Systematic assessment of microneedle injection into the mouse cornea.
Systematic assessment of microneedle injection into the mouse cornea.
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DOI：
10.1186/2047-783x-17-19
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发表时间：
2012-06-20
影响因子：
4.2
通讯作者：
Jun AS
中科院分区：
文献类型：
--
作者：
Matthaei M;Meng H;Bhutto I;Xu Q;Boelke E;Hanes J;Jun AS
角膜基质内注射是基因载体应用于上皮下的一种重要方式。在小鼠模型中，由于角膜尺寸减小，该过程基本上变得复杂。此外，可能难以估计单次注射的体积所达到的角膜面积。本研究旨在研究使用不同微针向小鼠角膜进行基质内注射，并量化注射不同体积的效果。描述了一种可重复的注射技术。对20只129 Sv/J小鼠的40只眼睛进行了测试。使用30°斜面33 G针、三表面25°斜面35 G针或手拉和25°斜面玻璃针进行层内注射印度墨水。每只眼睛接受1或2 μL体积的单次注射。角膜巩膜扣固定并平放，用于受影响角膜面积的计算机辅助定量。进行组织学评估以研究注射染料的基质内位置。基质内注射1 μ L和2 μL分别覆盖的平均角膜面积为5.0 ±1.4 mm 2（平均值± SD）和7.7 ±1.4 mm 2。对于1 μL注射液，达到的总角膜面积的平均百分比范围为39%-53%，对于2 μL注射液，为65%-81%。使用33 G针头的注射往往提供最高的分布面积。30°斜面33 G针的穿孔率为8%，三面斜面35 G针的穿孔率为44%。玻璃针未观察到穿孔;然而，在这些病例中，25%的病例发现针尖基质内断裂。使用30°斜面33 G针头进行角膜内注射安全有效。使用三面斜面35 G针大大增加了角膜穿孔的数量。玻璃针可能在角膜基质内破裂。1 μL和2 μL的注射导致总平均值分别为49%和73%的总角膜面积受累。
Corneal intrastromal injection is an important mode of gene-vector application to subepithelial layers. In a mouse model, this procedure is substantially complicated by the reduced corneal dimensions. Furthermore, it may be difficult to estimate the corneal area reached by the volume of a single injection. This study aimed to investigate intrastromal injections into the mouse cornea using different microneedles and to quantify the effect of injecting varying volumes. A reproducible injection technique is described. Forty eyes of 20 129 Sv/J mice were tested. India ink was intrastromally injected using 30° beveled 33 G needles, tri-surface 25° beveled 35 G needles, or hand-pulled and 25° beveled glass needles. Each eye received a single injection of a volume of 1 or 2 μL. Corneoscleral buttons were fixed and flat mounted for computer-assisted quantification of the affected corneal area. Histological assessment was performed to investigate the intrastromal location of the injected dye. A mean corneal area of 5.0 ±1.4 mm2 (mean ± SD) and 7.7 ±1.4 mm2 was covered by intrastromal injections of 1 and 2 μL, respectively. The mean percentage of total corneal area reached ranged from 39% to 53% for 1 μL injections, and from 65% to 81% for 2 μL injections. Injections using the 33 G needles tended to provide the highest distribution area. Perforation rates were 8% for 30° beveled 33 G needles and 44% for tri-surface beveled 35 G needles. No perforation was observed with glass needle; however, intrastromal breakage of needle tips was noted in 25% of these cases. Intracorneal injection using a 30° beveled 33 G needle was safe and effective. The use of tri-surface beveled 35 G needles substantially increased the number of corneal perforations. Glass needles may break inside the corneal stroma. Injections of 1 μL and 2 μL resulted in an overall mean of 49% and 73% respectively of total corneal area involved.