The formation of human auricular cartilage from microtic tissue: An in vivo study
| Autor: | M F Ishak, Asma bt Abdullah, Goh Bee See, Idrus Ruszymah Bt Haji, Aminuddin Bin Saim, Chua Kien Hui, Lokman Saim |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Pathology
medicine.medical_specialty Cell Culture Techniques Mice Nude Matrix (biology) Real-Time Polymerase Chain Reaction Extracellular matrix Mice Chondrocytes Ear Cartilage medicine Animals Humans Aggrecan Congenital Microtia Tissue Engineering biology business.industry Cartilage General Medicine Anatomy Chondrogenesis Immunohistochemistry Staining medicine.anatomical_structure Otorhinolaryngology Pediatrics Perinatology and Child Health biology.protein business Elastin |
| Zdroj: | International Journal of Pediatric Otorhinolaryngology. 79:1634-1639 |
| ISSN: | 0165-5876 |
| DOI: | 10.1016/j.ijporl.2015.06.034 |
| Popis: | Objectives 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. Study design Cell culture experiment with the use of microtic chondrocytes. Design 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. Conclusion Chondrocytes isolated from microtia cartilage has the potential to be used as an alternative cell source for external ear reconstruction in future clinical application. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect