Search

Aim: To evaluate the growth of Human Gingival Fibroblasts (HGFs) cultured onto sample discs of CAD/CAM zirconia and veneering ceramic for zirconia by means of Scanning Electron Microscope (SEM) analysis at different experimental times., Methods: A total of 26 experimental discs, divided into 2 groups, were used: Group A) CAD/CAM zirconia (3Y-TZP) discs (n=13); Group B) veneering ceramic for zirconia discs (n=13). HGFs were obtained from human gingival biopsies, isolated and placed in culture plates. Subsequently, cells were seeded on experimental discs at 7,5×10(3)/cm(2) concentration and cultured for a total of 7 days. Discs were processed for SEM observation at 3h, 24h, 72h and 7 days., Results: In Group A, after 3h, HGFs were adherent to the surface and showed a flattened profile. The disc surface covered by HGFs resulted to be wider in Group A than in Group B samples. At SEM observation, after 24h and 72h, differences in cell attachment were slightly noticeable between the groups, with an evident flattening of HGFs on both surfaces. All differences between Group A and group B became less significant after 7 days of culture in vitro., Conclusions: SEM analysis of HGFs showed differences in terms of cell adhesion and proliferation, especially in the early hours of culture. Results showed a better adhesion and cell growth in Group A than in Group B, especially up to 72h in vitro. Differences decreased after 7 days, probably because of the rougher surface of CAD/CAM zirconia, promoting better cell adhesion, compared to the smoother surface of veneering ceramic.

Objectives: The present research has been performed to evaluate whether a commercial magnesium-enriched hydroxyapatite (MgHA)/collagen-based scaffold engineered with ovine amniotic fluid mesenchymal cells (oAFMC) could improve bone regeneration process in vivo., Materials and Methods: Bilateral sinus augmentation was performed on eight adult sheep in order to compare the tissue regeneration process at 45 and 90 days after implantation of the oAFMC-engineered scaffold (Test Group) or of the scaffold alone (Ctr Group). The process of tissue remodeling was analyzed through histological, immunohistochemical, and morphometric analyses by calculating the proliferation index (PI) of oAFMC loaded on the scaffold, the total vascular area (VA), and vascular endothelial growth factor (VEGF) expression levels within the grafted area., Results: MgHA/collagen-based scaffold showed high biocompatibility preserving the survival of oAFMC for 90 days in grafted sinuses. The use of oAFMC increased bone deposition and stimulated a more rapid angiogenic reaction, thus probably supporting the higher cell PI recorded in cell-treated sinuses. A significantly higher VEGF expression (Test vs. Ctr Group; p = 0.0004) and a larger total VA (p = 0.0006) were detected in the Test Group at 45 days after surgery. The PI was significantly higher (p = 0.027) at 45 days and became significantly lower at 90 days (p = 0.0007) in the Test Group sinuses, while the PI recorded in the Ctr Group continued to increase resulting to a significantly higher PI at day 90 (CTR day 45 vs. CTR day 90; p = 0.022)., Conclusions: The osteoinductive effect of a biomimetic commercial scaffold may be significantly improved by the presence of oAFMC., Clinical Relevance: The amniotic fluid mesenchymal cell (AFMC) may represent a novel, largely and easily accessible source of mesenchymal stem cells to develop cell-based therapy for maxillofacial surgery.

The purpose of this study is to evaluate macroscopic and microscopic appearance of a new implant design, with particular emphasis given to the type of prosthesis connection. Two dental implants of the same type (Torque Type(®), WinSix(®), BioSAFin. S.r.l. - Ancona, Italy), with sandblasted and acid etched surfaces (Micro Rough Surface(®)), but differing from each other for the prosthesis connection system, were examined by scanning electron microscope (SEM) analysis at different magnifications: TTI implant, with a hexagonal internal connection, and TTX implant, with a hexagonal external connection. SEM analysis showed that the Torque Type(®) implant is characterized by a truncated cone shape with tapered tips. The implant body showed a double loop thread and double pitch with blunt tips. For both types of connection, the implant neck was 0.7 mm in height with a 3% taper. This implant design may be able to guarantee osteotomic properties at the time of insertion in a surgical site suitably prepared, a facilitated screwing, thanks to the thread pitch and to the broad and deep draining grooves, thereby ensuring a good primary stability. The different connection design appears defined and precise, in order to ensure a good interface between the fixture and the prosthetic components. Therefore, this design appears to be particularly suitable in cases where a good primary stability is necessary and a precise coupling between endosseous and prosthetic components, as it allows an easy insertion of the fixture even in conditions of reduced bone availability, and in cases of immediately loaded full-arch rehabilitations.

The aim of this study was to analyze histologic and immunohistochemical modifications taking place in maxillary sites reconstructed by iliac crest bone grafts, 4 months (T1) and 10 years (T2) after grafting, in comparison to native iliac crest bone (T0).By light microscopy, T1 samples showed well-organized osteoblasts and extracellular matrix (ECM), and surrounding areas of connective tissue where a discrete number of blood vessels can be distinguished. Samples obtained from reconstructed areas after a long period of follow-up (T2) were characterized by uniform mineralized ECM with cavities containing osteocytes, whereas T0 samples disclosed both mineralized matrix and bone marrow. Matrix metalloproteinase 2 (MMP-2), vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), and bone sialoprotein (BSP) expressions were investigated by immunohistochemical analysis. Both MMP-2 and VEGF showed significantly increased expression in T1, with respect to T0, and lowered in T2. Otherwise, BSP expression, which was elevated in native iliac crest bone (T0), was deeply decreased in T1 and T2 samples. Moreover, changes in iNOS expression and in apoptotic cell nuclei percentages (TUNEL analysis) seemed to have a similar trend, increasing in T1 and lowering in T2.After a period of 4 months, iliac crest bone graft-reconstructed sites show mineralizing nuclei not strongly represented, as suggested by BSP lower expression, whereas new blood vessel proliferation and active remodeling phenomena are developing. After a long period from the grafting (T2), BSP expression decreases along with MMP-2, VEGF, and iNOS, suggesting the presence of only weak arrangement phenomena of the graft.

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' count, showed a significant (P < 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).

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, was found lower in all autologous samples (eAB and iAB) compared to hB, at statistically significant values. Moreover, the expression of the pro-apoptotic molecule, Bax, resulted significantly lower (p < 0.05) in eAB than in iAB and hB samples. These values, together with the low number of apoptotic cells detected in autologous samples, suggest a good regenerative response when extraoral autologous bone graft is used in comparison to the response from the other grafts, and also suggest the use of calvaria graft as a predictable therapeutic procedure for repairing severe bone defects in oral and maxillofacial surgery, not only by clinical and biomechanical criteria, but also from a biomolecular aspect.

The aim of this study is to evaluate and compare the response of bone tissue after osteotomy carried out with either rotating cutters or with piezoelectric terminals.Bioptic samples of bone tissue were taken during operations with rotating burs and piezoelectric terminals to increase bone volume before implantology. Samples first underwent histomorphometric analysis. Subsequently, osteoblastic cells, obtained from different samples, were placed in culture and allowed to proliferate to in vitro evaluate the time to initiate growth and to reach confluence. Finally, a molecular biologic study by reverse transcription polymerase chain reaction was performed to evaluate the expression of typical osteoblastic molecular markers, such as osteoprotegerin and osteopontin.Histomorphometric analysis showed that the width of necrotic line on the osteotomic margins from samples taken using different techniques did not vary significantly. Moreover, the times of initial growth and of confluence in cells from the 2 groups did not show any statistically significant differences. However, a highly significant correlation was revealed between the age of the patient and the initial growth time and the confluence. Similarly, reverse transcription polymerase chain reaction showed that the osteoprotegerin and osteopontin expression levels did not change significantly according to the surgical technique used.In conclusion, osteotomies carried out with either instrument do not seem to substantially influence the vitality of the bone tissue. The variability of the expression levels of typical osteoblastic markers seems to be linked more to other factors than to the surgical technique used.

Introduction: The dental pulp undergoes age-related changes that could be ascribed to physiological, defensive, or pathological irritant-induced changes. These changes are regulated by pulp cell activity and by a variety of extracellular matrix (ECM) macromolecules, playing important roles in growth regulation, tissue differentiation and organization, formation of calcified tissue, and defense mechanisms and reactions to inflammatory stimuli. The aim of this research was to better understand the genetic changes that underlie the histological modification of the dental pulp in aging., Methods: The gene expression profile of the human dental pulp in young and older subjects was compared by RNA microarray analysis that allowed to simultaneously analyze the expression levels of thousands of genes. Data were statistically analyzed by Significance Analysis of Microarrays (SAM) Ingenuity Pathway Analysis (IPA) software. Semiquantitative and real-time reverse-transcriptase polymerase chain reaction analyses were performed to confirm the results., Results: Microarray analysis revealed several differentially expressed genes that were categorized in growth factors, transcription regulators, apoptosis regulators, and genes of the ECM. The comparison analysis showed a high expression level of the biological functions of cell and tissue differentiation, development, and proliferation and of the immune, lymphatic, and hematologic system in young dental pulp, whereas the pathway of apoptosis was highly expressed in older dental pulp., Conclusions: Expression profile analyses of human dental pulp represent a sensible and useful tool for the study of mechanisms involved in differentiation, growth and aging of human dental pulp in physiological and pathological conditions.

Purpose: To evaluate in vivo collagen fiber behavior around two different dental implant necks placed in the mandibular bone of adult pigs., Materials and Methods: 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., Results: 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 values that were lower than those seen around the titanium necks. Histologic observation indicated that collagen fiber orientation was similar for both types of implants. The majority of fibers showed a parallel or parallel-oblique orientation to the implant surface for all samples. Implants that were not osseointegrated, as determined by clinical evaluation, showed inflammatory infiltrate, whereas healthy connective tissue was found around all the other implant necks., Conclusions: Collagen fiber orientation was similar, regardless of implant material, demonstrating a predominantly parallel or parallel-oblique pattern. Moreover, zirconia, which is used as a transgingival collar on some implants, showed connective tissue adhesion that was similar to that seen on the machined titanium surface, but demonstrated limited plaque formation and may provide better esthetics.

The purpose of this study is to characterise the expression of matrix extracellular phosphoglycoprotein (MEPE) in cultured mesenchymal cells isolated from human dental papilla (PaMCs) of impacted third molars either before or during differentiation of these cells into osteo/odontoblasts. PaMCs, like mesenchymal cells deriving from human dental pulp (DPMCs), resulted positive for a number of mesenchymal markers including CD146 and STRO-1. During the first week in culture they showed a faster proliferation rate than DPMCs, coupled to an earlier down-regulation of MEPE. Also when the cells were further cultured in osteogenic medium (containing beta-glycerophosphate, ascorbic acid and dexamethasone) for 40 days, MEPE down-regulation coupled to an increased expression of osteogenic markers, such as osteocalcin and alkaline phosphatase, occurred earlier in PaMCs than in DPMCs. Thus, our data, indicating that also in PaMCs MEPE expression is higher when cells proliferate, whereas it is downregulated as cells differentiated, are in favour of a role of MEPE as an early regulator of odontogenic differentiation. We also confirm the superior proliferative potential of PaMCs in comparison with DPMCs, coupled to a more rapid induction of osteogenic differentiation. Therefore, these cells represent an optimal source to be conveniently used for dental tissue engineering and tooth regeneration.

Cancer of the oral cavity is a frequent worldwide neoplasia, averaging 4% in men and 2% in women. Cancer of the oral cavity is often preceded by particular epithelial lesions, called precancerous lesions. The aim of this study is to evaluate, by SEM microscope analysis, before and after treatment with 0.1% 13-cis-retinoic, the morphostructural variation of the surface of the oral mucous affected by lichen planus in respect to healthy oral mucous. Following topical therapy with 0.1% 13-cis-retinoic acid for 8 weeks, a restoration of the physiological morphostructure was observed by SEM, demonstrating the efficaciousness of the treatment and the SEM analysis for identifying the lesions of oral epithelium.