Effects of extracorporeal fucosylation of CD44 on the homing ability of rabbit bone marrow mesenchymal stem cells.

BACKGROUND:The aim of the study was to investigate the effects of extracorporeal fucosylation of CD44 on the homing ability of rabbit bone marrow mesenchymal stem cells (BMSCs). METHODS:The rabbit BMSCs were extracorporeal fucosylated using alpha-(1,3)-fucosyltransferase VI (FTVI), then the positive rate of sialyl-LewisX (sLeX) and the binding rate of E-selectin were detected by flow cytometry, as well as the fluid adhesion of rabbit BMSCs were detected by the parallel flow chamber adhesion test. Then BMSCs were constructed to stably express enhanced green fluorescent protein (EGFP) and were injected intravenously into the model rabbits with tibial fractures. After 6 weeks of injection, the levels of stromal cell-derived factor (SDF-1) and monocyte chemoattractant protein-1 (MCP-1) in rabbit serum and damaged bone tissues were detected. The positive rate of EGFP expressions was detected by immunohistochemistry staining. RESULTS:After fucosylation, the positive rate of sLeX and the binding rate of E-selectin were significantly higher than those in the no fucosylated group. The results of fluorescence microscopy showed that BMSCs with stable expression of EGFP were successfully constructed. The results of ELISA and Western Blot showed that the secretion of SDF-1 and MCP-1 and the expression of SDF-1 and MCP-1 protein in BMSCs treatment group processed by fucosylated were significantly higher than those in BMSCs treatment group processed by no fucosylated. The results of immunohistochemical staining showed that the positive rate of EGFP expression was also significantly increased, which indicated that the BMSCs at the injured bone tissues were significantly increased and helpful in the repair of bone injury. CONCLUSIONS:Extracorporeal fucosylation of CD44 molecules can significantly enhance the homing ability of rabbit BMSCs, which may be achieved by SDF-1 and MCP-1 regulation.