Biodegradable gelatin hydrogel potentiates the angiogenic effect of fibroblast growth factor 4 plasmid in rabbit hindlimb ischemia
Biodegradable gelatin hydrogel potentiates the angiogenic effect of fibroblast growth factor 4 plasmid in rabbit hindlimb ischemia
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DOI：
10.1016/s0735-1097(02)03007-3
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发表时间：
2003-03-19
影响因子：
24
通讯作者：
Mori, H
中科院分区：
文献类型：
--
作者：
Kasahara, H;Tanaka, E;Mori, H
我们研究了通过将质粒脱氧核糖核酸（DNA）与可生物降解的明胶水凝胶（GHG）结合使用成纤维细胞生长因子4（FGF 4）基因的基因治疗的增强作用。背景病毒载体有效地转移基因但具有生物危害性，而裸DNA更安全但效率较低。脱氧核糖核酸（DNA）带负电荷，GHG带正电荷，可生物降解，可植入体内; FGF 4具有促血管生成作用。用~（125）I标记的GHG-DNA复合物检测基因降解。用逆转录巢式聚合酶链反应和X-Gal组织染色评价转染效率。结果明胶水凝胶能有效地维持质粒的结构，延长基因降解时间，直至肌肉注射后28 d，并能提高转染效率。基因转移后4周，GHG-FGF 4组的后肢肌肉坏死比裸FGF 4基因组和GHG-β-半乳糖苷酶（对照）组更显著地改善（p < 0.05，Kruskal-Wallis检验）。同步辐射微血管造影术（空间分辨率，20 μ m）和微球流量测定证实了GHG-FGF 4组对腺苷给药的显著血管反应性，而在FGF 4-基因组和对照组中则无表达。FGF 4复合物可能通过延长基因降解和提高转染效率促进新生血管的血管生成和血流调节，而无生物危害与病毒载体有关。(C)2003年由美国心脏病学会基金会。
OBJECTIVES We investigated the potentiation of gene therapy using fibroblast growth factor 4 (FGF4)-gene by combining plasmid deoxyribonucleic acid (DNA) with biodegradable gelatin hydrogel (GHG).BACKGROUND Virus vectors transfer genes efficiently but are biohazardous, whereas naked DNA is safer but less efficient. Deoxyribonucleic acid charges negatively, GHG has a positively charged structure and is biodegradable and implantable; FGF4 has an angiogenic ability.METHODS The GHG-DNA complex was injected into the hindlimb muscle (63 mice and 55 rabbits). Gene degradation was evaluated by using I-125-labeled GHG-DNA complex in mice. Transfection efficiency was evaluated with reverse-transcription nested polymerase chain reaction and X-Gal histostaining. The therapeutic effects of GHG-FGF4-gene complex (GHG-FGF4) were evaluated in rabbits with hindlimb ischemia.RESULTS Gelatin hydrogel maintained plasmid in its structure, extending gene degradation temporally until 28 days after intramuscular delivery, and improving transfection efficiency. Four weeks after gene transfer, hindlimb muscle necrosis was ameliorated more markedly in the GHG-FGF4 group than in the naked FGF4-gene and GHG-beta-galactosidase (control) groups (p < 0.05, Kruskal-Wallis test). Synchrotron radiation microangiography (spatial resolution, 20 Am) and flow determination with microspheres confirmed significant vascular responsiveness to adenosine administration in the GHG-FGF4 group, but not in the naked FGF4-gene and the control.CONCLUSIONS The GHG-FGF4 complex promoted angiogenesis and blood flow regulation of the newly developed vessels possibly by extending gene degradation and improving transfection efficiency without the biohazard associated with viral vectors. (C) 2003 by the American College of Cardiology Foundation.