Comparing modified papanicolaou stain with ayoub-shklar and haematoxylin-eosin stain for demonstration of keratin in paraffin embedded tissue sections

Department of Oral Pathology, Navodaya Dental College, Raichur, Karnataka, India.
Journal of Oral and Maxillofacial Pathology 03/2013; 17(1):23-30. DOI: 10.4103/0973-029X.110698
Source: PubMed


The aim of the present study was to stain the known keratin containing tissues by Haematoxylin and eosin stain (H-E), ayoub-shklar (A/S) and modified Papanicolaou (PAP) stain and to compare the efficacy of modified PAP staining procedure with that of A/S stain and H-E staining technique, so as to device a staining procedure which is easy and effective for keratin.
A Total Number of 60 paraffin embedded tissue sections of known keratin containing tissues including normal keratinized oral mucosa (NKOM), Keratinized Odontogenic Keratocyst (OKC), Verrucous Carcinoma (VC) and Well differentiated Squamous Cell Carcinoma (WDSCC) were taken and 3 sections of 4 microns thickness of each block were cut and stained with above mentioned three stains.
Surface keratin was stained distinctly and uniformly in all the three staining techniques in NKOM, OKC, and VC and WDSCC. But results were statistically significant in WDSCC when Amount of keratin pearls and Pattern of staining were compared in all 3 staining procedures and p value was p=0.000 and p=0.001 respectively.
Based on the above findings we conclude that the efficacy of modified PAP is comparable with that of H-E stain and A-S stain for surface keratin and thus be used effectively to stain Surface keratin. Whereas to know the exact pattern of cytokeratin expression in SCC a more sensitive tool like immunohistochemical method can be applied.

59 Reads
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: This study evaluated the fluoride release and uptake of five common dental restoratives mainly glass ionomer formulations, including a conventional glass ionomer, a relatively new caries stabilization glass ionomer and resin-modified glass ionomer (Fuji II, Fuji VII and Fuji II LC); one compomer (F2000); and one fluoride releasing composite resin (tetric ceram). A total of 12 cylindrical specimens for each of the five materials were prepared following manufacturer's instructions for manipulation and immersed independently in 25 ml of artificial saliva and stored as five groups Group I-V. Each group was further divided into three sub Groups A, B, C. The saliva was changed every day in all the specimens. No treatment was carried out for the specimens in subgroup A. The specimens were immersed in 2% sodium fluoride for 1 min before changing saliva in sub group B and the specimens were treated by brushing with a fluoridated dentifrice for 2 min before changing saliva in sub Group C. The fluoride release was evaluated on the 1(st), 7(th) and 28(th) day using a fluoride ion specific electrode. The results demonstrated that the conventional glass ionomer and the recently introduced caries stabilizing glass ionomer showed similar patterns and quantity of fluoride release, which was significantly higher than the resin-modified glass ionomer, the compomer and the composite resin. The resin-modified glass ionomer showed higher fluoride release than the compomer and the composite resin. All the formulations of glass ionomers showed fluoride uptake from the neutral sodium fluoride and the fluoridated dentifrice, by releasing increased amounts of fluoride after treatment, in comparison with the untreated group. However, the compomer and the composite resin showed no fluoride uptake. The fluoride released by the glass ionomer cements (GICs) was found to be highest during the first 24 h and decreased significantly over the 1(st) week with lower levels obtained on the 7(th) and 28(th) day, thus demonstrating the phenomenon of "initial burst." The composite resin and compomer used in this study did not show this phenomenon of the initial burst. The resin-modified GICs released more fluoride than the compomer, and the composite resin.
    Full-text · Article · Feb 2015
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Keratins are the most abundant proteins and are characteristic findings in many epithelial pathologies, making it a diagnostically important marker, both histopathologically and immunohistochemically. Since, immunohistochemistry is an expensive diagnostic tool, special stains to detect the degree of keratinization could serve as a faster and economic option. The aim of the present study was to compare the efficacy of special stains for keratin with standard hematoxylin and eosin stain (H and E). Objectives include: (i) To subject the diagnosed cases of keratin disorders to the selected special stains: Ayoub-shklar method, Dane-Herman method, Alcian blue -periodic acid Schiff 's (PAS), rapid papanicolaou (PAP) and Gram's stain. (ii) To compare the staining specificity and staining intensity of special stains with respect to routine hematoxylin and eosin (H and E) stain. (iii) To compare the efficacy of special stains to routine H and E stain in identification of the type of keratin present in the selected cases. A total of 80 cases of known pathology for keratin were retrieved from the department archive, which included 10 each of normal gingiva, hyperkeratosis, squamous papilloma, verrucous hyperplasia, verrucous carcinoma, well-differentiated squamous cell carcinoma, orthokeratinized odontogenic cyst and keratocystic odontogenic tumors. Six sections of 4 ยต each from the paraffin blocks were made, stained with H and E and the special stains and these were evaluated by 2 pathologists based on the modified scoring criteria from Rahma Al-Maaini and Philip Bryant 2008. The results were tabulated using Chi square and kappa statistics. The statistical values for identification of the type of keratinization was insignificant showing that ortho and parakeratinized epithelia could be correctly identified by both H and E as well as all the special stains. Furthermore, all the special stains showed a positive result and statistical significance (P < 0.001) with respect to the staining of keratin. To conclude, though the special stains distinctly stained the keratin with a higher intensity, H and E proves to be overall better stain with respect to specificity.
    Full-text · Article · Mar 2015
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The aim of the study is to compare hematoxylin and eosin (H&E) stained tissue sections with Papanicolaoustained (PAP-stained) tissue sections. Paraffin block tissue sections of five oral pathologies were prepared and stained routinely with H&E and PAP stains. The results of the stained slides revealed a marked difference in staining with respect to color balance, contrast and intensity between H&E and PAP stains. Areas of keratinization were stained from shades of orange to pink depending on the degree of keratinization. In moderately differentiated squamous cell carcinoma, areas of keratin pearl formation and individual cell keratinzation were clearly demonstrated; these were not apparent in the H&E stained slides. The review of this study suggest that the PAP stain more clearly demonstrated key morphological features, associated with keratinization, and it may be considered as a supplemental staining procedure, aiding in diagnosis.
    Full-text · Article · Mar 2015 · Journal of histotechnology