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Tissue engineering of auricular cartilage with human chondrocytes and nanoPGA scaffolds
Hitomi Nakao, Mark Shasti, Robin Jacquet, Josh Bundy, Seika Matsushima, Noritaka Isogai, Ananth Murthy, and William Landis
Akron Children's Hospital
2014-03-14
Presenter: Ananth Murthy
Affidavit:
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Director Name: Ananth Murthy
Author Category: Other Specialty Resident
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Tissue engineering represents an approach to augmentation and replacement of microtia and other auricular cartilage impairments. A previous study obtained microtia and normal auricular cartilage (from otoplasty) on surgery from young patients (4-14 years old) and investigated for 5 and 10 weeks in vivo auricular cartilage secreted extracellular matrix molecules. This study reports results from 20 and 40 week samples. Isolated chondrocytes from specimens were cultured one week in Dulbecco's Modified Eagle Medium: Ham's F-12 (50:50) supplemented with 10 ng/ml FGF-2 and antibiotic/antimycotic. Chondrocyte suspensions (108 cells/ml) were applied to thin (80 µm) polymer scaffolds consisting of biodegradable nano-polyglycolic acid (nPGA; nanofibers ≤ 1 µm in diameter) and cultured an additional week. Chondrocyte/scaffold constructs (n = 6 for each cell type) implanted in athymic mice for 20 and 40 weeks were harvested and the length, width, and thickness of constructs were measured at three equally spaced locations along these parameters. Constructs were sectioned (5 µm thick) and stained histologically with toluidine blue for cell and extracellular matrix structure and with Safranin-O red and Verhoeff solution for detection of proteoglycans and elastin, respectively. After 20 and 40 weeks of implantation, light microscopy demonstrated chondrocytes, abundant matrix, proteoglycans and elastin in sections of constructs for both microtia and normal auricular cells. No significant differences were found in construct dimensions or histology comparing the two types of neocartilage. Data provide support for use of nPGA for extensive (to 40 weeks) development of both cartilage types and usefulness of scaffolds of degradable nPGA.
