Mouth and Tooth Diseases

Skin and Connective Tissue Diseases

All Conditions

Neoplasms

Symptoms and General Pathology

Rare Diseases

Lichen Planus, Oral

Lichen Planus

Precancerous Conditions

Leukoplakia

Leukoplakia, Oral

Lichenoid Eruptions

Exanthema

Skin Diseases

Skin Diseases, Papulosquamous

Mouth Diseases

Stomatognathic Diseases

Pathological Conditions, Anatomical

Mouth Neoplasms

Head and Neck Neoplasms

Oral Leukoplakia

Oral Cancer

Coagulants

All Drugs and Chemicals

Fibrinolytic Agents

Anticoagulants

Tolonium Chloride

Hemostatics

Heparin

Calcium heparin

Lesions that were stained positive with rose bengal were biopsied and assessed for dysplasia

Lesions that were stained positive with toluidine blue were biopsied and assessed for dysplasia

rose bengal stain and toluidine is applied over a premalignant lesion and positively stained lesions are subjected to biopsy.

Biopsy

the lesions stained positive were subjected to biopsy

Rose Bengal

toluidine blue

Abstract

Objective:

To study the diagnostic efficiency of Rose Bengal with Toluidine blue in detecting the biopsy sites and thus establish an accurate diagnosis in oral premalignant lesions.

Materials and method:

In our study 27 patients with 41 lesions were included. Since one patient had not quit the habit in the two weeks following initial examination and another lesion disappeared in the waiting period, 2 patients (3 lesions) were not included in the study. Out of 38 lesions diagnosed based on clinical criteria, 32 were leukoplakia, 5 lichen planus and 1 SCC. After initial examination they were subjected to Rose Bengal and Toluidine blue stain. If stained positive they were subjected to biopsy.

Introduction Toluidine blue staining is the most common technique used for the early detection of dysplastic changes in patients with premalignant lesions. One meta-analysis of Rosenberg et al. previously published reported sensitivity ranged from 93.5% to 97.8% and the specificity ranged from 73.3% to 92.9%. Zang et al reported that TB not only detects high-grade dysplasia but detects OPLs with minimal or no dysplasia with high-risk clinical and molecular attributes.4 But, studies have shown as high as 30% risk of false-positive staining.

Rose Bengal (RB) has been widely used to diagnose various ocular surface disorders including delineation of the extent of corneal and conjunctival neoplasms. It has been believed to stain desquamated ocular epithelial cells, dead or degenerated cells, or wherever there is poor protection of the surface epithelium by the preocular tear ﬁlm rather than lack of cell vitality. These characteristic features of RB lead the researchers to apply it in oral premalignant lesions.5 In none of the studies, reliability of RB stains was not compared with existing or previously practiced methods in oral premalignant lesions. Hence this study was undertaken with the aim of comparing the RB and TB stain and for early detection of dysplasia in oral premalignant lesions.

Materials and Methods:

Study group consists of 41 oral premalignant lesions in patients visiting the Department of Oral Medicine and Radiology of The oxford dental college and hospital, Bangalore. Patients with OSMF, patients with bleeding disorders, patient with other systemic diseases were excluded from the study.To perform the present study, ethical clearance was obtained from the Institutional Ethical Board. Study procedure was explained and informed consent was taken from the selected patients with premalignant lesions. The patients with lesions were subjected to detailed case history, intra oral examination and photographs of the lesions were recorded. Patients with habits were counselled to quit the habit and recalled after 2 weeks for staining. Since one patient had not quit the habit in the two weeks from first visit, in another patient the lesion disappeared in two weeks 2 patients (3 lesions) were not included in the study. (Graph1)

Initially patients were asked to rinse their mouth with distilled water for 1 minute. 1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed. Following this, patients were asked to rinse their mouth with 1% acetic acid for 1 min to remove the remaining RB stain from the lesion and were prepared for TB staining. 1%Toluidineblue is applied over the lesion and after 30 seconds patients were made to swish with 1% acetic acid and the area stained was recorded photographically.

These two photographs were assessed and if the stained area was similar in both the procedures single biopsy was taken. If the stained areas were different two different biopsies were taken.

Biopsy specimen of the lesion was retained.

Comparison or Rose Bengal and Toluidine Blue Staining for Lesion Detection Efficacy

Efficacy of Rose Bengal in Comparison With Toluidine Blue in Detection of Premalignant Lesions: a Preliminary Study

sensitivity: Percentage of lesions stained positive with Toluidine blue stain that were also stained positive with Rose Bengal stain specificity: Percentage of lesions stained negative with Toluidine blue stain that were also stained negative with Rose Bengal stain

this outcome measure was limited to the biopsied lesions.additionally, the intent of this outcome measure was only to study early detection of dysplasia in oral premalignant lesions using the Rose bengal stain. (no comparison between the two stains were intended )

The Oxford Dental College, Hospital and Research Center, Bangalore, India

doctor

lesions were subjected to rose bengal staining and positive lesions were subjected to biopsy

Rose Bengal Positive Lesion

lesions were subjected to toluidine blue staining and positive lesions were subjected to biopsy

Toluidine Blue Positive Lesions and Biopsy

All study participants received Rose Bengal and Toulidine Blue. Patients were asked to rinse their mouth with distilled water for 1 minute. 1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed. Following this, patients were asked to rinse their mouth with 1% acetic acid for 1 min to remove the remaining RB stain from the lesion and were prepared for TB staining. 1% Toluidine blue was applied over the same lesion and after 30 seconds patients were made to swish with 1% acetic acid and the area stained was photographed.

All Study Participants

a proforma was used to record the baseline details

Age, Categorical

Age, Continuous

Sex: Female, Male

patients were asked regarding the usage of tobacco products and was recorded in proforma by investigator

tobacco habit

location of the lesion

all the patients who had premalignant lesions and had stopped using tobacco in the past two weeks

1. small sample size
2. only lesions stained positive were selected for biopsy procedure hence interpretation of the staining was limited to certain lesions only
3. a colorimetric guide was not used measure the intensity of staining

Dr. Anisha Yaji

Initially patients were asked to rinse their mouth with distilled water for 1 minute. 1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed. positively stained lesions were subjected for biopsy.

Rose Bengal Staining

Following RB staining, patients were asked to rinse their mouth with 1% acetic acid for 1 min to remove the remaining RB stain from the lesion and were prepared for TB staining. 1% Toluidine blue was applied over the same lesion and after 30 seconds patients were made to swish with 1% acetic acid and the area stained was photographed. positively stained lesions were subjected to biopsy.

Toluidine Blue Staining

patients diagnosed with premalignant lesions who consented to participate in the study

Number and Percentage of Lesions That Were Stained Positive

sensitivity

specificity

1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed.8 Following this, patients were asked to rinse their mouth with 1% acetic acid for 1 min to remove the remaining RB stain from the lesion.

1% toluidine blue applied over the lesion and after 30 seconds were made to swish with 1% acetic acid. the area retaining the stain was photographed and compared with that of rose bengal. only positively stained areas were subjected to biopsy procedure.

Sensitivity and Specificity (Percentage of True Positives and True Negatives)

dysplasia present

dysplasia absent

squamous cell carcinoma

Patients were asked to rinse their mouth with distilled water for 1 minute. 1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed. Positively stained lesions were biopsied.

Rose Bengal Staining and Biopsy

Presence of Dysplasia in Biopsied Lesions That Were Stained by Rose Bengal

All patients received Rose Bengal and Toluidine Blue. Initially patients were asked to rinse their mouth with distilled water for 1 minute. 1% RB solution was applied with a cotton tip for 2 minutes. Again patients were asked to rinse their mouth for 1 minute with distilled water to remove excess RB solution and the area which had taken up the stain was photographed. Following this, patients were asked to rinse their mouth with 1% acetic acid for 1 min to remove the remaining RB stain from the lesion and were prepared for TB staining. 1% Toluidine blue was applied over the same lesion and after 30 seconds patients were made to swish with 1% acetic acid and the area stained was photographed.

Overall Study

Carcinoma

Carcinoma, Squamous Cell

Squamous Cell Carcinoma of Head and Neck

Neoplasms, Glandular and Epithelial

Neoplasms by Histologic Type

Neoplasms, Squamous Cell

Oral Squamous Cell Carcinoma

Hana'a H Algadi, Msc student

The purpose of this study is to test the accuracy of toluidine blue in the assessment of intraoperative tumor margin after excision of oral squamous cell carcinoma(OSCC)in comparison to H\&E stain on frozen section.

The incidence of oral cancer remains high, it is ranked to be the sixth most common cancer worldwide, oral squamous cell carcinoma represent more than 90% of all oral malignances.

Standard treatment for oral squamous cell carcinoma is surgical resection with free margin. Negative margin status is an important for control local recurrence and improve outcome in squamous cell carcinoma of the head and neck. Hematoxylin and Eosin (H\&E) Frozen section biopsy is useful tool for intraoperative assessment of resected margins and control of local recurrence, allowing for additional resection intraoperatively in case of positive margins. However in the least developed countries with limited resource, the frozen section is not available in a majority of centers, in addition to that if present in developed countries, it somewhat considered time and cost consumer.

Toluidine blue(TB) is metachromatic stain, easily available, economical, with high affinity for DNA and RNA. It is rapidly uptaken by malignant epithelium which contain quantitatively more nucleic acid than normal tissue. It has been widely used as screening tool for malignant and premalignant lesion.

The purpose of this study is to test the accuracy of toluidine blue in the assessment of intraoperative tumor margin after excision of oral squamous cell carcinoma(OSCC)in comparison to H\&E stain on frozen section.

Toluidine Blue Versus Frozen Sections for Assessment of Tumor Margins in Oral Squamous Cell Carcinoma

The diagnostic accuracy will be assessed by comparing the results of each technique with the final routine histopathological results.

Study title	NCT ID	Conditions	Interventions	Eligibility	Organization
Comparison or Rose Bengal and Toluidine Blue Staining for Lesion Detection Efficacy	NCT03031899	Oral Leukoplaki... Oral Lichen Pla... Premalignant Le...	Biopsy Rose Bengal toluidine blue	18 Years -	The Oxford Dental College, Hospital and Research Center, Bangalore, India
Toluidine Blue Versus Frozen Sections for Assessment of Tumor Margins in Oral Squamous Cell Carcinoma	NCT03554967	Oral Squamous C...		-	Cairo University
Comparison or Rose Bengal and Toluidine Blue Staining for Lesion Detection Efficacy	NCT03031899	Oral Leukoplaki... Oral Lichen Pla... Premalignant Le...	Biopsy Rose Bengal toluidine blue	18 Years -	The Oxford Dental College, Hospital and Research Center, Bangalore, India
Toluidine Blue Versus Frozen Sections for Assessment of Tumor Margins in Oral Squamous Cell Carcinoma	NCT03554967	Oral Squamous C...		-	Cairo University

Spots Global Cancer Trial Database for toluidine blue