Intrafibrillar mineralization of type I collagen by micelle-loaded amorphous calcium phosphate nanoparticles

Mineralization of type I collagen fibrils is highly desired for artificial bone preparation and teeth repairing. Generally, amorphous calcium phosphate (ACP) combined with non-collagenous protein analogue (NCPA) were used for biomimetic remineralization of collagen fibrils. However, the ACP was likely to aggregate to form larger particles that could not infiltrate into the gaps of the collagen for intrafibrillar mineralization, and the poor storage stability of ACP has challenged its practical applications. To address this question, here we assembled ACP that was stabilized by carboxylated polyamidoamine (CPAMAM) at a pH of 6.5 to form dispersed nanoparticles of 25 nm in size, which was named as ACP/CPAMAM. The ACP/CPAMAM nanoparticles were further loaded into micelles composed of polysorbate and polyethylene glycol (PEG) to further improve their storage stability. The micelle-loaded ACP/CPAMAM particles could maintain their amorphous phase after storage for 12 months. During the mineralization of collagen fibrils, isopropanol (IPA) was introduced to dissolve the micelles and release the ACP/CPAMAM nanoparticles. By using micelle-loaded ACP/CPAMAM, good intrafibrillar mineralization of type I collagen was demonstrated. This work provides novel methods for preparing ACP nanoparticles with good storage stability and controllable release for intrafibrillar mineralization.