Evaluation of the Levels of Salivary Paxillin Oral Premalignant and Malignant Lesions
Condition: Comparison of Salivary Paxillin Levels in OPMLs and OSCC to Healthy Subjects
MeSH terms: Precancerous Conditions
Study type: Observational

Summary: 
      This investigation was carried out to analyze and compare the salivary paxillin levels
      between oral premalignant and malignant lesions (OPMLs), OSCC and the healthy controls in
      order to assess its potential role as a biomarker of oral cancer aiming for early diagnosis
      and better prognosis of OSCC.

      Methods: Forty-five patients, ranging in age from thirty to seventy-five, were divided into
      three groups: fifteen patients with OPMLs (lichen planus, leukoplakia), fifteen patients with
      OSCC, and fifteen controls who were in general well. Paxillin was identified in saliva
      samples by using an ELISA kit.
    

Detailed description: 
      The aim of this study is: comparison of salivary paxillin levels in oral premalignant lesions
      and OSCC to its levels in healthy subjects.

      Subjects and Methods Forty-five participants between the ages of 30 and 70 were accepted for
      this study. They were registered in the outpatient's clinic of Oral Medicine and
      Periodontology department in Faculty of Dentistry,Cairo university. Three sets of subjects
      were assigned: fifteen patients with premalignant lesions, fifteen patients with OSCC, and
      fifteen systemically healthy control subjects. A complete medical history was obtained for
      each individual using a modified Cornell Medical Index questionnaire. Upon receiving
      clarification regarding the scope of the inquiry, each participant completed an informed
      consent form.

      Patients with OSCC were clinically diagnosed, and biopsy specimens from the lesions were
      taken for histology to confirm the diagnosis.

      Collection of saliva samples:

      Unstimulated Salivary samples were collected once preoperatively before any intervention All
      participants were asked to refrain from drinking, eating, or chewing gum for at least 60
      minutes before sampling. Samples were collected by asking the patient to swallow first, then
      lean the head forward for 5 minutes and spit the entire saliva into 50-ml sterile centrifuge
      tubes. After being assembled, all specimens were immediately stored at -20 ºC until tested.

      Quantitation of Human Paxillin salivary level:

      Saliva was collected from patients and controls. After centrifugation, the supernatant was
      separated for detection of Paxillin by using the ELISA technique provided by Cloud Clone
      Corp, Houston, USA, Catalogue Number: E-01184hu. This kit's microtiter plate has been
      pre-coated with a paxillin-specific antibody. The relevant microtiter plate wells are then
      treated with a biotin-conjugated antibody specific to paxillin. Then, Avidin conjugated to
      Horseradish Peroxidase (HRP) is added and incubated in each microplate well. After adding the
      TMB substrate solution, only the wells containing paxillin, biotin-conjugated antibody, and
      enzyme-conjugated Avidin will change colour. The enzyme-substrate reaction is stopped by
      adding a solution of sulphuric acid, and the colour change is measured spectrophotometrically
      at 450nm 10nm. The samples' optical density (OD) is then compared to the reference curve to
      determine their paxillin concentration.

      Statistical analysis:

      Categorical data were given as frequency and percentage values, and Fisher's exact test was
      used to assess them. Mean and standard deviation (SD) numbers were used to depict numerical
      data. The Shapiro-Wilk test was used to determine their normalcy. A one-way ANOVA test
      followed by Tukey's post hoc test was used to analyse normally distributed data (age and
      Paxillin (ng/mL) levels). ROC curve analysis was done to determine diagnostic accuracy. A
      z-test was used to compare ROC curves. The significance level was set at p<0.05 for all
      tests. R statistical analysis software version 4.3.1 for Windows was used for the statistical
      analysis [18].
    

Intervention name(s): Paxillin biomarker
Intervention type(s): Diagnostic Test
Intervention description(s): Paxillin used as a salivary biomarker
Observational model: Other
Time perspective: Retrospective
Sampling method: Non-Probability Sample
Study population: 
        Forty-five participants between the ages of 30 and 70 were accepted for this study. They
        were registered in the outpatient's clinic of Oral Medicine and Periodontology department
        in Faculty of Dentistry,Cairo university. Three sets of subjects were assigned: fifteen
        patients with premalignant lesions, fifteen patients with OSCC, and fifteen systemically
        healthy control subjects
      
Keywords: oral potentially malignant lesions; oral squamous cell carcinoma; saliva; paxillin