PLGA microcarrier as an injectable scaffold for NT-3 overexpressing bone marrow stem cells: a potential tool for treatment of Parkinson's disease

摘要

Parkinson's disease (PD) is a neurodegenerative disorder that characterized by destruction of substantia nigrostriatal pathway due to the loss of dopaminergic (DA) neurons. However, the progression of this disease can be prevented by delivering a neurotrophin-3 (NT-3) to substantia nigra. The aim of this study is production of genetically modified BMSCs which overexpressed NT-3 to stimulate the neuronal-like differentiation of BMSCs and promote neuronal survival. Moreover, a PLGA microcarrier was designed to deliver the NT-3 overexpressing BMSCs into substantia nigra as an injectable scaffold. The NT-3 gene was isolated from brain tissue of mouse using RT-PCR method. The resultant NT-3 gene was inserted into pIRES-EGFP plasmid and transfected into BMSCs using PEI-grafted multi-walled carbon nanotubes (PEI-g-MWCNTs). The overexpression of NT-3 into BMSCs was analyzed using RT-PCR and the expression of tyrosine hydroxylase (TH) was analyzed using Immunocytochemistry (ICC). PLGA microcarrier was synthetized by double emulsion method. The surface of PLGA microcam'ers was functionalized using collagen in order to increase the attachment of cells onto the microcarriers. The cell attachment was observed using scanning electron microscopy (SEM) and DAPI staining techniques. The results demonstrated that PEI-g-MWCNTs effectively delivered NT-3 containing plasmid into BMSCs. the expression of TH in transfected BMSCs showed that NT-3 promote the intracellular signaling pathway of DA neuron differentiation. The results of SEM and DAPI staining showed that transfected BMSCs was successfully attached to the surface of microcarriers. In conclusion, the result of this study showed that combination of gene therapy and PLGA microcarriers can be used as a potential tool for transfection and transferring engineered BMSCs for treatment of Parkinson disease.