Microdialysis: current applications in clinical pharmacokinetic studies and its potential role in the future

Microdialysis is a probe-based sampling method, which, if linked to analytical devices, allows for the measurement of drug concentration profiles in selected tissues. During the last two decades, microdialysis has become increasingly popular for preclinical and clinical pharmacokinetic studies. The advantage of in vivo microdialysis over traditional methods relates to its ability to continuously sample the unbound drug fraction in the interstitial space fluid (ISF). This is of particular importance because the ISF may be regarded as the actual target compartment for many drugs, e.g. antimicrobial agents or other drugs mediating their action through surface receptors. In contrast, plasma concentrations are increasingly recognised as inadequately predicting tissue drug concentrations and therapeutic success in many patient populations. Thus, the minimally invasive microdialysis technique has evolved into an important tool for the direct assessment of drug concentrations at the site of drug delivery in virtually all tissues. In particular, concentrations of transdermally applied drugs, neurotransmitters, antibacterials, cytotoxic agents, hormones, large molecules such as cytokines and proteins, and many other compounds were described by means of microdialysis. The combined use of microdialysis with non-invasive imaging methods such as positron emission tomography and single photon emission tomography opened the window to exactly explore and describe the fate and pharmacokinetics of a drug in the body. Linking pharmacokinetic data from the ISF to pharmacodynamic information appears to be a straightforward approach to predicting drug action and therapeutic success, and may be used for decision making for adequate drug administration and dosing regimens. Hence, microdialysis is nowadays used in clinical studies to test new drug candidates that are in the pharmaceutical industry drug development pipeline.     

Microdialysis for the evaluation of penetration through the human skin barrier — a promising tool for future research?

The direct measurement of local drug concentration levels at discreet skin locations with minor trauma has recently become possible with the introduction of cutaneous microdialysis. Cutaneous microdialysis is an in vivo sampling technique for measuring solutes in the extracellular fluid of the dermis. When used in combination with other experimental approaches, for example with a variety of non-invasive techniques to describe the functional status of the skin (bioengineering methods), it may help investigators to gain new insights into the fields of skin diseases, metabolism and drug absorption/penetration. An important parameter to describe the efficacy of microdialysis is the relative recovery. This is the ratio between the concentration of a substance in the dialysate and the true extracellular concentration. Several methods are in common use to describe the relative recovery (no-net-flux method or retrodialysis). Parameters such as probe design, depth of the probe in the dermis, physico-chemical properties of the compound of interest, and analytical aspects are important factors influencing microdialysis. Microdialysis has been used to investigate the influence of penetration enhancers, vehicles or iontophoresis on percutaneous absorption, performed by in vivo studies in rats. In human volunteers, most of the experiments have been performed to study the kinetics of fast penetrating substances, e.g. nicotine, non-steroidal antiinflammatory drugs, local anaesthetics, or solvents. Problems have been encountered in the detection of lipophilic and highly protein-bound substances. Further, dermal metabolism and the influence of barrier perturbation on percutaneous absorption have been analyzed. Investigations suggest that microdialysis, in combination with traditional techniques, might give valuable information regarding the assessment of the penetration of drugs and other exogenous agents through the skin. In spite of the clearly defined and accepted advantages of microdialysis technology for studies of transdermal drug delivery, to date no standardized test procedure exists nor has the reproducibility of the results been evaluated. In the future, these problems have to be solved to enable this method to find its place in standard research.     

Microdialysis as a tool in local pharmacodynamics

In many cases the clinical outcome of therapy needs to be determined by the drug concentration in the tissue compartment in which the pharmacological effect occurs rather than in the plasma. Microdialysis is an in vivo technique that allows direct measurement of unbound tissue concentrations and permits monitoring of the biochemical and physiological effects of drugs throughout the body. Microdialysis was first used in pharmacodynamic research to study neurotransmission, and this remains its most common application in the field. In this review, we give an overview of the principles, techniques, and applications of microdialysis in pharmacodynamic studies of local physiological events, including measurement of endogenous substances such as acetylcholine, catecholamines, serotonin, amino acids, peptides, glucose, lactate, glycerol, and hormones. Microdialysis coupled with systemic drug administration also permits the more intensive examination of the pharmacotherapeutic effect of drugs on extracellular levels of endogenous substances in peripheral compartments and blood. Selected examples of the physiological effects and mechanisms of action of drugs are also discussed, as are the advantages and limitations of this method. It is concluded that microdialysis is a reliable technique for the measurement of local events, which makes it an attractive tool for local pharmacodynamic research.     

Methodological issues in microdialysis sampling for pharmacokinetic studies

Microdialysis is an in vivo technique that permits monitoring of local concentrations of drugs and metabolites at specific sites in the body. Microdialysis has several characteristics, which makes it an attractive tool for pharmacokinetic research. About a decade ago the microdialysis technique entered the field of pharmacokinetic research, in the brain, and later also in peripheral tissues and blood. Within this period much has been learned on the proper use of this technique. Today, it has outgrown its child diseases and its potentials and limitations have become more or less well defined. As microdialysis is a delicate technique for which experimental factors appear to be critical with respect to the validity of the experimental outcomes, several factors should be considered. These include the probe; the perfusion solution; post-surgery interval in relation to surgical trauma, tissue integrity and repeated experiments; the analysis of microdialysate samples; and the quantification of microdialysate data. Provided that experimental conditions are optimized to give valid and quantitative results, microdialysis can provide numerous data points from a relatively small number of individual animals to determine detailed pharmacokinetic information. An example of one of the added values of this technique compared with other in vivo pharmacokinetic techniques, is that microdialysis reflects free concentrations in tissues and plasma. This gives the opportunity to assess information on drug transport equilibration across membranes such as the blood-brain barrier, which already has provided new insights. With the progress of analytical methodology, especially with respect to low volume/low concentration measurements and simultaneous measurement of multiple compounds, the applications and importance of the microdialysis technique in pharmacokinetic research will continue to increase.     

微透析技术在靶组织局部药物浓度检测中的应用

微透析技术是一项越来越被广泛使用的在体研究技术,其遵循透析原理,以探针为基础取样,可连续检测局部组织细胞外液的药物浓度,能满足常规的药代动力学/药效学(PK-PD)研究。本文就其在靶组织局部药物浓度检测中的应用作一简要介绍。     

Microdialysis for pharmacokinetic analysis of drug transport to the brain

The intracerebral microdialysis technique represents an important tool for monitoring free drug concentrations in brain extracellular fluid (brain(EcF)) as a function of time. With knowledge of associated free plasma concentrations, it provides information on blood-brain barrier (BBB) drug transport. However, as the implantation of the microdialysis probe evokes tissue reactions, it should be established if the BBB characteristics are maintained under particular microdialysis experimental conditions. Several studies have been performed to evaluate the use of intracerebral microdialysis as a technique to measure drug transport across the BBB and to measure regional pharmacokinetics of drugs in the brain. Under carefully controlled conditions, the intracerebral microdialysis data did reflect passive BBB transport under normal conditions, as well as changes induced by hyperosmolar opening or by the presence of a tumor in the brain. Studies on active BBB transport by the mdr1a-encoded P-glycoprotein (Pgp) were performed, comparing mdr1a(-/-) with wild-type mice. Microdialysis surgery and experimental procedures did not affect Pgp functionality, but the latter did influence in vivo concentration recovery, which was in line with theoretical predictions. It is concluded that intracerebral microdialysis provides meaningful data on drug transport to the brain, only if appropriate methods are applied to determine in vivo concentration recovery.     

Microdialysis as an Important Technique in Systems Pharmacology—a Historical and Methodological Review

Microdialysis has contributed with very important knowledge to the understanding of target-specific concentrations and their relationship to pharmacodynamic effects from a systems pharmacology perspective, aiding in the global understanding of drug effects. This review focuses on the historical development of microdialysis as a method to quantify the pharmacologically very important unbound tissue concentrations and of recent findings relating to modeling microdialysis data to extrapolate from rodents to humans, understanding distribution of drugs in different tissues and disease conditions. Quantitative microdialysis developed very rapidly during the early 1990s. Method development was in focus in the early years including development of quantitative microdialysis, to be able to estimate true extracellular concentrations. Microdialysis has significantly contributed to the understanding of active transport at the blood-brain barrier and in other organs. Examples are presented where microdialysis together with modeling has increased the knowledge on tissue distribution between species, in overweight patients and in tumors, and in metabolite contribution to drug effects. More integrated metabolomic studies are still sparse within the microdialysis field, although a great potential for tissue and disease-specific measurements is evident.     

Microdialysis Sampling for the Investigation of Dermal Drug Transport

Microdialysis perfusion in vivo has the potential to be a powerful sampling technique in dermal and transdermal drug delivery studies. Characterization of a commercially available microdialysis probe in vitro considering relevant physiological parameters is a vital first step in the evaluation of microdialysis as a dermal sampling technique. In previous microdialysis studies, analyte concentration and neutrality have been implicated in altering microdialysis recovery. The recovery of a model compound 5-fluorouracil (5-FU) was investigated at several pH values and donor concentrations. The relative recovery of 5-FU by the microdialysis probe was affected by pH but not by donor concentration. To confirm further that the changing concentration and pH profile presented by the flux of 5-FU was not significantly altering microdialysis recovery, an experiment comparing direct and microdialysis sampling of a Franz diffusion cell receptor compartment was performed. Although the 5-FU concentration (0-686 ng/ml) and pH (7.40-7.24) changed substantially, the recovery of 5-FU was not adversely affected. To demonstrate the feasibility of dermal microdialysis, the flux of a commercial preparation of 5-fluorouracil was monitored utilizing a microdialysis probe implanted in excised rat skin in vitro. The results from the dermally implanted probe demonstrate the potential of the technique while establishing the limitations of the current microdialysis system.     

Microdialysis in peripheral tissues

The objective of this review is to survey the recent literature regarding the applications of microdialysis in pharmacokinetic studies and facilitating many other studies in peripheral tissues such as muscle, subcutaneous adipose tissue, heart, lung, etc. It has been reported extensively that microdialysis is a useful technique for monitoring free concentrations of compounds in extracellular fluid (ECF), and it is gaining popularity in pharmacokinetic and pharmacodynamic studies, both in experimental animals and humans. The first part of this review discusses the use of microdialysis technique for ECF sampling in peripheral tissues in animal studies. The second part of the review describes the use of microdialysis for ECF sampling in peripheral tissues in human studies. Microdialysis has been applied extensively to measure both endogenous and exogenous compounds in ECF. Of particular benefit is the fact that microdialysis measures the unbound concentrations in the peripheral tissue fluid which have been shown to be responsible for the pharmacological effects. With the increasing number of applications of microdialysis, it is obvious that this method will have an important place in studying drug pharmacokinetics and pharmacodynamics.     

微透析技术在抗菌药物药动学和药效学中的应用

微透析技术是一项新兴的体内药物分析技术。通过微透析技术与药动学和药效学模型结合,实现对组织或细胞外游离态药物浓度及其相应药理效应的同时研究,不仅有利于进一步明确药物的剂量-效应关系,制定临床最佳给药方案,而且也为个体化给药提供了科学依据。本文综合近年文献,对微透析技术的基本原理以及在抗菌药物药动学和药效学研究中的应用作一综述。     

The use of microdialysis for the determination of plasma protein binding of drugs.

Microdialysis sampling was used for the determination of the protein binding and the free therapeutic drug concentration of drugs in plasma in vitro. Several drugs with varying extent of protein binding and for which the plasma monitoring is important were studied. To mimic the in vivo situation, an artificial blood vessel was constructed and filled with spiked plasma circulating at the flow rate of human blood at 37 degrees C. The microdialysis probe (16 mm membrane length, 20000 MW cut off) was placed in the vessel and perfused with 0.9% NaCl at 5 microliters min-1. Dialysates were collected every 10 min and were analysed by reversed-phase LC with UV detection. The free concentration of the drug was calculated by correcting the concentration in the dialysate for the recovery of the probe, which was also determined in the artificial blood vessel after the experiment. The data confirm that microdialysis is a valid alternative technique for the determination of protein binding or free therapeutic plasma concentration of drugs on a comparative basis. Reference to literature values indicates that the results of the proposed method correspond reasonably well with accepted values.     

The Use of Microdialysis in Pharmacokinetics and Pharmacodynamics

Microdialysis is a new in vivo sampling technology applied to the study of pharmacokinetics and drug metabolism in the blood and soft tissues of living systems. A small-diameter probe containing a dialysis membrane is implanted into tissue and perfused with a suitable fluid. Low-molecular-weight substances passively diffuse through the semipermeable membrane along a concentration gradient, resulting in the collection of purified dialysate samples. The advantage of this approach over blood sampling and dissection of tissues is the ability to sample blood and extracellular fluid with minimal tissue damage or alteration of fluid balance. Sampling several tissues simultaneously and continuously in animal models allows data to be obtained that more directly reflect interactions of drugs at their sites of activity and detoxification. Techniques such as this will have a tremendous impact on preclinical and clinical pharmacologic research.     

Application of microdialysis to characterize drug disposition in tumors

Microdialysis is an in vivo sampling technique that was initially developed to measure endogenous substances in the field of neurotransmitter research. In the past decade, microdialysis has been increasingly applied to study the pharmacokinetics and drug metabolism in the blood and various tissues of both animals and humans. This paper describes the general aspects of this in vivo sampling technique followed by the survey of the recent papers regarding the application of microdialysis to characterize anticancer drug disposition in solid tumors. It can be concluded that microdialysis is a very suitable method to obtain drug concentration-time profiles in the interstitial fluid of solid tumors as well as of other variety of tissues.     

Muscle distribution of the neuromuscular blocker gallamine using microdialysis

Measurement of drug concentrations in target tissue has the potential to provide insight into the pharmacokinetics and pharmacodynamics of a drug. In this study, the distribution of the neuromuscular blocker, gallamine, into muscle tissue was investigated in urethane-anesthetized rats after an intravenous bolus dose (6 mg/kg). Microdialysis sampling was used to continuously determine gallamine concentrations in muscle interstitial fluid (MIF). In vivo microdialysis recovery of gallamine was determined as the relative loss of gallamine from the perfusate into muscle tissue after perfusion with gallamine (2 microg/mL). Recovery was determined in each rat before the pharmacokinetic studies. Terminal muscle sampling followed by homogenization was also performed to examine gallamine distribution within muscle tissue. All samples were assayed for gallamine using a validated high-performance liquid chromatography assay. Gallamine was rapidly distributed into MIF with a MIF-plasma partition coefficient of 0.9 +/- 0.1 (n = 6). By contrast, the estimated gallamine concentration in muscle tissue homogenate was only 23 +/- 5% (n = 5) of the concentration in MIF as estimated by microdialysis sampling at the terminal sampling time. These findings suggest that gallamine is not distributed uniformly within muscle but selectively distributes into MIF. Simulations using a hybrid physiologically based pharmacokinetic model which describes uptake of drug only into the interstitial space showed good agreement between predicted and observed concentration data obtained from microdialysis sampling, supporting the findings that gallamine selectively distributes into MIF. These studies demonstrate microdialysis combined with conventional terminal tissue sampling provides valuable information on intra-tissue drug distribution.     

In Vivo Microdialysis Sampling for Pharmacokinetic Investigations

In vivo microdialysis sampling coupled to liquid chromatography was used to study acetaminophen disposition in anesthetized rats. The pharmacokinetics of acetaminophen and its sulfate and glucuronide metabolites were determined using both microdialysis sampling and collection of whole blood. For microdialysis, samples were continuously collected for over 5 hr without fluid loss using a single experimental animal. Microdialysis sampling directly assesses the free drug concentration in blood. The pharmacokinetic results obtained with microdialysis sampling were the same as those obtained from blood collection. The administration of heparin, necessary when collecting blood samples, was found to double the elimination half-life of acetaminophen. Microdialysis sampling is a powerful tool for pharmacokinetic studies, providing accurate and precise pharmacokinetic data.     

Pitfalls of the application of microdialysis in clinical oncology: controversial findings with docetaxel

Microdialysis is a novel and minimally invasive sampling technique, based on the diffusion of analytes from the interstitial compartment through a semi-permeable membrane, and enables direct assessment of tissue disposition and penetration of drugs. Variable antitumor responses may be associated with differences in tumor vascularity, capillary permeability or tumor interstitial pressure resulting in variable delivery of anticancer agents. In preparation of pharmacokinetic studies, aimed at measuring docetaxel concentrations in healthy and malignant tissues in vivo, in pre-clinical as well as clinical studies, in vitro recovery experiments were performed. In contrast to published data, the recovery experiments suggest that docetaxel has a very low recovery as a result of non-specific binding to currently available microdialysis catheters. Here we discuss our findings with docetaxel in a historical perspective and we report on our experience using polysorbate 80 to eliminate the non-specific binding and its effects on the recovery of docetaxel.     

微透析技术在药物靶组织分布和代谢研究中的应用

微透析技术是一项新兴的在体研究技术，近年来已广泛应用于药物在靶组织分布和代谢研究，对于阐明药物体内过程、疗效和安全性有重要意义，为新药研发与临床合理用药提供科学依据。现通过检索分析相关文献，就微透析技术的概况、微透析探针、透析液的分析及微透析技术在药物靶组织分布和代谢研究中的应用作一综述。     

The use of microdialysis in CNS drug delivery studies. Pharmacokinetic perspectives and results with analgesics and antiepileptics

Microdialysis can give simultaneous information on unbound drug concentration-time profiles in brain extracellular fluid (ECF) and blood, separating the information on blood-brain barrier (BBB) processes from confounding factors such as binding to brain tissue or proteins in blood. This makes microdialysis suitable for studies on CNS drug delivery. It is possible to quantify influx and efflux processes at the BBB in vivo, and to relate brain ECF concentrations to central drug action. The half-life in brain ECF vs. the half-life in blood gives information on rate-limiting steps in drug delivery and elimination from the CNS. Examples are given on microdialysis studies of analgesic and antiepileptic drugs.     

Microdialysis as a tool to determine free kidney levels of voriconazole in rodents: a model to study the technique feasibility for a moderately lipophilic drug

Microdialysis has been employed for the in vivo measurement of endogenous compounds and a variety of drugs in different tissues. The applicability of this technique can be limited by drug lipophilicity which can impair the diffusion through dialysis membrane. The objective of this study was to evaluate the feasibility of using microdialysis to study kidney penetration of voriconazole, a moderately lipophilic antifungal triazolic agent (LogD7.4=1.8). Microdialysis probe recoveries were investigated in vitro by dialysis and retrodialysis using four different drug concentrations (0.1-2 microg/mL) at five flow rates (1-5 microL/min). Recoveries were dependent on the method used for the determination as well as on the flow rate, but independent of drug concentration. The average apparent recoveries determined by dialysis and retrodialysis, at flow rate of 2 microL/min, were 21.1+/-1.5% and 28.7+/-2.0%, respectively. Recovery by retrodialysis was bigger than the recovery by dialysis. The average apparent dialysis/retrodialysis recovery ratio in vitro was 0.73 for all concentrations investigated. The differences between retrodialysis and dialysis recoveries were attributed to the drug's binding to the plastic tubing before and after the dialysis membrane which was experimentally evaluated and mathematically modeled. The in vivo apparent recovery determined by retrodialysis in healthy Wistar rats' kidney was 38.5+/-3.5%, similar to that observed in vitro using the same method (28.7+/-2.0%). The in vivo apparent recovery after correcting for plastic tubing binding (25.1+/-2.8%) was successfully used for determining free kidney levels of voriconazole in rats following 40 and 60mg/kg oral dosing. The results confirmed that microdialysis can be used as sampling technique to determine free tissue levels of moderately lipophilic drugs once the contribution of tubing binding and membrane diffusion on the apparent recovery are disentangled.     

微透析-高效液相色谱法联用研究吴茱萸提取物的经皮吸收药动学特性

目的:利用在体皮肤微透析技术研究吴茱萸提取物经皮吸收的特性。方法:以裸鼠为研究对象,建立在体经皮微透析采样技术,以吴茱萸碱和吴茱萸次碱为指标成分,采用高效液相色谱测定吴茱萸提取物经皮给药后透析液中的药物浓度,通过相对损失率的校正,计算皮肤药物浓度,并利用Kinetica 5.0软件对皮肤药物浓度和时间进行非房室模型拟合,计算相关统计参数。结果:吴茱萸提取物中吴茱萸碱的达峰时间为(150±15.3)min,半衰期(t_1/2)(263.7±41.6)min;吴茱萸次碱的达峰时间为(90±2.1)min,半衰期(t_1/2)(194±17.3)min;二者均能较快地达到峰值,并在较长的时间内保持稳定释放,使皮下组织中的药物浓度保持在一个相对恒定的水平。结论:本研究中所建立的微透析方法可用于吴茱萸提取物的皮肤药动学研究,吴茱萸碱和吴茱萸次碱可透过皮肤吸收而发挥临床药效。