The Performance Of Complex PEN Loading ODN MT01and Its Effect On The Proliferation Of Osteoblast

The ODN MT01is a self design of single stranded DNA sequencethat contains27base pairs of simulation mitochondrial nucleotidesequence. Research shows that, ODN MT01can promote osteoblast ofrat maturation and activation; reduce rat alveolar bone absorption thatcaused by periodontitis; regulate expression levels of osteogenic relatedfactors like intracellular Runx-2, SP7and Collagen-I, promote bonemarrow mesenchymal stem cells differentiating into osteoblasts.Theabove research suggests, ODN MT01may have regulatory effects onbone remodeling during orthodontic tooth movement.ODN MT01as an oligonucleotide,affected by the short half-life andintracellular DNA enzymatic degradation, leading to the uptake rate ofcell effective cannot reach the threshold level, in addition, because theunmethylated ODN MT01surface is negatively charged, which makesthe combination with cell membrane that is negative difficult. In order tosolve these problems, it is often carried out on the ODN phosphorothioatemodified to improve their stability at present.But the modification ofphosphorothioation failed to change the negative charge and can notincrease the cell uptake rate, and literature reported chemicalmodification on the ODN backbone produces a decrease in immuneresponse, damage lymph follicles and organs increased sideeffects.Therefore, a variety of presentation system is designed and used,including cationic liposome, cationic polymers, cationic peptide, cationicpolymer, microspheres and nanoparticles.PEI has already become a kindof non viral gene vector transmission carrier class in one of the fastest-growing and most widely used.This study used PEN thatderivatives from cationic polymer polyethylenimine (Polyetherimine PEI)as a carrier for ODN MT01, to investigate whether high efficiency ratioand presenting a ODN MT01into MG63cells promote the proliferationof the best quality PEN loading ODN MT01, to provide the experimentalbasis for the follow-up study.Objective: To determinate the performance of PEI derivatives ofPEN loading ODN MT01and discussion the proliferation of MG63cellstransfected with PEN/ODN MT01complex．Methods: The following methods use Nanodrop to determinate theloading efficiency and Agarose gel electrophoresis to determinate thecapability of PEN with ODN MT01by different mass ration．Using theMalvin particle potential to test the particle size and potential ofPEN/ODN MT01complex.By FCM to ensure which is the best ration atthe same time by the fluorescence observation of PEN loading ODNMT01-Cy5which transfects MG63．Detemined the cells of proliferationrate of PEN/MT01complex by MTT detection．Results: The surface of PEN/MT01complex is positivelycharged,particle size is90-110nm，low cytotoxicity,and the quality rationof ODN MT01and PEN is1:6，the loading ration is77.67%，what ismore，the transfection of MG63cell florescence intensity is the largest．Conclusion:Loading PEN ODN MT01can can change the surfaceelectricity compound, beneficial to the cell uptake; PEN load ODNMT01particle size suitable for cellular uptake; PEN loading ODN MT01mass ratio was1:6, can efficiently transfer ODN MT01into MG63cells;at the same time can obviously promote the proliferation of MG63cells.