Research On Mesoporous Millimeter Nanomaterials Carrying SiRNA For Gene Therapy

Among large amounts of studies on gene therapy vector, mesoporous silicananomaterials (MSNs) as a gene vector gets widespread attention in recent years.MSNs has drawn a great deal of research interest for the potential biomedicalapplications.It has become one of the hottest areas in gene therapy research field.This is mainly due to MSNs’ ordered pore structure,easy preparation,super highspecific pore volume, tunable pore size, and easily targeted modification of surface.In this paper,firstly we used scanning electron microscope(SEM) andtransmission electron microscope(TEM) to acquaint the morphologicalcharacteristics of D-MSNs, N-MSNs which20%was amino-modified, Tf-MSNswhich4%was transferrin targeting modified.After that X-ray diffraction, Zetapotential, Fourier transform infrared(FTIR) spectroscopy were used to characterizethe particles’ composition, structure and important physical and chemicalproperties.Agarose gel electrophoresis and micro-UV spectrophotometer combinedtogether to take a research of MSNs’ adsorption, release and protection on genes.Ultimately, confocal laser scanning microscope(CLSM) and flow cytometry(FCM)were utilized to detect MSNs-siRNA to transfection Hela cells in vitro.We can stillsee if the survivin gene silencing resulted in apoptosis of Hela cells.MSNs are uniform spherical particles with pore structure.Its’ size is about(200±10) nm. FTIR analysis showes that the amino and transferrin modification areaccomplished.After adsorption of DNA, N-MSNs-DNA and Tf-MSNs-DNA have alittle aggregation.In a2mg MSNs’ system, D-MSNs, N-MSNs and Tf-MSNs’adsorption rate of20bp DNA fragment (160μg/mL×100μL) was12.1%,93.7%,90%.N-MSNs and Tf-MSNs’s release rate was72.8%,60.5%. The differencebetween MSNs-DNA dealing with and without nuclease was quite tiny.The cellularuptake efficiency of D-MSNs、N-MSNs and Tf-MSNs’ were34.25%,95.86%,99.33%.The rate of apoptosis detected by FCM was4.14%,8.45%,16.98%.This research successfully accomplished amino modification which was fortransfection in vitro and transferrin modification which was for targeted transfection.Modified MSNs especially Tf-MSNs had an excellent ability on adsorption，releaseand protection of the target genes.The cellular uptake efficiency of MSNs by Helacells was very high.Tf-MSNs’ average intake was quite impressive.There was alsoan ideal trend in apoptosis of T-MSNs.The performance shows the targeting isfavorable and Tf-MSNs gets the potential to become an ideal gene vector.