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Tissue engineering of auricular cartilage with human chondrocytes and nanoPGA scaffolds
Hitomi Nakao, MD, Mark Shasti, BS, Robin Jacquet, MS, Josh Bundy, Seika Matsushima, MD, PhD, Noritaka Isogai, MD, PhD, Ananth Murthy, MD, and William Landis, PhD
University of Akron
2013-03-01
Presenter: Hitomi Nakao
Affidavit:
I do do certify.
Director Name: William J. Landis, Ph.D.
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: Craniomaxillofacial
Tissue engineering represents a potential approach to augmentation and replacement of microtia and other auricular cartilage impairments. In that context, microtia and normal auricular cartilage (from otoplasty) were obtained surgically from young patients (4-15 years old) and investigated in vivo for extracellular matrix structure and auricular cartilage secreted molecules. Isolated chondrocytes from specimens were cultured one week in Dulbecco's Modified Eagle Medium: Hams 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 scaffold sheets consisting for the first time of biodegradable nano-polyglycolic acid (nPGA; nanofibers ≤ 1 µm in diameter; and cultured an additional week. Chondrocyte/scaffold constructs (n = 6) were implanted in athymic (nude) mice for 5 or 10 weeks. On harvest from mice, constructs were paraffin-embedded, 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. Light microscopy demonstrated numerous chondrocytes and abundant matrix in tissue sections and presence of proteoglycans and elastin in nPGA- engineered constructs for both microtia and normal auricular cells after 5 or 10 weeks implantation. Results provide evidence supporting nPGA for extensive growth and development of both cell types and usefulness of scaffolds of degradable nPGA. Further studies will utilize nPGA with a variety of other more resilient polymers for auricular cartilage tissue engineering.
