biostudies-literatureUnknown33Ling CExtremely low actual biological effect of insoluble small molecule drugs in ischemia region is a pain point and aporia in Ischemia Stroke (IS) therapy, Although there are studies on single, double-ligands modified liposomes or biomimetic exogenous carriers for directly targeting IS so far, but they often have off-target effects due to they were swallowed, degraded directly (instability) and sabotaged without the help of endogenous cell in the systemic circulation. DSPE-PEG₃₄₀₀-PGP (PGP) and DSPE-PEG₃₄₀₀-cRGD (cRGD) were synthesized via michael addition reaction of maleimide (-Mal) with sulfhydryl (-SH), succinimidyl ester (-NHS) with active primary amine group (-NH₂) respectively. The cRGD and PGP were modified on liposomes by thin film hydration method. Optimal modified ratio of cRGD and PGP were achieved by cellular uptake of HL-60 cells and THP-1 cells <i>in vitro</i>. The precise targeting effects of cRGD/PGP-Lips were examined in a nude MCAO model by an in a vivo imaging system. Puerarin (Pue) was cleverly encapsulated using a calcium acetate gradient to construct cRGD/PGP-Pue-Lips, and its therapeutic efficiency were assessed by rat MCAO model of IS. Optimal modification ratio for both cRGD and PGP were 3 %. The cRGD/PGP-Lips had significant synergetic targeting efficiency <i>in vitro</i> and <i>in vivo</i>, and the encapsulation efficiency of Pue were greater than 80 % through calcium acetate gradient. The cRGD/PGP-Pue-Lips could effectively penetrate BBB and enhance Pue retention on the brain ischemia region <i>in vivo</i>, resulting in a nearly two-fold reduction significantly in cerebral infarction area and edema in rats. In addition, cRGD/PGP-Pue-Lips didn't cause systemic toxicity in major organ tissues. Precise dual-ligands modified nanocarrier targeting endogenous cells is highly competitive as a novel anti-stroke and perspective for treatment of IS.Materials today. Bio102077https://www.ebi.ac.uk/biostudies/studies/S-EPMC12281153biostudies-literatureNeutrophil/monocyte-targeted dual-ligands modified liposomes delivering puerarin for ischemia stroke treatment.PMC12281153Li XLing CYu CChen LLiang JWei HWang JfalseNeutrophil/monocyte-targeted dual-ligands modified liposomes delivering puerarin for ischemia stroke treatment.Extremely low actual biological effect of insoluble small molecule drugs in ischemia region is a pain point and aporia in Ischemia Stroke (IS) therapy, Although there are studies on single, double-ligands modified liposomes or biomimetic exogenous carriers for directly targeting IS so far, but they often have off-target effects due to they were swallowed, degraded directly (instability) and sabotaged without the help of endogenous cell in the systemic circulation. DSPE-PEG₃₄₀₀-PGP (PGP) and DSPE-PEG₃₄₀₀-cRGD (cRGD) were synthesized via michael addition reaction of maleimide (-Mal) with sulfhydryl (-SH), succinimidyl ester (-NHS) with active primary amine group (-NH₂) respectively. The cRGD and PGP were modified on liposomes by thin film hydration method. Optimal modified ratio of cRGD and PGP were achieved by cellular uptake of HL-60 cells and THP-1 cells <i>in vitro</i>. The precise targeting effects of cRGD/PGP-Lips were examined in a nude MCAO model by an in a vivo imaging system. Puerarin (Pue) was cleverly encapsulated using a calcium acetate gradient to construct cRGD/PGP-Pue-Lips, and its therapeutic efficiency were assessed by rat MCAO model of IS. Optimal modification ratio for both cRGD and PGP were 3 %. The cRGD/PGP-Lips had significant synergetic targeting efficiency <i>in vitro</i> and <i>in vivo</i>, and the encapsulation efficiency of Pue were greater than 80 % through calcium acetate gradient. The cRGD/PGP-Pue-Lips could effectively penetrate BBB and enhance Pue retention on the brain ischemia region <i>in vivo</i>, resulting in a nearly two-fold reduction significantly in cerebral infarction area and edema in rats. In addition, cRGD/PGP-Pue-Lips didn't cause systemic toxicity in major organ tissues. Precise dual-ligands modified nanocarrier targeting endogenous cells is highly competitive as a novel anti-stroke and perspective for treatment of IS.2025-01-01T00:00:00Z2025 Aug2026-03-27T15:14:21.272Z2025-08-23T03:09:04.791ZS-EPMC122811534069732310.1016/j.mtbio.2025.102077