Assessment of collagen crosslinking and denaturation for the design of regenerative scaffolds.
Crosslinking and denaturation were two variables that deeply affected the performance of collagen-based scaffolds designed for tissue regeneration. If crosslinking enhances the mechanical properties and the enzymatic resistance of collagen, while masking or reducing the available cell binding sites, denaturation has very opposite effects, as it impairs the mechanical and the enzymatic stability of collagen, but increases the number of exposed cell adhesive domains. The quantification of both crosslinking and denaturation was thus fundamental to the design of collagen-based scaffolds for selected applications. The aim of this work was to investigate the extents of crosslinking and denaturation of collagen-based films upon dehydrothermal (DHT) treatment, that is, one of the most commonly employed methods for zero-length crosslinking that shows the unique ability to induce partial denaturation. Swelling measurements, differential scanning calorimetry, Fourier transform infrared spectroscopy, colorimetric assays for the quantification of primary amines, and mechanical tests were performed to analyze the effect of the DHT temperature on crosslinking and denaturation. In particular, chemically effective and elastically effective crosslink densities were evaluated. Both crosslinking and denaturation were found to increase with the DHT temperature, although according to different trends. The results also showed that DHT treatments performed at temperatures up to 120°C maintained the extent of denaturation under 25%. Coupling a mild DHT treatment with further crosslinking may thus be very useful not only to modulate the crosslink density, but also to induce a limited amount of denaturation, which shows potential to partially compensate the loss of cell binding sites caused by crosslinking.
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.
Marta
M
Madaghiele
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.|Dhitech Scarl, Distretto Tecnologico High Tech, Lecce, 73100, Italy.
Emanuela
E
Calò
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.
Luca
L
Salvatore
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.
Valentina
V
Bonfrate
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.|Dhitech Scarl, Distretto Tecnologico High Tech, Lecce, 73100, Italy.
Deborah
D
Pedone
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.
Mariaenrica
M
Frigione
Department of Engineering for Innovation, University of Salento, Lecce, 73100, Italy.
Alessandro
A
Sannino
D000818:Animals; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D055119:Elastic Modulus; D011474:Prosthesis Design; D011489:Protein Denaturation; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D054457:Tissue Scaffolds / Q000737:chemistry*; D014302:Trinitrobenzenesulfonic Acid; D014867:Water
D003432:Cross-Linking Reagents; D014867:Water; D014302:Trinitrobenzenesulfonic Acid; D003094:Collagen
collagen; crosslinking; dehydrothermal treatment; denaturation; scaffolds