Evaluation of Cartilage Engineering Using PKH26 and Molecular Light Imaging System / 天津医药

ABSTRACT

Objective To investigate the application of PKH26 and molecular light imaging system in cartilage en?gineering. Methods Canine chondrocyte was labeled by fluorescent dye PKH26 and seeded into the porous cartilage acel?lular matrix scaffold. The cells/scaffold constructs were cultured in vitro for 1 week. Then the constructs were implanted into the dorsal pocket of nude mice. We utilized a molecular light imaging system to macroscopically observe cells/scaffold con?structs in vivo with fluorescence at the 4th weeks, and compared with X-rays taken at the same position. The fluorescence im?ages were compared with the immunohistochemical and immunofluorescent results of cartilage-like tissue in vivo. Results Luminescent images were acquired at the 4th weeks, a red color enhanced overlay of the luminescent image over X-ray photo?graphic image demonstrated the location of the implants and the cell viability and cell growth on porous CACM scaffold in vivo were very well. Histological results show that the safranin O, anti-collagenⅡimmunohistochemistry and toluidine blue stain of cartilage-like tissue is positive. Immunofluorescence examination demonstrated chondrocytes in the constructs whitch is showen red fluorescence, and anti-collagenⅡimmunofluorescent staining was showen in green while the overlap?ping image is showen in yellow. Conclusion This study outlines an applicable non-destructive method to evaluate cell growth in tissue engineering constructs in vivo using PKH26 and molecular light imaging system.