title: Short-term retention of labeled chondrocyte subpopulations in stratified tissue-engineered cartilaginous constructs implanted in vivo in mini-pigs.
abstract: It is likely that effective application of cell-laden implants for cartilage defects depends on retention of implanted cells and interaction between implanted and host cells. The objectives of this study were to characterize stratified cartilaginous constructs seeded sequentially with superficial (S) and middle (M) chondrocyte subpopulations labeled with fluorescent cell tracking dye PKH26 (*) and determine the degree to which these stratified cartilaginous constructs maintain their architecture in vivo after implantation in mini-pigs for 1 week. Alginate-recovered cells were seeded sequentially to form stratified S*/M (only S cells labeled) and S*/M* (both S and M cells labeled) constructs. Full-thickness defects (4 mm diameter) were created in the patellofemoral groove of adult Yucatan mini-pigs and filled with portions of constructs or left empty. Constructs were characterized biochemically, histologically, and biomechanically, and stratification visualized and quantified, before and after implant. After 1 week, animals were sacrificed and implants retrieved. After 1 week in vivo, glycosaminoglycan and collagen content of constructs remained similar to that at implant, whereas DNA content increased. Histological analyses revealed features of an early repair response, with defects filled with tissues containing little matrix and abundant cells. Some implanted (PKH26-labeled) cells persisted in the defects, although constructs did not maintain a stratified organization. Of the labeled cells, 126 +/- 38% and 32 +/- 8% in S*/M and S*/M* constructs, respectively, were recovered. Distribution of labeled cells indicated interactions between implanted and host cells. Longer-term in vivo studies will be useful in determining whether implanted cells are sufficient to have a positive effect in repair.
authors.affiliation: Department of Bioengineering, University of California, San Diego, La Jolla, California 92093-0412, USA.
authors.forename: Kanika
authors.initials: K
authors.lastname: Chawla
authors.forename: Travis J
authors.initials: TJ
authors.lastname: Klein
authors.forename: Barbara L
authors.initials: BL
authors.lastname: Schumacher
authors.forename: Kyle D
authors.initials: KD
authors.lastname: Jadin
authors.forename: Bansari H
authors.initials: BH
authors.lastname: Shah
authors.forename: Koichi
authors.initials: K
authors.lastname: Nakagawa
authors.forename: Van W
authors.initials: VW
authors.lastname: Wong
authors.forename: Albert C
authors.initials: AC
authors.lastname: Chen
authors.forename: Koichi
authors.initials: K
authors.lastname: Masuda
authors.forename: Robert L
authors.initials: RL
authors.lastname: Sah
mesh_terms: D000818:Animals; D002358:Cartilage, Articular; D002478:Cells, Cultured; D019902:Chondrocytes; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D019736:Prostheses and Implants; D013194:Staining and Labeling; D013552:Swine; D013556:Swine, Miniature; D023822:Tissue Engineering
chemical_list: D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26