Study on 5-carboxyfluorescein N-succinimidyl ester in detecting the degree of cross-linked allogenic tissue / 生物医学工程学杂志

This study was designed to creat a new method for detecting the degree of cross-linked allogenic tissue based on fluorescent technique. The thoracic aorta of New Zealand rabbits were divided randomly into four groups according to the concentration of Glutaraldehyde (GA), which were group A (control group-with no GA), group B (cross-linked with GA of 0.625%), group C (1.25%), and group D (2.5%). Each group was cross-linked with GA and reacted with 5-FAMSE, and then the fluorescence intensity was observed via fluorescence microscopy (analyzed with Image-Pro Plus 6.0, a professional image analysis software). The differences between groups in order of fluorescence intensity were found to be: group A > roup B > group C > group D (P < 0.01). Meanwhile, the tissue proteins extracted from aorta in each group were submitted to conventional polyacrylamide gel electrophoresis (PGE) after being cross-linked with GA; the result from this method was compared with that from the method of 5-FAMSE. In group A, the tissue proteins extracted from the aorta cross-linked with GA were obviously less than those not cross-linked with GA. However, this phenomenom was not clearly seen among the B, C and D groups. Nevertheless, 5-FAMSE can detect the degree of cross-linkage more conveniently and directly.

Research on short-term biological stability and cytotoxicity of allogenic arteries cross-linked by dye-mediated photooxidation in vivo / 生物医学工程学杂志

This study evaluated the biological stability and cytotoxicity of rabbit thoracic aorta cross-linked by dye-mediated photooxidation in vivo environment of blood flow. Rabbits, whose allogenic thoracic aorta was cross-linked by dye-mediated photooxidation (DMP, n=6) and glutaraldehyde (GA, n=6), were in the DMP group and GA group, respectively; rabbits, whose homogenic abdominal aorta was orthotopic transplanted (OT, n=6), were in the OT group. Then the donor arteries were transplanted into the position of the abdominal arteries of rabbits. After operation, the animals were fed for 1 month, then each of graft arteries was removed for observation. Biological stability was evaluated through histological analysis under optical microscope. Cytotoxicity was evaluated through ratio of coverage of endothelial cell (ES) under scanning-electron microscope. The results showed that no statistically significant difference was noted between DMP group and OT group (P>0.05); however, there was a significant difference between DMP group and GA group (P<0.01). The stability index in DMP group was much higher than that in GA group. There was a statistically significant difference between DMP group and GA group (P<0.01). Ratio was much higher in DMP group than in GA group. In conclusion, the arteries cross-linked by dye-mediated photooxidation treatment appeared to have higher biological stability and lower cytotoxicity in rabbit allogenic transplation model.