Biomimetically Mineralized Salmon Collagen Scaffolds for Application in Bone Tissue Engineering

Abstract

Biomimetic mineralization of collagen is an advantageous method to obtain resorbable collagen/hydroxy-apatite composites for application in bone regeneration. In this report, established procedures for mineralization of bovine collagen were adapted to a new promising source of collagen from salmon skin challenged by the low denaturation temperature. Therefore, in the first instance, variation of temperature, collagen concentration, and ionic strength was performed to reveal optimized parameters for fibrillation and simultaneous mineralization of salmon collagen. Porous scaffolds from mineralized salmon collagen were prepared by controlled freeze-drying and chemical cross-linking. FT-IR analysis demonstrated the mineral phase formed during the preparation process to be hydroxyapatite. The scaffolds exhibited interconnecting porosity, were sufficiently stable under cyclic compression, and showed elastic mechanical properties. Human mesenchymal stem cells were able to adhere to the scaffolds, cell number increased during cultivation, and osteogenic differentiation was demonstrated in terms of alkaline phosphatase activity.

Preparation and characterization of monomers to tetramers of a collagen-like domain fromStreptococcus pyogenes

“…4 C was obtained ( Table 1), some 2.9 C higher than when the CysCys was not present. However, the T m obtained after removal of the V-domain, removing the additional stabilizing at the N-terminal, gave a product CLcc that had essentially the same stability, 37.5 C, as the construct lacking the CysCys sequence.…”

“…For the V-(CL) [1][2][3][4] series, the longest construct V-(CL) 4 was very insoluble and not enough could be dissolved to get a good signal for a reliable T m determination; the www.landesbioscience.comsame was the case after the V-domain was removed. For all samples with the C-terminal Cys-Cys sequence (Fig.…”

“…2) lack of solubility was more of an issue, with possible aggregation associated with disulphide interchange between individual molecules. For the V-(CL) [1][2][3][4] cc series, the longest construct V-(CL) 4 cc was too insoluble for a reliable T m determination; after the V-domain was removed the (CL) 4 cc sample was too insoluble to allow any T m determination.…”

“…The solubility of the proteins lacking the V-domain seemed less than for those that included the Vdomain, but this was not quantified. Thus, there were more difficulties getting good CD spectra for (CL) 3 and particularly for (CL) 4 . The T m for (CL) 3 was 0.5 C lower than for V-(CL) 3 , while the estimated T m for (CL) 4 was similar to that for (CL) 3 and was slightly higher than for the V-(CL) 4 (Table 1).…”

“…There have also been concerns that these animalderived collagens may also lead to the transmission of diseases, particularly spongiform encephalopathies including "mad cow disease." Other non-mammalian tissue sources have been proposed to minimize risks of disease, including avian, 3 piscine 4 and cnidarian (jellyfish) 5 sources. The alternative, which also addresses variability and quality issues, is to examine production of recombinant collagens.…”