Laryngopharyngeal Reflux Scoring in a Pediatric Population

Abstract

In recent years, the prevalence of laryngopharyngeal reflux has risen, especially among pediatric patients. The diagnosis of laryngopharyngeal reflux relies on patient history and clinical assessment using the Reflux Finding Score and Reflux Symptom Index as crucial diagnostic tools. Some studies have proposed a link between pepsin and laryngopharyngeal reflux, potentially triggering palatine tonsil hypertrophy. Our study aimed to investigate the correlation between laryngeal and pharyngeal manifestations of laryngopharyngeal reflux through two questionnaires and the presence of pepsin in saliva and palatine tonsils in a pediatric population. Pepsin in saliva was detected using a Western blot method, while immunohistochemistry assessed its presence in palatine tonsils. Although no statistically significant differences in Reflux Finding Score and Reflux Symptom Index were found between the immunohistochemistry-positive (IHC-positive) and immunohistochemistry-negative (IHC-negative) groups, median reflux symptom index and Reflux Finding Score values consistently trended higher in the IHC-positive group. This suggests a potential connection between elevated index values and pepsin presence in tonsillar tissue. Further investigations are essential to fully comprehend the clinical implications of these findings.

Keywords: laryngopharyngeal reflux; palatine tonsil; pepsin; reflux finding score; reflux symptom index; saliva; tonsillectomy.

Figure 1

(A) Anatomical position of the palatine tonsils: 1—hard palate; 2—uvula; 3—anterior faucial pillar; 4—posterior faucial pillar; 5—palatine tonsil. (B) Histological section through the palatine tonsil: 1—crypt; 2—lymphoepithelium; 3—germinal center of lymphatic follicles; 4—mantle zone; 5—area between lymphatic follicles. Magnification 200×; scale 50 µm.

Figure 2

Immunohistochemistry of tonsil tissue stained against pepsin and visualized with DAB-HRP reaction. Positive staining was mostly visualized around germinal centers, connective tissue, and marginal zones. (A) Human stomach as positive control; (B) tonsil tissue—positive staining; (C) tonsil tissue—negative staining. Magnification 200×; scale 50 µm. A positive control was a human stomach in which positive reaction was much more pronounced in all cellular and muscular structures.

Figure 3

Box and whisker plots representing (A) the Reflux Finding Score (RFS) and (B) the Reflux Symptom Index (RSI). Statistical analysis was performed using the Mann–Whitney U test to assess potential significant differences, with a significance level (α) set at 0.05. The results of the Mann–Whitney U test revealed the following: for RFS, the U statistic (URFS) = 594.5, p-value (p) = 0.20174; for RSI, the U statistic (URSI) = 598.5, p-value (p) = 0.21686.

Figure 4

Quantitation of immunoblotting assays of pepsin in saliva. (A) Box and whisker plots representing the percentage of pepsin signal calculated against 1 µg of pepsin standard loaded in the first well. Statistical analysis was performed using the Mann–Whitney U test to assess potential significant differences, with a significance level (α) set at 0.05. The results of the Mann–Whitney U statistic (Uwb) = 716.5, p-value (p) = 0.99584. (B) Representative Western blot membrane of immunoblotting against pepsin in saliva. The first two samples are porcine pepsin standards, followed by saliva samples. Signal intensity normalization was performed using stain-free blots.

Figure 4

Quantitation of immunoblotting assays of pepsin in saliva. (A) Box and whisker plots representing the percentage of pepsin signal calculated against 1 µg of pepsin standard loaded in the first well. Statistical analysis was performed using the Mann–Whitney U test to assess potential significant differences, with a significance level (α) set at 0.05. The results of the Mann–Whitney U statistic (Uwb) = 716.5, p-value (p) = 0.99584. (B) Representative Western blot membrane of immunoblotting against pepsin in saliva. The first two samples are porcine pepsin standards, followed by saliva samples. Signal intensity normalization was performed using stain-free blots.