title: Cartilage engineering using chondrocyte cell sheets and its application in reconstruction of microtia.
abstract: The imperfections of scaffold materials have hindered the clinical application of cartilage tissue engineering. The recently developed cell-sheet technique is adopted to engineer tissues without scaffold materials, thus is considered being potentially able to overcome the problems concerning the scaffold imperfections. This study constructed monolayer and bilayer chondrocyte cell sheets and harvested the sheets with cell scraper instead of temperature-responsive culture dishes. The properties of the cultured chondrocyte cell sheets and the feasibility of cartilage engineering using the chondrocyte cell sheets was further investigated via in vitro and in vivo study. Primary extracellular matrix (ECM) formation and type II collagen expression was detected in the cell sheets during in vitro culture. After implanted into nude mice for 8 weeks, mature cartilage discs were harvested. The morphology of newly formed cartilage was similar in the constructs originated from monolayer and bilayer chondrocyte cell sheet. The chondrocytes were located within evenly distributed ovoid lacunae. Robust ECM formation and intense expression of type II collagen was observed surrounding the evenly distributed chondrocytes in the neocartilages. Biochemical analysis showed that the DNA contents of the neocartilages were higher than native human costal cartilage; while the contents of the main component of ECM, glycosaminoglycan and hydroxyproline, were similar to native human costal cartilage. In conclusion, the chondrocyte cell sheet constructed using the simple and low-cost technique is basically the same with the cell sheet cultured and harvested in temperature-responsive culture dishes, and can be used for cartilage tissue engineering.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Libin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Ruiying
authors.initials: R
authors.lastname: Ding
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Baowei
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Haolun
authors.initials: H
authors.lastname: Han
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Hongnan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Bingxin
authors.initials: B
authors.lastname: Xu
authors.affiliation: Institute of Otolaryngology, Chinese PLA General Hospital Beijing 100853, P.R China.
authors.forename: Suoqiang
authors.initials: S
authors.lastname: Zhai
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, 306 Hospital of PLA Beijing 100101, P.R China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
mesh_terms: D000293:Adolescent; D000818:Animals; D002356:Cartilage; D018929:Cell Culture Techniques; D019902:Chondrocytes; D065817:Congenital Microtia / Q000601:surgery*; D005109:Extracellular Matrix; D005260:Female; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D023822:Tissue Engineering / Q000379:methods*
keywords: Cell sheet; cartilage; chondrocyte; extracellular matrix; tissue engineering; type II collagen