Eriksson E, Yao F, Svensjö T, Winkler T, Slama J, Macklin M D, Andree C, McGregor M, Hinshaw V, Swain W F
Division of Plastic Surgery, Brigham and Women's Hospital, Boston, Massachusetts, 02115, USA.
J Surg Res. 1998 Aug;78(2):85-91. doi: 10.1006/jsre.1998.5325.
Gene transfer to skin has many potential applications but lacks a safe, practical delivery method. This report presents a new technique, microseeding, for in vivo gene transfer to skin and wounds and for DNA-mediated vaccination. The plasmid DNA solution was delivered directly to the target cells of the skin by a set of oscillating solid microneedles driven by a modified tattooing device.
Skin and partial-thickness excisional wounds in pigs were microseeded with either hEGF expression plasmid or beta-galactosidase expression plasmid. Human EGF was also delivered by single injection or particle bombardment. hEGF expression in wound fluid and in target tissue was determined by ELISA with anti-hEGF-specific antibodies. Additionally, weanling pigs were microseeded with a hemagglutinin of swine influenza virus expression plasmid and production of anti-HA-specific antibodies was determined by blocking ELISA.
hEGF expression in microseeded partial thickness wounds (5664 pg/site) and skin sites (969 pg/site) peaked 2 days after transfection being four- to seven-fold higher than gene transfer by a single intradermal injection and two- to three-fold higher than particle-mediated gene transfer. The beta-galactosidase-expressing cells were detected in dermis and epidermis. Pigs microseeded with HA expression plasmid were protected from infection by the Swine influenza virus.
These results demonstrate that microseeding is a simple and effective method for in vivo gene transfer to skin and wounds and is more efficient than single injection and particle-mediated gene transfer.
基因转移至皮肤有许多潜在应用，但缺乏一种安全、实用的递送方法。本报告介绍了一种新技术——微播种，用于体内基因转移至皮肤和伤口以及DNA介导的疫苗接种。通过改良纹身装置驱动的一组振荡固体微针将质粒DNA溶液直接递送至皮肤的靶细胞。
用hEGF表达质粒或β-半乳糖苷酶表达质粒对猪的皮肤和部分厚度切除伤口进行微播种。人表皮生长因子（hEGF）也通过单次注射或粒子轰击递送。用抗hEGF特异性抗体通过酶联免疫吸附测定（ELISA）法测定伤口液和靶组织中的hEGF表达。此外，用猪流感病毒血凝素表达质粒对断奶仔猪进行微播种，并通过阻断ELISA法测定抗HA特异性抗体的产生。
微播种的部分厚度伤口（5664 pg/部位）和皮肤部位（969 pg/部位）中的hEGF表达在转染后2天达到峰值，比单次皮内注射基因转移高4至7倍，比粒子介导的基因转移高2至3倍。在真皮和表皮中检测到表达β-半乳糖苷酶的细胞。用HA表达质粒微播种的猪受到猪流感病毒感染的保护。
这些结果表明，微播种是一种简单有效的体内基因转移至皮肤和伤口的方法，比单次注射和粒子介导的基因转移更有效。
