Research On Influence Of Seed Cells Adhesion By Galectin-8Protein In Tissue Engineering Heart Valve

Objective:To detect the influence of seed cells adhesion by Galectin-8protein in differentconditions,find a new method to improve the adhesion of seed cells for tissueengineering heart valve.Methods:The fourth generation of human umbilical vein endothelial cells were dividedinto four groups: Group I was recombinant plasmid-Gal-8transfection group;GroupII was empty plasmid transfection group;Group III was Gal-8protein pre layingfixation group; Group IV is no treatment control group.Identification of Gal-8generecombined with plasmid vector to transfect into endothelial cells and proteinexpression in Group I. The same empty vector was transfected into endothelial cellsin Group II.Gal-8protein is laid on the culture plate, and then the endothelial cellswere seeded on it in Group III.Group IV was not any of the above processing.The4groups of cells were planted on96well plate and set a contrastive hole,for group IV，Galectin-8protein was Pre-laying on the plate，detect the adhesion rateunder a microscope in200times respectively and do statistical comparison after thecells attached to the wall at30、60、90minute，ten view for each observation hole.The4groups of cells were planted on the decellularized scaffolds, for groupIV，Galectin-8protein was Pre-laying on the decellularized scaffolds, observed thecells on the decellularized scaffolds after1weeks and compared them.Results:After transfection, Western and RT-PCR methods respectively test in the levelof protein and gene to identify Gal-8, Gal-8protein and mRNA were found a largeincrease ultimately, compared with the negative control group, the results werestatistically significant (P<0.05).After planted for30,60,90min, the adhesion rate of group I(48.50±7.86，67.90±1.67，84.90±4.25Each hole)was significantly lower than group II(79.10±6.28，90.10±5.78，118.40±8.62Each hole)、III(76.90±6.52，88.40±7.50，117.60±6.67Each hole)and VI(209.60±16.91，230.10±17.52，272.30±12.40Eachhole)at the same time, but the adhesion rate of group II was close to group III at thesame time, the adhesion rate of group VI was higher than group I、II、III at the sametime.After cells were seeded on the decellularized scaffolds for1week,HE stainingand electron microscope was used，the cell number on the scaffold of group I was theleast, the cell number of group II and group III was similar, group IV had the largestcell number, the cells were in good condition.Conclusions:(1)Gal-8protein in a dissolved state which was secreted by the endothelial cellsafter Gal-8gene was transfected into them can inhibit endothelial cell adhesion toextracellular matrix.(2) while in the fixed state after coated with Gal-8protein, Gal-8can promoteendothelial cell adhesion to extracellular matrix.