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对乙肝疫苗进行体外经皮实验以评价疫苗在被动扩散和离子导入情况下的经皮渗透特性。体外透皮研究采用Franz扩散池,以SD大鼠的腹部皮肤为渗透屏障,以酶联免疫法测定药物累积渗透量和在皮肤中的滞留量。乙肝疫苗(质量浓度为23μg·mL-1与46μg·mL-1)经完整皮肤被动扩散的经皮渗透量与皮肤滞留量均极少,24 h累积渗透量仅(2.1±0.1)ng.cm-2和(2.3±0.1)ng.cm-2。去除角质层后,经皮渗透量与皮肤滞留量分别提高至(383.7±86.2)ng.cm-2和(16.8±4.6)ng.cm-2,是完整皮肤的171.6倍与2.1倍(46μg·mL-1)。离子导入对于乙肝疫苗具有明显的经皮渗透促进作用:完整皮肤经皮离子导入6 h,乙肝疫苗的累积渗透量是被动扩散6 h的2.7倍(23μg·mL-1)和6.6倍(46μg·mL-1);去角质层皮肤离子导入,乙肝疫苗的累积渗透量是被动扩散6 h的1.6倍(23μg·mL-1)和1.8倍(46μg·mL-1)。离子导入也能显著增加疫苗在皮肤中的滞留量。离子导入6 h疫苗在完整皮肤中的滞留量和去角质层皮肤中的滞留量均与被动扩散24 h的皮肤滞留量接近[完整皮肤:(16.8±4.6)ng.cm-2 vs(13.3±5.4)ng.cm-2;去角质层皮肤:(36.7±14.1)ng.cm-2 vs(26.8±11.2)ng.cm-2](P>0.05)。研究结果表明,离子导入是促进乙肝疫苗等生物活性大分子经皮渗透的有效手段,有希望应用于乙肝疫苗的经皮免疫领域。
An in vitro transdermal experiment was conducted on Hepatitis B vaccine to evaluate the transdermal permeation characteristics of the vaccine under passive diffusion and iontophoresis. In vitro transdermal study using Franz diffusion cell, the abdominal skin of SD rats as a permeable barrier, enzyme-linked immunosorbent assay drug cumulative penetration and retention in the skin. Transdermal permeation and skin retention of HBsAg (mass concentrations of 23μg · mL-1 and 46μg · mL-1) after passive skin diffusion were minimal, with a cumulative infiltration of only (2.1 ± 0.1) ng · cm -2 and (2.3 ± 0.1) ng.cm-2. After removing the stratum corneum, the transdermal penetration and skin retention increased to (383.7 ± 86.2) ng.cm-2 and (16.8 ± 4.6) ng.cm-2, respectively, which were 171.6 times and 2.1 times mL-1). The iontophoresis had a significant transdermal permeation promotion effect on hepatitis B vaccine. The intact percutaneous iontophoresis was 6 h, and the cumulative infiltration of hepatitis B vaccine was 2.7 times (23 μg · mL -1) and 6.6 times (46 μg · mL-1). Exfoliant skin iontophoresis resulted in 1.6 times (23μg · mL-1) and 1.8 times (46μg · mL-1) cumulative permeation of hepatitis B vaccine over 6 hours. Ion introduction can also significantly increase the amount of vaccine retained in the skin. The retention of intact skin in the intact skin and the retention in the exfoliated skin of the 6 h iontophoresis vaccine were similar to those of the passive diffusion for 24 h [whole skin: (16.8 ± 4.6) ng.cm-2 vs (13.3 ± (36.7 ± 14.1) ng.cm-2 vs (26.8 ± 11.2) ng.cm-2] (P> 0.05). The results show that iontophoresis is an effective means to promote transdermal penetration of biologically active macromolecules such as hepatitis B vaccine, which is promising for the transcutaneous immunization of hepatitis B vaccine.