[A preliminary study on the preparation of perfusion-decellularized laryngeal scaffold and the feasibility of laryngeal muscle reconstruction].
OBJECTIVE
To prepare a decellularized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct decellularized laryngeal muscles.


METHODS
Perfusion decellularized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day's adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed.


RESULTS
Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfusion method showed better decullularized effect. More vintages and collagen fibers but no intact cell or nuclei were retained in the decellularized matrix. Porosity measured by Image pro plus 6.0 was 80.4% +/- 3.2% (x +/- s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86.9% +/- 1.5%. After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistochemical examination indicated that sarcomeric-alpha actin expressed positively in corresponding areas.


CONCLUSIONS
It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.
Hou
Nan
N
Department of Otorhinolaryngology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
Cui
Peng-Cheng
PC
Chen
Wen-Xian
WX
Luo
Jia-Sheng
JS
Ma
Rui-Na
RN
D000818:Animals; D005109:Extracellular Matrix; D005240:Feasibility Studies; D007821:Laryngeal Muscles / Q000166:cytology / Q000502:physiology*; D007831:Larynx, Artificial; D011817:Rabbits; D012038:Regeneration; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds