This study describes the characterization of pharmacokinetic (PK) properties of acetaminophen (APAP) in the Two-Organ-Chip platform (2-OC), a two-chamber device able to cultivate 3D tissues under flow. The APAP intestinal absorption and hepatic metabolism were emulated by human intestine and liver equivalents respectively. The intestinal barrier was produced using Caco-2 and HT-29 cells. The liver spheroids were produced with HepaRG and HHSTeC cells. Cell viability and toxicity were assessed by MTT assay, histology, confocal immunohistochemistry, and multiparametric high content analysis. Gene expression of intestine and liver equivalents were assessed by real-time PCR. Three assemblies of Microphysiological System (MPS) were applied: Intestine 2-OC, Liver 2-OC, and Intestine/Liver 2-OC. The oral administration was emulated by APAP placement over the apical side of the intestinal barrier and the intravenous routes were mimic by the application in the medium. Samples were analyzed by HPLC/UV. APAP 12 μM or 2 μM treatment did not induce cytotoxicity for the intestinal barrier (24 h time-point) or for the liver spheroids 12 h time-point), respectively. All preparations showed slower APAP absorption than reported for humans: Peak time (Tmax) = 12 h for Intestine 2-OC and 6 h for Intestine/Liver 2-OC in both static and dynamic conditions, against reported Tmax of 0,33 to 1,4 h after oral administration to humans. APAP metabolism was also slower than reported for humans. The APAP half-life (T_1/2) was 12 h in the dynamic Liver 2-OC, against T_1/2 = 2 ± 0,4 h reported for humans. Samples taken from the Liver 2-OC static preparation did not show APAP concentration decrease. These findings show the MPS capability and potential to emulate human PK properties and highlight the critical role of mechanical stimulus over cell functionality, especially by demonstrating the clear positive influence of the microfluidic flow over the liver equivalents metabolic performance.

这项研究描述了对乙酰氨基酚（APAP）在两个器官芯片平台（2-OC）中的药代动力学（PK）特性的表征，该平台是一种能够在流动条件下培养3D组织的两腔设备。APAP的肠道吸收和肝脏代谢分别由人体的肠道和肝脏等效物模拟。肠屏障是使用Caco-2和HT-29细胞产生的。肝球体由HepaRG和HHSTeC细胞产生。通过MTT测定，组织学，共聚焦免疫组织化学和多参数高含量分析评估细胞活力和毒性。通过实时PCR评估肠和肝等同物的基因表达。应用了三个微生理系统（MPS）组件：肠2-OC，肝2-OC和肠/肝2-OC。通过将APAP放置在肠屏障的顶侧来模拟口服，通过在培养基中的应用模拟静脉内途径。通过HPLC / UV分析样品。APAP 12μM或2μM处理分别不会对肠屏障（24小时时间点）或肝球体12小时时间点产生细胞毒性。所有制剂的APAP吸收均比人类报告的慢：在静态和动态条件下，肠2-OC的峰值时间（Tmax）= 12小时，肠/肝2-OC的峰值时间（Tmax）= 6小时，而报告的Tmax为0.33至1口服后，4 h。APAP的代谢也比人类报告的要慢。APAP半衰期（T APAP 12μM或2μM处理分别不会对肠屏障（24小时时间点）或肝球体12小时时间点产生细胞毒性。所有制剂的APAP吸收均比人类报告的慢：在静态和动态条件下，肠2-OC的峰值时间（Tmax）= 12小时，肠/肝2-OC的峰值时间（Tmax）= 6小时，而报告的Tmax为0.33至1口服后，4 h。APAP的代谢也比人类报告的要慢。APAP半衰期（T APAP 12μM或2μM处理分别不会对肠屏障（24小时时间点）或肝球体12小时时间点产生细胞毒性。所有制剂均显示出比人类报告的更慢的APAP吸收：在静态和动态条件下，肠2-OC的峰值时间（Tmax）= 12小时，肠/肝2-OC的峰值时间= 6小时，而报告的Tmax为0.33至1口服后，4 h。APAP的代谢也比人类报告的要慢。APAP半衰期（T 对人类口服后的Tmax为0.33至1.4h。APAP的代谢也比人类报告的要慢。APAP半衰期（T 口服给人后的Tmax为0.33至1.4h。APAP的代谢也比人类报告的要慢。APAP半衰期（T在动态肝脏2-OC中_1/2）是12 h， 而人类报告的T _1/2 = 2±0.4 h。从肝脏2-OC静态制剂中提取的样品未显示APAP浓度降低。这些发现显示了MPS模仿人类PK特性的能力和潜力，并突出了机械刺激对细胞功能的关键作用，特别是通过证明微流对肝脏等效代谢性能的明显积极影响。