Enhanced bone morphogenic protein adenoviral gene delivery to bone marrow stromal cells using magnetic nanoparticle
이정태, Jung Jae-Whan, 최재용, 권태건,
소속 상세정보
이정태 ( Lee Jung-Tae ) - Kyungpook National University School of Dentistry Department of Oral and Maxillofacial Surgery
( Jung Jae-Whan ) - Kyungpook National University School of Medicine Department of Biochemistry
최재용 ( Choi Jae-Yong ) - Kyungpook National University School of Medicine Department of Biochemistry
권태건 ( Kwon Tae-Geon ) - Kyungpook National University School of Dentistry Department of Oral and Maxillofacial Surgery
KMID : 0355620130390030112
Abstract
Objectives: This study investigated the question of whether adenoviral magnetofection can be a suitable method for increasing the efficacy of gene delivery into bone marrow stromal cell (BMSC) and for generation of a high level of bone morphogenic protein (BMP) secretion at a minimized viral titer.
Materials and Methods: Primary BMSCs were isolated from C57BL6 mice and transduced with adenoviral vectors encoding ? galactosidase or BMP2 and BMP7. The level of BMP secretion, activity of osteoblast differentiation, and cell viability of magnetofection were measured and compared with those of the control group.
Results: The expression level of ? galactosidase showed that the cell transduction efficiency of AdLacZ increased according to the increased amount of magnetic nanoparticles. No change in cell viability was observed after magnetofection with 2 ?L of magnetic nanoparticle. Secretion of BMP2 or BMP7 was accelerated after transduction of AdBMP2 and 7 with magnetofection. AdBMP2 adenoviral magnetofection resulted in up to 7.2-fold higher secretion of BMP2, compared with conventional AdBMP2-transduced BMSCs. Magnetofection also induced a dramatic increase in secretion of BMP7 by up to 10-fold compared to the control. Use of only 1 multiplicity of infection (moi) of magnetofection with adenoviral transduction of AdBMP2 or AdBMP7 resulted in significantly higher transgene expression compared to 20 moi of conventional adenoviral transduction.
Conclusion: Magnetic particle-mediated gene transudation is a highly efficient method of gene delivery to BMSCs. Magnetofection can lower the amount of viral particles while improving the efficacy of gene delivery.
키워드
Magnetics; Nanoparticles; Adenovirus; Mesenchymal stromal cells
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