic effect of CaS hemi-hydrate (CaSO4), commonly known as plaster of Paris, in endodontic surgery. METHODS: Twenty-four patients with 31 periradicular lesions were enrolled in this study. The apical roots were exposed, and the bleeding would have made it difficult to correctly fill the root-end cavities. To avoid such an inconvenience, the teeth were divided into 3 groups. Hemostasis was attempted by using CaS in 11 teeth (group I), gauze tamponade in another 10 teeth (group II), or 20% ferric sulfate in the last 10 teeth (group III). RESULTS: Control of the bleeding was achieved in all teeth of group I, whereas in group II adequate hemostasis was achieved in 3 of 10 cases and in group III in 6 of 10 cases. CONCLUSIONS: The use of CaS completely eliminated the bleeding, with a very good level of hemostasis.