First evidence of a functional spiracle in stem chondrichthyan acanthodians, with the oldest known elastic cartilage.

Spiracles are a general character of gnathostomes (jawed fishes), being present in antiarch placoderms, commonly regarded as the most basal gnathostome group. The presence of spiracular tubes in acanthodians has been deduced from grooves on the neurocranium of the derived acanthodiform Acanthodes bronni from the Permian of Germany, but until now these tubes were presumed to lack an external opening, rendering them non-functional. Here we describe the external spiracular elements in specimens of the Middle Devonian acanthodiforms Cheiracanthus murchisoni, Cheiracanthus latus and Mesacanthus pusillus from northern Scotland, and the internal structure of these elements in C. murchisoni, demonstrating that the spiracle in acanthodiforms differed from all known extant and extinct fishes in having paired cartilage-pseudobranch structures. This arrangement represents a transitional state between the presumed basal gnathostome condition with an unconstricted first gill slit (as yet not identified in any fossil) and the derived condition with a spiracle and a single pseudobranch derived from the posterior hemibranch of the mandibular arch. We identify the main tissue forming the pseudobranch as elastic cartilage, a tissue previously unrecorded in fossils.

Development of scaffold-free elastic cartilaginous constructs with structural similarities to auricular cartilage.

External ear reconstruction with autologous cartilage still remains one of the most difficult problems in the fields of plastic and reconstructive surgery. As the absence of tissue vascularization limits the ability to stimulate new tissue growth, relatively few surgical approaches are currently available (alloplastic implants or sculpted autologous cartilage grafts) to repair or reconstruct the auricle (or pinna) as a result of traumatic loss or congenital absence (e.g., microtia). Alternatively, tissue engineering can offer the potential to grow autogenous cartilage suitable for implantation. While tissue-engineered auricle cartilage constructs can be created, a substantial number of cells are required to generate sufficient quantities of tissue for reconstruction. Similarly, as routine cell expansion can elicit negative effects on chondrocyte function, we have developed an approach to generate large-sized engineered auricle constructs (≥3 cm(2)) directly from a small population of donor cells (20,000-40,000 cells/construct). Using rabbit donor cells, the developed bioreactor-cultivated constructs adopted structural-like characteristics similar to native auricular cartilage, including the development of distinct cartilaginous and perichondrium-like regions. Both alterations in media composition and seeding density had profound effects on the formation of engineered elastic tissue constructs in terms of cellularity, extracellular matrix accumulation, and tissue structure. Higher seeding densities and media containing sodium bicarbonate produced tissue constructs that were closer to the native tissue in terms of structure and composition. Future studies will be aimed at improving the accumulation of specific tissue constituents and determining the clinical effectiveness of this approach using a reconstructive animal model.