Effect of hyaluronidase on tissue-engineered human septal cartilage

Abstract

Objectives Structural properties of tissue-engineered cartilage can be optimized by altering its collagen to sulfated glycosaminoglycan (sGAG) ratio with hyaluronidase. The objective was to determine if treatment of neocartilage constructs with hyaluronidase leads to increased collagen:sGAG ratios, as seen in native tissue, and improved tensile properties. Study Design Prospective, basic science Methods Engineered human septal cartilage from 12 patients were treated with hyaluronidase prior to culture. Control and treated constructs were analyzed at 3, 6, or 9 weeks for their biochemical, biomechanical, and histological properties. Results Levels of sGAG were significantly reduced in treated compared to control constructs at 3, 6 and 9 weeks. Treated constructs had higher collagen:sGAG ratios when compared with control constructs at 3, 6, and 9 weeks. Treated constructs had greater tensile strength, strain at failure as well as increased stiffness as measured by the equilibrium and ramp tensile moduli when compared with the untreated control constructs. Continued time in culture improved tensile strength in both treated and control constructs. Conclusions Hyaluronidase treatment of engineered septal cartilage decreased total sGAG content without inhibiting expansive growth of the constructs. Decreased sGAG in treated constructs resulted in increased collagen to sGAG ratios and was associated with an increase in tensile strength and stiffness. With additional culture time, sGAG increased modestly in depleted constructs, and some initial gains in tensile properties were dampened. Alterations in the dosage of hyalurondiase during neocartilage fabrication can create constructs that have improved biomechanical properties for eventual surgical implantation. Level of Evidence NA

Toward tissue-engineering of nasal cartilages

“…The choice of stimulus is equally as important as the cell type used. Culture supplementation with GDF-5 and IGF-I, as well as bFGF and TGF-β2 was shown to increase the histological staining intensity of GAG and type II collagen content in scaffold-free human nasal chondrocyte-derived neocartilage [89,106,117]. The use of IGF-I and GDF-5 increased the thickness of engineered neocartilage 12-fold over the untreated control [106] in scaffold-free nasal neocartilage engineered from human septal chondrocytes.…”

“…However, similar to their prominence in articular cartilage engineering, scaffold-free techniques are the most commonly used tissue formation strategy for nasal cartilage. Passaged chondrocytes frequently undergo culture in alginate, dissociation, and seeding into Transwell plates to form neocartilage [72,90,99,106,107,117].…”

“…For example, it has been reported to exhibit a flexure stiffness of 0.014 N/mm [99], a flexure modulus of 0. [117]. Other techniques, such as pellet cultures and multilayered cell sheets are also used [88,118].…”

Tissue engineering applications in otolaryngology—The state of translation

“…Heatmap representing the proportion of publications by year that utilized specific translational research methodologies from construct characterization to human trial to investigate nasal tissue engineering 8‐24,98‐106 …”

Computational technology for nasal cartilage-related clinical research and application

“…For the first nasal cartilage target, with the development of tissue engineering, four elements have gradually been developed: cells, scaffolds, bioactive factors and physical features. 145 Different cells, 146 – 148 scaffolds from different materials, 149 – 153 and different bioactive factors 154 , 155 have been applied to the tissue engineering of nasal cartilage and have achieved good results, but tissue engineering related to physical factors is more common in other types of cartilage. 156 , 157 Nevertheless, the 3D bioprinting of nasal cartilage remains in the experimental stage because it is limited by the clinical application of related bioactive factors.…”