Bioactive hydrogel scaffolds reinforced with alkaline-phosphatase containing halloysite nanotubes for bone repair applications.

Alkaline phosphatase (ALP), biomineralization promoting enzyme, was immobilized in halloysite (HAL) nanotubes and used as a bioactive component of the chitosan (CH) and chitosan-collagen (C-CH) hydrogel scaffolds for bone regeneration. The influence of HAL-ALP and collagen on the properties of the obtained materials was studied. 30 wt% of HAL-ALP increased significantly the swelling ratio of chitosan-based scaffolds. The presence of both: collagen and HAL-ALP had positive effect on the scaffolds' porosity, which was considerably higher for C-CH. The presence of HAL has improved the mechanical properties of both types of scaffolds, while the addition of 20% of collagen to the chitosan hydrogels have considerably decreased their storage modulus. Biomineralization tests showed that although mineral was formed in both CH and C-CH scaffolds with HAL-ALP, the process was more effective for collagen-containing hydrogels. Biological studies, performed using MG-63 osteoblast-like cell line showed that C-CH scaffolds, especially those after biomineralization, were a better material for cell adhesion and growth. Both types of scaffolds degraded slowly in physiological pH. C-CH scaffolds containing 30% of HAL-ALP have the highest potential as bioactive material for bone regeneration.