High-speed spinning of collagen microfibers comprising aligned fibrils for creating artificial tendons.

Bundles of engineered collagen microfibers are promising synthetic tendons as substitutes for autogenous grafts. The purpose of this study was to develop high-speed and continuous spinning of collagen microfibers that involves stretching of collagen stream. Our study revealed the 'critical fibrillogenesis concentration (CFC)' of neutralized collagen solutions, which is defined as the upper limit of the collagen concentration at which neutralized collagen molecules remain stable as long as they are cooled (⩽10 °C). Neutralized collagen solutions at collagen concentrations slightly below the CFC formed cord-like collagen gels comprising longitudinally aligned fibrils when extruded from nozzles into an ethanol bath. Dry collagen microfibers with a controlled diameter ranging from 122 ± 2-31.2 ± 1.7 µm can be spun from the cord-like gels using nozzles of various sizes. The spinning process was improved by including stretching of collagen stream to further reduce diameter and increase linear velocity. We extruded a collagen solution through a 182 µm diameter nozzle while simultaneously stretching it in an ethanol bath during gelation and fiber formation. This process resembles the stretching of a melted thermoplastic resin because it solidifies during melt spinning. The mechanical properties of the stretched collagen microfibers were comparable to the highest literature values obtained using microfluidic wet spinning, as they exhibited longitudinally aligned fibrils both on their surface and in their core. Previous wet spinning methods were unable to generate collagen microfibers with a consistent tendon-like fibrillar arrangement throughout the samples. Although the tangent modulus (137 ± 7 MPa) and stress at break of the swollen bundles of stretched microfibers (13.8 ± 1.9 MPa) were lower than those of human anterior cruciate ligament, they were within the same order of magnitude. We developed a spinning technique that produces narrow collagen microfibers with a tendon-like arrangement that can serve as artificial fiber units for collagen-based synthetic tendons.

Fabrication and Properties of Electrospun Collagen Tubular Scaffold Crosslinked by Physical and Chemical Treatments.

Tissue engineered scaffold was regarded as a promising approach instead of the autograft. In this study, small diameter electrospun collagen tubular scaffold with random continuous smooth nanofibers was successfully fabricated. However, the dissolution of collagen in concentrated aqueous (conc. aq.) acetic acid caused to the serious denaturation of collagen. A novel method ammonia treatment here was adopted which recovered the collagen triple helix structure according to the analysis of IR spectra. Further dehydrothermal (DHT) and glutaraldehyde (GTA) treatments were applied to introduce the crosslinks to improve the properties of collagen tube. The nanofibrous structure of collagen tube in a wet state was preserved by the crosslinking treatments. Swelling ratio and weight loss decreased by at least two times compared to those of the untreated collagen tube. Moreover, tensile strength was significantly enhanced by DHT treatment (about 0.0076 cN/dTex) and by GTA treatment (about 0.075 cN/dTex). In addition, the surface of crosslinked collagen tube kept the hydrophilic property. These results suggest that DHT and GTA treatments can be utilized to improve the properties of electrospun collagen tube which could become a suitable candidate for tissue engineered scaffold.

Effect of the Application of a Dehydrothermal Treatment on the Structure and the Mechanical Properties of Collagen Film.

Dehydrothermal (DHT) treatment was used to improve the properties of collagen casings because of its non-cytotoxicity. Understanding the effects of DHT treatment on the structure and mechanical properties of collagen films is beneficial to developing satisfying collagen casings. Herein, DHT treatment with various temperatures (85-145 °C) and timescales (1-7 days) were investigated. It was clarified that the chemical crosslinking covalent bond between collagen molecules was formed after the DHT treatment. Crosslinking density increased with increasing DHT treatment temperatures, contributing to the increase of tensile strength up to over three times of that of the untreated collagen film. The increased crosslinking density was also found when increasing the DHT treatment time, and the maximum was obtained in 3 days. Further DHT treatment time did not change the crosslinking density. The damage in the triple helix structure and the self-assembly of collagen molecules were observed from IR and SAXS. The extent of denaturation increased with increasing DHT treatment temperature and time, although the effect of the DHT treatment time on the denaturation was more moderate. When the DHT treatment temperature was as high as 145 °C or the DHT treatment time exceeded 5 days, serious denaturation occurs, leading to the deterioration of mechanical properties.