Differences between human septal and alar cartilage with respect to biomechanical features and biochemical composition.

Cartilage grafts have become popular in facial plastic surgery to reconstruct defects or to improve aesthetic outcomes in various applications. But there is a considerable rate of graft failure like resorption or deformation. To improve graft survival and function, accurate understanding of the properties of the recipient site is indispensable. Therefore 10 noses of human cadavers were meticulously dissected and specimens of alar and septal cartilage subjected to confined compression and tensile tests. Furthermore, cell number, glycosaminoglycan and hydroxyproline content were measured. RESULTS: showed a significant difference (p < 0.05) of alar and septal cartilage regarding Equilibrium Modulus, cell number and glycosaminoglycan but not hydroxyproline content. Tensile tests showed a significant difference (p < 0.001) between alar and septal cartilage (vertical vector of force) for E-modulus, maximal force and maximal strain but not for horizontal vector of force. There was a significant difference (p < 0.05) within septal cartilage samples depending on vector of force (vertical vs. horizontal). Finally multifactorial linear regression allowed an estimation of Equilibrium Modulus depending on compression, glycosaminoglycan content and cell number with statistical significance (p < 0.05). In conclusion, nasal cartilage differs in function and composition depending on anatomical location and the prevalent forces. Therefore further research will be necessary to evaluate if graft failure depends on a mismatch of functional properties and if grafts can be adapted to the recipient site.

Cartilage regeneration using decellularized cartilage matrix: Long-term comparison of subcutaneous and intranasal placement in a rabbit model.

Autologous cartilage as donor tissue for various surgical reconstructions such as nasal septum regeneration is limited and associated with donor site morbidity. Our goal was to evaluate a new resorbable chondroconductive biomaterial made of decellularized porcine nasal septum cartilage compared with autologous native auricular cartilage as the gold standard. In order to examine the material and determine its long-term outcome further, we used subcutaneous implantation and septal implantation in an orthotopic rabbit model. In addition to non-seeded decellularized xenogenic cartilage, chondrocyte-seeded decellularized xenogenic cartilage was implanted as a septal replacement. After a three- or six-month period, the formation of newly synthesized cartilage extracellular matrix was evaluated immunohistochemically, whereas septal integrity and biocompatibility were evaluated histologically. The formation of the implanted neoseptum and form stability was analyzed by using 7-Tesla Magnetic Resonance Imaging. Good biocompatibility with no excessive rejection was demonstrated in all groups. Long-term stable and reliable septal reconstruction could be achieved in the study groups with or without cell seeding with autologous auricular chondrocytes. Autologous cell seeding was advantageous only with regard to septal perforations. Thus, cell seeding provides a benefit regarding long-term stability. However, because of slightly better biocompatibility, less pronounced septum deviation and the markedly lower effort involved, the non-seeded scaffold is favoured for possible clinical application.

Acoustic Properties of Collagenous Matrices of Xenogenic Origin for Tympanic Membrane Reconstruction.

HYPOTHESIS: The acoustic properties of scaffolds made from decellularized extracellular cartilage matrices of porcine origin are comparable to those of the human tympanic membrane. BACKGROUND: Currently, the reconstruction of tympanic membrane in the context of chronic tympanic membrane defects is mostly performed using autologous fascia or cartilage. Autologous tissue may be associated with lack of graft material in revision patients and requires more invasive and longer operative time. Therefore, other materials are investigated for reconstruction. An increasingly important role could be played by scaffolds from different materials, which are known to induce constructive tissue remodeling. METHODS: To analyze the acoustic properties, the vibrations of the scaffolds, cartilage, perichondrium and tympanic membrane were measured by laser scanning doppler vibrometry under different static pressures. RESULTS: The analysis of volume velocities serves as an indicator for sound transmission. The results of the average volume velocities at atmospheric pressure show a similar frequency response of the tympanic membrane and the scaffolds with a peak at about 800 Hz. After changing the artificial ear-canal pressure from atmospheric pressure to negative pressure (-100, -200, and -300 daPa) the vibration characteristics of the different membranes remain fairly constant, whereas the results of the perichondrium show a decrease after changing the pressure into the negative range in the frequencies 1 to 3 kHz. CONCLUSION: The present study showed that the vibration characteristics of the scaffolds under atmospheric and negative pressure can be interpreted as similar to those of thin cartilage (<0.5 mm) and human tympanic membranes. However, in relation to the behavior of these scaffolds made from decellularized extracellular cartilage matrices in vivo, further investigations should be carried out.

Processed xenogenic cartilage as innovative biomatrix for cartilage tissue engineering: effects on chondrocyte differentiation and function.

One key point in the development of new bioimplant matrices for the reconstruction and replacement of cartilage defects is to provide an adequate microenvironment to ensure chondrocyte migration and de novo synthesis of cartilage-specific extracellular matrix (ECM). A recently developed decellularization and sterilization process maintains the three-dimensional (3D) collagen structure of native septal cartilage while increasing matrix porosity, which is considered to be crucial for cartilage tissue engineering. Human primary nasal septal chondrocytes were amplified in monolayer culture and 3D-cultured on processed porcine nasal septal cartilage scaffolds. The influence of chondrogenic growth factors on neosynthesis of ECM proteins was examined at the protein and gene expression levels. Seeding experiments demonstrated that processed xenogenic cartilage matrices provide excellent environmental properties for human nasal septal chondrocytes with respect to cell adhesion, migration into the matrix and neosynthesis of cartilage-specific ECM proteins, such as collagen type II and aggrecan. Matrix biomechanical stability indicated that the constructs retrieve full stability and function during 3D culture for up to 42 days, proportional to collagen type II and GAG production. Thus, processed xenogenic cartilage offers a suitable environment for human nasal chondrocytes and has promising potential for cartilage tissue engineering in the head and neck region.

In vitro cytotoxicity and in vivo effects of a decellularized xenogeneic collagen scaffold in nasal cartilage repair.

Tissue engineering is considered a promising future option for nasal cartilage repair. However, until now, an optimal material has not been identified for this specific purpose. Therefore, the aim of this study was to analyze a recently developed decellularized collagen matrix, which has promising material properties for septal cartilage repair. A tetrazolium dye based cytotoxicity assay using rat nasal septum chondrocytes was performed to examine the cytotoxic effects of decellularized cartilage matrices. Unseeded scaffolds as well as scaffolds seeded with chondrocytes were implanted in nasal septum defects in Lewis rats to investigate the cellular and humoral inflammatory responses in the surrounding tissue as well as the effect on the formation of nasal septum perforations. Samples were analyzed histochemically and immunohistochemically after 1, 4, and 12 weeks. Although cells for the cytotoxicity assay were cultured under serum-free conditions for 24 h to increase sensitivity, no cytotoxic effects were detected. Histological and immunohistochemical evidence displayed that the implanted scaffolds induced minor macrophage and lymphocyte infiltration and were well integrated at the contact site to native cartilage and between the mucosal membranes. The biocompatibility index revealed only slightly irritating effects during the study period. Septal perforations were prevented efficiently. In summary, our results provide evidence that decellularized xenogeneic collagen scaffolds are suitable for cartilage tissue engineering. The scaffolds were integrated well into septal cartilage defects without causing a strong inflammatory reaction and prevented the development of nasal septum perforations. Therefore, we envision the possibility to use them in nasal cartilage repair in the future.

Decellularized cartilage matrix as a novel biomatrix for cartilage tissue-engineering applications.

Damage of cartilage structures in the head and neck region as well as in orthopedic sites are frequently caused by trauma, tumor resection, or congenital defects. Despite a high demand in many clinical fields, until today, no adequate cartilage replacement matrix is available for these fields of application. Materials that are clinically applied for joint cartilage repair still need optimization due to difficult intraoperative handling and risk of early mechanical damage. We have developed and applied a novel chemical process to completely decellularize and sterilize human and porcine cartilage tissues (meniscus cartilage and nasal septum) to generate a new type of bioimplant matrix. To characterize this matrix and to determine the effect of the decellularization process, the content of denatured collagen (w(D)) and the content of glycosaminoglycans (GAGs) (w(G)) were determined. Possible cytotoxic effects and cellular compatibility of the matrix in vitro have been examined by seeding processed cartilage biomatrices with human primary chondrocytes as well as murine fibroblasts (L929). Vitality and state of metabolism of cells were measured using MTS assays. Both cell types adhered to scaffold surfaces and proliferated. No areas of growth inhibition or cytotoxic effects were detected. New synthesis of cartilage-specific extracellular matrix was observed. By histological staining, electron microscopy, and µCT analysis, an increase of matrix porosity, complete cell elimination, and high GAG removal were demonstrated. Being from natural-origin, processed xenogenic and allogeneic cartilage biomatrices are highly versatile with regard to shape, size, and biomechanics, making them promising candidates for various biomedical applications.