80 Chapter 3 (title, abstract and full text), discrepancies between PPGM and BKHLB were reviewed, and studies were included in case of disagreement. Study characteristics Independently, PPGM, MV and BKHLB extracted the study characteristics, including animal details (species, strain, age, sex and weight), burn method (burn agent, burn size, burn temperature, burn time, burn depth and injury site) and experimental set up (type of control, anesthesia, pain medication, resuscitation, inflammatory mediators and detection method). All of the extracted data was checked by at least one of the other reviewers. Study quality and risk of bias assessment The reporting of any form of randomization or blinding and the presence of a conflict-ofinterest statement was scored for all of the included studies by PPGM and BKHLB who both assessed half of the studies and checked at least 10% of those of the other reviewer. Full risk of bias (RoB) assessment was conducted independently by PPGM and BKHLB using SYRCLE’s tool [81] on 25 randomly selected studies (random number generator in Excel, Microsoft, Redmond, WA). We evaluated the reporting of the following baseline characteristics: animal sex, age, or weight (reporting of a range <10% was considered as a low RoB). To check the completeness of outcome reporting, we evaluated the number of animals in the method and results section for each experiment and outcome. In the case of discrepancies between the two reviewers that were not dismissible, MV was consulted as decisive third reviewer. This assessment provided an indication of the RoB of all included studies. All baseline-controlled studies (n = 29) were scored separately, as only items 7, 8, and 9 of the SYRCLE’s RoB tool applied these studies. Outcome data extraction All quantitative outcome measures related to inflammatory mediators, such as cytokines, chemokines and growth factors, in either blood or wound tissue were collected in a database, which is available upon request. PPGM, MV and BKHLB independently extracted the outcome measures (mean outcome and SD, unit of measurement, number of animals). Extracted data was checked by at least one of the other reviewers. As a secondary outcome measure, data on the wound area (re-epithelization rate, wound closure time) was extracted and recalculated to percentage of remaining wound area. Data from graphs were extracted using the digital ruler feature in ImageJ (version 1.53j, National Institutes of Health, Bethesda, MD) [82]. Data presented as SEM were transformed to SD with the following formula: SD = SEM * . When studies used a relative expression (e.g., protein or mRNA expression level compared to uninjured animals) and no SD/SEM was available, the SD/SEM of the burn-injured group was used as imputation
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