Regulation of Cell Signaling Factors Using PLGA Nanoparticles Coated/Loaded with Genes and Proteins for Osteogenesis of Human Mesenchymal Stem Cells.

Transfection of specific genes and transportation of proteins into cells have been a focus of stem cell differentiation research. However, it is not easy to regulate codelivery of a gene and a protein into cells. For codelivery into undifferentiated cells (human mesenchymal stem cells (hMSCs)), we used biodegradable carriers loaded with Runt-related transcription factor 2 (RUNX2) protein and coated with bone morphogenetic protein 2 (BMP2) plasmid DNA (pDNA) to induce osteogenesis. The released gene and protein were first localized in the cytosol of transfected hMSCs, and the gene then moved into the nucleus. The levels of internalized PLGA nanoparticles were tested using different doses and incubation durations. Then, transfection of BMP2 pDNA was confirmed by determining mRNA and protein levels and acquiring cell images. The same techniques were used to assess osteogenesis of hMSCs both in vitro and in vivo upon internalization of PLGA NPs carrying the BMP2 gene and RUNX2 protein. Detection of specific genes and proteins demonstrated that cells transfected with PLGA NPs carrying both the BMP2 gene and RUNX2 protein were highly differentiated compared with other samples. Histological and immunofluorescence analyses demonstrated that transfection of PLGA nanoparticles carrying both the BMP2 gene and RUNX2 protein dramatically enhanced osteogenesis of hMSCs.