Beliefs regarding pain drive pain experiences, including attitudes and beliefs regarding treatment options. The purpose of this study was to determine if a brief, one-time pain neuroscience education (PNE) lecture to middle school students can alter their beliefs regarding pain and pain medication. Two hundred and sixty-three middle school students attended a 30-minute PNE lecture. Prior to and immediately following the lecture measurements regarding pain beliefs and pain medication were completed. Pain beliefs were measured with an adapted Health Care Provider’s Pain and Impairment Relationship Scale (HC-PAIRS) and pain medication beliefs were measured using the Beliefs about Medicines Questionnaire (BMQ). Immediately following the lecture, all pain beliefs shifted positively, with two reaching significance - “You can control how much pain you feel” (p < 0.001) and “Your brain decides if you feel pain, not your tissues” (p < 0.001). Three pain medication beliefs shifted post-PNE, with students agreeing more with “People who take pain medicines should stop their treatment for a while every now and again” (p = 0.002) and “All pain medicines are poisons” (p = 0.027), whereas they disagreed more with “Most pain medicines are addictive” (p = 0.015). This study concluded that a one-time PNE lecture to middle school students positively influences student beliefs regarding pain, as well as some shifts in their beliefs about the use of pain medication. This is the first study to explore if a lecture on pain can alter beliefs in middle school children regarding pain medication. More research is needed to determine if these changes remain intact over time and can be replicated in other student populations.