Details

Autor(en) / Beteiligte

• Wang, Yufu, MD, PhD
• Mostafa, Nesrine Z., PhD
• Hsu, Charlie Y.M., PhD
• Rose, Laura, PhD
• Kucharki, Cezary, MD
• Yan, Jinglong, MD, PhD
• Jiang, Hongxing, MD, PhD
• Uludağ, Hasan, PhD

Titel

Modification of human BMSC with nanoparticles of polymeric biomaterials and plasmid DNA for BMP-2 secretion

Ist Teil von

• The Journal of surgical research, 2013-07, Vol.183 (1), p.8-17

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2013

Quelle

MEDLINE

Beschreibungen/Notizen

• Abstract Background Genetic modification of human bone marrow stem cells (hBMSCs) before administration to a patient is emerging as a viable approach to creating tailored cells that perform effectively in a clinical setting. To this end, safe delivery systems are needed that can package therapeutic genes into nanoparticles for cellular delivery. Methods We evaluated different plasmids on gene expression and compared the effective plasmids directly in hBMSCs. Then, we evaluated the transfection efficiencies of the polymeric carriers linoleic acid-substituted polyethylenimine (PEI-LA), polyethylenimine (PEI)-25, and PEI-2 using flow cytometry. We used 3-(4,5-dimethylthiazol-2- yl )-2,5-diphenyltetrazolium bromide to compare the toxicity of PEI-LA and PEI-25 on hBMSCs. We further assessed bone morphogenetic protein-2 (BMP-2) secretion and the osteogenic activity of hBMSCs transfected with the polymeric (PEI-LA and PEI-25) gWIZ-BMP-2 complex. Results Unlike the transformed cells that gave robust (>50%) transfection, only a few percent (<10%) of hBMSCs was transfected by the developed nanoparticles in culture. The plasmid DNA design was critical for expression of the transgene product, with the choice of the right promoter clearly enhancing the efficiency of transgene expression. Using the in-house designed PEI-LA, hBMSCs secreted BMP-2 in culture (∼4 ng BMP-2/106 cells/d), which indicates the feasibility of using PEI-LA as a delivery system. Furthermore, we demonstrated an increased osteogenic activity in vitro for hBMSCs transfected with the PEI-LA containing the BMP-2 expression system. Conclusions These results provide encouraging evidence for the potential use of a low toxic PEI-LA to genetically modify hBMSC.