Although Negative Pressure Wound Therapy (NPWT) has been increasingly used in wound care to improve impaired healing, there is little scientific evidence supporting its role and underlying biomolecular mechanisms. Aims of the present study are to provide a quantitative analysis of recent literature investigating NPWT in diabetic wound healing focusing on healing duration, wound closure, hospitalisation period and complications, and qualitative insight into studies analysing biomolecular mechanisms. The systematic review and meta-analysis were conducted following PRISMA guidelines (PROSPERO: CRD42024524813). 21 studies published in PubMed, Cochrane Library, EMBASE between 2019 and 2024 were included. Clinical studies indicated NPWT was superior to standard care dressings (SCD), promoting faster wound healing with significantly reduced hospitalisation times by 7.8 days (95% CI: −14.2 to −1.4, p = 0.017), and significantly reduced complications rates, particularly major and minor amputations (95% CI: −10.2 to −1.3, p = 0.01). Mechanistic in vitro and animal studies highlighted NPWT can reduce local inflammation, oxidative stress, support angiogenesis and improve scarring, essential components of normal healing. Although studies suggest NPWT is more effective than SCD for diabetic wound healing, the paucity of studies, small cohorts and scarce outcomes consistency make defining clear conclusions challenging. There is still more evidence required to fully understand NPWT's role in the complex diabetic wound healing.