IADR Abstract Archives
MicroRNA-26a-5p Delivered by Core-Cone Mesoporous Silica Nanoparticles Promotes Osteogenic Differentiation
Objectives: RNA mediated bone regeneration is a novel, safe, and effective modality which has been studied in recent years. Despite there being many microRNAs, little work has been done on their use for promoting osteogenesis. Moreover, microRNA needs a safe, affordable, and efficient transfection method. The present study explored the osteogenic effect of rno-miRNA-26a-5p delivery on rat bone marrow mesenchymal stem cells (rBMSCs) using polyethyleneimine coated core-cone mesoporous silica nanoparticles (CC-MSNs).
Methods: The novel microRNA delivery system was based on CC-MSNs (45 nm pore size, surface area 2.59 cm3/g). Nanoparticles were fabricated by an oil and water system. Some batches of nanoparticles were coated by polyethyleneimine (10 KDa) to increase the zeta potential, for greater nucleic acid interaction. MSNs were characterized by SEM and TEM. Cell viability assessed were assessed after 1, 3, and 7 days by the MTT assay. Rhodamine-labeled MSNs were loaded with fluorescent-tagged microRNAs, and their transfection efficiency for rBMSCs was evaluated by confocal laser scanning microscopy and flow cytometry analysis. CC-MSNs wereloaded with 1. MicroRNA-26a-5p mimic, 2. MicroRNA-26a-5p inhibit, or 3. Negative control microRNA. Expression of osteogenic genes (Runx-2, OCN, collagen 1, and BMP-2) was assessed at 7 days using the real-time quantitative polymerase chain reaction. In addition, osteogenic differentiation was assessed at 14 days by alkaline phosphate activity and alizarin red S staining.
Results: Polyethyleneimine coated CC-MSNs effectively transfected rBMSCs with miRNA-26a. A low concentration of these nanocomplexes significantly intensified the expression of osteogenic genes compared to controls (p <0.05). Moreover, both matrix mineralization and ALP activity were enhanced significantly by miRNA treatment.
Conclusions: Overall, these findings support the concept that microRNA-26a-5p mimic can be effectively delivered into mesenchymal stem cells using CC-MSNs as the vector for gene therapy, to promote osteogenic differentiation of stem cells.
Methods: The novel microRNA delivery system was based on CC-MSNs (45 nm pore size, surface area 2.59 cm3/g). Nanoparticles were fabricated by an oil and water system. Some batches of nanoparticles were coated by polyethyleneimine (10 KDa) to increase the zeta potential, for greater nucleic acid interaction. MSNs were characterized by SEM and TEM. Cell viability assessed were assessed after 1, 3, and 7 days by the MTT assay. Rhodamine-labeled MSNs were loaded with fluorescent-tagged microRNAs, and their transfection efficiency for rBMSCs was evaluated by confocal laser scanning microscopy and flow cytometry analysis. CC-MSNs wereloaded with 1. MicroRNA-26a-5p mimic, 2. MicroRNA-26a-5p inhibit, or 3. Negative control microRNA. Expression of osteogenic genes (Runx-2, OCN, collagen 1, and BMP-2) was assessed at 7 days using the real-time quantitative polymerase chain reaction. In addition, osteogenic differentiation was assessed at 14 days by alkaline phosphate activity and alizarin red S staining.
Results: Polyethyleneimine coated CC-MSNs effectively transfected rBMSCs with miRNA-26a. A low concentration of these nanocomplexes significantly intensified the expression of osteogenic genes compared to controls (p <0.05). Moreover, both matrix mineralization and ALP activity were enhanced significantly by miRNA treatment.
Conclusions: Overall, these findings support the concept that microRNA-26a-5p mimic can be effectively delivered into mesenchymal stem cells using CC-MSNs as the vector for gene therapy, to promote osteogenic differentiation of stem cells.