Mineralization potential of cellulose-nanofibrils reinforced gelatine scaffolds for promoted calcium deposition by mesenchymal stem cells

Abstract

Cellulose-nanofibrils (CNFs) enriched gelatine (GEL) scaffolds were fabricated in-situ by the combined freeze-thawing process and carbodiimide crosslinking chemistry. The original- and variously surface anionised CNFs (carboxylated/CNF-COOH/, and phosphonated with 3-AminoPropylphosphoric Acid/CNF-COOH-ApA/) were used in order to tune the scaffolds' biomimetic structure towards a more intensive mineralization process. The pore size reduction (from 208±35μm to 91±35μm) after 50% v/v of CNFs addition to GEL was identified, while separated pore-walls' alignment vs. shorter, dense and elongated pores are observed when using 80% v/v of original-CNFs vs. anionised-CNFs, all of them possessed osteoid-like compressive strength (0.025–0.40MPa) and elasticity (0.04–0.15MPa). While randomly distributed Ca2+-deficient hydroxyapatite/HAp/(Ca/P≈1.4) aggregates were identified in the case of original-CNF prevalent scaffolds after four weeks of incubation in SBF, the more uniform and intensified deposition with HAp-like (Ca/P≈1.69) structures were established using CNF-COOH-Apa. The growth of Mesenchymal Stem Cells (MSCs) was observed on all CNF-containing scaffolds, resulting in more extensive Ca2+ deposition compared to the positive control or pure GEL scaffold. Among them, the scaffold prepared with the 50% v/v CNF-COOH-ApA showed significantly increased mineralization kinetic as well as the capacity for bone-like patterning in bone tissue regeneration.