Assessment of the Influence of Acetic Acid Residue on Type I Collagen during Isolation and Characterization

Abstract

Various methods for isolation of type I collagen using acids, bases, enzymes, and their combinations have been applied. However, a lack of standardization exists among type I collagens isolated by various approaches. Consequently, in this study, we assessed the influence of acetic acid residue on type I collagen isolated by pepsin-acetic acid treatment, the fabrication of collagen-based porous scaffolds, and the seeded cells on collagen scaffolds. Unlike the isolated collagen dialyzed by deionized water (DDW), collagen dialyzed by 0.5 M acetic acid (DAC) exhibited structural and thermal denaturation. Both DDW- and DAC-based porous scaffolds at all collagen concentrations (0.5, 1 and 2% w/v) showed the high degree of porosity (>98%), and their pore morphologies were comparable at the same concentrations. However, the DDW- and DAC-based collagen scaffolds displayed significant differences in their physical properties (weight, thickness, and volume) and swelling behaviors. In particular, the weight losses induced by mechanical stimulation reflected the high degradation of DAC-collagen scaffolds. In cell culture experiments using adipose-derived stem cells (ADSCs), the characteristics of mesenchymal stem cell (MSC) did not change in both DDW- and DAC-collagen scaffolds for 10 days, although cells proliferated less in the DAC-collagen scaffolds. Our results suggest that the elimination of acetic acid residue from isolated collagen is recommended to produce collagen scaffolds that provide a stable environment for cells and cell therapy-related applications.