Gene delivery by surface immobilization of plasmid to tissue-engineering scaffolds.
Gene delivery by surface immobilization of plasmid to tissue-engineering scaffolds.
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作者：
生物材料支架作为基因传递载体，促进诱导因子的表达，在再生医学领域有着广泛的应用。在这份报告中，我们使用表面固定化策略研究了生物材料支架中的质粒传递。以聚(丙交酯-乙交酯)共聚物(PLG)为载体，通过干燥法制备了多孔支架。体外载体释放实验表明，大多数(~gt；70%)吸附的载体在24小时内释放，~gt；98%在3天内释放，而体内植入支架后，转基因表达至少持续28周，并定位于植入部位。DNA吸附支架的组织学分析表明，支架上的巨噬细胞在植入后的前两周开始表达，而植入物周围的肌肉细胞在4周时主要表达转基因。除了局部的基因表达外，一种分泌蛋白(人凝血因子IX)保留在植入部位，3天后无法系统获得，这表明偏离靶点的影响最小。这些发现表明，将载体固定在微孔PLG支架上可以在体内产生局部和长期的基因表达，这可能用于提高用于再生医学的支架的生物活性。
Biomaterial scaffolds that serve as vehicles for gene delivery to promote expression of inductive factors have numerous regenerative medicine applications. In this report, we investigate plasmid delivery from biomaterial scaffolds using a surface immobilization strategy. Porous scaffolds were fabricated from poly(lactide-co-glycolide) (PLG), and plasmids were immobilized by drying. In vitro plasmid release indicated that the majority (>70%) of adsorbed plasmid was released within 24 hours and >98% within 3 days; however, in vivo implantation of the scaffolds at the subcutaneous site yielded transgene expression that persisted for at least 28 weeks and was localized to the site of implantation. Histological analysis of DNA-adsorbed scaffolds indicated that macrophages at the scaffold were transfected in the first two weeks following implantation, whereas muscle cells adjacent to the implant primarily expressed the transgene at 4 weeks. In addition to localized gene expression, a secreted protein (human factor IX) was retained at the implant site and not available systemically after 3 days, indicating minimal off-target effects. These findings demonstrate that surface immobilization of plasmid onto microporous PLG scaffolds can produce localized and long-term gene expression in vivo, which may be employed to enhance the bioactivity of scaffolds used for regenerative medicine.
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DOI：
10.1002/jbm.1251
发表时间：
2002-02-01
期刊：
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH
影响因子：
--
作者：
Nof, M;Shea, LD
通讯作者：
Shea, LD
影响因子：
12.4
作者：
Jang, JH;Rives, CB;Shea, LD
通讯作者：
Shea, LD
影响因子：
2.9
作者：
Chandler, LA;Gu, DL;Phillips, ML
通讯作者：
Phillips, ML
影响因子：
3.5
作者：
Murphy, WL;Mooney, DJ
通讯作者：
Mooney, DJ
影响因子：
3
作者：
Sarkar, N;Blomberg, P;Islam, KB
通讯作者：
Islam, KB