15502-m-pleumeekers

MATERIALS AND METHODS Chemicals were obtained from Sigma-Aldrich, USA unless stated otherwise. Sample harvesting and preparation Cadaveric auricles were harvested by Science Care (Phoenix, Arizona, USA, n =4) and Erasmus Medical Center (Rotterdam, the Netherlands, n =11) according to ethical guidelines of the respective institution. Additionally ear (EC) and nasoseptal (NC) cartilage was obtained from patients ( n =30, EC; n =69, NC) undergoing middle ear or cholesteatoma surgery (EC) and functional septo- or septorhinoplasties (NC) at University Hospital Zurich (Zurich, Switzerland), and Ulm University Medical Center (Ulm, Germany) according to the ethics regulations of the respective institution. EC samples were harvested from 15 male and 12 female donors, NC samples were harvested from 40 male and 12 female donors. Samples were pooled according to anthropomorphic age (child, <20 years; young adult, 20–34; middle adult, 35–49; and old adult, ≥50). [152] All samples were shipped at 4°C in phosphate buffered saline (PBS) supplemented with antibiotic/antimycotic (Gibco, Invitrogen Corporation, California, USA) to ETH Zurich (Zurich, Switzerland). The perichondrium was removed from EC samples, and cylindrical plugs (5 mm, 1–2 mm thick) were cut perpendicular to the surface. Six harvesting regions were defined (anti-helix, AH; anti-tragus, AT; concha, CO; helix, HE; scapha, SC; tragus, TR). (Figure 1A) NC plugs were similarly prepared, where samples originated from the center of the nasal septum. (Figure 1B) Differences in sample number for biomechanical and biochemical assays is due to sample loss during processing, unusual sample shape preventing mechanical analysis or limitations of biochemical assays. Figure 1. Anatomy of human ear and nasal septum. (A) Map of the human auricle. Six harvesting regions are identified based on the ear morphology: anti-helix, AH, anti-tragus, AT, concha, CO, helix, HE, scapha, SC and tragus TR. Adapted from Atlas der Anatomie des Menschen, B.N. Tillmann, Springer–Lehrbuch. [153] (B) Harvesting site for nasoseptal cartilage. Adapted from Gray’s Anatomy of the Human Body, Henry Gray. [154] 30 CHAPTER 2