去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized 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 perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. 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. Immunohistoehemical examination indicated that sarcomeric-α 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.

A preliminary study on the preparation of perfusion-decellularized laryngeal scaffold and the feasibility of laryngeal muscle reconstruction