17 General Introduction is a valuable method to synthesize an overview of empirical evidence from separate investigations. These overviews provide insights that will advance experimental design contributing to the reduction and refinement of animal experimentation and will support evidence-based clinical practice [63,64]. BLOOD AND WOUND TISSUE FROM BURN PATIENTS Alternatively, valuable insights into the burn-induced immune response can be generated by investigating patient specimens. Phenotypic characterization and quantification of cells, and analysis of inflammatory mediators in blood and wound tissue inform us of the specific immune cells and factors that are actively involved in inflammation. Patient studies are limited by restrictions in sampling and absence of baseline values. However, valuable information can be generated by using leftover blood and burn tissue specimen originating from routine blood withdrawals and surgeries as part of clinical practice. Laboratory techniques such as flow cytometry, immunoassays and microscopy can uncover cellular activity and processes that are involved in the burn-induced immune response. Moreover, by analyzing patient samples from different time intervals after burn injury, time-dependent effects can be investigated. MODELING THE POST-BURN IMMUNE RESPONSE Growing ethical and scientific concerns drive scientists to search for animal-free approaches to study burn injury. An appealing alternative to animal experimentation is the use of in vitro skin models. Such skin models mimic the tissue architecture of native human skin. The information collected from literature and patient studies can be used to develop and adjust in vitro skin models. Such models can be used to study aspects of wound healing and inflammation after burn injury in a standardized and controlled setting. Since in vitro skin models are not connected to a blood circulation, the influx of immune cells and factors is missing and should be simulated. Therefore, there is a need for more sophisticated in vitro skin models to mimic defined aspects of the burn-induced immune response. Ultimately, the information from animal, patient and skin model studies can spark the design of therapeutic interventions that will improve recovery speed and reduce the side effects of a hyper-inflammatory response such as excessive scarring. Early safety and efficacy tests of promising therapeutic candidates can be performed using the in vitro skin models before progressing to burn patients. 1
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