Various
growth factor delivery systems were used in the treatment
of glottal insufficiency; however, relatively little attention has
been paid to a gene delivery system for aspects of active vocal fold
(VF) regeneration. Herein, we present a plasmid DNA (pDNA; bFGF gene
encoding) complex-loaded alginate (ALG)/hyaluronic acid (HA) mixture
hydrogel dispersed with polycaprolactone (PCL) microspheres that can
enhance simultaneous regeneration of VF muscle and lamina propria,
as well as have a bulking effect on atrophied VF. We have demonstrated
long-term efficacy of bFGF synthesized from pDNA complex-transfected
cells in vitro. PCL microspheres–dispersed
ALG/HA hydrogel (with or without pDNA complex loading) are injected
into rabbit VFs with recurrent laryngeal nerve denervation. The PCL
microspheres dispersed in the hydrogel bulking agents remain stable
at the applied site, leading to constant medialization of the paralyzed
VF without significant initial volume loss even after 24 weeks. A
regenerative effect for collagen deposition and HA synthesis around
the injected site, which are major components of VF tissue, is well
confirmed in the pDNA-complex-loaded hydrogel group. Moreover, the
compensation of atrophied VFs also leads to the contact of bilateral
VF and the remarkable recovery of voice function in the pDNA-complex-loaded
group. Based on the results, pDNA (bFGF encoding) complex-loaded hydrogel
dispersed with PCL microspheres may be employed as a bioactive bulking
agent for the treatment of glottal insufficiency.