Molecular weight and concentration of poly (acrylic acid) dual-responsive homogeneous and intrafibrillar collagen mineralization using an in situ co-organization strategy

Intrafibrillar mineralization is crucial in the synthesis of collagen-hydroxyapatite (HAP) composites with hierarchical structure resembling natural bone. The polymer-induced liquid precursor method, which involves immersing the matrix into polymer-stabilized amorphous calcium phosphate (ACP) solutions, has been widely used to regulate collagen mineralization; however, heterogeneous infiltration and crystallization of ACP precursors from the surface to the inner matrix remain a concern. In this study, the polymer-mediated co-organization strategy in which poly (acrylic acid) (PAA)-stabilized ACP solutions were initially introduced to collagen solutions before sol-gel transformation was employed to achieve homogeneous mineralization. Moreover, the effects of PAA with various molecular weights (2, 10, and 450 kDa) and concentrations (40, 80, and 120 mg/L) on collagen mineralization were evaluated. Increasing concentration and decreasing molecular weight both improved the stability of ACP precursors, which affected collagen mineralization. A lower concentration (40 mg/L) of 2 kDa PAA and higher concentration (80 and 120 mg/L) of 100 and 450 kDa PAA effectively retarded ACP crystallization, thereby inducing intrafibrillar mineralization. Extremely unstable ACP precursors failed to infiltrate the interior of fibrils using 100 and 450 kDa of PAA at 40 mg/L, resulting in extrafibrillar mineralization. Inversely, collagen fibrils were unmineralized until 7 days owing to excessive stability of ACP precursors using 2 kDa of PAA at higher concentrations. The present results further explain the mechanisms of intrafibrillar mineralization, and may provide a facile method for producing biomimetic materials.