Thymoquinone Case Study
Background and Rationale:
In patients with oral premalignant lesions, does the use of Thymoquinone (TQ) prevent malignant transformation compared to a placebo?
Justification for undertaking the trial:
Over the past three decades oral cancer –although can be avoided- possess a low survival rate nearly around five years. Oral cancer arises from potentially malignant conditions as leukoplakia, erythroplakia, oral lichen planus and other disorders including oral submucous fibrosis (Jainkittivong et al. 2002). All stages of dysplasia the mild, the moderate and the severe dysplasia as well as carcinoma in situ may lead to oral potentially malignant disorders (PMD) and to end with invasive oral squamous cell carcinoma (OSCC). New studies object …show more content…
Composition of capsules
The hard gelatin capsules (size: 2, color: green/white) will consist of: either 100 or 200 mg Nigella Sativa extract together with the following excipients: F-Melt (88 mg) and Crospovidone (5 mg). Both excipients will serve as superdisintegrants.
Group B (Experimental group) Thymoquinone group:
Subjects of this group will receive a capsule containing 200 mg of Nigella Sativa extract twice a day for 3 months.
Group C (Control group) Placebo group:
Subjects of this group will receive the same form of the capsule without active ingredient twice daily for 3 months.
Criteria for discontinuing or modifying intervention:
Subjects will be discontinued from the study if they show evidence of histological or clinical progression of disease, non-hepatic unacceptable toxicity, or a current illness requiring premature termination, pregnant or unwilling to comply with protocol requirements and follow up.
Strategies to improve adherence to intervention …show more content…
Sections will be stained with hematoxylin and eosin (H&E). According to WHO 2005 classification, histopathological grading of epithelial dysplasia and squamous cell carcinoma will be performed [20]. Immunohistochemical analysis:
Onto positively charged slides, sections of 4 μm thick will be mounted and deparaffinized incubation with xylene overnight. The sections will be rehydrated in gradual descending concentrations of ethanol will be followed by a wash with a phosphate-buffered saline (PBS). Blockade of endogenous peroxidase activity will be performed using 3% hydrogen peroxide (H2O2) for 5 minutes at room temperature. For antigen repossession, tissue sections will be retained into glass jars having 0.01M sodium citrate buffer (pH 6.0) and will be boiled in a microwave oven for 5 minutes 2 times to boost immunoreactivity. The slides will cool and will be rinsed with PBS, pH 7.2. Immunohistochemical staining for caspase-3 and ki-67 antibodies will be done according to the manufacturer’s instructions (Thermo Scientific, Thermo Fisher Scientific, Anatomical pathology, Fremont, CA 94538, USA). Detection will be performed using a universal kit (DAKO, Denmark) through washing the slides in PBS for 5 minutes and incubation with secondary antibody (biotinylated goat serum conjugated rabbit
In patients with oral premalignant lesions, does the use of Thymoquinone (TQ) prevent malignant transformation compared to a placebo?
Justification for undertaking the trial:
Over the past three decades oral cancer –although can be avoided- possess a low survival rate nearly around five years. Oral cancer arises from potentially malignant conditions as leukoplakia, erythroplakia, oral lichen planus and other disorders including oral submucous fibrosis (Jainkittivong et al. 2002). All stages of dysplasia the mild, the moderate and the severe dysplasia as well as carcinoma in situ may lead to oral potentially malignant disorders (PMD) and to end with invasive oral squamous cell carcinoma (OSCC). New studies object …show more content…
Composition of capsules
The hard gelatin capsules (size: 2, color: green/white) will consist of: either 100 or 200 mg Nigella Sativa extract together with the following excipients: F-Melt (88 mg) and Crospovidone (5 mg). Both excipients will serve as superdisintegrants.
Group B (Experimental group) Thymoquinone group:
Subjects of this group will receive a capsule containing 200 mg of Nigella Sativa extract twice a day for 3 months.
Group C (Control group) Placebo group:
Subjects of this group will receive the same form of the capsule without active ingredient twice daily for 3 months.
Criteria for discontinuing or modifying intervention:
Subjects will be discontinued from the study if they show evidence of histological or clinical progression of disease, non-hepatic unacceptable toxicity, or a current illness requiring premature termination, pregnant or unwilling to comply with protocol requirements and follow up.
Strategies to improve adherence to intervention …show more content…
Sections will be stained with hematoxylin and eosin (H&E). According to WHO 2005 classification, histopathological grading of epithelial dysplasia and squamous cell carcinoma will be performed [20]. Immunohistochemical analysis:
Onto positively charged slides, sections of 4 μm thick will be mounted and deparaffinized incubation with xylene overnight. The sections will be rehydrated in gradual descending concentrations of ethanol will be followed by a wash with a phosphate-buffered saline (PBS). Blockade of endogenous peroxidase activity will be performed using 3% hydrogen peroxide (H2O2) for 5 minutes at room temperature. For antigen repossession, tissue sections will be retained into glass jars having 0.01M sodium citrate buffer (pH 6.0) and will be boiled in a microwave oven for 5 minutes 2 times to boost immunoreactivity. The slides will cool and will be rinsed with PBS, pH 7.2. Immunohistochemical staining for caspase-3 and ki-67 antibodies will be done according to the manufacturer’s instructions (Thermo Scientific, Thermo Fisher Scientific, Anatomical pathology, Fremont, CA 94538, USA). Detection will be performed using a universal kit (DAKO, Denmark) through washing the slides in PBS for 5 minutes and incubation with secondary antibody (biotinylated goat serum conjugated rabbit