Noninvasive Evaluation of Tissue-Engineered Cartilage with Time-Resolved Laser-Induced Fluorescence Spectroscopy

作者：Kutsuna Toshiharu; Sato Masato^*; Ishihara Miya; Furukawa Katsuko S; Nagai Toshihiro; Kikuchi Makoto; Ushida Takashi; Mochida Joji

来源：Tissue Engineering - Part C: Methods , 2010, 16(3): 365-373.

DOI：10.1089/ten.tec.2009.0008

Regenerative medicine requires noninvasive evaluation. Our objective is to investigate the application of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) using a nano-second-pulsed laser for evaluation of tissue-engineered cartilage (TEC). To prepare scaffold-free TEC, articular chondrocytes from 4-week-old Japanese white rabbits were harvested, and were inoculated at a high density in a mold. Cells were cultured for 5 weeks by rotating culture (RC) or static culture (SC). The RC group and SC group at each week (n = 5), as well as normal articular cartilage and purified collagen type II (as controls), were analyzed by TR-LIFS. The peak wavelength was compared with those of type II collagen immunostaining and type II collagen quantification by enzyme-linked immunosorbent assay and tensile testing. The fluorescence peak wavelength of the TEC analyzed by this method shifted significantly in the RC group at 3 weeks, and in the SC group at 5 weeks (p<0.01). These results correlated with changes in type II collagen (enzyme-linked immunosorbent assay) and changes in Young's modulus on tensile testing. The results were also supported by immunohistologic findings (type II collagen staining). Our findings show that TR-LIFS is useful for evaluating TEC.

• 出版日期2010-6