Hyaluronan in vocal folds and false vocal folds in patients with recurrent respiratory papillomatosis

Figures & data

Table 1. Sample characteristics of RRP patients, biopsy site: 1: false vocal folds, 2: vocal folds.

Histochemistry
Patient	Biopsy site	HPV type	Onset	Vaccine	# of surgeries	Age at biopsy surgery	gender
1	1,2	6	Adult		4	57	Male
2	1,2	6	Adult	Yes (finished 2013)	15	46	Male
3	1,2	6	Adult	Yes (finished 2014)	10	37	Male
4	1,2	6	Adult		9	74	Male
5	1,2	6	Adult		2	34	Male
6	1,2	6	Adult		2	40	Female
7	1,2	6	Adult		2	33	Male
8	1,2	6	Adult		2	67	Male
9	1,2	6	Adult		1	26	Male
10	1,2	6	Adult		13	37	Male
11	1,2	6	Adult		1	33	Male
12	1,2	6	Adult	Yes (finished 2016)	10	45	Male
Gemma
Patient	Biopsy site	HPV type	Onset	Vaccine	# of surgeries	Age at biopsy surgery	gender
13	1,2	6	Juvenile		1	2	Male
14	1,2	11	Adult		2	26	Female
15	1,2	11	Adult		5	50	Female
16	1,2	6	Adult		6	43	Male
17	1,2	6	Adult		5	51	Male
18	1	6	Juvenile		6	6	Female
19	1	6	Adult		12	67	Male
20	1	16	Adult		6	45	Male
21	1,2	6	Juvenile		2	6	Female
22	1,2	6	Adult		4	43	Male
23	1,2	6	Adult		1	51	Female
24	1,2	6	Juvenile		51	39	Male

Figure 1. Flowchart for histochemistry and GEMMA (gas-phase electrophoretic molecular mobility analyzer analysis), FVF: false vocal folds, VF: vocal folds.

Figure 2. Mass distribution analysis of hyaluronan (HA) using GEMMA (gas-phase electrophoretic molecular mobility analyzer). LMHA: Low mass Hyaluronan. HMHA: high mass hyaluronan. Two major peaks of HA were detected ranging from about 50 kDa to larger than HA standards (>10 MDa). There was a trend of relatively more of the very high mass HA (vHMHA) in the vocal folds compared to false vocal folds. HA from skin biopsy analyses at our laboratory did not show vHMHA.

Table 2. Scoring of HA and CD44 throughout the epithelium, stroma and RRP lesion in vocal folds and false vocal folds.

HA staining intensity in false and true vocal folds
	Epithelium		Stroma		RRP lesion
Patient	False vocal folds	Vocal folds	False vocal folds	Vocal folds	False vocal folds	Vocal folds
1	1	0	1	0	1	No RRP lesion
2	0	0	1	2	0	0
3	0	0	1	1	0	0
4	0	0	0	1	No RRP lesion	No RRP lesion
5	0	1	2	1	No RRP lesion	1
6	0	0	1	1	No RRP lesion	0
7	0	0	2	1	No RRP lesion	1
8	0	0	1	0	0	0
9	Stain absent	1	Stain absent	2	Stain absent	No RRP lesion
10	0	0	1	2	0	0
11	0	0	1	2	No RRP lesion	No RRP lesion
12	0	0	1	2	No RRP lesion	No RRP lesion
∅	0.09	0.17	1.09	1.25	0.2	0.28
0 = no staining, 1 = faint staining, 2 = intense staining
CD44 staining intensity in false and true vocal folds
1	1-	1-	1-	1-	1-	No RRP lesion
2	1-	1-	1-	1-	1-	1-
3	1-	1-	1-	1-	1-	1+
4	1-	1-	1-	1-	No RRP lesion	No RRP lesion
5	1-	1-	1-	1-	No RRP lesion	1-
6	Stain absent	Stain absent	Stain absent	Stain absent	Stain absent	Stain absent
7	1-	1-	1-	1-	No RRP lesion	1-
8	1-	Stain absent	1-	Stain absent	1-	Stain absent
9	Stain absent	1-	Stain absent	1-	Stain absent	No RRP lesion
10	1-	1-	1-	1-	No RRP lesion	1-
11	1-	1-	1-	1-	No RRP lesion	No RRP lesion
12	1-	1-	1-	1-	No RRP lesion	No RRP lesion

0: no staining, 1: staining, +: CD44 congruent with HA staining, −: CD44 not congruent with HA staining.

Figure 3. Area under curve (AUC) from GEMMA (gas-phase electrophoretic molecular mobility analyzer) mass distribution analysis. AUC equals relative amount of hyaluronan in tissue. Low mass hyaluronan (LMHA) and high mass hyaluronan (HMHA) showed little variation between true and false vocal folds. High variation was observed of very high mass hyaluronan (vHMHA) in true vocal folds compared to false vocal folds. Levene’s test for equality of variances, vHMHA p = .046