Mesenchymal Stem Cell Membrane‐Camouflaged Liposomes for Biomimetic Delivery of Cyclosporine A for Hepatic Ischemia‐Reperfusion Injury Prevention

Abstract Hepatic ischemia‐reperfusion injury (HIRI) is a prevalent issue during liver resection and transplantation, with currently no cure or FDA‐approved therapy. A promising drug, Cyclosporin A (CsA), ameliorates HIRI by maintaining mitochondrial homeostasis but has systemic side effects due to its low bioavailability and high dosage requirements. This study introduces a biomimetic CsA delivery system that directly targets hepatic lesions using mesenchymal stem cell (MSC) membrane‐camouflaged liposomes. These hybrid nanovesicles (NVs), leveraging MSC‐derived proteins, demonstrate efficient inflammatory chemotaxis, transendothelial migration, and drug‐loading capacity. In a HIRI mouse model, the biomimetic NVs accumulated at liver injury sites entered hepatocytes, and significantly reduced liver damage and restore function using only one‐tenth of the CsA dose typically required. Proteomic analysis verifies the protection mechanism, which includes reactive oxygen species inhibition, preservation of mitochondrial integrity, and reduced cellular apoptosis, suggesting potential for this biomimetic strategy in HIRI intervention.


Figure S1 .
Figure S1.Inverted fluorescence microscopic images of (a) MSCs after freeze-thaw, (b) pellets of cell nuclei after centrifugation at 700 g for 10 min, and (c) pellets of MSC membranes after centrifugation at 15000 g for 30 min.Cell nuclei were pre-stained by DAPI.Scale bar: 100 µm.

Figure S2 .
Figure S2.Tyndall effect of CLs and MMCLs after 30 days of storage at 4 o C in PBS.

Figure S3 .
Figure S3.Colocalizations of CLs and MMs at phospholipid-to-membrane protein ratios of 1:0.05 and 1:0.025 after incubation with AML12 cells for 2 h and observed by CLSM.Scale bar: 50 µm.

Figure S4 .Figure S5 .
Figure S4.Functional enrichment analysis of the differently expressed proteins in MMs.

Figure S7 .
Figure S7.(a) In vivo fluorescence imaging of the supernatant and pellet of DiR-labeled MMCLs (MMCLs-DiR) in a HIRI mouse model at 6 h after injection.(b) Ex vivo tissue distribution of the supernatant of MMCLs-DiR in the main organs.Excitation wavelength: 750 nm; emission wavelength: 780 nm.

Figure S8 .
Figure S8.Immunofluorescence images of liver sections at 2 h after intravenous administration

Figure S9 .
Figure S9.In vivo fluorescence imaging of DiR-labeled MMCLs or CLs in a HIRI mouse model at 2 h after reperfusion.Red arrows: spleen.Excitation wavelength: 750 nm; emission wavelength: 780 nm.

Figure S10 .
Figure S10.Evaluation of the protection effect of MMs on H/R-injured AML12 cells in vitro.(a) Cell viability of H/R-injured cells after treated with MMs equivalent to protein dose of MMCLs at CsA concentrations of 0.1, 1, and 10 ng mL −1 as measured by CCK-8 assay (n = 5).(b) Flow cytometry analysis of cell apoptosis of control, H/R-injured, and MMs-treated (equivalent to MMCLs at CsA concentrations of 0.1 ng mL −1 ) groups by annexin V-FITC/PI

Figure S11 .
Figure S11.Evaluation of liver functions of AST, ALT, and LDH after administration of (a) CsA or (b) MMCLs at different CsA dosage in HIRI mouse model.Data are presented as mean ± SD.The statistical significance was analyzed using one-way ANOVA following Tukey's multiple comparisons test (n = 4, *p < 0.05; **p < 0.01; ***p < 0.001, ns = no significance).

Figure S12 .
Figure S12.Evaluation of the protective effect of MMs.(a) Liver functions of AST, ALT, and LDH and (b) H&E staining of liver sections after administration with MMs at a protein dosage equivalent to 0.1mg kg −1 of MMCLs.(c) Suzuki scores of the H&E staining images.Data are presented as mean ± SD.The statistical significance was analyzed using one-way ANOVA following Tukey's multiple comparisons test (n = 3, *p < 0.05; **p < 0.01; ***p < 0.001, ns = no significance).

Figure S13 .
Figure S13.H&E staining of major organs including the liver, kidney, heart, and lung of healthy mice after treatment with PBS, CsA or CLs at CsA dose of 1 mg kg −1 , or MMCLs at CsA dose of 0.1 mg kg −1 .

Figure S15 .
Figure S15.Volcano diagram showing the differently expressed proteins between the Sham

Figure S16 .
Figure S16.Functional enrichment analysis of the differently expressed proteins (a)