Abstract: Objective  To prepare a nano medicine(MMC-ATS-@PLGA) with poly lactic-co-glycolic acid (PLGA) as the carrier, mitomycin C (MMC) loading on the PLGA, and analyze the biological safety and anti-inflammatory effect of MMC-ATS-@PLGA on inhibiting the fiber proliferation after glaucoma surgery from an in vitro experimental perspective. Design  Experimental research. Participants  Rabbit conjunctival fibroblasts (RCF). Methods  The thin film dispersion hydration ultrasound method was used to prepare nano drug MMC-ATS-@PLGA. The physical and chemical properties of MMC-ATS-@PLGA were detected. The effect of MMC-ATS-@-PLGA on the activity of rabbit conjunctival fibroblasts (RCF)was evaluated using CCK-8 and Cam PI cell viability staining. The flow cytometry, cell immunofluorescence, q-PCR, and other methods from multiple perspectives were used to compare the anti-inflammatory effects of flumiolone eye drops (FML), MMC, and MMC-ATS-@PLGA nanoparticles on inflamed RCF cells in vitro research. Main Outcome Measures  Physical and chemical properties, RCF activity, anti-inflammatory effects. Results  The average particle size and zeta potential of MMC-ATS-@PLGA are (128.78±2.54) nm and 36.49±4.25) mV, with encapsulation efficiency and drug loading rate of (78.49±2.75) % and (30.86±1.84) %, respectively. Under the condition of 33℃ (ocular surface temperature), the cumulative release of MMC-ATS-@PLGA nanoparticles can reach (76.58±2.68) % after 600 minutes. The cell morphology and flow cytometry results showed that MMC-ATS-@PLGA can effectively promote the apoptosis of inflammatory RCF cells, resulting in smaller cell morphology and a pyknotic appearance of the nucleus. Further research confirmed from the perspectives of cell immunofluorescence and Q-PCR experiments that after inflammatory RCF cells co-incubation with FML, MMC, and MMC-ATS-@PLGA for 24 hours, the expression intensity of α-SMA, CTGF, and type III collagen fiber showed a gradually decreasing trend, with the MMC-ATS-@PLGA group showing the most significant decrease , and the difference was statistically significant (all P<0.05). Conclusion  This study successfully synthesized a nanomedicine MMC-ATS-@PLGA that can inhibit the scar proliferation after glaucoma surgery. The cytotoxicity of this nano-drug was significantly reduced, and the anti-inflammatory effect in vitro was enhanced. This research provided a new treatment approach for anti-scar proliferation and improving surgical success rate after glaucoma surgery.

Key words:  Glaucoma, Mitomycin C, Poly lactic-co-glycolic acid, Nanomedicine, Anti-inflammation