Targeted gene transfer into rat facial muscles by nanosecond pulsed laser-induced stress waves
Targeted gene transfer into rat facial muscles by nanosecond pulsed laser-induced stress waves
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通过纳秒脉冲激光诱导应力波将目标基因转移到大鼠面部肌肉中
DOI：
10.1117/1.3628313
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发表时间：
2011
影响因子：
3.5
通讯作者：
Akihiro Shiotani
中科院分区：
文献类型：
--
作者：
Akihiro Kurita;Takeshi Matsunobu;Yasushi Satoh;Takahiro Ando;Shunichi Sato;Minoru Obara;Akihiro Shiotani
研究了纳秒脉冲激光诱导应力波（LISWs）用于大鼠面肌基因转移的可行性。LISW是通过用来自Q开关Nd：YAG激光器的二次谐波（532 nm）的纳秒脉冲激光照射放置在靶组织上的黑色天然橡胶盘来产生的，该激光器广泛用于头颈外科手术并且被证明是安全的。将编码Lac Z的质粒脱氧核糖核酸（DNA）注射到大鼠面部肌肉中后，在不切开或暴露肌肉的情况下，用脉冲激光照射皮肤表面的激光靶。通过对切除的大鼠面部皮肤和肌肉进行X-gal染色来检测Lac Z表达。在基因转移后7天观察到强Lac Z表达，并持续长达14天。基因转移是在距离表面几毫米深的面部肌肉中实现的。基因表达定位于暴露于LISW的组织。未观察到LISW造成的组织损伤。LISW是一种很有前途的非病毒靶基因转移方法，因为它具有高度的空间可控性，易于应用，和微创。使用LISW产生治疗性蛋白质（如生长因子）的基因转移可用于治疗神经损伤和瘫痪。
We investigate the feasibility of using nanosecond pulsed laser-induced stress waves (LISWs) for gene transfer into rat facial muscles. LISWs are generated by irradiating a black natural rubber disk placed on the target tissue with nanosecond pulsed laser light from the second harmonics (532 nm) of a Q-switched Nd:YAG laser, which is widely used in head and neck surgery and proven to be safe. After injection of plasmid deoxyribose nucleic acid (DNA) coding for Lac Z into rat facial muscles, pulsed laser is used to irradiate the laser target on the skin surface without incision or exposure of muscles. Lac Z expression is detected by X-gal staining of excised rat facial skin and muscles. Strong Lac Z expression is observed seven days after gene transfer, and sustained for up to 14 days. Gene transfer is achieved in facial muscles several millimeters deep from the surface. Gene expression is localized to the tissue exposed to LISWs. No tissue damage from LISWs is observed. LISW is a promising nonviral target gene transfer method because of its high spatial controllability, easy applicability, and minimal invasiveness. Gene transfer using LISW to produce therapeutic proteins such as growth factors could be used to treat nerve injury and paralysis.