Journal: Science Advances

Article Title: Engineering bacterial outer membrane vesicles as transdermal nanoplatforms for photo-TRAIL–programmed therapy against melanoma

doi: 10.1126/sciadv.aba2735

Figure Lengend Snippet: Part I: Preparation of I-P-OMVs. (i) Transformation of E. coli with pDNA-TRAIL (T– E. coli ). (ii) Isolation of OMVs from T– E. coli . (iii) Detoxification of OMVs with lysozyme. (iv) Modification of OMVs with RGP forming P-OMVs. (v) Conjugation of ICG to P-OMVs forming I-P-OMVs. Part II: Topical application of I-P-OMVs induces photo-TRAIL treatment in skin melanoma. (i) I-P-OMVs penetrate through skin and target to melanoma. (ii) NIR irritation triggers ICG to induce hyperthermia effect and secret singlet oxygen that clears primary melanoma spheroids promptly. (iii) Photothermal effect induces the deformation of OMVs that release TRAIL, followed by their binding to death receptors in melanoma cells surface, activating the apoptosis in residual melanoma cells. (iv) I-P-OMVs+NIR treatments prevents the metastatic potential of melanoma through interfering the relevant genes and proteins.

Article Snippet: The TRAIL protein in OMVs was determined by ELISA (Boster, China).

Techniques: Transformation Assay, Isolation, Modification, Conjugation Assay, Binding Assay

Journal: Science Advances

Article Title: Engineering bacterial outer membrane vesicles as transdermal nanoplatforms for photo-TRAIL–programmed therapy against melanoma

doi: 10.1126/sciadv.aba2735

Figure Lengend Snippet: ( A ) TEM images of T– E. coli with their derived OMVs. Scale bars, 2 μm and 200 nm. ( B ) Confocal laser scanning microscopy (CLSM) image of Dil-OMVs. Scale bar, 2 μm. ( C ) Size distribution of OMVs measured by DLS. ( D ) WB analysis of S3 protein (a classical protein contained within E. coli ) and TRAIL protein in bacterial cells and their derived OMVs. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( E ) Size distribution and ( F ) zeta potential (ZP) of OMVs, R-OMVs, and P-OMVs. ( G ) TEM images of OMVs, R-OMVs, and P-OMVs. Scale bars, 200 nm.

Article Snippet: The TRAIL protein in OMVs was determined by ELISA (Boster, China).

Techniques: Derivative Assay, Confocal Laser Scanning Microscopy, Zeta Potential Analyzer

Journal: Science Advances

Article Title: Engineering bacterial outer membrane vesicles as transdermal nanoplatforms for photo-TRAIL–programmed therapy against melanoma

doi: 10.1126/sciadv.aba2735

Figure Lengend Snippet: ( A ) Photothermal response of I-P-OMVs (0 to 5 μg/ml) to NIR irritation (2 W/cm 2 for 3 min) ( n = 3). ( B ) Changes in the DPBF absorbance spectra in the presence of I-P-OMVs under NIR irradiation (0, 0.6, and 2 W/cm 2 ). ( C ) DPBF absorbance in the presence of I-P-OMVs at 412 nm under NIR irradiation (0, 0.6, and 2 W/cm 2 ) ( n = 3). ( D ) TRAIL release profiles at 37°C without or with NIR irradiation (2 W/cm 2 ) ( n = 3). ( E ) CLSM images of 3D tumor spheroids incubated with Dil-OMVs, Dil-R-OMVs, and Dil-P-OMVs. ( F and G ) CM-Dil fluorescence intensity in different depths of 3D tumor spheroids incubated with Dil-OMVs (F), Dil-R-OMVs (F), and Dil-P-OMVs (G). All data are represented as means ± SD. *** P < 0.001 and ** P < 0.01.

Article Snippet: The TRAIL protein in OMVs was determined by ELISA (Boster, China).

Techniques: Irradiation, Incubation, Fluorescence

Journal: Science Advances

Article Title: Engineering bacterial outer membrane vesicles as transdermal nanoplatforms for photo-TRAIL–programmed therapy against melanoma

doi: 10.1126/sciadv.aba2735

Figure Lengend Snippet: ( A ) The distribution of OMVs-GFP, R-OMVs-GFP, and P-OMVs-GFP in skin slice. Scale bars, 100 nm. ( B ) The accumulative transdermal amounts of TRAIL protein ( n = 3). ( C ) Scanning electron microscopy (SEM) (c 1 ) and TEM (c 2 , c 3 , and c 4 ) images of skin tissues after topical application of DiI-P-OMVs. Scale bars, 1 μm. DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate)–P-OMVs were indicated by red arrows. ( D ) Schematic illustration of the distribution of OMVs in skin and their transdermal routes. ( E ) In vivo fluorescence imaging of tumor-bearing mice after topically applied with Dil-P-OMVs. ( F ) Fluorescent images of tumors and major organs ( G ) Therapeutic regimen of I-P-OMVs+NIR in mice with B16F10 melanoma. ( H to J ) The tumors size, relapse rates, and body weights in mice during tested periods. ( K ) The TUNEL (terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling), cleaved caspase 3, Fontana-Masson (12 hours, day 6, and day 12), S100-β (day 12), and CD63 (day 12) staining of tumors. ( L to R ) Quantification of (K)-positive cells in the tumors ( n = 3). Scale bars, 100 nm. I to V represent the ICG+NIR, I-TRAIL+NIR, I-OMVs+NIR, I-R-OMVs+NIR, and I-P-OMVs+NIR groups, respectively. All data are represented as means ± SD. *** P < 0.001, ** P < 0.01, and * P < 0.05. n.s., not significant.

Article Snippet: The TRAIL protein in OMVs was determined by ELISA (Boster, China).

Techniques: Electron Microscopy, In Vivo, Fluorescence, Imaging, TUNEL Assay, End Labeling, Staining