Auricular and laryngeal chondritis in nursery and finishing pigs.

This work aimed to characterize the clinic-pathological presentation of an outbreak of auricular and laryngeal chondritis in pigs. Visits were made to pig farms, where the clinical history was obtained, and clinical and postmortem examinations were performed. In those farms, 3% to 4% of pigs presented otohematomas, which started in the nursery and extended to the finishing phase. Moreover, some finishing pigs presented with respiratory distress, initially characterized as inspiratory dyspnea, associated by an uncommon respiratory stridor and culminating in death. Grossly, nursery piglets had enlarged ears, and on the cut surface, the cartilage was fragmented and associated with blood clots. In the finishing phase, in addition to auricular lesions, the epiglottis and arytenoid cartilages were thickened and distorted, which partially occluded the lumen. Microscopically, the laryngeal and auricular cartilages were fragmented, displayed a loss of matrix basophilia, and were surrounded by lymphohistiocytic inflammatory infiltrate, with occasional multinucleated giant cells and fibrosis. The lesions exclusively affected elastic cartilages. The disease in finishing pigs led to increased mortality and was a differential diagnosis to respiratory challenges. It was not possible to determine the factor that triggered this condition; however, a nutritional association is suspected. To the authors' knowledge, this is the first report of primary auricular and laryngeal chondritis in pigs.

Remaining microtia tissue as a source for 3D bioprinted elastic cartilage tissue constructs, potential use for surgical microtia reconstruction.

The absence of ears in children is a global problem. An implant made of costal cartilage is the standard procedure for ear reconstruction; however, side effects such as pneumothorax, loss of thoracic cage shape, and respiratory complications have been documented. Three-dimensional (3D) printing allows the generation of biocompatible scaffolds that mimic the shape, mechanical strength, and architecture of the native extracellular matrix necessary to promote new elastic cartilage formation. We report the potential use of a 3D-bioprinted poly-ε-caprolactone (3D-PCL) auricle-shaped framework seeded with remaining human microtia chondrocytes for the development of elastic cartilage for autologous microtia ear reconstruction. An in vivo assay of the neo-tissue formed revealed the generation of a 3D pinna-shaped neo-tissue, and confirmed the formation of elastic cartilage by the presence of type II collagen and elastin with histological features and a protein composition consistent with normal elastic cartilage. According to our results, a combination of 3D-PCL auricle frameworks and autologous microtia remnant tissue generates a suitable pinna structure for autologous ear reconstruction.

Proximal ear hole injury heals by limited regeneration during the early postnatal phase in mice.

Ear pinna is a particular feature of mammals that shows several repair responses depending on age. Two millimeter hole made in the pinna of middle-aged female mice heals due to partial reconstitution of new tissues (limited regeneration), whereas a hole punched in the ear of young mice forms a scar tissue. In these studies, the injury is made in the center of the ear pinna, but little is known about the type of reparative response along the proximodistal polarity of the ear. This study evaluated the effect of pinna polarity, age, and sex in the ear hole-repairing response in Balb/c mice. Proximal injuries were repaired more efficiently by limited regeneration than wounds made in the middle region. Non-injured ear histological analysis revealed a higher presence of muscle, adipose tissue, cartilage, and larger blood vessels in the proximal ear area, which could influence ear hole closure by limited regeneration. To evaluate the healing response during ear growth, we punched a standard hole in the proximal area of the ear on postnatal day 21 and 8-month-old mice (adults). Thirty-five days after the wound, both groups reached the same wound closure, despite the greater proportional size of holes made in the younger mice. Ear growth also improved ear hole closure in male mice. These results suggest that ear growth accelerates hole closure, providing an example of enhanced regenerative abilities in growing structures. Finally, hole closure kinetics in the growing ear indicated an early re-differentiation phase exhibited at 14 days post-wound. In conclusion, ear topography and growth positively influenced the healing response to ear holes, making it a tractable model to study in mammals.

Characterization of pediatric microtia cartilage: a reservoir of chondrocytes for auricular reconstruction using tissue engineering strategies.

The external ear is composed of elastic cartilage. Microtia is a congenital malformation of the external ear that involves a small reduction in size or a complete absence. The aim of tissue engineering is to regenerate tissues and organs clinically implantable based on the utilization of cells and biomaterials. Remnants from microtia represent a source of cells for auricular reconstruction using tissue engineering. To examine the macromolecular architecture of microtia cartilage and behavior of chondrocytes, in order to enrich the knowledge of this type of cartilage as a cell reservoir. Auricular cartilage remnants were obtained from pediatric patients with microtia undergoing reconstructive procedures. Extracellular matrix composition was characterized using immunofluorescence and histological staining methods. Chondrocytes were isolated and expanded in vitro using a mechanical-enzymatic protocol. Chondrocyte phenotype was analyzed using qualitative PCR. Microtia cartilage preserves structural organization similar to healthy elastic cartilage. Extracellular matrix is composed of typical cartilage proteins such as type II collagen, elastin and proteoglycans. Chondrocytes displayed morphological features similar to chondrocytes derived from healthy cartilage, expressing SOX9, COL2 and ELN, thus preserving chondral phenotype. Cell viability was 94.6 % during in vitro expansion. Elastic cartilage from microtia has similar characteristics, both architectural and biochemical to healthy cartilage. We confirmed the suitability of microtia remnant as a reservoir of chondrocytes with potential to be expanded in vitro, maintaining phenotypical features and viability. Microtia remnants are an accessible source of autologous cells for auricular reconstruction using tissue engineering strategies.

Biocompatibility of human auricular chondrocytes cultured onto a chitosan/polyvynil alcohol/epichlorohydrin-based hydrogel for tissue engineering application / Biocompatibilidad de condrocitos humanos cultivados sobre un hidrogel a base de quitosano/alcohol polivnílico/epiclorhidrina para aplicación en ingeniería de tejidos

Tissue engineering (TE) has become an alternative for auricular reconstruction based on the combination of cells, molecular signals and biomaterials. Scaffolds are biomaterials that provide structural support for cell attachment and subsequent tissue development. Ideally, a scaffold should have characteristics such as biocompatibility and bioactivity to adequate support cell functions. Our purpose was to evaluate biocompatibility of microtic auricular chondrocytes seeded onto a chitosan-polyvinyl alcohol-epichlorohydrin (CS-PVA-ECH) hydrogel to propose this material as a scaffold for tissue engineering application. After being cultured onto CS-PVA-ECH hydrogels, auricular chondrocytes viability was up to 81%. SEM analysis showed cell attachment and extracellular matrix formation that was confirmed by IF detection of type II collagen and elastin, the main constituents of elastic cartilage. Expression of elastic cartilage molecular markers during in vitro expansion and during culture onto hydrogels allowed confirming auricular chondrocyte phenotype. In vivo assay of tissue formation revealed generation of neotissues with similar physical characteristics and protein composition to those found in elastic cartilage. According to our results, biocompatibility of the CS-PVA-ECH hydrogel makes it a suitable scaffold for tissue engineering application aimed to elastic cartilage regeneration.

La ingeniería de tejidos (TE) es una alternativa para la reconstrucción auricular basada en la combinación de células, señales moleculares y biomateriales. Los andamios fabricados con biomateriales brindan un soporte estructural que favorece la adhesión cellular y el desarrollo del tejido. Un andamio debe poseer características como biocompatibilidad y bioactividad para soportar adecuadamente funciones celulares. Nuestro objetivo fue evaluar la biocompatibilidad de condrocitos auriculares de microtia cultivados sobre un hidrogel a base de quitosano-alcohol polivinílico-epiclorhidrina (CS-PVA-ECH) y proponerlo como andamio con aplicaciones en ingeniería de tejidos. La viabilidad de los condrocitos auriculares es superior al 81% después de ser cultivados sobre el hidrogel. El análisis por SEM reveló la unión celular y formación de matriz extracellular sobre el hidrogel; confirmada mediante detección por IF de colágena tipo II y elastina. La expresión de marcadores moleculares durante la expansión in vitro y el cultivo sobre los hidrogeles confirmaron el fenotipo condral. El ensayo de formación de tejido in vivo demostró la generación de neotejidos con características físicas y composición similar al cartílago elástico. Nuestros resultados indican que la biocompatibilidad del hidrogel de CS-PVA-ECH lo hace un andamio adecuado para aplicaciones en ingeniería de tejidos enfocadas a regeneración de cartílago elástico.