Currently, only oral formulations of acetylcholine analogues (e.g. pilocarpine) are used to treat patients suffering from dry mouth pharmacologically. However, these can produce systemic cardiovascular life-threatening effects, particularly in the elderly. Therefore, a topical pilocarpine formulation to locally target and stimulate salivary glands (SG) is mandatory. To address this, we developed pilocarpine-loaded poly (lactic-co-glycolic acid) (PLGA) nanofiber mats by electrospinning. Firstly, nanofiber mats were characterised morphologically and chemically. Cumulative pilocarpine release studies (4.5h28 days) were then performed. These were followed by ex vivo SG organ treatment with pilocarpine-loaded nanofiber mats to assess biocompatibility and normal glandular homeostasis and epithelial growth. Results showed that loaded PLGA nanofiber mats with 300nm of mean diameter were homogeneously distributed, and the drug encapsulation efficiency was 81.1%. Pilocarpine release was 25.7% at initial burst and 51.8% at 28 days supporting a sustained long-term release. Lastly, pilocarpine-loaded nanofiber mats (diameter:​ 0.5-1mm) supported the homeostasis of SG organs, suggesting the potential for future in vivo use.