There is an increase in expression of Matrix Metalloproteinase-9 (MMP-9) during numerous pathologic conditions, including excitotoxicity and its inhibition is considered as a potential therapeutic target. Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) is endogenous inhibitor of MMP-9 that can play an important role in neuroprotection. Here, we show that TIMP-1 and TIMP-1 loaded PLGA nanoparticles (NPs) have neuroprotective effects against Kainic Acid (KA) induced excitotoxicity in organotypic hippocampal slice cultures. Moreover, TIMP-1/TIMP-1 NPs decreases LDH release and further supporting its neuroprotective effect. For blood brain barrier (BBB) penetration the NPs were coated with polysorbate 80 (Ps80). We used rat brain capillary endothelial cell culture to study their uptake/binding, toxicity and BBB penetration. BBB penetration studies showed that in the group treated with TIMP-1 NPs coated with Ps80, 11.21% ± 1.35% of TIMP-1 was detected in lower compartment of endothelial cells. To summarize, TIMP-1 loaded NPs showed neuroprotective effects in vitro and they have BBB penetration properties.