Diagnostic criteria for provoked vestibulodynia (PVD) rely on mucosal pain in the vulvar vestibule, with less emphasis on pain from pelvic floor muscles. It is unknown how psychosocial variables associated with PVD are differentially associated with mucosal versus muscle pain. Analysis of data from the National Vulvodynia Registry (n=202) revealed several factors associated with increased mucosal pain: pain duration (p=0.043), the McGill sensory subscore (p=0.0086) and the Gracely pain scale (p<0.001). Increased mucosal pain was also associated with decreased arousal (p=0.036). On the other hand, factors significantly associated with greater muscle pain included number of comorbid pain conditions (p=0.001), decreased intercourse frequency post PVD onset (p=0.02) and higher scores on the McGill sensory (p=0.0001) and affective (p=0.0002) subscores, the Gracely pain scale (p=0.0012), and state anxiety (p<0.001). Sexual function was also significantly impacted by high pelvic floor muscular pain, with lower scores for arousal (p=0.046), orgasm (p=0.0014) and satisfaction (p=0.013), and higher pain (p=0.01). Significant differences in the relationship between muscle and mucosal pain for pain duration (p= 0.005), McGill affective score (p=0.001), orgasm (p=0.049), change in intercourse frequency (p=0.027), and state anxiety (p=0.030) suggest the possibility of mucosal or muscle pain predominant PVD subtypes. Perspective: Patients with higher pelvic floor muscle pain scores than mucosal pain scores may represent different subgroups or characteristics of patients with provoked vestibulodynia. This research highlights the importance of assessment of the pelvic floor muscles in addition to the cotton swab test of the vestibule.Copyright © 2020. Published by Elsevier Inc.