Objective: To quantify the distribution of hyaluronic acid (HA) within the
normal human vocal fold and evaluate gender- and age-related differences. S
tudy Design: Prospective, cross-sectional study by age and gender groups us
ing cadaveric vocal folds to study hyaluronic acid differences. Methods:Twe
nty-five vocal fold halves obtained at autopsy were evaluated from nine adu
lt male (age range, 34-52 y), seven adult female (age range, 21-41 y), four
geriatric male (age range, 65-77 y), and five geriatric female (age range
65-82 y) subjects, Midmembranous vocal fold sections were stained for HA us
ing the Muller-Mowry colloidal iron (acid mucopolysaccharide) stain with an
d without hyaluronidase, Digital imaging software was used to quantify the
distribution of HA across the lamina propria at the vocal fold edge from su
perficial (subepithelial) to deep (vocalis muscle). Results: A significant
gender difference (P =.03) was noted in distribution when controlled for ag
e. Combined, male subjects had a relatively constant distribution pattern t
hroughout the depth of lamina propria, Female subjects showed relatively le
ss HA in the first 15% of depth (most superficial) but showed more HA in th
e deeper 40% to 100% (toward vocalis muscle) than in male subjects. Conclus
ions: Important clinical implications may be extrapolated from our findings
. First, relatively less HA in the most superficial area implies less prote
ction from vibratory trauma and overuse and may explain in part why more fe
male than male patients have phonotrauma to phonotraumatic lesions. Second,
evaluating the distribution of HA in the normal human vocal fold allows mo
re direction in our application of HA as a therapeutic lamina propria subst
itute in patients with voice disorders.