Gray Andrea, Marrero-Berrios Ileana, Ghodbane Mehdi, Maguire Timothy, Weinberg Jonathan, Manchikalapati Devasena, SchianodiCola Joseph, Schloss Rene S, Yarmush Joel
Department of Biomedical Engineering, Rutgers, The State University of New Jersey, 599 Taylor Road, Piscataway, New Jersey 08805, USA.
Department of Anesthesiology, New York Methodist Hospital, Brooklyn, New York 11215, USA.
Nano Life. 2015 Jun 1;5(2):1550001-1550014. doi: 10.1142/S1793984415500014.
Anti-fibrotic and tissue regenerative mesenchymal stromal cell (MSC) properties are largely mediated by secreted cytokines and growth factors. MSCs are implanted to augment joint cartilage replacement and to treat diabetic ulcers and burn injuries simultaneously with local anesthetics, which reduce pain. However, the effect of anesthetics on therapeutic human MSC secretory function has not been evaluated. In order to assess the effect of local anesthetics on the MSC secretome, a panel of four anesthetics with different potencies - lidocaine, procaine, ropivacaine and bupivacaine - was evaluated. Since injured tissues secrete inflammatory cytokines, the effects of anesthetics on MSCs stimulated with tumor necrosis factor (TNF)- and interferon (IFN)- were also measured. Dose dependent and anesthesia specific effects on cell viability, post exposure proliferation and secretory function were quantified using alamar blue reduction and immunoassays, respectively. Computational pathway analysis was performed to identify upstream regulators and molecular pathways likely associated with the effects of these chemicals on the MSC secretome. Our results indicated while neither lidocaine nor procaine greatly reduced unstimulated cell viability, ropivacaine and bupivacaine induced dose dependent viability decreases. This pattern was exaggerated in the simulated inflammatory environment. The reversibility of these effects after withdrawal of the anesthetics was attenuated for TNF-/IFN--stimulated MSCs exposed to ropivacaine and bupivacaine. In addition, secretome analysis indicated that constitutive secretion changes were clearly affected by both anesthetic alone and anesthetic plus TNF/IFN cell stimulation, but the secretory pattern was drug specific and did not necessarily coincide with viability changes. Pathway analysis identified different intracellular regulators for stimulated and unstimulated MSCs. Within these groups, ropivacaine and bupivacaine appeared to act on MSCs similarly via the same regulatory mechanisms. Given the variable effect of local anesthetics on MSC viability and function, these studies underscore the need to evaluate MSC in the presence of medications, such as anesthetics, that are likely to accompany cell implantation.
抗纤维化和组织再生的间充质基质细胞（MSC）特性很大程度上由分泌的细胞因子和生长因子介导。将MSC植入以增强关节软骨置换，并与局部麻醉剂同时用于治疗糖尿病溃疡和烧伤，局部麻醉剂可减轻疼痛。然而，麻醉剂对治疗性人MSC分泌功能的影响尚未得到评估。为了评估局部麻醉剂对MSC分泌组的影响，评估了一组四种不同效力的麻醉剂——利多卡因、普鲁卡因、罗哌卡因和布比卡因。由于受伤组织会分泌炎性细胞因子，还测量了麻醉剂对用肿瘤坏死因子（TNF）和干扰素（IFN）刺激的MSC的影响。分别使用alamar蓝还原法和免疫测定法定量了对细胞活力、暴露后增殖和分泌功能的剂量依赖性和麻醉特异性影响。进行了计算通路分析，以确定可能与这些化学物质对MSC分泌组的影响相关的上游调节因子和分子通路。我们的结果表明，虽然利多卡因和普鲁卡因都没有显著降低未刺激的细胞活力，但罗哌卡因和布比卡因诱导了剂量依赖性的活力下降。在模拟的炎症环境中，这种模式更为明显。对于暴露于罗哌卡因和布比卡因的TNF-/IFN-刺激的MSC，麻醉剂撤除后这些影响的可逆性减弱。此外，分泌组分析表明，组成性分泌变化明显受到单独麻醉剂以及麻醉剂加TNF/IFN细胞刺激的影响，但分泌模式具有药物特异性，不一定与活力变化一致。通路分析确定了刺激和未刺激的MSC的不同细胞内调节因子。在这些组中，罗哌卡因和布比卡因似乎通过相同的调节机制对MSC产生类似的作用。鉴于局部麻醉剂对MSC活力和功能的影响各不相同，这些研究强调了在可能伴随细胞植入的药物（如麻醉剂）存在的情况下评估MSC的必要性。
