title: The formation of human auricular cartilage from microtic tissue: An in vivo study.
abstract: OBJECTIVE
  This study aimed to isolate, culture-expand and characterize the chondrocytes isolated from microtic cartilage and evaluate its potential as a cell source for ear cartilage reconstruction. Specific attention was to construct the auricular cartilage tissue by using fibrin as scaffold.
  
  
  METHODS
  Cell culture experiment with the use of microtic chondrocytes.
  
  
  METHODS
  Cell culture experiment with the use of microtic chondrocytes.
  
  
  METHODS
  After ear reconstructive surgery at the Universiti Kebangsaan Malaysia Medical Center, chondrocytes were isolated from microtic cartilage. Chondrocytes isolated from the tissue were cultured expanded until passage 4 (P4). Upon confluency at P4, chondrocytes were harvested and tissue engineered constructs were made with human plasma polymerized to fibrin. Constructs formed later is implanted at the dorsal part of nude mice for 8 weeks, followed by post-implantation evaluation with histology staining (Hematoxylin and Eosin (H&E) and Safranin O), immunohistochemistry and RT-PCR for chondrogenic associated genes expression level.
  
  
  RESULTS
  Under gross assessment, the construct after 8 weeks of implantation showed similar physical characteristics that of cartilage. Histological staining showed abundant lacunae cells embedded in extracellular matrix similar to that of native cartilage. Safranin O staining showed positive staining which indicates the presence of proteoglycan-rich matrix. Immunohistochemistry analysis showed the strong positive staining for collagen type II, the specific collagen type in the cartilage. Gene expression quantification showed no significant differences in the expression of chondrogenic gene used which is collagen type I, collagen type II, aggrecan core protein (ACP), elastin and sox9 genes when compared to construct formed from normal auricular tissue.
  
  
  CONCLUSIONS
  Chondrocytes isolated from microtia cartilage has the potential to be used as an alternative cell source for external ear reconstruction in future clinical application.
authors.affiliation: Department of Otorhinolaryngology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors.forename: Mohamad Fikeri bin
authors.initials: MF
authors.lastname: Ishak
authors.affiliation: Department of Otorhinolaryngology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia. Electronic address: irenegbs@yahoo.com.
authors.forename: Goh Bee
authors.initials: GB
authors.lastname: See
authors.affiliation: Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors.forename: Chua Kien
authors.initials: CK
authors.lastname: Hui
authors.affiliation: Department of Otorhinolaryngology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors.forename: Asma bt
authors.initials: Ab
authors.lastname: Abdullah
authors.affiliation: Department of Otorhinolaryngology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors.forename: Lokman bin
authors.initials: Lb
authors.lastname: Saim
authors.affiliation: Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Ear, Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Kuala Lumpur, Malaysia.
authors.forename: Aminuddin bin
authors.initials: Ab
authors.lastname: Saim
authors.affiliation: Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors.forename: Ruszymah bt Haji
authors.initials: Rb
authors.lastname: Idrus
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D065817:Congenital Microtia / Q000628:therapy*; D004425:Ear Cartilage / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D060888:Real-Time Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*
keywords: Auricular chondrocytes; Ear; Fibrin; Microtia; Tissue engineering