Auricular Scaffold Research Articles

Bioengineering Full-scale auricles using 3D-printed external scaffolds and decellularized cartilage xenograft

Reconstruction of the human auricle remains a formidable challenge for plastic surgeons. Autologous costal cartilage grafts and alloplastic implants are technically challenging, and aesthetic and/or tactile outcomes are frequently suboptimal. Using a small animal “bioreactor”, we have bioengineered full-scale ears utilizing decellularized cartilage xenograft placed within a 3D-printed external auricular scaffold that mimics the size, shape, and biomechanical properties of the native human auricle. The full-scale polylactic acid ear scaffolds were 3D-printed based upon data acquired from 3D photogrammetry of an adult ear. Ovine costal cartilage was processed either through mincing (1 mm3) or zesting (< 0.5 mm3), and then fully decellularized and sterilized. At explantation, both the minced and zested neoears maintained the size and contour complexities of the scaffold topography with steady tissue ingrowth through 6 months in vivo. A mild inflammatory infiltrate at 3 months was replaced by homogenous fibrovascular tissue ingrowth enveloping individual cartilage pieces at 6 months. All ear constructs were pliable, and the elasticity was confirmed by biomechanical analysis. Longer-term studies of the neoears with faster degrading biomaterials will be warranted for future clinical application. Statement of significanceAccurate reconstruction of the human auricle has always been a formidable challenge to plastic surgeons. In this article, we have bioengineered full-scale ears utilizing decellularized cartilage xenograft placed within a 3D-printed external auricular scaffold that mimic the size, shape, and biomechanical properties of the native human auricle. Longer-term studies of the neoears with faster degrading biomaterials will be warranted for future clinical application.

Current Status of Auricular Reconstruction Strategy Development.

Microtia has severe physical and psychological impacts on patients, and auricular reconstruction offers improved esthetics and function, alleviating psychological issues. Microtia is a congenital disease caused by a multifactorial interaction of environmental and genetic factors, with complex clinical manifestations. Classification assessment aids in determining treatment strategies. Auricular reconstruction is the primary treatment for severe microtia, focusing on the selection of auricular scaffold materials, the construction of auricular morphology, and skin and soft tissue scaffold coverage. Autologous rib cartilage and synthetic materials are both used as scaffold materials for auricular reconstruction, each with advantages and disadvantages. Methods for achieving skin and soft tissue scaffold coverage have been developed to include nonexpansion and expansion techniques. In recent years, the application of digital auxiliary technology such as finite element analysis has helped optimize surgical outcomes and reduce complications. Tissue-engineered cartilage scaffolds and 3-dimensional bioprinting technology have rapidly advanced in the field of ear reconstruction. This article discusses the prevalence and classification of microtia, the selection of auricular scaffolds, the evolution of surgical methods, and the current applications of digital auxiliary technology in ear reconstruction, with the aim of providing clinical physicians with a reference for individualized ear reconstruction surgery. The focus of this work is on the current applications and challenges of tissue engineering and 3-dimensional bioprinting technology in the field of ear reconstruction, as well as future prospects.

Proteoglycan 4 (PRG4) treatment enhances wound closure and tissue regeneration

The wound healing response is one of most primitive and conserved physiological responses in the animal kingdom, as restoring tissue integrity/homeostasis can be the difference between life and death. Wound healing in mammals is mediated by immune cells and inflammatory signaling molecules that regulate tissue resident cells, including local progenitor cells, to mediate closure of the wound through formation of a scar. Proteoglycan 4 (PRG4), a protein found throughout the animal kingdom from fish to elephants, is best known as a glycoprotein that reduces friction between articulating surfaces (e.g. cartilage). Previously, PRG4 was also shown to regulate the inflammatory and fibrotic response. Based on this, we asked whether PRG4 plays a role in the wound healing response. Using an ear wound model, topical application of exogenous recombinant human (rh)PRG4 hastened wound closure and enhanced tissue regeneration. Our results also suggest that rhPRG4 may impact the fibrotic response, angiogenesis/blood flow to the injury site, macrophage inflammatory dynamics, recruitment of immune and increased proliferation of adult mesenchymal progenitor cells (MPCs) and promoting chondrogenic differentiation of MPCs to form the auricular cartilage scaffold of the injured ear. These results suggest that PRG4 has the potential to suppress scar formation while enhancing connective tissue regeneration post-injury by modulating aspects of each wound healing stage (blood clotting, inflammation, tissue generation and tissue remodeling). Therefore, we propose that rhPRG4 may represent a potential therapy to mitigate scar and improve wound healing.

Fabrication of chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold for microtia reconstruction.

Fibrin gel-based scaffolds have promising potential for microtia reconstruction. Autologous chondrocytes and chondrocyte cell sheets are frequently used seed cell sources for cartilage tissue engineering. However, the aesthetic outcome of chondrocyte-based microtia reconstruction is still not satisfactory. In this study, we aimed to fabricate the chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold for microtia reconstruction. We designed a unique auricular mold that could fabricate a fibrin gel scaffold resembling human auricle anatomy. Primary chondrocytes were harvested from rabbit auricular cartilage, and chondrocyte cell sheets were developed. Chondrocyte-microtissues were prepared from the cell sheets. The mixture of chondrocytes/chondrocyte-microtissues was laden in fibrin gel during the auricular scaffold fabrication. The protrusions and recessed structure in the auricular scaffold surface were still clearly distinguishable. After a one-week in vitro culture, the 3 D structure and auricular anatomy of the scaffold were retained. And followed by eight-week subcutaneous implantation, cartilaginous tissue was regenerated in the artificial auricular structure as indicated by the results of H&E, Toluidine blue, Safranin O, and type II collagen (immunohistochemistry) staining. Protrusions and depressions of the auricular scaffold were slightly deformed, but the overall auricular anatomy was maintained after 8-week in vivo implantation. Extracellular matrix components content were similar in artificial auricular cartilage and rabbit native auricular cartilage. In conclusion, the mixture of chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold showed a promising potential for cartilaginous tissue regeneration, suggesting this as an effective approach for autologous chondrocyte-based microtia reconstruction.

Mechanical study of polyurethane elastomer and Medpor as the material of artificial auricular scaffold

By comparing the mechanics of human auricular cartilage, polyurethane elastic material, and high density polyethylene material (Medpor), to produce theoretical proof on choosing optimal artificial auricular scaffold materials. The experimental materials were divided into 3 groups with 6 samples in each: the auricular cartilage group (group A), the polyurethane elastic material group (group B), and the Medpor group (group C). With an Instron5967 mechanical testing machine, compression and tensile testing were performed to respectively measure values of compression parameters (including yield stress, yield load, elastic modulus, yield compressibility, compressibility within 2 MPa, and compression stress within 10% strain) and values of tensile parameters (including yield stress, yield load, elastic modulus, yield elongation, elongation within 2 MPa, tensile stress within 1% strain) for comparison. Compression testing: no obvious yield points were observed in the whole process in samples of group B, while obvious yield points were observed in samples of groups A and C. There was no significant difference between groups A and C with respect to yield stress and yield load ( P>0.05); while the yield compressibility in group C was significantly lower than that in group A ( P<0.05) and the elastic modulus in group C was significantly higher than that in group A ( P<0.05). There was a significant difference with respect to compressibility within 2 MPa of materials among the 3 groups ( P<0.05), the high, medium, and low values go to groups B, A, and C respectively. The compression stress within 10% strain in group C was significantly higher than that in groups A and B ( P<0.05), and there was no significant difference between that in groups A and B ( P>0.05). Tensile testing: the materials in group B had extremely high tensile strength. The yield stress in groups A and B was significantly higher than that in group C ( P<0.05), and the elastic modulus and tensile stress within 1% strain were significantly lower than those in group C ( P<0.05); but no significant difference was found between those in groups A and B ( P>0.05). There was no significant difference with respect to yield load among the 3 groups ( P>0.05); but there was significant difference with respect to yield elongation among the 3 groups ( P<0.05), and the high, medium, and low values go to groups B, A, and C respectively. The elongation within 2 MPa in group B was significantly higher than that in groups A and C ( P<0.05), and there was no significant difference between that in groups A and C ( P>0.05). Compared with the Medpor, the polyurethane elastic material is a more ideal artificial auricular scaffold material.

Pore architecture effects on chondrogenic potential of patient-specific 3-dimensionally printed porous tissue bioscaffolds for auricular tissue engineering

ObjectiveThis study aims to determine the effect of auricular scaffold microarchitecture on chondrogenic potential in an in vivo animal model. MethodsDICOM computed tomography (CT) images of a human auricle were segmented to create an external anatomic envelope. Image-based design was used to generate 1) orthogonally interconnected spherical pores and 2) randomly interspersed pores, and each were repeated in three dimensions to fill the external auricular envelope. These auricular scaffolds were then 3D printed by laser sintering poly-l-caprolactone, seeded with primary porcine auricular chondrocytes in a hyaluronic acid/collagen hydrogel and cultured in a pro-chondrogenic medium. The auricular scaffolds were then implanted subcutaneously in rats and explanted after 4 weeks for analysis with Safranin O and Hematoxylin and Eosin staining. ResultsAuricular constructs with two micropore architectures were rapidly manufactured with high fidelity anatomic appearance. Subcutaneous implantation of the scaffolds resulted in excellent external appearance of both anterior and posterior auricular surfaces. Analysis on explantation showed that the defined, spherical micropore architecture yielded histologic evidence of more robust chondrogenic tissue formation as demonstrated by Safranin O and Hematoxylin and Eosin staining. ConclusionsImage-based computer-aided design and 3D printing offers an exciting new avenue for the tissue-engineered auricle. In early pilot work, creation of spherical micropores within the scaffold architecture appears to impart greater chondrogenicity of the bioscaffold. This advantage could be related to differences in permeability allowing greater cell migration and nutrient flow, differences in surface area allowing different cell aggregation, or a combination of both factors. The ability to design an anatomically correct scaffold that maintains its structural integrity while also promoting auricular cartilage growth represents an important step towards clinical applicability of this new technology.

Ear Reconstruction and 3D Printing: Is It Reality?

Autologous reconstruction of microtia, the most common congenital external ear deformity, is one of the more challenging plastic surgical procedures, causing obligatory donor site morbidity and often resulting in suboptimal aesthetic outcomes. Recent advances in the fields of tissue engineering and 3D printing promise to profoundly affect the practice of reconstructive surgery. 3D printed guides are already used by reconstructive surgeons during correction of complex anatomic defects. Similarly, the prosthetics industry has benefited from the ability to rapidly prototype customized pieces. Bioprinting, the ability to 3D print living tissue, is an emerging field that may soon allow the possibility of creating autologous cartilage in specific shapes. In this review, we explore the numerous ways 3D printing is being employed to address external ear deformity and how, when used in combination with cutting-edge tissue engineering technology, it may finally help us achieve the holy grail of ear reconstruction—an “off-the-shelf” auricular scaffold.

Design and development of nanocomposite scaffolds for auricular reconstruction

Auricular reconstruction using sculpted autologous costal cartilage is effective, but complex and time consuming and may incur donor site sequelae and morbidity. Conventional synthetic alternatives are associated with infection and extrusion in up to about 15% of cases. We present a novel POSS-PCU nanocomposite auricular scaffold, which aims to reduce extrusion rates by mimicking the elastic modulus of human ears and by encouraging desirable cellular interactions. The fabrication, physicochemical properties (including nanoscale topography) and cellular interactions of these scaffolds were compared to Medpor®, the current synthetic standard. Our scaffold had a more similar elastic modulus (5.73±0.17MPa) to ear cartilage (5.02±0.17MPa) compared with Medpor®, which was much stiffer (140.9±0.04MPa). POSS-PCU supported fibroblast ingrowth and proliferation; significantly higher collagen production was also produced by cells on the POSS-PCU than those on Medpor®. This porous POSS-PCU nanocomposite scaffold is therefore a promising alternative biomaterial for auricular surgical reconstruction. From the Clinical EditorIn this paper, a novel POSS-PCU nanocomposite auricular scaffold is described to reduce extrusion rates by having a much closer elastic modulus of human ears than the currently available synthetic standard. Enabling desirable cellular interactions may lead to the successful clinical application of these novel scaffolds.

Tissue engineering auricular reconstruction: in vitro and in vivo studies

Although investigators have demonstrated that neocartilage can be constituted in a predetermined shape and in complex three-dimensional structures, such as a human ear, by using cell transplantation on polymer constructs, many unsolved problems still remain. The crucial issues for auricular tissue engineering consisted of optimal cell culture environment, choice of polymers, behavior of chondrocytes, study of cell–polymer constructs in an acceptable animal model, and long-term structural integrity. Here we describe our tissue engineering approaches for auricular reconstruction including auricular scaffold fabrication, in vitro chondrogenesis, in vivo immunocompromized xenograft and immunocompetent autologous animal models, and long-term follow-up. Though many current obstacles regarding auricular tissue engineering still exist, we demonstrate techniques of auricular scaffold fabrication with promising in vitro and in vivo neocartilage formation, optimal selection and application of animal models, and, to the best of our knowledge, the first report of different biodegradable biomaterial trials and the longest in vivo results (10 months) for auricular tissue engineering.