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One of the greatest challenges in responding to the COVID-19 pandemic is preventing staff exposure and infection by ensuring consistent and effective use of personal protective equipment (PPE). This study explored health care workers' experience of prolonged PPE use in clinical practice settings and their concerns regarding PPE supply, effectiveness and training needs.A descriptive cross-sectional design was adopted in this study.Health care workers (N = 592) from an acute care hospital completed an online survey from July to September 2020 assessing: (i) usage frequencies, side effects and interference with patient care; and (ii) perceptions of access to PPE, likelihood of exposure to infection and adequacy of PPE training.PPE-related side effects were reported by 319 (53.8%) participants, the majority being nurses (88.4%) and those working in high-risk areas such as the emergency department (39.5%), respiratory wards (acute 22.3% and non-acute 23.8%) and COVID-19 isolation ward (13.8%). The average time wearing PPE per shift was 6.8 h (SD 0.39). The most commonly reported symptoms were from donning N95 masks and included: pressure injuries (45.5%), mask-induced acne (40.4%) and burning/pain (24.5%). Some 31.3% expressed that PPE-related side effects had negatively affected their work. The odds of having PPE-associated side effects was higher in women (OR 2.10, 95% CI [1.29-03.42], p = .003) and those working in high-risk wards (OR 3.12, 95% CI [2.17-4.60], p .001]. Most (90.1%) agreed that PPE supplies were readily available, sufficient for all (86.1%) and there was sufficient training in correct PPE use (93.6%). Only 13.7% of participants reported being 'highly confident' of overall PPE protection.Prevention and management of PPE-related adverse effects is vital to: preserve the integrity of PPE, improve adherence and minimize viral transmission.The high incidence of PPE-associated pressure injuries and perception that PPE use can interfere with clinical care should inform future development of PPE products, and strategies to better equip health care workers to prevent and manage PPE-related side effects.