Influence of chitosan and Cissus quandrangularis coating on osseointegration in titanium implants in rabbits: A preclinical in vivo study.

BACKGROUND: Titanium (Ti) implants has been criticized for the tiring wait for osseointegration, often making the patient reconsider implant treatment. Surface treated Ti implants are emerging as a promising solution with superior osseointegration, early loading protocols and shortened period of edentulousness. The aim of this study is to assess the osseointegration of Ti surface coated with novel Cissus quandrangularis Chitosan Hydrogel (CqChH) compared to Commercially pure (Cp) implants. METHODS: 24 Cp Ti implants were divided into 2 subgroups (n = 12). The test group consisted of Ti implants surface treated with the novel hydrogel and control group consisted of Cp Ti implants. 3 % CqChH was prepared and was coated on the Ti implants prior to placement in the femur and tibial heads of rabbits. Implant Stability Quotient (ISQ) was recorded at the 6th and 12th week. Animals were sacrificed and subjected to Removal Torque Quotient (RTQ). The samples were retrieved en bloc and stained for histopathologic analysis. The collected data was subjected to statistical analysis using Unpaired student t-Test. RESULTS: At the end of 6th week CqChH coated implants did not show any statistically significant difference in both ISQ and RTQ values compared to Cp ones. However, at the end of the 12th week CqChH coated implants demonstrated significantly higher ISQ (73.91 ± 4.39) and RTQ (75.96 ± 14.10) compared to Cp ones. CONCLUSIONS: This study demonstrated that the novel hydrogel coating applied to the implant's surface exhibited not only enhanced bone regeneration but also elicited a new bone formation.

Remineralising potential of Ocimum basilicum varnish and fluoride varnish on initial enamel caries: An in vitro microscopic study.

Background: The focus of caries research has switched to early identification and non-invasive treatment of carious lesions. Aim: This study aimed to evaluate and compare the remineralising potential of Ocimum (O.) basilicum varnish and fluoride varnish on initial enamel caries. Method: The authenticated O. basilicum seeds were procured from a repository, and the extract was prepared using the Soxhlet method, which was vortexed with Indian Pharmaceutical (IP)-graded chemicals to obtain varnish. Extracted premolar tooth samples were divided into three groups of 33 each after demineralisation with a pH of 4.5 for 48 hours at 37°C. Each group was subjected to remineralisation twice daily with respective agents for 4 minutes for 30 consecutive days. Each sample was ground-sectioned through an enamel window. The lesion depth was measured using a light microscope (Leica™ DM2500) and ImageJ software. The data were evaluated using analysis of variance (ANOVA) and post hoc analysis. Results: The mean (± SD) pre-treatment lesion depth across the groups ranged from 242.11 ± 26.144 µm to 352.66 ± 34.531 µm. The highest lesion depth recovery rate of 45.938% was recorded for the fluoride varnish group, followed by 36.015% in the O. basilicum varnish group, which was statistically significant by Tukey's post hoc analysis (p < 0.001). The gingival fibroblast cells were viable by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Conclusion: The O. basilicum varnish demonstrated a homogenous layer of mineral deposition. However, the remineralising efficacy was slightly lesser than that of the fluoride varnish. Hence, the novel O. basilicum-based remineralisation agent appears to have potential as a non-invasive alternative to topical fluorides in the therapy of early caries lesions.

Histochemical and Immunohistochemical Analysis of Collagen Fibers and Microvascular Density in Various Grades of Oral Submucous Fibrosis.

BACKGROUND AND OBJECTIVE: This study was aimed to evaluate the collagen fibers qualitatively and its correlation with microvascular density in various grades of oral submucous fibrosis (OSMF). MATERIAL AND METHODS: The present study comprised of total 40 cases of oral submucous fibrosis. Picrosirius red staining was done on all the specimens' sections. They were analyzed for the colour and orientation of collagen fibers. Morphometric measurements were done using image analysis on immunohistochemical stained sections for Factor VIII-related antigen and analyzed for microvascular density. RESULTS: Picrosirius red polarizing microscopy results revealed that there was a shift in the colour of collagen fibers from greenish yellow to orange red and red colour as the severity of the oral submucous fibrosis increased. The collagen fibers showed mixed orientation in early oral submucous fibrosis and parallel orientation in advanced oral submucous fibrosis. There was a significant decrease in microvascular density from early to advanced oral submucous fibrosis. CONCLUSION: The change in the colours and orientation of collagen fibers in early and advanced oral submucous fibrosis could be attributed to the fibre thickness, type of collagen, alignment and packing, cross-linking of the fibers and the section thickness. However, in advanced cases the vascularity is reduced which may predispose to epithelial atrophy and subsequent malignant changes.

Fibrinogen Demonstration in Oral Lichen Planus: An Immunofluorescence Study on Archival Tissues.

BACKGROUND: Lichen planus is a premalignant condition with minimal diagnostic aids. This study is an attempt to use paraffin embedded sections of lichen planus with immunofluorescein stain and to evaluate the immunofluorescent sections to establish pattern of fibrinogen deposition. MATERIALS AND METHODS: Thirty-five paraffin embedded sections of old and new cases of oral lichen planus (study group) and five normal oral mucosa (control group) were chosen. Two sections of each (H & E) case were taken, one was stained with hematoxylin and eosin and another with fluorescein isothiocynate conjugate (FITC) polyclonal rabbit antibody against fibrinogen. Fluorescent findings were examined with a fluorescent microscope. RESULTS: A high statistical significant correlation was found in respect to fluorescence positivity, intensity of fluorescence and distribution of fluorescence each with p < 0.0001 and fluorescence at blood vessel walls (p = 0.0003). CONCLUSION: This study suggested that paraffin embedded sections can be successfully used in direct immunofluorescence staining in routine set up where only formalin fixed tissues are received. CLINICAL SIGNIFICANCE: Paraffin embedded sections can be successfully used in direct immunofluorescence staining when only formalin fixed tissues are received.

An immunohistochemical study of basement membrane heparan sulfate proteoglycan (perlecan) in oral epithelial dysplasia and squamous cell carcinoma.

BACKGROUND: Basement membrane heparan sulfate proteoglycan (perlecan) has been demonstrated in precancer lesions and carcinomas of oral cavity. It helps in malignant transformation of epithelial cells. The aim of our study was to understand the immuno-localization of perlecan in oral dysplastic epithelium and oral carcinomas. MATERIALS AND METHODS: A total of 50 cases comprising 10 normal mucosa, 20 dysplastic mucosa, and 20 oral squamous cell carcinomas (OSCC) were included in the retrospective study. They were examined for the presence of perlecan protein core by immunohistochemistry using monoclonal antibody. Interpretation of the pattern of staining was done, and majority of the observations were taken for statistical analysis. RESULTS: In normal epithelium, perlecan was found to be present in basal layer at the cell border. In dysplastic epithelium, it was present in suprabasal layers also. With the increase in severity of dysplasia, its expression was more in suprabasal layers, and the immuno-localization was found to be at cell border and cytoplasm. In OSCC cases, perlecan was present in stroma and tumor islands. CONCLUSION: It was deduced from the above results that perlecan helps potentially in dysplastic changes of epithelial cells. It gets accumulated within the cell and intercellular spaces and serves as a reservoir for various growth factors. In OSCC, it breaks down and releases growth factors, which help in tumor progression, angiogenesis, and metastasis of the carcinoma.

Diabetes mellitus accelerates cartilaginous metaplasia and calcification in atherosclerotic vessels of LDLr mutant mice.

BACKGROUND: Vascular calcification is highly prevalent in patients with type II diabetes mellitus (T2DM). Little is known about whether T2DM is causative. METHODS: Low-density lipoprotein receptor mutant (LDLr(-/-)) mice were fed with customized diabetogenic and/or procalcific diets to induce atherosclerosis, cartilaginous metaplasia and calcification, along with obesity, hyperglycemia, hyperinsulinemia, and hypercholesterolemia at various levels, and euthanized for study after 18-24 weeks on diet. RESULTS: We found that T2DM accelerated cartilaginous and calcific lesion development by ~3- and 13-fold as determined by incidence of vascular cartilaginous metaplasia and calcification in LDLr(-/-) mice. Lowering dietary fat from ~60% to ~40% kcal reduced body weight and serum glucose and insulin levels, leading to a 2-fold decrease in aortic calcium content. Correlation analysis of calcium content with a calculated insulin resistance index, homeostasis model assessment of insulin resistance, showed a positive correlation of insulin resistance with vascular calcification. Finally, we used genetic fate mapping strategy to trace cells of SM origin in these animals. Vascular smooth muscle cells (SMCs) were found to be a major cell source contributing to osteochondrogenic differentiation and calcification. Receptor for advanced glycation end-products (RAGE) was up-regulated, co-localizing with osteochondrogenic SMCs. CONCLUSIONS: Through quantitative measure of aortic calcium content, we provided experimental findings that LDLr(-/-) mice, like T2DM patients, are predisposed to vascular calcification. Our study is also the first to establish a distinct role of hyperglycemia and hypercholesterolemia in osteochondrogenic differentiation of SMCs and determined these cells as a major source contributing to cartilaginous and calcifying lesions of T2DM blood vessels, possibly mediated by RAGE.

Perianal Paget's disease-Report of a rare case.

INTRODUCTION: Perianal extra-mammary Paget's disease is a rare skin disorder of unknown aetiology, which is frequently associated with malignancy. This case report draws attention to this rare condition and comments upon its diagnosis and treatment. PRESENTATION OF CASE: A 64-year-old otherwise fit man, presented to us in 2006 with one-year-long history of perianal irritation. On examination there was an erythematous discoid skin lesion in the right perianal area. The lesion was excised with wide margins and the defect closed with a local transposition flap. Histology confirmed extra-mammary Paget's disease (EMPD) with a focus of invasion showing a well-differentiated mucinous adenocarcinoma. Adjuvant therapy was not advised. On follow-up in 2011, a small irregular skin lesion, well away from the previous excision site was noted on the left perianal area. Biopsies from this lesion confirmed EMPD with no focus of invasion. Once again wide local excision with closure using local transposition flap was undertaken. Long term follow up has been advised. DISCUSSION: The optimal treatment for Perianal Paget's disease (PPD) remains controversial. Surgery is the commonest modality used with wide local excision being the treatment of choice for resectable disease. We report herein a short review of various therapies reported so far in the management of this rare disorder. CONCLUSION: A thorough initial evaluation and long-term follow-up is essential to identify recurrence and the development of other related malignancies.

Sources of cells that contribute to atherosclerotic intimal calcification: an in vivo genetic fate mapping study.

AIMS: Vascular cartilaginous metaplasia and calcification are common in patients with atherosclerosis. However, sources of cells contributing to the development of this complication are currently unknown. In this study, we ascertained the origin of cells that give rise to cartilaginous and bony elements in atherosclerotic vessels. METHODS AND RESULTS: We utilized genetic fate mapping strategies to trace cells of smooth muscle (SM) origin via SM22α-Cre recombinase and Rosa26-LacZ Cre reporter alleles. In animals expressing both transgenes, co-existence within a single cell of ß-galactosidase [marking cells originally derived from SM cells (SMCs)] with osteochondrogenic (Runx2/Cbfa1) or chondrocytic (Sox9, type II collagen) markers, along with simultaneous loss of SM lineage proteins, provides a strong evidence supporting reprogramming of SMCs towards osteochondrogenic or chondrocytic differentiation. Using this technique, we found that vascular SMCs accounted for ~80% of Runx2/Cbfa1-positive cells and almost all of type II collagen-positive cells (~98%) in atherosclerotic vessels of LDLr-/- and ApoE-/- mice. We also assessed contribution from bone marrow (BM)-derived cells via analysing vessels dissected from chimerical ApoE-/- mice transplanted with green fluorescence protein-expressing BM. Marrow-derived cells were found to account for ~20% of Runx2/Cbfa1-positive cells in calcified atherosclerotic vessels of ApoE-/- mice. CONCLUSION: Our results are the first to definitively identify cell sources attributable to atherosclerotic intimal calcification. SMCs were found to be a major contributor that reprogrammed its lineage towards osteochondrogenesis. Marrow-derived cells from the circulation also contributed significantly to the early osteochondrogenic differentiation in atherosclerotic vessels.

Oral hyaline ring granuloma: a case report.

An unusual granulomatous inflammatory lesion characterised by the presence of ring shaped hyaline bodies accompanied by foreign body giant cell reaction has been mentioned in the literature. A lack of agreement exists on the nature of these hyaline rings. Opinions ranging from their being hyaline degenerative blood vessels to remains of leguminous cells have been postulated. Although the nature and origin of these bodies is uncertain, there is now considerable evidence that they represent, at least in part, vegetable material, especially pulses, that have been implanted in the tissues. An inflammatory oral lesion in a 17-years-old male patient caused due to traumatic implantation of the vegetable matter (sugar cane) has been reported in this paper. Based on the histological finding of a granulation tissue surrounding a hyaline ring the lesion was diagnosed as hyaline ring granuloma.

Outcome of breast lesions diagnosed as lesion of uncertain malignant potential (B3) or suspicious of malignancy (B4) on needle core biopsy, including detailed review of epithelial atypia.

AIMS: To provide updated evidence of the outcome of breast lesions of uncertain malignant potential (B3) and suspicious of malignancy (B4) diagnosed on needle core biopsy (NCB) and analyse the outcome of the different types of intraductal epithelial atypia. METHODS AND RESULTS: One-hundred and forty-nine B3 and 26 B4 NCBs diagnosed over a 2-year period (2007-2008) were compared with those diagnosed over a previous 2-year period (1998-2000). The proportion of B3 diagnoses increased from 3.1% to 4.5%, and the positive predictive value (PPV) of malignancy of a B3 core decreased from 25% to 10%. Increased diagnosis of radial scar and reductions in the PPV of lobular neoplasia and of atypical intraductal proliferation may explain the reduction in the PPV of the B3 group as a whole. There were no significant changes in the proportion of B4 diagnosis (1.1% and 0.8%) or the PPV of B4 (83% and 88%). Review of cores with intraductal atypia showed a wide range of PPVs, from 100% for suspicious of ductal carcinoma in situ, to 40% for atypical ductal hyperplasia categorized as B3, and 14% for isolated flat epithelial atypia. CONCLUSION: The study has found a decrease in the PPV for a B3 diagnosis and suggests possible explanations.

Immunohistochemical expression of Tenascin in embryogenesis, tumorigenesis and inflammatory oral mucosa.

OBJECTIVE: Tenascin is a large extracellular matrix glycoprotein that plays specific role in cell matrix interaction. This protein is mainly attracted because of its oncofetal predominance expression at epithelial-mesenchymal interaction and also been associated with inflammatory response. Thus the aim was to study the expression of Tenascin within the oral cavity in a developing tooth, normal oral mucosa, squamous cell carcinoma and inflammatory mucosa and further to compare its expression in inflammatory mucosa with that of squamous cell carcinoma. DESIGN: A total numbers of 92 cases were included, with 22 being all morphological stages of developing tooth, 10 cases of normal oral mucosa, 30 cases each of inflammatory gingival hyperplasia and oral squamous cell carcinoma. The intensity and pattern of expression was assessed immunohistochemically using anti-human mouse monoclonal Tenascin antibody. RESULTS AND CONCLUSION: Tenascin expression in developing tooth was seen mainly at epithelial-mesenchymal junctions, but temporally reduced at cap stage. In normal mucosa TN expression was restricted only at basement membrane zone. Inflammatory gingival hyperplasia intensity of expression was enhanced at the juxtraepithelial stroma and showed reticular pattern of expression. In oral squamous cell carcinoma, intensity of expression was seen in superficial front of the stroma and also around tumour islands with intraepithelial expression and predominantly showed fibrillar pattern of expression. Furthermore, Tenascin expression was noticed around neovascularization. Hence, there is a regulatory system in Tenascin expression and plays a vital role in embryogenesis, tumerogenesis and inflammation in remodelling the stroma for cell migration and also for healing.

Perlecan (basement membrane heparan sulfate proteoglycan) and its role in oral malignancies: an overview.

Perlecan means pearl-like structures. Perlecan is a large proteoglycan (400-500 kDa) present in virtually all vascularized tissues with a distribution that is primarily confined to basement membranes including those of oral mucosa. It is a basement membrane-type heparan sulfate proteoglycan. Perlecan is synthesized by basal cells and fibroblasts adjacent to the basal lamina . Perlecan is also synthesized by vascular endothelial and smooth muscle cells present in the extracellular matrix. It has been demonstrated in recent years that perlecan is distributed in the stromal space of various pathophysiological conditions. The complex pleiotropy of perlecan suggests that this gene product is involved in several developmental processes, at both early and late stages of embryogenesis, as well as in cancer and diabetes. In the oral cavity, perlecan expression is reported to basal cells in normal mucosa and its expression increases in precancer and cancerous conditions. It is also expressed in various odontogenic tumors such as ameloblastoma, keratocyst odontogenic tumor, and also salivary gland tumors such as adenoid cystic carcinoma, mucoepidermoid carcinoma, etc.

Eccrine angiomatous hamartoma: a rare multifocal variant with features suggesting trauma.

Eccrine angiomatous hamartoma (EAH) is a rare, benign cutaneous tumor characterized by proliferation of the eccrine gland elements closely associated with capillary angiomatosis and proliferation of other dermal elements, such as adipose tissue, hair and epidermis. Patients usually present with a solitary nodule on the extremities appearing at birth or during the prepubertal years. However multifocal lesions or late onset of this condition may occur. Eccrine angiomatous hamartoma is usually sporadic, but one familial case of the multifocal variant has been reported. The clinical presentation ranges from a simple angiomatous nodule to erythematous--purpuric plaques. Eccrine angiomatous hamartoma is generally asymptomatic but may occasionally be associated with pain and hyperhidrosis. We report a rare case of the multifocal variant of EAH in a 13-year-old girl, with histological features suggesting trauma. Clinically, this condition must be differentiated from other angiomatoses and a definitive diagnosis is based upon histology. Eccrine angiomatous hamartoma is a benign slowly growing lesion for which aggressive treatment is not indicated. Simple excision is reserved for painful or cosmetically disfiguring examples.

Trichilemmal cyst with carcinoma in situ within an atypical fibroxanthoma.

A 91-year-old lady presented with an 8-month history of a slow growing keratotic nodule on the forehead, which was excised. Histological examination showed a well-defined lesion with marked cellular pleomorphism. Numerous mitoses were present. The cells stained positively with CD10 and CD68 and weakly with smooth muscle actin. However, the tumor was negative for cytokeratins (AE1/AE3), epithelial membrane antigen (EMA), S100, Melan A, HMB45, leukocyte common antigen (LCA), desmin, and CD31. Therefore, the lesion was designated as atypical fibroxanthoma (AFX) by exclusion. Within the AFX, there was also a cyst lined by squamous epithelium showing pilar keratinization. The epithelium showed full-thickness dysplasia with increased mitotic activity and was stained positively with AE1/AE3, thus supporting our view of carcinoma in situ within the wall of the cyst. To the best of our knowledge, this article describes the first case of trichilemmal cyst with carcinoma in situ arising within an AFX.

Dentin dysplasia: single-tooth involvement?

Dentin dysplasia is a genetic defect of dentin formation inherited as an autosomal dominant trait. It is characterized by normal enamel but atypical dentin formation with abnormal pulpal morphology. Once thought to be a single entity, dentin dysplasia has now been divided into type I (radicular) and II (coronal). Type I is by far the more common. Both types include multiple/generalized involvement of primary and permanent dentition. Combinations of both types have also been described in the literature. Four distinct forms of dentin dysplasia type I and 1 form of dentin dysplasia type II are identified. Although there seems to be no need to identify more than 2 distinct types of this relatively rare inherited defect of human dentin, the possible existence of additional forms of the disease cannot be ruled out. Here is a case report of dentin dysplasia in a single tooth, with crown and roots of normal dimensions, associated with severe pain and mobility and histologically involving both coronal and radicular dentin. Focal odontoblastic dysplasia or dentin dysplasia type III could be the new entity.

Ameloblastic carcinoma: a case report.

Ameloblastic carcinoma is a rare malignant lesion with characteristic histologic features and behavior that dictate a more radical surgical approach than does a simple ameloblastoma. Clinically, ameloblastic carcinoma is more aggressive than most typical ameloblastomas with extensive local destruction, perforation of the cortical plate, extension into surrounding soft tissues, numerous recurrent lesions, and metastasis, usually to cervical lymph nodes. The radiographic appearance of ameloblastic carcinoma is consistent with that of ameloblastoma except for occasional presence of some focal radiopacities, apparently reflecting dystrophic calcification. Histologically, the tumor cells resemble cells seen in ameloblastoma but show cytologic atypia, cellular pleomorphism, nuclear hyperchromatism, mitoses, and vascular and neural invasion. These identifying features of ameloblastic carcinoma must be known and recognized by dental practitioners. It is probable that ameloblastoma, like other tumors (such as carcinoid tumors and epithelial tumors of the ovary), shows a spectrum of histologic and biologic behavior ranging from benignity at one end to frank malignancy at the other. A case of ameloblastic carcinoma of the maxilla in a 70-year-old man is reported.