Experimental nano calcium aluminate/tri calcium silicate root repair: Synthesis, physical and mechanical properties compared to mineral trioxide aggregate and Biodentine / Reparo radicular experimental de nano aluminato de cálcio/silicato de tri cálcio: Síntese, propriedades físicas e mecânicas em comparação com agregado de trióxido mineral e Biodentine

Objective: Endodontic perforation is a challenging mishap that should be repaired with a biocompatible material, Mineral trioxide aggregate (MTA) and Biodentine are the most commonly used repair materials. However, these materials are expensive, (MTA) has prolonged setting time and difficult manipulation. The purpose of this study is to prepare the experimental nano calcium-aluminate/tri-calcium-silicate (CA/C3S) material and comparing its physical properties with biodentine and MTA, to evaluate the experimental material eligibility to compete the commercial repair materials. And to perform part two (animal study) that will evaluate the cytotoxicity, the biocompatibility and the efficacy of (CA/C3S) in furcal perforation repair compared to diode laser. Material and Methods: A mixture of calcium carbonate and aluminum oxide was used to formulate calcium aluminate phase (CA), tri-calcium-Silicate phase (C3S) was formulated by firing of calcium carbonate and quartz. The produced powders were investigated by X-ray diffraction, then (CA) and (C3S) mixed with water.(CA/ C3S) compared with MTA and biodentine for setting-time, micro-hardness, dimensional-stability and solubility. Results: Mean setting time of (CA/C3S) was (32.70±0.75min) which is significantly higher than MTA and Biodentine. The Mean microhardness of (CA/C3S) was (56.50±7.41VHN) which has no statical difference with MTA and Biodentine. Solubility results showed weight increase for (CA/C3S) as following (6.29±3.05)and loss of weight for MTA and Biodentine. The percentage of change in dimensions for(CA/C3S) increased as following (0.64±0.78) while decreased for MTA and Biodentine. Conclusion: The experimental (CA/C3S) material showed good microhardness, dimensional stability and acceptable setting time that could be improved in further work (AU)

Objetivo: A perfuração endodôntica é um percalço desafiador que deve ser reparado com um material biocompatível, Agregado de trióxido mineral (MTA) e Biodentina são os materiais de reparo mais comumente usados. No entanto, esses materiais são caros, (MTA) tem tempo de presa prolongado e difícil manipulação. O objetivo deste estudo é preparar o material experimental de nano aluminato de cálcio/silicato tricálcico (CA/C3S) e comparar suas propriedades físicas com biodentina e MTA, para avaliar a elegibilidade do material experimental para competir com os materiais de reparo comerciais. E realizar a segunda parte (estudo animal) que avaliará a citotoxicidade, a biocompatibilidade e a eficácia do (CA/C3S) no reparo de perfuração de furca em comparação ao laser de diodo.Material e Métodos: Uma mistura de carbonato de cálcio e óxido de alumínio foi usada para formular a fase de aluminato de cálcio (CA), a fase tri-cálcio-silicato (C3S) foi formulada por queima de carbonato de cálcio e quartzo. Os pós produzidos foram investigados por difração de raios X, em seguida (CA) e (C3S) misturados com água. (CA/ C3S) comparados com MTA e biodentina para tempo de presa, microdureza, estabilidade dimensional e solubilidade. Resultados: O tempo médio de presa de (CA/C3S) foi (32,70±0,75min) que é significativamente maior que MTA e Biodentine. A microdureza média de (CA/C3S) foi (56,50±7,41VHN) que não tem diferença estática com MTA e Biodentine. Os resultados de solubilidade mostraram aumento de peso para (CA/C3S) conforme a seguir (6,29±3,05) e perda de peso para MTA e Biodentine. A porcentagem de mudança nas dimensões para (CA/C3S) aumentou como segue (0,64±0,78), enquanto diminuiu para MTA e Biodentine. Conclusão: O material experimental (CA/C3S) apresentou boa microdureza, estabilidade dimensional e aceitável tempo de presa, que pode ser melhorado em trabalhos futuros (AU)

Candida Albicans Alcohol Dehydrogenase 1 gene in oral dysplasia and oral squamous cell carcinoma.

This study was designed to examine the prevalence of Candida albicans alcohol dehydrogenase 1 (CaADH1) gene in oral dysplasia as well as oral squamous cell carcinoma (OSCC) with and without lymph node metastasis (LNM) using reverse transcription polymerase chain reaction (RT-PCR) in order to determine its role in initiation, propagation and metastasis of oral dysplasia and carcinoma. Formalin-fixed parafin-embedded specimens were grouped into four groups: 7 control, 16 oral dysplasia, 16 OSCC without LNM and 15 OSCC with LNM. All specimens were examined by periodic acid Schiff (PAS) stain to detect Candida hyphae, while CaADH1 gene was detected using RT-PCR. Candida hyphae were detected by PAS stain in one specimen of oral dysplasia group, 5 OSCC without LNM and 5 OSCC with LNM. CaADH1 gene was detected in 29 specimens (one case of sever dysplasia, 16 OSCC without LNM and 12 OSCC with LNM), with a highly statistically significant between the groups. CaADH1 gene is associated with OSCC with and without metastasis whereas in oral dysplasia it could not be estimated. Further studies with larger sample size are needed to confirm the role of CaADH1 in oral dysplasia and OSCC.

Histomorphometric evaluation of a nano-sized eggshell-containing supplement as a natural alloplast: An animal study.

BACKGROUND: Limitations of autogenous grafts have dictated the need for development of new biomaterials that can serve as allografts. A paradigm shift directed manufacturers to revert to nature in the search for such allografts. This study aimed to evaluate an eggshell-based supplement, Membrell's® BONEhealth Plus D3 & K2, indicated to support bone mineral density, as a natural bone graft material. METHODS: Twelve 5 × 10 × 1 mm full-thickness cranial bone defects were created in six adult male New Zealand rabbits. Six defects were filled with Membrell's® BONEhealth™ Plus D3 & K2, and the others were left empty as control. The animals were sacrificed 14 days postoperatively. The defects were dissected and prepared for histological assessment. Bone formation was compared both qualitatively and quantitatively. The area percent of newly-formed bone was evaluated in five successive regions using image analysis. Statistical analysis was performed using unpaired t-test. Differences between the two groups were considered significant at p ≤ 0.05. RESULTS: Cranial bone defects filled with the nano-sized eggshell powder "Membrell's® BONEhealth™ Plus D3 & K2" revealed significantly higher levels of osteoid, newly-formed, regularly-arranged bone trabeculae in the center of the defects (47.37% ± 1.12) compared to the control defects (21.6% ± 4.92), which revealed no bone formation. A rapid rate of resorption of the nano-sized eggshell powder and consequently a rapid osteogenic effect was evident. CONCLUSIONS: The eggshell-based graft powder, Membrell's® BONEhealth™ Plus D3 & K2, is a biocompatible material which has the potential to enhance new bone formation.

Development of field cancerization in the clinically normal oral mucosa of shisha smokers.

OBJECTIVES: This study aimed to investigate the changes that occur in the oral mucosa of shisha smokers and compare them to cigarette smokers, nonsmokers and smokers of both shisha and cigarettes and to detect whether these changes could be detected biochemically in saliva. METHODS: Four groups of subjects were included in our study (16 in each group); cigarette smokers, shisha smokers, nonsmokers and smokers of both. A salivary sample and a mucosal biopsy were obtained from each subject. Salivary detection of CYFRA 21-1 was performed on each salivary sample. The mucosal biopsy was examined by routine H&E techniques. Furthermore, immunohistochemical expression of p53 was studied in all cases. RESULTS: A statistically significant difference in p53 expression was present between nonsmokers and the three smoker groups. However, no statistically significant variation was present between the 3 smoker groups. Furthermore, no statistically significant changes were found in the CYFRA 21-1 levels among the four groups. CONCLUSION: Shisha smoking has the same damaging effect on the oral mucosa as cigarette smoking. Furthermore, no correlation was detected between the histological findings and the salivary biomarker used (CYFRA 21-1).

Comparative study between laser and conventional techniques for class V cavity preparation in gamma-irradiated teeth (in vitro study).

OBJECTIVE: The purpose of this study was to compare laser with conventional techniques in class V cavity preparation in gamma-irradiated teeth. METHODS: Forty extracted human teeth with no carious lesions were used for this study and were divided into two main groups: Group I (n = 20) was not subjected to gamma radiation (control) and Group II (n=20) was subjected to gamma radiation of 60 Gray. Standard class V preparation was performed in buccal and lingual sides of each tooth in both groups. Buccal surfaces were prepared by the Er,Cr:YSGG laser (Waterlase iPlus) 2780 nm, using the gold handpiece with MZ10 Tip in non-contact and the "H" mode, following parameters of cavity preparation - power 6 W, frequency 50 Hz, 90% water and 70% air, then shifting to surface treatment laser parameters - power 4.5 W, frequency 50 Hz, 80% water and 50% air. Lingual surfaces were prepared by the conventional high-speed turbine using round diamond bur. Teeth were then sectioned mesio-distally, resulting in 80 specimens: 40 of which were buccal laser-treated (20 control and 20 gamma-irradiated specimens) and 40 were lingual conventional high-speed bur specimens (20 control and 20 gamma-irradiated specimens). RESULTS: Microleakage analysis revealed higher scores in both gamma groups compared with control groups. Chi-square test revealed no significant difference between both control groups and gamma groups (p=1, 0.819, respectively). A significant difference was revealed between all 4 groups (p=0.00018). CONCLUSION: Both laser and conventional high-speed turbine bur show good bond strength in control (non-gamma) group, while microleakage is evident in gamma group, indicating that gamma radiation had a dramatic negative effect on the bond strength in both laser and bur-treated teeth.

Comparative study between laser and conventional techniques for class V cavity preparation in gamma-irradiated teeth (in vitro study)

Abstract Objective: The purpose of this study was to compare laser with conventional techniques in class V cavity preparation in gamma-irradiated teeth. Methods: Forty extracted human teeth with no carious lesions were used for this study and were divided into two main groups: Group I (n = 20) was not subjected to gamma radiation (control) and Group II (n=20) was subjected to gamma radiation of 60 Gray. Standard class V preparation was performed in buccal and lingual sides of each tooth in both groups. Buccal surfaces were prepared by the Er,Cr:YSGG laser (Waterlase iPlus) 2780 nm, using the gold handpiece with MZ10 Tip in non-contact and the "H" mode, following parameters of cavity preparation - power 6 W, frequency 50 Hz, 90% water and 70% air, then shifting to surface treatment laser parameters - power 4.5 W, frequency 50 Hz, 80% water and 50% air. Lingual surfaces were prepared by the conventional high-speed turbine using round diamond bur. Teeth were then sectioned mesio-distally, resulting in 80 specimens: 40 of which were buccal laser-treated (20 control and 20 gamma-irradiated specimens) and 40 were lingual conventional high-speed bur specimens (20 control and 20 gamma-irradiated specimens). Results: Microleakage analysis revealed higher scores in both gamma groups compared with control groups. Chi-square test revealed no significant difference between both control groups and gamma groups (p=1, 0.819, respectively). A significant difference was revealed between all 4 groups (p=0.00018). Conclusion: Both laser and conventional high-speed turbine bur show good bond strength in control (non-gamma) group, while microleakage is evident in gamma group, indicating that gamma radiation had a dramatic negative effect on the bond strength in both laser and bur-treated teeth.

Lymphangiogenesis in Oral Squamous Cell Carcinoma: Correlation with VEGF-C Expression and Lymph Node Metastasis.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common oral malignancy that preferentially spreads to the cervical lymph node which, when involved, complicates the anticancer therapy and threatens the patient life. It was suggested that lymph node metastasis may be facilitated by lymphangiogenesis. VEGF-C is one of the most important lymphangiogenic inducers that promotes the lymphatic vessels growth and supports the survival of adult lymphatic endothelial cells. METHODS: Lymphatic vessels density (LVD) and LV morphometry were digitally evaluated using D2-40. The expression of VEGF-C was also assessed using immunohistochemistry and real-time polymerase chain reaction in 6 normal oral mucosa cases and 72 cases of OSCC. The correlation between LVD and LV morphometry, VEGF-C, and lymph node metastasis was statistically assessed. RESULTS: A positive cytoplasmic expression of VEGF-C was detected in both epithelial and connective tissue cells in 97% of OSCC, while all normal tissues reacted negatively. A greater expression of VEGF-C was associated with larger and more dilated LV and lymph node metastasis but not with LVD. CONCLUSION: VEGF-C is actively involved in the invasion and metastasis of OSCC via inducing morphological changes in LV. VEGF-C may be a promising target for anticancer therapy.

Effect of Transplantation of Bone Marrow Derived Mesenchymal Stem Cells and Platelets Rich Plasma on Experimental Model of Radiation Induced Oral Mucosal Injury in Albino Rats.

Normal tissue damage following radiotherapy is still a major problem in cancer treatment. Therefore, the current work aimed at exploring the possible role of systemically injected bone marrow derived mesenchymal stem cells (BM-MSCs) and/or locally injected platelet rich plasma (PRP) in ameliorating the side effects of ionizing radiation on the rat's tongue. Twelve rats served as control group (N) and 48 rats received a single radiation dose of 13 Gy to the head and neck region; then, they were equally divided into 4 experimental groups: irradiated only (C), irradiated + MSCs (S), irradiated + (PRP) (P), and combined group (PS). Animal scarification occurred in 3 and 7 days after radiation. Then, tongues were dissected and examined histologically and for expression of bcl-2 by RT-PCR. Histological examination of the treated groups (S), (P), and (PS) revealed an obvious improvement in the histological structure of the tongue, compared to group (C), in addition to upregulated expression of bcl-2, indicating decreased apoptotic activity. Conclusion. BM-MSCs and PRP have shown positive effect in minimizing the epithelial atrophy of normal oral mucosa after regional radiotherapy, which was emphasized by decreasing apoptotic activity in these tissues. Nevertheless, combined use of BM-MSCs and PRP did not reveal the assumed synergetic effect in oral tissue protection.

Correlation between ploidy status using flow cytometry and nucleolar organizer regions in benign and malignant epithelial odontogenic tumors.

OBJECTIVE: Differentiation between the aggressive benign odontogenic tumors and their malignant counterparts is controversial and difficult. While flow cytometry (FCM) allowed DNA analysis in neoplasia, argyrophilic organizer regions (AgNORs) number and/or size in a nucleus are correlated with the ribosomal gene activity and therefore with cellular proliferation. The aim of this research was to study the diagnostic accuracy of FCM and AgNORs staining in differentiating between benign and malignant epithelial odontogenic tumors and to correlate between these two interventions. DESIGN: Sixteen benign cases [8 cases of ameloblastoma (AB) and 8 cases of keratocystic odontogenic tumor (KCOT)] and 13 malignant epithelial odontogenic tumors [8 cases of ameloblastic carcinoma (ABC) and 5 cases of clear cell odontogenic carcinoma(CCOC)] were included in the current study. For FCM analysis, a single cell suspension from Formalin fixed paraffin-embedded (FFPE) tumors was prepared according to a modified method described by Hedley (1989) and AgNORs staining were performed in accordance to the Ploton protocol (1986). Analysis of AgNORs was performed using both quantitative and qualitative methods. RESULTS: The work revealed that all the examined tumors were diploid, except for 40% of CCOC cases. The S-phase fraction (SPF) value, AgNORs count and AgNORs area/cell showed statistically significant difference on comparing benign and malignant groups. A weak positive correlation was observed between SPF and AgNORs count. CONCLUSION: The SPF value was considered to be more sensitive and specific in differentiation between aggressive benign and malignant epithelial odontogenic tumors in comparison to AgNORs counting.

Assessment of the effect of low-energy diode laser irradiation on gamma irradiated rats' mandibles.

OBJECTIVE: The purpose of the present study was to evaluate the biostimulative and regenerative effects of low intensity laser irradiation (LILT) (applied before or after initiation of radiotherapy) on gamma irradiated rats' jaw bones. METHODS: Forty eight male Albino rats were equally divided into two groups: group 1, in which the left side of the mandible was subjected to three successive sessions of laser (LILT) prior to whole body gamma radiation (2Gy/3 fractions/week) and group 2, received whole body gamma radiation (2Gy/3 fractions/week) prior to three successive sessions of laser applied to left side. The right side of both groups was used as gamma irradiated non-lased control group. Each group was then subdivided into four equal subgroups (a, b, c, d) according to the time of scarification (3, 7, 14, 21 days respectively). Specimens were subjected to histological, histomorphometric and scanning electron microscopic examinations. RESULTS: Thin irregular bone trabeculae and widened marrow spaces were identified in the control group. The lased sides of groups 1 and 2 demonstrated regular, thick and continuous bone trabeculae. Ultrastructurally, collagen fibres of the control group appeared irregularly arranged and more spaced compared to groups 1 and 2. Normal-sized osteocytic lacunae were seen in the lased groups, as compared to the wide lacunar spaces noted in the control group. Histomorphometric analysis showed a significant increase in the area of bone trabeculae, as well as the width of compact bone, for the lased groups. CONCLUSIONS: LILT seemed to attenuate the radiation-related damage in alveolar bones.

Is human papilloma virus associated with salivary gland neoplasms? An in situ-hybridization study.

OBJECTIVE: HPV can infect cells of epithelial origin and is closely associated with carcinomas. Studies investigating its presence in salivary gland neoplasms are few and conflicting. METHODS: Detection of HPV types 16 & 18 was done on 34 formalin-fixed, paraffin-embedded archival material of different salivary gland neoplasms using Digene HPV types 16 & 18 probe using in situ hybridization technique. RESULTS: Eight of neoplastic salivary gland specimens were positively infected by HPV types 16 & 18. Seven of them were benign (4 Warthin's tumour, 2 pleomorphic adenoma and one myoepithelioma), in addition to one malignant specimen (lymphoma). Correlation was found between the incidence of HPV infection and histological differentiation of salivary gland neoplasms. CONCLUSIONS: An association exists between HPV infections and salivary gland neoplasms. However, given the sparse pattern of reactive cells, it cannot be confirmed that this virus is implicated in the aetiology of this group of tumours.

Histological and immunohistochemical evaluation of the chemopreventive role of lycopene in tongue carcinogenesis induced by 4-nitroquinoline-1-oxide.

OBJECTIVE: Increasing attention has been given to the potential protective roles of specific antioxidant nutrients found in fruits and vegetables. However, there are relatively few reports on the cancer chemopreventive effects of lycopene or tomato carotenoids in animal models. Therefore, the chemopreventive efficacy of lycopene with regard to oral carcinogenesis was investigated using 4-nitroquinoline-1-oxide (4-NQO) induced tongue squamous cell carcinoma. MATERIALS AND METHODS: Twenty albino rats were equally divided into 2 groups. 4-NQO was dissolved in the drinking water (20 ppm) for rats of both groups. Rats in group 2 were administered lycopene at a dose of 2.5mg/kg body weight by intragastric intubation once a day. Incidence of oral neoplasms and histopathological changes were microscopically evaluated after 32 weeks of administration of the specific treatments. Moreover, immunohistochemical expressions of proliferating cell nuclear antigen (PCNA), E-cadherin and ß-catenin were analysed in tongue specimens using an image analyser computer system. RESULTS: Lycopene treatment significantly decreased the incidence of 4-NQO induced tongue carcinogenesis. A decreased percentage of PCNA-positive nuclei was associated with lycopene treatment. Proliferating cells were mainly confined to the basal and parabasal epithelial cell layers. Increased E-cadherin and ß-catenin immunoexpression was recorded in the lycopene treated group in comparison to the carcinogen group. CONCLUSION: Results of the present study indicate that lycopene can exert protective effects against 4-NQO induced tongue carcinogenesis through reduction in cell proliferation and enhanced cellular adhesion, suggesting a new mechanism for the anti-invasive effect of lycopene. Further studies are needed to provide more definitive answers to the question of the anticancer effects of lycopene.

Association of macrophages with angiogenesis in oral verrucous and squamous cell carcinomas.

BACKGROUND: Tumor-associated macrophages (TAMs) are a major cellular component of human cancers, yet there is still no consensus as to their role in cancer growth and angiogenesis. METHODS: The association between TAMs and angiogenesis was investigated in formalin-fixed, paraffin-embedded archival material of oral squamous cell carcinoma (OSCC) and oral verrucous carcinoma (OVC). TAMs shown by immunohistochemistry for CD68 and microvessels demonstrated by immunohistochemistry for CD31 were quantified using an image analyzer computer system. RESULTS: TAMs were observed in all studied specimens. The area percentage of CD68 immunoreactivity and microvessel density (MVD) were significantly lower in OVC compared with the different grades of OSCC (P = 0.0009), (P = 0.0045). Both parameters increased in high-grade malignancy of OSCC. Linear regression analysis revealed a positive correlation between the area percentage of CD68 immunoreactivity and the MVD in the studied tumors. CONCLUSIONS: Increased TAMs is associated with angiogenesis and higher histopathological grades in oral cancer.

The effect of lathyrism on dentin structure of the rat incisors: a morphometric and scanning electron microscopic investigation.

BACKGROUND: The present study was designed to study the effect of beta-aminopropionitrile (beta-APN), present in Lathyrus sativus grass pea consumed in drought prone areas, on dentin of the continuously erupting rat incisors. METHODS: Eighteen adult male rats were used. In the experimental group (18 rats), lathyrism was induced by a once daily subcutaneous administration of beta-APN for 40 days. The maxillary and mandibular incisors were examined ultrastructurally and morphometrically. RESULTS: The mean number of patent tubules, the mean area, perimeter and the area percent of the tubules were analyzed. Ultrastructurally, the dentinal tubules of both coronal and radicular dentin in the lathyritic group were narrower or even obliterated compared with those in the control. The coronal and radicular dentin of the lathyritic group exhibited an irregular lattice of non-mineralized small branching collagen fibrils obliterating the dentinal tubules. The mean number of patent tubules in the control and lathyritic groups revealed an insignificant difference. The mean area of the tubules showed a statistically significant difference in lathyritic radicular dentin (P = 0.0353). The percentage of the total surface area of the dentinal tubules significantly decreased in the radicular dentin of the lathyritic group (P = 0.024). CONCLUSIONS: These findings indicated a deleterious effect of lathyrism on dentin, with a possible negative impact on developing teeth integrity.

Ultrastructural and histomorphometric alterations of rat jaw bones after experimental induction of lathyrism.

OBJECTIVE: The aim of this study was to investigate the effect of beta-aminopropionitrile (beta-APN), present in Lathyrus sativus (L. sativus) seeds consumed in drought prone areas, on the maxillary and mandibular bones of rats. DESIGN: Eighteen rats were used; they were divided into experimental group (12 rats) and a control group (6 rats). In the experimental group, lathyrism was induced by a once daily subcutaneous administration of beta-APN at a dose of 5mg in 0.4ml distilled water per 100g of body weight for 40 days. The specimens were examined histologically, histomorphometrically and ultrastructurally. RESULTS: Histopathological examination of lathyritic rats showed that both mandible and maxillae were significantly affected. Irregular bone trabeculae with wide medullary cavities and abundant marrow spaces were observed. Focal resorptive areas lined with osteoclasts were also present. Histomorphometric analysis showed a decrease in the area percentage of bone in lathyritic rats as compared to the control. This decrease was statistically significant only in the mandible (p=0.018). Moreover, a significant increase in the number of osteoclasts (p=0.014, 0.012) and resorptive bays (p=0.013, 0.002) was detected in the maxillae and mandible of the experimental group, respectively. Ultrastructurally, there was a significant difference in the microarchitecture of the collagen fibres in lathyritic rats compared to the control. The collagen fibres in the lathyritic group appeared disorganized, fewer in number and more spaced. CONCLUSIONS: This study provided evidence for lathyritic disturbances in bone architecture. These disturbances are likely to affect mineralization and bone strength as well.