RESUMO
Detection and characterization of particles in the visible and subvisible size range is critical in many fields of industrial research. Commercial particle analysis systems have proliferated over the last decade. Despite that growth, most systems continue to be based on well-established principles, and only a handful of new approaches have emerged. Identifying the right particle-analysis approach remains a challenge in research and development. The choice depends on each individual application, the sample, and the information the operator needs to obtain. In biopharmaceutical applications, particle analysis decisions must take product safety, product quality, and regulatory requirements into account. Biopharmaceutical process samples and formulations are dynamic, polydisperse, and very susceptible to chemical and physical degradation: improperly handled product can degrade, becoming inactive or in specific cases immunogenic. This article reviews current methods for detecting, analyzing, and characterizing particles in the biopharmaceutical context. The first part of our article represents an overview about current particle detection and characterization principles, which are in part the base of the emerging techniques. It is very important to understand the measuring principle, in order to be adequately able to judge the outcome of the used assay. Typical principles used in all application fields, including particle-light interactions, the Coulter principle, suspended microchannel resonators, sedimentation processes, and further separation principles, are summarized to illustrate their potentials and limitations considering the investigated samples. In the second part, we describe potential technical approaches for biopharmaceutical particle analysis as some promising techniques, such as nanoparticle tracking analysis (NTA), micro flow imaging (MFI), tunable resistive pulse sensing (TRPS), flow cytometry, and the space- and time-resolved extinction profile (STEP®) technology.
RESUMO
BACKGROUND: Soft tissue trauma induces an local inflammatory response and yields a microvascular perfusion failure due to trauma-induced oxidative stress. Using high-resolution multifluorescence microscopy, we herein report on the efficiency of treatment with the oxygen radical scavenger ebselen to improve compromised perfusion of traumatized muscle tissue and to minimize secondary tissue damage. METHODS: By using a pneumatically driven computer-controlled impact device, closed soft tissue trauma of the left hind limb was induced in pentobarbital-anesthetized rats that received either ebselen (30 mg/kg body weight, intraperitoneally) or equal volumes of the vehicle dimethyl sulfoxide (DMSO). In an additional series of animals, ebselen or DMSO were applied without soft tissue trauma. RESULTS: Ebselen restored microcirculatory impairment within the injured muscle, as given by values of nutritive perfusion (763 +/- 44 cm/cm2), nicotinamide adenine dinucleotide levels (56 +/- 3 aU) and inflammatory cell interaction (leukocytes: 226 +/- 31 mm(-2)) at 24 hours after trauma, being not different to those found in noninjured muscle tissue of controls. In contrast, skeletal muscle in DMSO-treated animals revealed persistent perfusion failure (564 +/- 32 cm/cm2) with tissue hypoxia (nicotinamide adenine dinucleotide 75 +/- 11 aU) and enhanced endothelial interaction of leukocytes (383 +/- 18 mm(-2)) at 24 hours after trauma. CONCLUSIONS: Treatment of skeletal muscle soft tissue trauma with the glutathione peroxidase mimic ebselen is highly effective in restoration of disturbed microcirculation. Moreover, reduced inflammatory cell response helps to prevent leukocyte-dependent secondary tissue injury.
