title: Tissue-engineered larynx using perfusion-decellularized technique and mesenchymal stem cells in a rabbit model.
abstract: CONCLUSIONS
  Reseeding mesenchymal stem cells (MSCs) into the decellularized laryngeal muscle matrix for construction of a tissue-engineered larynx is feasible. This in vivo maturation in the omentum could be the first step before in situ implantation of the construct. This construct could facilitate a tissue-engineered method for laryngeal reconstruction.
  
  
  OBJECTIVE
  The extracellular matrix (ECM) and MSCs have been widely used for organ reconstruction. Our study aimed to prepare a soft tissue decellularized laryngeal scaffold with intact laryngeal cartilages utilizing a perfusion-decellularized technique, reseeding cells on it, and then construct a recellularized larynx.
  
  
  METHODS
  Perfusion-decellularized larynges were obtained from 20 rabbits by perfusion of the common carotid arteries with detergents. Twelve perfused larynges were observed by macroscopic visualization, histological examination, scanning electron microscopy (SEM), and cartilage viability. The remaining eight perfusion constructs were reseeded with induced MSCs aspirated from eight receptor rabbits. Composites were transferred into greater omentums of receptor rabbits after adherence for 1 day in vitro. Rabbit larynges were harvested after 4 weeks and 8 weeks, respectively. Macroscopic visualization, histological examination, and immunohistochemistry were performed.
  
  
  RESULTS
  Larynges perfused by sodium dodecyl sulfate became transparent after 2 h of perfusion. Histology and SEM indicated that the perfusion method showed a better decullularized effect. Almost no intact cells or nuclei were found, while more pores and collagen fibers were retained in the decellularized matrix. The chondrocyte vitality assay indicated that chondrocyte vitality was high. Vascularization was clearly seen by 4 weeks and relatively integrated cartilage frameworks remained by 8 weeks. Histological and immunohistochemical examinations clearly showed muscle bundles and vessels.
authors.affiliation: Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital, Chengdu Medical College, Sichuan Province, PR China. bihaichaoxin.hn@163.com
authors.forename: Nan
authors.initials: N
authors.lastname: Hou
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Cui
authors.forename: Jiasheng
authors.initials: J
authors.lastname: Luo
authors.forename: Ruina
authors.initials: R
authors.lastname: Ma
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000502:physiology; D019902:Chondrocytes / Q000166:cytology; D005109:Extracellular Matrix; D007817:Laryngeal Cartilages / Q000166:cytology; D007821:Laryngeal Muscles / Q000166:cytology; D007830:Larynx / Q000098:blood supply / Q000166:cytology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D018919:Neovascularization, Physiologic / Q000502:physiology; D009852:Omentum; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds