Ferreira Luiz Eduardo Nunes, Muniz Bruno Vilela, Burga-Sánchez Jonny, Volpato Maria Cristina, de Paula Eneida, Rosa Edvaldo Antonio Ribeiro, Groppo Francisco Carlos
Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
Department of Biochemistry, Biology Institute, University of Campinas - UNICAMP - Campinas, São Paulo, Brazil.
J Pharm Pharmacol. 2017 Feb;69(2):161-171. doi: 10.1111/jphp.12680. Epub 2016 Dec 29.
Modified drug delivery systems have been developed to improve pharmacological properties of local anaesthetics. However, the inflammatory potential of these formulations was not investigated. This study compared the in-vitro effects of ropivacaine (ropi) in plain, liposomal (MLV) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) formulations on cell viability, apoptosis and cytokine (IL-1α, TNF-α, IL-6 and IL-10) release.
Human immortalized keratinocytes (HaCaT) and human immortalized gingival fibroblasts (HGF) were exposed to 1-100 μm ropi concentrations. The cell viability was measured by XTT and LIVE/DEAD assay. Apoptosis was performed by flow cytometry, and cytokine release was measured by ELISA assay.
Human immortalized keratinocyte viability was reduced by ropi and both drug delivery systems. However, none of the formulations induced apoptosis. Results showed a differential regulation of IL-1α TNF-α, IL-6 and IL-10 by HaCaT and HGF. Ropi-HP-β-CD increased twofold the IL-6 release by HGF in comparison with the control, while 100 μm ropi-MLV led to an increased release of all pro-inflammatory cytokines by HGF.
The loss in cell viability was not related to cellular apoptosis. Ropi complexed with HP-β-CD showed a similar cytokine release pattern when compared to the plain formulation. Thus, the HP-β-CD form was a better drug carrier than the MLV form for ropivacaine drug delivery.
已开发出改良的药物递送系统以改善局部麻醉药的药理特性。然而，这些制剂的炎症潜力尚未得到研究。本研究比较了罗哌卡因（ropi）普通制剂、脂质体（MLV）制剂或2-羟丙基-β-环糊精（HP-β-CD）制剂在体外对细胞活力、细胞凋亡和细胞因子（IL-1α、TNF-α、IL-6和IL-10）释放的影响。
将人永生化角质形成细胞（HaCaT）和人永生化牙龈成纤维细胞（HGF）暴露于浓度为1-100μm的罗哌卡因中。通过XTT和活/死检测法测量细胞活力。通过流式细胞术检测细胞凋亡，并通过ELISA检测法测量细胞因子释放。
罗哌卡因及两种药物递送系统均降低了人永生化角质形成细胞的活力。然而，所有制剂均未诱导细胞凋亡。结果显示，HaCaT和HGF对IL-1α、TNF-α、IL-6和IL-10有不同的调节作用。与对照组相比，罗哌卡因-HP-β-CD使HGF释放的IL-6增加了两倍，而100μm的罗哌卡因-MLV导致HGF释放的所有促炎细胞因子增加。
细胞活力的丧失与细胞凋亡无关。与普通制剂相比，与HP-β-CD复合的罗哌卡因显示出相似的细胞因子释放模式。因此，对于罗哌卡因的药物递送，HP-β-CD形式是比MLV形式更好的药物载体。
