Journal: Nature Communications

Article Title: Local microRNA delivery targets Palladin and prevents metastatic breast cancer

doi: 10.1038/ncomms12868

Figure Lengend Snippet: ( a ) Live imaging of BALB/c mice with 4T1 (expressing mCherry) breast tumours induced in the mammary fat pad and implanted with hydrogels embedded with targeted gold NPs carrying miR-96, miR-182 or scrambled (Ctrl) miRNAs with and without cisplatin. ( b ) Ex vivo fluorescent images of breast tumours to evaluate mCherry expression and NPs (FITC) accumulation. ( c ) Primary tumour volume following treatment. Quantitative PCR (qPCR) determination of ( d ) miR-96, ( e ) miR-182, and ( f ) Palladin mRNA expression levels in mice treated with hydrogels embedded with targeted gold NPs carrying miR-96, miR-182 or scrambled (Ctrl) miRNAs with and without cisplatin. ( g ) H&E and immuno-histochemistry for Palladin, Vinculin and Ki-67 of resected tumours from treated mice. All values are presented as mean±s.e.m. ( n =5, Student's t -test, * P <0.05, ** P <0.01, *** P <0.005).

Article Snippet: Histological sections of the tumours ( n =5) were stained with haematoxylin and eosin, and for IHC analysis the tumours ( n =5) were stained with anti-Ki67 antibody (Abcam ab15580, dilution 1:200), anti-Vinculin (Sigma cat#v4139, dilution 1:50) or anti-Palladin (Proteintech, cat#10853-1-AP, dilution 1:50) primary antibodies.

Techniques: Imaging, Expressing, Ex Vivo, Real-time Polymerase Chain Reaction, Immunohistochemistry

Journal: Advanced Science

Article Title: Renal Endothelial Cell‐Targeted Extracellular Vesicles Protect the Kidney from Ischemic Injury

doi: 10.1002/advs.202204626

Figure Lengend Snippet: Endothelial cell targeting capacity of PBP‐EVs. a) H/R‐injured HUVECs (Alexa Fluor 488‐labeled phalloidin, green, actin) demonstrated enhanced internalization of PBP‐EVs (Cy5.5, yellow) in vitro. Scale bars, 100 µm, n = 3. b) Diagram of a modified Transwell assay to detect penetration of EVs or PBP‐EVs into an H/R‐injured endothelial monolayer barrier (endothelial cells, HUVECs) and infiltration into renal parenchyma cells (tubular epithelial cells, HK2). c) Gluc signals of EVs and PBP‐EVs in the upper and lower chambers of the modified Transwell system were evaluated by BLI, n = 3. d) Quantitative analysis of EVs and PBP‐EVs accumulated in the injured kidney by Gluc imaging. The average radiance of the Gluc signals was expressed as photons/s/cm 2 /steradian, n = 3. e) The renal targeting ability of PBP‐EVs was traced in real time in vivo by Gluc imaging. f) Organ distributions of EVs and PBP‐EVs in severe IRI mice were detected by Cy5.5 radiant efficiency after a single intravenous injection of 100 µg EVs or PBP‐EVs. H: heart, Lu: lung, S: spleen, K: kidney, Li: liver. g) Quantitative analysis of EVs and PBP‐EVs accumulated in the injured kidney by Cy5.5 signals at 2, 6, 12, and 24 h after injection. The radiant efficiency of Cy5.5 was expressed as [photons/s/cm 2 /steradian]/[µW cm −2 ], n = 3. h) Quantitative analysis of Cy5.5 radiances in the indicated organs of severe IRI mice at 12 h after injection. The radiant efficiency of Cy5.5 was expressed as [photons/s/cm 2 /steradian]/[µW cm −2 ], n = 3. H: heart, Lu: lung, S: spleen, K: kidney, Li: liver. i) Representative images and quantitative analysis of EVs and PBP‐EVs (Cy5.5, yellow) accumulated in injured renal tissues 12 h postinjection. Scale bar, 25 µm, n = 3. j) Representative images showing the accumulation of PBP‐EVs (Cy5.5, yellow) in endothelial cells (CD31 + , red), tubular epithelial cells (E‐cadherin + , red), fibroblasts ( α ‐SMA + , red), and macrophages (F4/80 + , red) in injured renal tissues 12 h postinjection. The bottom was a higher magnification of the boxed region. Scale bar, 25 µm. All data are expressed as the mean ± s.d. For a), c), d), and g), statistical analysis was performed using two‐way ANOVA with Tukey's multiple comparison tests. For h) and i), statistical analysis was performed using two‐tailed unpaired Student's t ‐tests. * P < 0.05. The nuclei were counterstained with DAPI (blue).

Article Snippet: For immunofluorescence staining, cryosections were incubated with primary antibodies against P‐selectin (1:100; sc8419, Santa Cruz Biotechnology), CD31 (1:200; 550 274, BD Transduction Laboratories), E‐cadherin (1:200; ab76055, Abcam), α ‐SMA (1:200; ab7817, Abcam), F4/80 (1:50; ab16911, Abcam), Ki67 (1:200; ab16667, Abcam), Kim1 (1:200; ab47635, Abcam), and Collagen IV (1:200; ab6586, Abcam), followed by incubation with secondary antibodies labeled Alexa Fluor 488, Alexa Fluor 594, or Alexa Fluor 647 (Life Technologies, Carlsbad, CA).

Techniques: Labeling, In Vitro, Modification, Transwell Assay, Imaging, In Vivo, Injection, Two Tailed Test

Journal: ACS Applied Materials & Interfaces

Article Title: Acceleration of Oral Wound Healing under Diabetes Mellitus Conditions Using Bioadhesive Hydrogel

doi: 10.1021/acsami.2c17424

Figure Lengend Snippet: Fe-TA@P(AM-AA) improves the inflammatory microenvironment of diabetic oral wounds in vivo. (A) Scheme for the creation and covering of a diabetic rat palatal mucosal defect model to assess the wound-healing potential of gels. (B) H&E staining showing the successful creation of mucosal defects of the same size in the epithelium and lamina propria (labeled with dashed arrows) on both sides of the palatal mucosa. (C) CD16-stained neutrophils in the defect regions, with higher-magnification images of the regions marked by red boxes shown on the right. The percentage of CD16-positive cells was measured. (D) CD206-stained M2 macrophages in the defect regions, with higher-magnification images of the regions marked by red boxes shown on the right. The percentage of CD206-positive cells was measured. (E) CD86-stained M1 macrophages in the defect regions, with higher-magnification images of the regions marked by red boxes shown on the right. The percentage of CD86-positive cells was measured.

Article Snippet: Subsequently, they were blocked with 5% bovine serum albumin (BSA) for 1 h, followed by incubation with primary antibodies anti-CD86 (ABclonal, A1199, 1:200) and anti-CD206 (Proteintech, 60143-1-Ig, 1:200), respectively, at 4 °C overnight.

Techniques: In Vivo, Staining, Labeling