Abstract Objective: To assess the impact of optimized pulsed technology (OPT) on the morphological and functional changes of meibomian glands in patients with meibomian gland dysfunction (MGD). Methods: This prospective case-control study enrolled 60 MGD patients (60 eyes, right eye) treated at Weifang Eye Hospital from September 2023 to February 2024. Patients were categorized into mild, moderate, and severe groups based on the extent of meibomian gland loss, with 20 individuals per group. Treatments consisted of bilateral OPT combined with meibomian gland massages, administered biweekly over four sessions. We monitored ocular surface function indicators including the ocular surface disease index (OSDI), corneal fluorescein staining (CFS), non-invasive average tear break-up time (NIBUTav), and non-invasive tear meniscus height (NITMH), as well as meibomian gland function scores (MGES and MGYSS). Cellular-level assessments using in vivo confocal microscopy (IVCM) examined meibomian gland acinar unit density (MGAUD), inflammatory cell density (ICD), and acinar dimensions (MGALD and MGASD). Results: At baseline, no significant differences were found in NITMH across groups (P>0.05). Significant disparities were observed in NIBUTav, MGES, MGYSS, MGAUD, MGALD, and MGASD (P<0.05). Compared to the mild group, the moderate and severe groups showed significant differences in OSDI, CFS, and ICD (P<0.05), though no significant differences existed between moderate and severe groups (P>0.05). After three months of treatment, all groups showed no significant differences in NITMH (P>0.05). All parameters improved significantly in the mild group (P<0.05); all but MGASD improved in the moderate group (P>0.05 for MGASD); significant improvements were noted in OSDI, CFS, and NIBUTav in the severe group (P<0.05), while MGES and MGYSS did not differ significantly (P>0.05). IVCM parameters (MGAUD, ICD, MGALD, and MGASD) showed no significant change in the severe group post-treatment (P>0.05). Conclusion: OPT effectively enhances various ocular surface functions and improves gland expressibility and secretion quality in mild to moderate MGD cases, while also positively impacting certain cellular parameters. In severe cases, where most acinar functions are lost and structural reversibility is limited, OPT can still mitigate MGD symptoms and decelerate disease progression.