To investigate the efficacy of eyelid margin cleansing with lid hygiene detergent in patients with obstructive meibomian gland dysfunction (MGD).As a pilot study, we investigated the safety and efficacy of lid hygiene using tap water and/or lid hygiene shampoo in fourteen eyes of 7 normal subjects. All subjects were instructed to cleanse the eyelid margin with tap water and/or lid hygiene shampoo. As a main prospective clinical study, thirty-four eyes of 34 patients with obstructive MGD were enrolled. All subjects were instructed to cleanse the eyelid margin with lid hygiene shampoo at least once daily for 1 month. Ocular surface conditions were observed before and 1 month after study initiation.A significant exacerbating change (p < 0.05) was not detected after either method in the pilot study. In the main study, significant improvements were observed in tear break-up time (TBUT), lid margin lissamine green staining scores, vascular dilatation, and meibum status (p < 0.05). No significant improvements in corneal or conjunctival fluorescein staining scores, the mucocutaneous junction, lid margin deformation, or plugging were observed (p ≥ 0.05). Subjective improvements were observed in 27 subjects. Meibography revealed that 28 subjects had normal meibomian glands, and 6 subjects had meibomian gland atrophy. Significant improvements were observed in TBUT, vascular dilatation, and meibum status only in the group with normal meibomian glands (p < 0.05), but subjective symptoms and lid margin lissamine green staining scores improved in both groups (p < 0.05). No subjects experienced any problems throughout the study.Daily routine use of lid hygiene detergent can alleviate the symptoms of MGD regardless of meibomian gland atrophy.