title: Evaluation of nanoarchitectured collagen type II molecules on cartilage engineering.
abstract: Scaffold architecture, including the geometry and dimension of scaffolds, is an important parameter in cell adhesion, migration, proliferation, and differentiation. Following the characterization of collagen type II nanoarchitectured molecules, collagen fibrils (CNFs) and collagen spheres (CNPs) prepared using a high-voltage electric field in our laboratory, we proposed to use these nanoarchitectured molecules to assess their influence on the culturing of chondrocytes in stirred bioreactors. The results demonstrate that chondrocytes rapidly formed more and larger chondrocyte pellets (spheroids) after the addition of nanoarchitectured molecules into the culture medium. The maintenance of chondrocytes with round morphology and increased glycosaminoglycan secretion indicated that these spheroids contained viable and un-dedifferentiated chondrocytes. No significant increases in DNA content were detected. These results show that the introduction of these molecules did not affect chondrocyte proliferation during a 3-day culture period. After the addition of CNPs and CNFs into the culture medium, the expression levels of collagen type II and aggrecan genes in chondrocytes increased significantly as demonstrated by real-time PCR analysis. Interestingly, chondrocytes exhibited distinct collagen type II and aggrecan gene expression profiles in culture with CNPs and CNFs. The aggrecan gene expression level of the chondrocytes was 2.5-fold greater following CFN addition than following the addition of CNPs. In contrast, the collagen type II expression level of the chondrocytes was 2.2-fold greater following the addition of CNPs than following the addition of CNFs. The chondrocyte pellets rapidly restored defects in articular cartilage during a 1-month implantation period in a rabbit model.
authors: Department of Biomedical Engineering, I-Shou University, Kaohsiung, Taiwan.
authors: Shyh Ming
authors: SM
authors: Kuo
authors: Ming Yu
authors: MY
authors: Chiang
authors: Cheng Wen
authors: CW
authors: Lan
authors: Gregory Cheng-Chie
authors: GC
authors: Niu
authors: Shwu Jen
authors: SJ
authors: Chang
mesh_terms: D000423:Alcian Blue / Q000378:metabolism; D000818:Animals; D002356:Cartilage / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D024043:Collagen Type II / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D054118:Nanospheres / Q000737:chemistry* / Q000648:ultrastructure; D011817:Rabbits; D013194:Staining and Labeling; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D000423:Alcian Blue