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Substance use is a risk factor for being both a perpetrator and a victim of violence. The aim of this systematic review was to report the prevalence of acute pre-injury substance use in patients with violence-related injuries. Systematic searches were used to identify observational studies that included patients aged ≥15 years presenting to hospital after violence-related injuries and used objective toxicology measures to report prevalence of acute pre-injury substance use. Studies were grouped based on injury cause (any violence-related, assault, firearm, and other penetrating injuries including stab and incised wounds) and substance type (any substance, alcohol only, drugs other than alcohol only), and they were summarized using narrative synthesis and meta-analyses. This review included 28 studies. Alcohol was detected in 13%–66% of any violence-related injuries (five studies), 4%–71% of assaults (13 studies), 21%–45% of firearm injuries (six studies; pooled estimate = 41%, 95% CI: 40%–42%, n = 9,190), and 9%–66% of other penetrating injuries (nine studies; pooled estimate = 60%, 95% CI: 56%–64%, n = 6,950). Drugs other than alcohol were detected in 37% of any violence-related injuries (one study), 39% of firearm injuries (one study), 7%–49% of assaults (five studies), and 5%–66% of penetrating injuries (three studies). The prevalence of any substance varied across injury categories: any violence-related injuries = 76%–77% (three studies), assaults = 40%–73% (six studies), firearms = n/a, other penetrating injuries = 26%–45% (four studies; pooled estimate = 30%, 95% CI: 24%–37%, n = 319).Overall, substance use was frequently detected in patients presenting to hospital for violence-related injuries. Quantification of substance use in violence-related injuries provides a benchmark for harm reduction and injury prevention strategies. [ABSTRACT FROM AUTHOR]