Evaluation of biosafe alternative to eosin in hematoxylin and eosin staining procedure: A comparative study.

Background: Hematoxylin and eosin staining is the globally accepted staining technique for histology and histopathology sections. Hematoxylin is a natural dye as compared to eosin which is a synthetic dye manufactured from chemicals. Eosin acts as a counterstain to hematoxylin giving a sharp contrast to its blue color. Eosin being a chemical, can cause health hazards and environmental pollution. Using eco-friendly materials and going organic is the demand of this era. Curcuma longa rhizomes are proven to contain curcuminoids that exhibit various medicinal properties and curcumin the bright yellow component of the plant, that is, being used as a colorant. This natural dye from C. longa provides a significant alternative to eosin as they are safer to use without health hazards and are biodegradable and have easy disposability. Aim: To evaluate and compare the staining qualities of C. longa (turmeric) with hematoxylin and eosin (H&E). Materials and Methods: A total of 100 paraffin-embedded tissue block specimens of various pathological tissues were stained with H and E and hematoxylin and curcumin (H&C). Evaluation and comparison of the staining efficacy were done for all the pathological tissues. The obtained data were subjected to statistical analysis. Results: H&C staining gave comparable results with H and E staining with a statistically insignificant P value (p > 0.05). Conclusion: Curcumin has shown equivalent staining to that of eosin. Thus, it can be used as a better alternative.

Salivary malondialdehyde in oral submucous fibrosis - A marker for oxidative damage.

BACKGROUND: Oral submucous fibrosis (OSMF) is a potentially malignant disorder of oral mucosa and its malignant transformation rate accounts to about 7%-13%. Oxidative damage and lipid peroxidation plays an important role in OSMF. Lipid peroxidation has not been widely investigated in OSMF patients with respect to clinical staging and histopathological grading. As human saliva is a diagnostic fluid which can be obtained in a noninvasive procedure as compared to the blood for serum analysis, the present study was aimed at evaluating the salivary malondialdehyde (MDA) levels in OSMF and comparison with respect to clinical staging and histopathological grading. AIM: This study aims to evaluate salivary MDA levels in OSMF and compare the levels with respect to clinical and histopathological grading systems. MATERIALS AND METHODS: Forty cases of clinically diagnosed and histopathologically proven cases of OSMF were included for the purpose of this study. As controls 40 age-matched individuals without any systemic disease were selected. Unstimulated whole saliva was collected from each individual, centrifuged and frozen at - 20°C until analysis. Lipid peroxidation products MDA were analyzed by thiobarbituric acid reaction. RESULTS: Salivary MDA levels were significantly increased in OSMF patients compared to controls. The progressively increased salivary MDA levels showed a positive correlation with the clinical stages and histopathological grades of OSMF and the results were statistically significant. CONCLUSION: The increased salivary MDA levels in OSMF patients compared to the control group suggests an increased oxidative stress levels in the potentially malignant disorders such as OSMF. The mean salivary MDA levels were increased significantly as the clinical stage and histopathological grade of OSMF advances, suggesting MDA to be used as a reliable biochemical marker and also a prognostic marker to assess the extent of oxidative damage in OSMF.

Analysis of collagen fibers in keratocystic odontogenic tumor and ameloblastoma: A polarizing microscopic study.

INTRODUCTION: Collagen forms an integral part of connective tissue and maintains its structural integrity. It has natural birefringence which is attributed to the arrangement of its fibers and is enhanced by special stains such as picrosirius red through polarizing microscopy. The polarization colors differ according to the fiber thickness and pattern of arrangement which in turn related to aggressiveness. Hence, the present study was conducted to evaluate collagen fibers in keratocystic odontogenic tumor (KCOT) and ameloblastoma using polarizing microscopy. AIM: This study aims to compare and correlate different types and patterns of collagen fibers in KCOT and ameloblastoma using picrosirius red stain under polarizing microscopy to delineate their aggressiveness. MATERIALS AND METHODS: The color, thickness, and orientation of collagen fibers in the KCOTs (n = 15) and ameloblastomas (n = 15) were studied histochemically by staining the sections with picrosirius red and examined under polarizing microscope using image analyzer software. RESULTS: When collagen fiber bundles in KCOT and ameloblastoma were compared, significant difference was noted between yellowish-orange collagen fiber bundles, but no significant difference was observed between greenish-yellow and orange-red collagen bundles. With respect to orientation and organization, the results are statistically significant (P < 0.05). CONCLUSION: The connective tissue stroma of KCOT could be regarded not just as a structural support but as a functional part of the lesion. In KCOT, the thin, parallel, and loosely arranged greenish-yellow collagen fibers may be attributed to its high recurrence rate and biological aggressiveness.

Solitary plasmacytoma of mandible: A rare entity.

Plasmacytoma is a discrete, unifocal, monoclonal neoplastic proliferation of the plasma cells. It may present as one of the three distinct clinical entities: multiple myeloma (MM), solitary plasmacytoma of bone and extramedullary plasmacytoma. Solitary bone plasmacytoma accounts for 3% of all plasma cell neoplasms with approximately 50% of the cases transforming into MM. It is frequently seen in vertebrae and secondarily in long bones. Its presence in jaws is extremely rare and when it is seen, retromolar area, angle and ramus of the mandible are most common sites of occurrence. A comprehensive clinical, radiological, histological and immunohistochemical features of solitary plasmacytoma of the mandible in a 46-year-old female patient is reported.

Basaloid squamous cell carcinoma: Report of two rare cases and review of literature.

Basaloid squamous cell carcinoma (BSCC) is a histologically distinct variant of SCC in the head-and-neck region it occurs most commonly in older men with a history of heavy smoking and alcohol abuse and usually presents as a high stage disease with widespread metastases and hence poor prognosis. BSCC is believed to arise from a totipotential primitive cell in the basal layer of the surface epithelium or from the salivary duct lining epithelium. BSCC is an uncommon tumor with a predilection for the upper aerodigestive tract, is a distinct variant of squamous carcinoma, due to its unique histological features and ominous clinical behavior. In the oral cavity, BSCC has a predilection for the tongue, followed by the floor of the mouth, palate, buccal mucosa, retromolar trigone and gingiva. In the oral cavity, retromolar trigone/gingiva is very rare sites and only a few cases have been reported in the literature. This paper reports an additional two cases of BSCC, one reported in the retromolar region and other reported on the gingiva.

Natural substitutes for formalin: A boon to histopathology!!

INTRODUCTION: In routine tissue processing, formalin has been proved as efficient as fixative since inception and hazards associated with it are of major safety and health concern. AIM: The aim of this study is to compare the efficacy of natural fixatives such as jaggery and Khandsari over formalin. MATERIALS AND METHODS: Ninety normal tissue specimen collected during minor oral surgical procedures were included in this study. Thirty specimen each were fixed in 30% jaggery (Group-A), 30% Khandsari (Group-B) and 10% formalin (Group-C). The slides obtained after tissue processing were analyzed for the quality of fixation. The tissue sections were assessed for cellular outline, cytoplasmic details, nuclear details, staining quality and overall morphology. Each criterion was rated on a scale of 1-4. (one for poor and four for excellent). The study was double-blinded and subjected to the Kruskal-Wallis ANOVA and Chi-square test. RESULTS: The cellular outline is excellent in 90% (Group-C) followed by 36.67% (Group-B) of specimens. With respect to cytoplasmic staining 83.33% (Group-C) of tissues showed excellent results followed by 60% (Group-B), 33.33% (Group-A). Nuclear details are excellent in 86.67% (Group-C) followed by 83.33% (Group-B), 36.67% (Group-A) of specimens. With respect to staining quality 93.33% (Group-C) followed by 50% (Group-B), 26.67% (Group-A). Overall morphology is excellent in 90% (Group-C) followed by 46.67% (Group-B). CONCLUSION: In the present study, Khandsari was on par with formalin and our effort of using Khandsari and jaggery for tissue fixation in human oral tissues yielded good results.

Evaluation of biosafe alternatives as xylene substitutes in hematoxylin and eosin staining procedure: A comparative pilot study.

BACKGROUND: Xylene is synthetic hydrocarbon produced from coal tar known for its wide usage as universal solvent which has many hazardous effects. The aim of this study is to compare the efficacy of xylene-free hematoxylin and eosin (H and E) sections with conventional H and E sections. MATERIALS AND METHODS: The study included ninety paraffin-embedded tissue blocks. Of these, sixty blocks were processed with sesame oil (xylene alternative) and thirty blocks with xylene. The study sample was divided into three groups. Sixty sections which are taken from sesame oil-processed blocks were stained with xylene-free H and E staining method. In xylene-free staining method, 95% diluted lemon water (Group A) and 1.7% dish washing solution (DWS, Group B) were used as deparaffinizing agents whereas the remaining 30 sections were processed with xylene and stained with conventional H and E staining method (Group C). Slides were scored for the following parameters: (i) nuclear staining (adequate = score 1, inadequate = score 0), (ii) cytoplasmic staining (adequate = score 1, inadequate = score 0), (iii) uniformity (present = score 1, absent = score 0), (iv) clarity (present = score 1, absent = score 0) and (v) intensity (present = score 1, absent = score 0). Score ≤2 was considered inadequate for diagnosis while scores 3-5 were considered adequate for diagnosis. RESULTS: Adequate nuclear staining was noted in 90% of sections of Group A and 100% each in Group B and Group C (P < 0.05); adequate cytoplasmic staining in 96.7% in Group A and 100% each in Group B and Group C (P > 0.05); adequate uniformity of staining in 53.3% of sections of Group A, 70% in Group B and 83.3% in Group C (P < 0.05); adequate clarity of staining in 73.3% sections of Group A, 80% in Group B and 83.3% in Group C (P > 0.05) and adequate intensity of staining in 76.7% sections of Group A, 93.3% in Group B and 100% in Group C (P < 0.05). Group C sections stained adequate for diagnosis (93.3%) followed by Group B (88.7%) and Group A (78%; P < 0.05). CONCLUSION: Tissues processed with sesame oil and stained using 1.7% DWS were found to be effective alternative to xylene.

Age and gender assessment through three-dimensional morphometric analysis of maxillary sinus using magnetic resonance imaging.

BACKGROUND: Age and sex determinations are important tools in forensic odontology which help in the identification of an individual. Radiographic method of sex and age estimation is a noninvasive simple technique. Measurements of the maxillary sinuses can be used for the estimation of age and gender when other methods are inconclusive. Maxillary sinus dimensions were used as an important tool in the identification of unknown. AIM: This study aims to estimate age and sex using the dimensions and volume of the maxillary sinus in magnetic resonance imaging (MRI). MATERIALS AND METHODS: This study included sixty patients visiting Department of Radiology Mamata General Hospital, Khammam requiring MRI of the brain and paranasal sinuses. Maxillary sinus dimensions were measured using Siemens software, and statistical analysis was done. RESULTS: The volume and dimensions of the maxillary sinus were more in males when compared to the females with a statistically significant difference. The highest percentage of sexual dimorphism was seen in the volume of left maxillary sinus. Age estimated using the volume of maxillary sinus showed no statistically significant difference from the actual age of the subjects. CONCLUSION: The dimensions and volume of the maxillary sinuses were larger in males than in females, in addition to that they tend to be less with the older age. MRI measurements of maxillary sinuses may be useful to support gender and age estimation in forensic radiology.

Oral adenosquamous carcinoma: Report of a rare entity with a special insight on its histochemistry.

Adenosquamous carcinoma (ASC) of the head and neck (H and N) is an aggressive variant of squamous cell carcinoma (SCC). They are described as SCC subtype with high infiltrative capacity and also presents with dual histomorphology, having both squamous and glandular cell components. ASC of the H and N region is considered as a controversial tumor, as it is similar to salivary gland mucoepidermoid carcinoma. It has been described in a variety of body sites, including uterine cervix, lung and pancreas. ASC rarely develops in the upper aerodigestive tract, particularly in the oral cavity. The affected sites in oral cavity include palate, tonsillar pillar areas and floor of the mouth. To the best of our knowledge in the literature, only 17 cases of ASC in the floor of the mouth have been reported. Hereby, we report an additional case of ASC occurring in the floor of the mouth in a 70-year-old male patient.

Mesiodistal odontometrics as a distinguishing trait: A comparative preliminary study.

INTRODUCTION: Sex determination is a vital step in reconstructing an individual profile from unidentified skeletal remnants. Variations in tooth size are influenced by genetic and environmental factors. Tooth size variations have been reported among different populations. AIM: To identify the sex by determining the mesiodistal (MD) dimensions of maxillary canines. OBJECTIVES: (1) To compare the MD diameter of all maxillary canines - (a) in the entire urban and tribal population, (b) in urban male and urban female populations, (c) in tribal male and tribal female populations, and (d) in the entire male and female populations and (2) To estimate the percentage of sexual dimorphism individually in urban and tribal populations. MATERIALS AND METHODS: Fifty subjects each from urban and tribal populations in equal gender ratio were selected in Khammam district, Telangana, for the purpose of this study. After obtaining informed consent, maxillary study models of the selected subjects were made. MD diameters of left and right maxillary canines were measured on casts using vernier calipers. The obtained data were subjected to statistical analysis. RESULTS: (1) The total tribal population showed a greater MD diameter of maxillary canines than the total urban population, (2) Urban males showed a greater MD diameter of maxillary canines than urban females, (3) Tribal males showed a greater MD diameter of maxillary canines than tribal females, (4) The entire male population showed a greater MD diameter of maxillary canines than the entire female population, and (5) The percentage of dimorphism between males and females in individual groups was found to be significant. CONCLUSION: The study showed maxillary canines exhibiting significant sexual dimorphism and can be used as a distinguishing trait for sex determination along with other procedures.

Immunohistochemical expression of budding uninhibited by benzimidazole related 1 in leukoplakia and oral squamous cell carcinoma.

BACKGROUND: Budding uninhibited by benzimidazole related 1 (BUBR1) is an important protein in the mitotic spindle assembly checkpoint. Alterations in expression of BUBR1 have been reported in many premalignant and malignant lesions. AIM: To compare the expression of BUBR1 with respect to the normal mucosa and degree of dysplasia in oral leukoplakia (OL) and also with respect to different histopathological grades of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Neutral buffered formalin-fixed and paraffin-embedded biopsy specimens 30 each of normal, OL and OSCC tissue were included in this study. The expression of BUBR1 was detected using immunohistochemistry (IHC). The scores obtained were subjected to ANOVA test. RESULTS: Significant correlation was found in immunostaining between normal, dysplasia and OSCC groups with a P value of 0.00001. The expression of BUBR1 was significant when compared with different degrees of dysplasia and in different histopathological grades of OSCC with a P value of 0.00001. CONCLUSION: Higher IHC scores were obtained with increased histopathological grades of OL and OSCC suggesting its role as a prognostic indicator.

Primary biphasic synovial sarcoma of gingiva: Report of a rare case.

Synovial sarcoma is a mesenchymal spindle cell tumor with variable epithelial differentiation. It is unrelated to the synovium as the name might suggest but arises in the soft tissues of the extremities around the knee joints and tendon sheaths. The tumor cells are thought to resemble normal synovial tissue histopathologically, hence named "synovial sarcoma" (SS). Head and neck lesions are less common and oral cavity involvement is extremely rare. Few cases in tongue, soft palate, mandible, buccal mucosa and floor of mouth have been described in the literature. Here, we probably report the first case of primary biphasic SS (BSS) involving gingiva in the retromolar area of the mandible in a 21-year-old male patient.