Feasibility of biolistic gene therapy in burns.
Feasibility of biolistic gene therapy in burns.
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烧伤基因枪基因治疗的可行性。
DOI：
10.1097/00024382-200115040-00004
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Wang,SC
中科院分区：
文献类型：
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作者：
Steinstraesser,L;Föhn,M;Klein,RD;Aminlari,A;Remick,DG;Su,GL;Wang,SC
不明,皮肤是一个特别有吸引力的基因操作目标，因为它很容易接近和容易监测插入基因的存在和表达。本研究旨在评估颗粒介导基因转移到烧伤皮肤的可行性，并比较正常皮肤和烧伤皮肤中转基因表达的转染效率、解剖分布和持续时间。在60[度]C的水中（10秒：浅层部分；20秒：深层部分；40秒：全层）烫伤后2天，使用基因枪在不同的氦压力（200-600 psi）下将报告基因（[β]-半乳糖苷酶）构建体递送到正常皮肤和烧伤皮肤。通过时间过程研究来检验转基因表达的动力学。动物接受浅表部分厚度烧伤，并在基因转移后12小时、1、3、5、7、14或21天处死。采用墨汁注射法和免疫组化法评估烫伤损伤深度。转染效率在基因转移24 h后的皮肤匀浆中通过形态测定和化学发光测定。我们发现，组织损伤的程度与热源暴露的时间直接相关。报告基因活性在浅表部分厚度烧伤中显著高于正常对照组，并随着组织损伤的增加而逐渐下降。全层烧伤组未见活性。[β]-半乳糖苷酶活性在基因转移后12小时在正常和浅表部分厚度烧伤皮肤中达到最高水平，在转染后5天未见水平。这些发现表明，颗粒介导的基因转移在热损伤皮肤中是可行的，并且可能提供一种将生物制剂引入损伤组织的方法，能够增强细菌清除和促进伤口愈合。
mdash;: Skin is an especially attractive target for genetic manipulation because it is readily accessible and easily monitored for both the presence and the expression of inserted genes. This study was designed to assess the feasibility of particle mediated gene transfer to burned skin and to compare the transfection efficiency, anatomic distribution, and duration of transgene expression achievable in normal versus burned skin. Two days following scald injury of varying depths in 60 [degrees] C water (10 s: superficial partial; 20 s: deep partial; 40 s: full thickness) reporter gene ([beta]-galactosidase) constructs were delivered using a gene gun at various helium pressures (200-600 psi) to normal and burned skin. A time course study was performed to examine the kinetics of transgene expression. Animals received a superficial partial thickness burn and were sacrificed 12 h, 1, 3, 5, 7, 14, or 21 days after gene transfer. India Ink injection and immunohistochemistry were used to assess the depth of the scald injury. Transfection efficiency was measured in skin homogenates 24 h after gene transfer by morphometric and chemoluminescent assays. We found that the extent of tissue damage was directly related to the duration of heat source exposure. Reporter gene activity was significantly higher in superficial partial thickness burns compared to normal controls and gradually declined with increasing tissue injury. No activity was seen in the full thickness burn group.[beta]-galactosidase activity reached a maximum level 12 h after gene transfer in both normal and superficial partial thickness burned skin with no levels seen after 5 days post-transfection. These findings indicate that particle-mediated gene transfer in thermally injured skin is feasible and may provide a means of introducing biologic agents into injured tissue capable of enhancing bacterial clearance and improving wound healing.