Publications (5)13.43 Total impact
- [Show abstract] [Hide abstract] ABSTRACT: Mesenchymal stem cells (MSCs) are a promising tool for delivering of therapeutic agents in cancer treatment. In the present study, our findings suggested that both i.v. and intratumoral injection of MSCs could favor tumor growth under physiologic conditions. However, the anti-tumor effects of MSC-IL-12 were achieved using our strategy. Unlike the previously reported method, the genetic engineering of MSCs was conducted by non-viral transfection using the new vector, spermine-pullulan. The transfection, cytotoxicity, and the cellular internalization of this vector were evaluated. Then, the therapeutical gene, IL-12, was delivered to the MSCs using this vector. The in vitro secretions of IL-12 by MSC-IL-12 confirmed the success of using spermine-pullulan/DNA nanoparticles for the gene transfection. We used the MSC-IL-12 for the in vivo treatment of both B16F10 metastasis tumor and the established subcutaneous B16BL6 tumor. For the B16F10 metastasis tumor, treatment with MSC-IL-12 significantly reduced lung metastases. For the established subcutaneous B16BL6 tumor, intratumoral injected MSC-IL-12 cells considerably retarded tumor growth. Prolonged survival was observed when MSC-IL-12 cells were injected through the tail vein or intratumorally, indicating that the MSCs engineered with the therapeutic gene could reverse the tumor-promoting effects of MSCs using the nonviral transduction method. However, the intravenous injected MSC-IL-12 did not prevent the tumor growth of the established subcutaneous B16BL6 tumor. Thus, we examined the the in vivo distribution of MSCs in different organs and it was found that MSCs were mainly distributed in the lungs, which may explain the inability of intravenously injected MSC-IL-12 to inhibit the growth of the established subcutaneous tumor.
- [Show abstract] [Hide abstract] ABSTRACT: The current study aimed to examine the transfection ability of chitosan-linked-polyethylenimine (PEI) (CP), a newly synthesized PEI derivative, in mesenchymal stem cells (MSCs). Firstly, series of CP/DNA complex with different charge ratio (N/P) were prepared, and the physiochemical properties, such as particle size and the zeta potential of this vector, were measured. Analysis of its physicochemical properties demonstrated that the modified PEI polymers were able to form nanoparticles with DNA whose particle size ranging from 110–140 nm. And their surface charge decreased to about 24 mV. Then, the in vitro cellular internalization was observed under a confocal laser scanning microscope, followed by the transfection efficiency examination in mesenchymal stem cells (MSCs). Also, its cytotoxicity was compared with that of PEI by MTT assay. It was found that the novel CP polymer exhibited higher transfection efficiency and relatively lower cytotoxicity in MSCs than the control. Finally, the transfection of TGF-β1 based on CP was proved to successfully induce the osteogenic differentiation of MSCs in vitro. In conclusion, that the newly synthesized CP can form a complex with DNA, and had compatible physicochemical properties for use as a gene delivery system. Our result demonstrated that the CP would be a very attractive non-viral vector which can be utilized in MSCs.
- [Show abstract] [Hide abstract] ABSTRACT: This study evaluated the potential of utilizing transfected pTGFβ-1 gene-engineered rat mesenchymal stem cells (MSCs) using nonviral vector to promote cartilage regeneration. Pullulan-spermine was used as the nonviral gene vector and gelatin sponge was used as the scaffold. MSCs were engineered with TGF-β1 gene with either the three-dimensional (3D) reverse transfection system or the two-dimensional (2D) conventional transfection system. For the 3D reverse transfection system, pullulan-spermine/pTGF-β1 gene complexes were immobilized to the gelatin sponge, followed by the seeding of MSCs. Pullulan-spermine/pTGF-β1 gene complexes were delivered to MSCs cultured in the plate to perform the 2D conventional transfection system, and then MSCs were seeded to the gelatin sponge. Then, TGF-β1 gene-transfected MSC seeded gelatin sponge was implanted to the full-thickness cartilage defect. Compared with the control group, both groups of TGF-β1 gene-engineered MSCs improved cartilage regeneration through optical observation and histology staining. So, with pullulan-spermine as the nonviral vector, TGF-β1-gene engineered MSCs can induce cartilage regeneration in vivo.
- [Show abstract] [Hide abstract] ABSTRACT: Current efforts had been made to undertake a three-dimensional (3-D) reverse transfection of bone marrow-derived mesenchymal stem cells (BM-MSCs) in PLGA scaffolds. As a kind of multipotent stem cells, BM-MSCs show great potential and tremendous capacity in the gene transfection field and PLGA 3-D scaffold has been shown to be a biomaterial that provides structural support to cells proliferation and tissue engineering. The objective of this study was to assess the transfection efficiency of BM-MSCs with a 3-D reverse transfection method by using PLGA scaffold and observe the SEM photographs of BM-MSCs cultured on PLGA scaffold. BM-MSCs were cultured in 3-D PLGA scaffold which was incorporated with pullulan-spermine/pGL3. It was shown that the gene expression duration of BM-MSCs transfected using 3D reverse method with pullulan-spermine/DNA in the presence of serum maintained 12 days at high levels as compared with the plasmid DNA in medium, and scanning electronic microscopy (SEM) photographs of BM-MSCs cultured on PLGA scaffold exhibited robust cell attachment and viability when cultured in PLGA scaffold in vitro. This study demonstrates that the 3-D reverse transfection method of BM-MSCs using PLGA scaffold could achieve long gene expression in a relatively high level, therefore this transfection system is promising in gene transfection and tissue engineering.
- [Show abstract] [Hide abstract] ABSTRACT: To enhance the level and prolong the duration of gene expression for gene-engineered rat mesenchymal stem cells (MSCs) using non-viral vector. A novel transfection system based on reverse transfection method and three-dimensional (3D) scaffold was developed. The reverse gene transfection system was evaluated for transfection efficiency compared to conventional methods. Collagen sponge and polyethylene terephthalate non-woven fabric were introduced as scaffolds to perform 3D culture with reverse transfection. pDNA coding TGFβ-1 was delivered to MSCs to assess its ability in inducing chondrogenesis with the 3D non-viral reverse transfection system. The reverse transfection method induced higher transgene levels than the conventional transfection in the presence of serum. The electric charge of the anionic gelatin plays an important role in this system by affecting the release pattern of the gene complexes and through the adsorption of serum protein to the substrate. During a long-time in vitro culture, MSCs cultured on 3D scaffolds exhibited a higher transgene expression level and more sustained transgene expression than those cultured and transfected on the two-dimensional substrate. The combination of reverse transfection system with 3D cell culture scaffold benefits the cell proliferation and long-time gene transfection of MSCs.