Characterization of intrauterine growth, proliferation and biomechanical properties of the murine larynx.

Current research approaches employ traditional tissue engineering strategies to promote vocal fold (VF) tissue regeneration, whereas recent novel advances seek to use principles of developmental biology to guide tissue generation by mimicking native developmental cues, causing tissue or allogenic/autologous progenitor cells to undergo the regeneration process. To address the paucity of data to direct VF differentiation and subsequent new tissue formation, we characterize structure-proliferation relationships and tissue elastic moduli over embryonic development using a murine model. Growth, cell proliferation, and tissue biomechanics were taken at E13.5, E15.5, E16.5, E18.5, P0, and adult time points. Quadratic growth patterns were found in larynx length, maximum transverse diameter, outer dorsoventral diameter, and VF thickness; internal VF length was found to mature linearly. Cell proliferation measured with EdU in the coronal and transverse planes of the VFs was found to decrease with increasing age. Exploiting atomic force microscopy, we measured significant differences in tissue stiffness across all time points except between E13.5 and E15.5. Taken together, our results indicate that as the VF mature and develop quadratically, there is a concomitant tissue stiffness increase. Greater gains in biomechanical stiffness at later prenatal stages, correlated with reduced cell proliferation, suggest that extracellular matrix deposition may be responsible for VF thickening and increased biomechanical function, and that the onset of biomechanical loading (breathing) may also contribute to increased stiffness. These data provide a profile of VF biomechanical and growth properties that can guide the development of biomechanically-relevant scaffolds and progenitor cell differentiation for VF tissue regeneration.

Standardizing the Delivery of 20 µL of Hapten During Patch Testing.

BACKGROUND: The current method for patch test tray assembly requires hand dispensing a small volume of hapten onto chambers. Because of human error, this technique produces inaccurate and inconsistent results. The recommended volume of hapten for patch testing using Finn Chambers is 20 µL. OBJECTIVE: The aims of this study were to create a device that standardizes the delivery of 20 µL and to compare it with the current hand dispensing technique. MATERIALS AND METHODS: A device, named the Revolution, was created using the SolidWorks program. Five nurses in our Contact Dermatitis Clinic were asked to load 10 Finn Chambers using the current technique and also using the Revolution. Assembly time, volume of petrolatum, and accuracy of placement were measured. After the 3 trials, the nurses completed a survey on the 2 methods. RESULTS: The amount of petrolatum dispensed using the current technique ranged from 16 to 85 µL, with an average amount of 41.39 µL. The Revolution design dispensed an average of 19.78 µL. CONCLUSIONS: The current hand dispensing technique does not allow for accurate and consistent dispensing of 20 µL for patch testing. In contrast, the Revolution is an accurate and consistent device that can help standardize the patch testing method.