CAJ | 학술논문

目的：探讨PKH26荧光标记和分子荧光活体成像技术在软骨组织工程中的应用。方法用PKH26荧光标记犬软骨细胞，种植到多孔支架上，体外培养1周后异位移植到裸鼠背部，4周后用分子荧光活体成像系统示踪，并与X线检查结果对比。然后处死裸鼠取材，与免疫组织化学染色和免疫荧光观察结果对比。结果4周分子荧光活体成像系统观察裸鼠背部标本处呈圆形强荧光，表明组织工程软骨在裸鼠体内生长良好。组织学切片结果显示番红O染色、Ⅱ型胶原免疫组化染色和甲苯胺蓝染色阳性，荧光显微镜下观察结果显示组织工程化软骨中细胞均呈红色荧光，Ⅱ型胶原免疫荧光染色呈绿色荧光，叠加后呈黄色荧光。结论 PKH26荧光标记和分子荧光活体成像2种方法结合应用于软骨组织工程中，能够较理想地且大体无创伤性评估组织工程化软骨组织在体内的生长情况。
목적：탐토PKH26형광표기화분자형광활체성상기술재연골조직공정중적응용。방법용PKH26형광표기견연골세포，충식도다공지가상，체외배양1주후이위이식도라서배부，4주후용분자형광활체성상계통시종，병여X선검사결과대비。연후처사라서취재，여면역조직화학염색화면역형광관찰결과대비。결과4주분자형광활체성상계통관찰라서배부표본처정원형강형광，표명조직공정연골재라서체내생장량호。조직학절편결과현시번홍O염색、Ⅱ형효원면역조화염색화갑분알람염색양성，형광현미경하관찰결과현시조직공정화연골중세포균정홍색형광，Ⅱ형효원면역형광염색정록색형광，첩가후정황색형광。결론 PKH26형광표기화분자형광활체성상2충방법결합응용우연골조직공정중，능구교이상지차대체무창상성평고조직공정화연골조직재체내적생장정황。
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.