Wuya College of Innovation, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, China.
College of Pharmaceutical Engineering, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, China.
Int J Biol Macromol. 2024 Oct;277(Pt 3):134408. doi: 10.1016/j.ijbiomac.2024.134408. Epub 2024 Aug 2.
Skin interstitial fluid (ISF) has emerged as a significant reservoir of biomarkers for disease diagnosis and prevention. Microneedle (MN) patches are regarded as an optimal platform for ISF extraction from the skin due to their non-invasive nature. However, challenges such as prolonged sampling durations and complex detection procedures impede timely metabolic analysis. In this investigation, we amalgamated MN technology with immobilized enzyme technology to fabricate a dual-layer MN patch integrating sampling and detection functionalities, thereby enabling in-situ colorimetric detection of hyperglycemia. The tip layer of the patch, comprising polyvinyl alcohol/carboxymethyl chitosan (PVA/CMCS) MN, was synthesized utilizing a chemical crosslinking approach for the first time, with glucose oxidase (GOx) being incorporated. The hydrophilicity of CMCS expedited the extraction process, facilitating the retrieval of approximately 10 mg of ISF within 10 min. The backing layer consisted of an immobilized polyvinyl alcohol-chitosan-horseradish peroxidase (PVA-CS-HRP) hydrogel film loaded with 3,3', 5,5'-tetramethylbenzidine (TMB). Incorporating macromolecular polymer PVA and CS for HRP immobilization addressed the issue of poor stability associated with traditional natural enzymes, thereby enhancing the sensitivity of the reaction system. The in-situ colorimetric sensor facilitated minimally invasive ISF extraction and swift conversion of glucose levels into detectable color changes.
皮肤间质液（ISF）已成为疾病诊断和预防的重要生物标志物来源。微针（MN）贴片由于其非侵入性而被认为是从皮肤中提取 ISF 的最佳平台。然而，延长的采样持续时间和复杂的检测程序等挑战阻碍了及时的代谢分析。在这项研究中，我们将 MN 技术与固定化酶技术相结合，制造出一种具有采样和检测功能的双层 MN 贴片，从而能够现场比色检测高血糖症。贴片的尖端层由聚乙烯醇/羧甲基壳聚糖（PVA/CMCS）MN 组成，首次利用化学交联方法合成，其中掺入了葡萄糖氧化酶（GOx）。CMCS 的亲水性加速了提取过程，在 10 分钟内可提取约 10mg 的 ISF。背衬层由负载 3,3',5,5'-四甲基联苯胺（TMB）的固定化聚乙烯醇-壳聚糖-辣根过氧化物酶（PVA-CS-HRP）水凝胶膜组成。将高分子聚合物 PVA 和 CS 用于 HRP 固定化解决了传统天然酶稳定性差的问题，从而提高了反应系统的灵敏度。现场比色传感器促进了微创 ISF 提取，并迅速将葡萄糖水平转化为可检测的颜色变化。
