Antonio Centofanti

Dentin Morphology of Root Canal Surface: A Quantitative Evaluation Based on a Scanning Electronic Microscopy Study

Dentin is a vital, hydrated composite tissue with structural components and properties that vary in the different topographic portions of the teeth. These variations have a significant implication for biomechanical teeth properties and for the adhesive systems utilized in conservative dentistry. The aim of this study is to analyse the root canal dentin going from coronal to apical zone to find the ratio between the intertubular dentin area and the surface occupied by dentin tubules varies. Observations were conducted on 30 healthy premolar teeth extracted for orthodontic reasons in patients aged between 10 and 14. A SEM analysis of the data obtained in different canal portions showed that, in the coronal zone, dentinal tubules had a greater diameter (4.32 μm) than the middle zone (3.74 μm) and the apical zone (1.73 μm). The average number of dentinal tubules (in an area of 1 mm2) was similar in coronal zone (46,798 ± 10,644) and apical zone (45,192 ± 10,888), while in the middle zone they were lower in number (30,940 ± 7,651). However, intertubular dentin area was bigger going from apical to coronal portion. The differences between the analysed areas must be considered for the choice of the adhesive system.

Effect of bisphosphonates on the mandibular bone and gingival epithelium of rats without tooth extraction

Osteonecrosis of the jaw (ONJ) is an adverse effect of bisphosphonate treatment that has become the subject of increasing investigations, in particular due to its poorly understood pathogenesis. Several experimental studies on animal models have been conducted; however, the majority of these replicate human ONJ following tooth extraction, and describe alterations in the bone and gingival epithelium when necrosis is manifested. The aim of the present study was to analyze the rat mandibular bone and gingival epithelium during 45 days of zoledronate treatment (which is a bisphosphonate agent), without tooth extraction. Intraperitoneal injections of zoledronate acid (0.1 mg/kg) were performed three times a week in normal male Wistar rats (n=20), while a control group of rats (n=20) was treated with saline solution for 45 days. After 7, 15, 30 and 45 days of drug treatment, all rats were sacrificed and hematoxilin and eosin staining, immunofluorescence and scanning electron microscopy analyses were performed. The results of the analyses after 7 and 15 days of treatment were similar in the treatment and control group. After 30 and 45 days of treatment, structural alterations were observed in the bone. No structural alterations to the gingival epithelium were observed. Based on these results, it was hypothesized that low doses of zoledronate act directly on the bone tissues to induce morphological alterations from bone to necrotic tissue following surgical procedures, although no cytotoxic effects were detected in the gingival epithelium.

Morphofunctional compensation of masseter muscles in unilateral posterior crossbite patients

Unilateral posterior crossbite is a widespread, asymmetric malocclusion characterized by an inverse relationship of the upper and lower buccal dental cusps, in the molar and premolar regions, on one side only of the dental arch. Patients with unilateral posterior crossbite exhibit an altered chewing cycles and the crossbite side masseter results to be less active with respect to the contralateral one. Few studies about morphological features of masticatory muscle in malocclusion disorders exist and most of these have been performed on animal models. The aim of the present study was to evaluate morphological and protein expression characteristics of masseter muscles in patients affected by unilateral posterior crossbite, by histological and immunofluorescence techniques. We have used antibody against PAX-7, marker of satellite cells, and against α-, β-, γ-, δ-, ε- and ζ-sarcoglycans which are transmembrane glycoproteins involved in sarcolemma stabilization. By statistical analysis we have evaluated differences in amount of myonucley between contralateral and ipsilateral side. Results have shown: i) altered fibers morphology and atrophy of ipsilateral muscle if compared to the contralateral one; ii) higher number of myonuclei and PAX-7 positive cells in contralateral side than ipsilateral one; iii) higher pattern of fluorescence for all tested sarcoglycans in contralateral side than ipsilateral one. Results show that in unilateral posterior crossbite hypertrophic response of contralateral masseter and atrophic events in ipsilateral masseter take place; by that, in unilateral posterior crossbite malocclusion masticatory muscles modify their morphology depending on the function. That could be relevant in understanding and healing of malocclusion disorders; in fact, the altered balance about structure and function between ipsilateral and contralateral muscles could, long-term, lead and/ or worsen skeletal asymmetries.

Sarcoglycan complex in masseter and sternocleidomastoid muscles of baboons: an immunohistochemical study

The sarcoglycan complex consists of a group of single-pass transmembrane glycoproteins that are essential to maintain the integrity of muscle membranes. Any mutation in each sarcoglycan gene causes a series of recessive autosomal dystrophin-positive muscular dystrophies. Negative fibres for sarcoglycans have never been found in healthy humans and animals. In this study, we have investigated whether the social ranking has an influence on the expression of sarcoglycans in the skeletal muscles of healthy baboons. Biopsies of masseter and sternocleidomastoid muscles were processed for confocal immunohistochemical detection of sarcoglycans. Our findings showed that baboons from different social rankings exhibited different sarcoglycan expression profiles. While in dominant baboons almost all muscles were stained for sarcoglycans, only 55% of muscle fibres showed a significant staining. This different expression pattern is likely to be due to the living conditions of these primates. Sarcoglycans which play a key role in muscle activity by controlling contractile forces may influence the phenotype of muscle fibres, thus determining an adaptation to functional conditions. We hypothesize that this intraspecies variation reflects an epigenetic modification of the muscular protein network that allows baboons to adapt progressively to a different social status.

The Effect of Different Cleaning Protocols on Post Space: A SEM Study

Aim. Purpose of the present paper is to analyze the efficiency of different post-space irrigation protocols. Methods. 28 single rooted teeth were endodontically treated. After post-space preparation every sample was assigned to one of three experimental groups and to one control group. In each group different irrigation protocols were performed as follows: EDTA (Group A), 37% orthophosphoric acid (Group B), and EDTA + 37% orthophosphoric acid with ultrasounds activation (Group C). In the control group (Group D) the irrigate association was not activated by ultrasounds. Three zones (coronal, middle, and apical) of each sample were analyzed by using Scan Electron Microscopy (SEM) without any metallization procedures. The presence of smear layer on the canal surface was qualitatively evaluated by applying Serafinos score with values included between 0 and 2. Results. The results of the research showed how Group C recorded the better results (0.81 ± 0.72). Group A and Group B showed lower mean scores (1.06 ± 0.69 and 1.08 ± 0.77); Group D showed the lowest mean score of 1.30 ± 0.69. The SEM observation analysis demonstrated how the smear layer presence decreased in the crown-apical direction. Conclusions. The different post-space treatments statistically determine significant differences on the dentinal surfaces cleansing. The absence of ultrasonic activation lowers the cleansing efficacy of endocanalar irrigants, showing sensible differences among each post-space zone.

Morpho-structural alterations of sub-chondral bone tissue in patients with osteoarthritis: a scanning electron microscopy study

Osteoarthritis focuses principally on the degeneration of articular cartilage as a primary cause of the disease. The pathophysiological process of osteoarthritis is characterized by alteration of chondrocytes and the increased bone formation by sub-chondral osteoblasts. Infiltration of macrophages and perivascular T and B lymphocytes is observed, and these infiltrates have been demonstrated in both early and advanced disease. The morphological and phenotypic characteristics of osteocytic cells attached to the normal and the osteoarthritic matrix differ from each other, suggesting that specific signalling pathways arise or are altered between matrix and cells. On this basis, we have examined biopsies of bone obtained by normal femur and by femur of subjects affected by osteoarthritis using techniques of scanning electron microscopy in order to identify the morphostructural alterations that occur in the sub-chondral bone. Our results have shown that the bone tissue of subjects not affected by any disease of bone presents a well-organized structure, while the bone tissue obtained by patients affected by osteoarthritis shows a derangement of tissue itself possibly correlated with altered function of the osteoblasts, that during the pathological process produce a less mineralized extracellular matrix with consequent loss of the normal bone structure. In our opinion, during the osteoarthritic process there would be a defective signalling between bone cells leading to the production of an irregular, amorphous extracellular matrix by osteoblasts, characteristic of the pathological condition.

Sarcoglycans and integrins in bisphosphonate treatment: immunohistochemical and scanning electron microscopy study.

Osteonecrosis of the jaw is an adverse outcome associated with bisphosphonate treatment. Bisphosphonates are used in conjunction with antineoplastic chemotherapy for the treatment of hypercalcaemia associated with malignancy, lytic bone metastasis and multiple myeloma. However, it is not known if the osteonecrosis of the jaw lesion originates in the bone or whether it initiates in the gingival epithelium. Two bisphosphonates are commonly used in cancer treatment. One of these is pamidronate disodium, a second-generation bisphosphonate that differs from the first-generation drug because it inhibits bone resorption at a dose that does not affect bone mineralization. The other widely used BP, zoledronate, is a third-generation drug that is the most potent bisphosphonate in clinical use, showing strong anti-osteoclastic activity, similar to pamidronate. The aim of the present study was to evaluate the modifications of human oral mucosa and underlying bone in patients after treatment with these nitrogen-containing bisphosphonates for 24 and 36 months. We analyzed the structural damage of the oral mucosa and damage of the perilesional mandibular bone observing possible correlations from them. Our results allow to express two hypotheses about the mechanism responsible for these results relating to mandible matrix necrosis; first, an increased skeletal microdamage associated with turnover suppression occurred early in treatment and progress with longer treatment duration, second, opening damage in osteonecrosis of the jaw modifies structural morphology of gingival epithelium.

An Immunofluorescence Study About Staining Pattern Variability of Sarcoglycans in Ratâs Cerebral and Cerebellar Cortex

Title: An Immunofluorescence Study About Staining Pattern Variability of Sarcoglycans in Rat’s Cerebral and Cerebellar Cortex. Background: Sarcoglycans are transmembrane glycoproteins which connect extracellular matrix components to cytoskeleton. This protein system has been long studied in muscle but there are few data about its localization in non-muscular tissues. Methods and Findings: In the present report, we have conducted an indirect immunofluorescence study on normal rat’s cerebral and cerebellar cortex. Our results show that in these districts each sarcoglycan is expressed by a “spot-like” staining pattern, with spots of 0.5-2 μm average diameter, extending mainly around the soma of neurons and glial cells. In cerebral cortex, although all sarcoglycans are present, a staining pattern variability for each sarcoglycan, in the different cerebral cortex areas, exists. Instead, the pattern distribution level of sarcoglycans in cerebellar cortex doesn’t change. We also performed a statistical analysis which confirms the immunofluorescence results. Conclusions: Then, the presence of a sarcoglycans variability in cerebral cortex, where it is known the existence of several synaptic network, and the absence of a sarcoglycans variability in cerebellar cortex, where the same synaptic networks are repeated unchanged, suggest that in brain sarcoglycans may be associated with synaptic networks. Moreover, the distribution of sarcoglycans, mainly in post-synaptic regions of the neurons, suggests a role of these proteins in cellular signalling, regulating membrane receptors assembly. We also support that sarcoglycans in glial cells could be associated with the regulation of the mechanism in the brain-blood-barrier.

Steam Sterilization of Equine Bone Block: Morphological and Collagen Analysis

Introduction The use of equine bone blocks is widely reported for bone augmentation techniques. The block must be shaped according to the form of the defect that should be regenerated. The shaping could be performed by hand before or during the surgery, in a sterile ambient, or using a CNC milling machine that could not be sterile. The aim of our study was to evaluate if a steam sterilization could provide a medical grade sterilization of the blocks and to evaluate if bone microstructure and collagen structures change after different steam sterilization protocols provided by mainstream autoclave. Materials and Method Two blocks of equine bone were divided into 16 samples. 1 sample was used as control and not submitted to any treatment. 15 samples were infected with a Streptococcus faecalis bacterial culture. The samples were singularly packed, randomly divided into 3 groups, and submitted to autoclave sterilization on the same device. The groups were submitted to a sterilization cycle (Gr. A: 121°C, 1,16 bar for 20′; Gr. B:134°C, 2,16 bar for 4′; Gr. C: 134°C, 2,16 bar for 3.30 min.). 2 samples for each group were evaluated for the sterility. 3 samples for each group were observed at SEM to notice the macro- and microstructure modification and to confocal microscope to observe the collagen. Results All samples were sterile. The SEM evaluation showed, in all groups, a preserved morphological structure. Confocal microscope evaluation shows that the collagen structure appears to be more uniform and preserved in group C. Conclusion Data show that autoclave steam sterilization could be reliable to obtain sterilization of equine bone blocks.

Plica neuropathica (matting hair) in an autistic patient

Dear Editor, Irreversible hair tangle, occurring suddenly after a shampoo, was first described in 1884 by La Page in a 14-year-old hysterical patient and named “plica neuropathica”. Additional observations suggested physical–chemical factors, psychiatric illnesses and drugs to be involved, and a new name was suggested: matting hair. In 1996, we published a study on matting hair occurring soon after an energetic brushing of wet hair. We describe now a relapsing “plica neuropathica” in an autistic girl.