Analysis of the TGF-b signaling in cartilage by nonlinear optics imaging combined with genetic engineering techniques

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Multi-photon microscopy enables to characterize and evaluate alternation of collagen matrix in vivo and in vitro based on second harmonic generation (SHG) signal. We performed SHG imaging of the cartilage degeneration induced by enzyme treatment. In the result, changes in SHG signal intensity were observed in the experimental samples. We also established an animal osteoarthritis model by syndesmectomy using H2B-GFP line mice. We successfully evaluated over time the changes in cartilage matrix and chondrocytes by SHG and two-photon excited fluorescence. Furthermore, the articular cartilages of aged mice were observed as the same manner. In comparison with those of young mice, it was possible to evaluate them as spontaneous knee osteoarthritis. Our technique that combined genetic engineering and nonlinear interaction between the molecules and photons to approach the mechanism of cartilage degeneration is promising as a translational research leading to clinical application.