Clin Exp Otorhinolaryngol.  2014 Sep;7(3):193-197. 10.3342/ceo.2014.7.3.193.

Hematoxylin and Eosin Staining for Detecting Biofilms: Practical and Cost-Effective Methods for Predicting Worse Outcomes After Endoscopic Sinus Surgery

• Hong SD
• Dhong HJ
• Chung SK
• Kim HY
• Park J
• Ha SY

• KMID: 1973469
• DOI: http://doi.org/10.3342/ceo.2014.7.3.193

Abstract

OBJECTIVES
Although biofilms have been implicated in poor prognosis after endoscopic sinus surgery (ESS), traditional methods detecting biofilm such as scanning electron microscope and confocal scanning laser microscope were rarely used in the practice. The aims of this study was to determine whether the presence or absence of a biofilm detected by hematoxylin and eosin (H&E) staining followed by light microscopy (LM) that is widely used in daily practice, predicts surgical outcomes after ESS.
METHODS
Retrospective analysis of prospectively collected data. Fifty-five consecutive adult patients (>18 years) who underwent ESS for chronic rhinosinusitis with a minimum of 12-months of follow-up were enrolled in this study. Random sinonasal mucosal samples were assessed for biofilm presence using H&E staining with LM. Three independent observers scored whether a biofilm was present or absent based on H&E staining/LM, and the interrater variability was calculated. Pre- and postoperative sinus symptoms and sinonasal mucosal grading were assessed.
RESULTS
Biofilms were present in 28 patients (51%), and the intraclass correlation coefficient according to H&E staining/LM was 0.731. The presence of a biofilm was associated with a higher preoperative Lund-MacKay computed tomography score (22.3 for biofilm-positive patients vs. 18.6 for biofilm-negative patients; P=0.021) and persistent inflammation (mucosal edema and discharge) after ESS (P<0.05).
CONCLUSIONS
The presence or absence of a biofilm based on H&E staining/LM is correlated with disease severity and surgical outcomes after ESS. H&E staining/LM for detecting biofilm could be practical and cost-effective methods for predicting prognosis of ESS.

Keyword

Biofilms; Chronic rhinosinusitis; Endoscopic sinus surgery; Hematoxylin-eosin staining; Light microscope; Nasendoscopy; Postsurgical outcomes

MeSH Terms

Adult
Biofilms*
Edema
Eosine Yellowish-(YS)*
Follow-Up Studies
Hematoxylin*
Humans
Inflammation
Microscopy
Prognosis
Prospective Studies
Retrospective Studies
Eosine Yellowish-(YS)
Hematoxylin

Figure

• 

  Fig. 1 Appearance of biofilms on H&E-stained sections (×100) from patients with chronic rhinosinusitis. (A) Irregularly-shaped groupings of small basophilic material one third of the size of the surrounding epithelial or inflammatory cells, present over the epithelial lining, not in or under the epithelial lining. (B) Tightly adherent to the surface epithelium or pulled away slightly. (C) Dense extracellular polysaccharide substance material with embedded basophillic substance coating the epithelial surface.

• 

  Fig. 2 Comparison of (A) preoperative symptom scores (Mann-Whitney test, P=0.075) and (B) Lund-MacKay computed tomography scores (Mann-Whitney test, P=0.021) between biofilm-positive (+) and biofilm-negative (-) patients.

• 

  Fig. 3 Comparison of postoperative ongoing persistent symptoms between biofilm-positive (+) and biofilm-negative (-) patients.

• 

  Fig. 4 Comparison of postoperative endoscopic scores including (A) discharge (Fisher exact test, P<0.001) and (B) edema (Fisher exact test, P=0.005) between biofilm-positive (+) and biofilm-negative (-) patients.

• 

  Fig. 5 Mucicarmine staining to distinguish a biofilm from mucin. The red stained area is a mucus-containing area (arrows in A and C). Detection of a biofilm based on H&E staining (arrowhead in B) and mucicarmine staining (arrowhead in D) (×100).

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