Formation of Hydroxyapatite-Mediated Three-Dimensional Structures by Mouse Fibroblasts in Response to Physical Stimulation

Abstract

Hydroxyapatite (HAP) ceramics are widely used as implant materials for periodontal bone defects because of their excellent biocompatibility. We demonstrated that physical stimulation, that is, (1) mechanical stimuli or (2) laser irradiation, causes HAP-mediated C3H10T1/2 mouse fibroblasts to form three-dimensional tissue-like structures. Trypsinized 10T1/2 cells were cultured simultaneously with 200 HAP granules on a rotator for 7 days in mechanical stimulation experiments. The cells were later transferred to a regular incubator. Cell reactions were observed by phase-contrast microscopy. The formation of three-dimensional structures around the HAP granules was observed in the third week of cultivation after stimulation. In laser irradiation experiments, trypsinized cells were irradiated with 1, 5, and 16 J/cm² at a wavelength of 1000 nm and cultured with 200 HAP granules for 10 weeks. The formation of three-dimensional structures, like those observed in the mechanical stimulation experiments, was observed in the third week after irradiation. The formation of these structures was most frequent at 1 J/cm², and the frequency of formation of these structures gradually decreased as the irradiation dose was increased. These results indicate that physical stimuli may stimulate cell proliferation, leading to the repair of damaged tissue. These results also indicate that mouse fibroblasts do not form these three-dimensional structures without HAP and that HAP alone is not sufficient to stimulate the formation of three-dimensional structures.