Journal: International Journal of Biological Sciences

Article Title: ANXA1-mediated mTOR/FABP4 Inhibition Drives Antifibrotic Macrophage Reprogramming in Lupus Nephritis

doi: 10.7150/ijbs.118613

Figure Lengend Snippet: Ac2-26 administration treats established nephritis in MRL/ lpr mice. (A) Ac2-26 treatment protocol for MRL/ lpr mice. Ac2-26 (2 mg/kg) or PBS treatments were administered i.p. every other day from weeks 10 to 20. n = 6 per group. (B) Levels of anti-dsDNA antibodies in different groups at 10 weeks and 20 weeks (n = 6). (C) Serum creatinine levels in different groups at 10 weeks and 20 weeks (n = 6). (D) Urinary protein/creatinine ratios in different groups at 10 weeks and 20 weeks (n = 6). (E) Representative sections of renal tissue stained with H&E, Masson's trichrome, PAS, and PASM staining and semi-quantitative evaluation of glomerulonephritis at 20 weeks. Bars = 50 μm. (F) Representative photomicrographs and quantitative analysis of Sirius Red staining at 20 weeks. n = 5 per group. Bars = 50 μm. (G) Immunohistochemical staining and quantitative analysis of F4/80 at 20 weeks. n = 5 per group. Bars = 50 μm. (H) Immunohistochemical staining and quantitative analysis of osteopontin (encoded by Spp1 ) at 20 weeks. n = 5 per group. Bars = 50 μm. (I) Representative photomicrographs and quantitative analysis of Oil Red O staining at 20 weeks. n = 5 per group. Bars = 50 μm. (J) Immunohistochemical staining and quantitative analysis of Fabp4 at 20 weeks. n = 5 per group. Bars = 50 μm. Data analyses were performed by Student's t -test for two groups. * P < 0.05; ** P < 0.01; *** P < 0.001. i.p.: intraperitoneally; HE: hematoxylin-eosin; PAS: periodic acid-Schiff; PASM: periodic acid-silver methenamine.

Article Snippet: The Ac2-26 peptide was synthesized by MedChemExpress LLC (China), and the purity of Ac2-26 was 99.14%.

Techniques: Staining, Immunohistochemical staining

Journal: bioRxiv

Article Title: Orally delivered biodegradable targeted inflammation resolving pectin-coated nanoparticles induce anastomotic healing post intestinal surgery

doi: 10.1101/2024.01.01.569918

Figure Lengend Snippet: The pectin coating of the NPs protects from the harsh pH environment of the stomach, small intestine and enzymes secreted for digestion. Once in the colon, pectin and the subsequent chitosan layers are fermented and degraded almost completely by the microflora in the gut and this leads to the local release of the Col-IV Ac2-26 NPs, which binds to wounds and releases the Ac2-26 peptide in situ .

Article Snippet: The Collagen IV targeting peptide (KLWVLPKGGGC) and the scrambled Ac2-26 peptide (WLKQKFQESVEQIAYVMENATEFEV) were purchased from Mimotopes.

Techniques: In Situ

Journal: bioRxiv

Article Title: Orally delivered biodegradable targeted inflammation resolving pectin-coated nanoparticles induce anastomotic healing post intestinal surgery

doi: 10.1101/2024.01.01.569918

Figure Lengend Snippet: Collagen IV-targeted (Col-IV) NPs encapsulating the Ac2-26 peptide (Ac2-26-NP) or controls including a randomly generated, isoelectric mismatched scrambled sequence (Scrm-NP), were developed using biodegradable polymers via a single-step nanoprecipitation method, C: chitosan, P: pectin. (A) Dynamic light-scattering of empty (NP), Ac2-26 loaded (Ac2-26-NP) and scrambled peptide (Scrm-NP) NPs were measured (mean ± SD, n = 3). (B) ζ-Potential of the NPs indicating surface charge. (C) In vitro cumulative release curve of Ac2-26 or scrambled peptide from NPs incubated at 37 °C is shown (mean ± SD, n = 3). (D) Representative TEM image of Ac2-26-NPs (scale bar = 500 nm)

Article Snippet: The Collagen IV targeting peptide (KLWVLPKGGGC) and the scrambled Ac2-26 peptide (WLKQKFQESVEQIAYVMENATEFEV) were purchased from Mimotopes.

Techniques: Generated, Sequencing, In Vitro, Incubation

Journal: bioRxiv

Article Title: Orally delivered biodegradable targeted inflammation resolving pectin-coated nanoparticles induce anastomotic healing post intestinal surgery

doi: 10.1101/2024.01.01.569918

Figure Lengend Snippet: (A) Timeline of the experimental procedure. Mice were treated by 2 % DSS in drinking water. After 7 days DSS was changed to normal drinking water and microsurgical colonic anastomosis was performed. Mice were treated every 3.5 days with oral gavage of pectin-coated P-C-Col IV-Ac2-26 NPs in different doses (1µg Ac2-26 or 10µg Ac2-26) or pectin-coated Scrambled-nanoparticles (P-C-Col IV-Scrm NPs at a dose of 10µg of Scrm peptide) throughout the whole procedure. (B) Body weight is shown as percentage of starting weight every day during the whole procedure. (C) Disease activity index (DAI) at 3 different time points is shown: immediately preoperatively (preOP), at postoperative day 3 (POD3) and at postoperative day 7 (POD7). Statistical differences determined by Mann-Whitney-U test for nonparametric results are indicated with *(P<0.05). N=23 (P-C-Col IV-Scrm NPs), N=21 (P-C-Col IV-Ac2-26 NPs 1µg), N=22 (P-C-Col IV-Ac2-26 NPs 10µg). Three animals were excluded from analyses for reaching abort criteria (1 per each treatment group), 3 animals were excluded for intraoperative technical problems (2 in P-C-Col IV-Ac2-26 NPs 1 µg, 1 in P-C-Col IV-Ac2-26 NPs 10µg).

Article Snippet: The Collagen IV targeting peptide (KLWVLPKGGGC) and the scrambled Ac2-26 peptide (WLKQKFQESVEQIAYVMENATEFEV) were purchased from Mimotopes.

Techniques: Activity Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Orally delivered biodegradable targeted inflammation resolving pectin-coated nanoparticles induce anastomotic healing post intestinal surgery

doi: 10.1101/2024.01.01.569918

Figure Lengend Snippet: (A) Rigid colonic endoscopy was performed to analyze luminal macroscopic wound closure. Representative anastomoses at postoperative day 7 are shown. Dehiscent areas are marked in red. (B) Endoscopic deshiscence score evaluates local wound healing from a luminal perspective. Anastomotic healing was scored as described before : no dehiscence = 0; sutures protruding into the lumen = 1; dehiscence less than 25% of circumference = 2; dehiscence more than 25% of circumference =3; complete dehiscence = 4. (C) Anastomotic leakage leads to adhesion formation with the adjacent tissue. Therefore grade of anastomotic leakage could be measured as follows: adhesion of mesenteric fat tissue, uterus, intestine or other organs was awarded with 1 point each. Abundance of adhesions was scored according to the following score and added: no adhesion = 0; completely bluntly removable = 1; partially bluntly removable = 2; not bluntly removable =3. Statistical differences determined by Mann Whitney U test for nonparametric results are indicated with *(P<0.05), **(P<0.01), ***(P<0.001). N=23 (P-C-Col IV-Scrm NPs), N=21 (P-C-Col IV-Ac2-26 NPs 1µg), N=22 (P-C-Col IV-Ac2-26 NPs 10µg). Three animals were excluded from analyses for reaching abort criteria (1 per each treatment group), 3 animals were excluded for intraoperative technical problems (2 in P-C-Col IV-Ac2-26 NPs 1 µg, 1 in P-C-Col IV-Ac2-26 NPs 10µg), while additionally 3 were removed from analysis in P-C-Col IV-Ac2-26 NPs 10µg at POD 3 for technical endoscopy problems.

Article Snippet: The Collagen IV targeting peptide (KLWVLPKGGGC) and the scrambled Ac2-26 peptide (WLKQKFQESVEQIAYVMENATEFEV) were purchased from Mimotopes.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Orally delivered biodegradable targeted inflammation resolving pectin-coated nanoparticles induce anastomotic healing post intestinal surgery

doi: 10.1101/2024.01.01.569918

Figure Lengend Snippet: (A) HE-stained longitudinal sections of the anastomosis were analyzed by an experienced pathologist according to the established Phillips-Score. A score of 0 (not detectable) – 4 (highly detectable) was awarded to the following items and summed up: blood vessel ingrowth, fibroblast ingrowth, collagen formation and absence of inflammatory cells. (B) Microscopical wound closure was scored by adding additional items to the Phillips-score, focusing on closure of the individual intestinal layers . (C) Representative HE-stained sections of the different groups are shown. Statistical differences determined by Mann Whitney U test for nonparametric results are indicated with *(P<0.05), **(P<0.01). N=23 (P-C-Col IV-Scrm NPs), N=21 (P-C-Col IV-Ac2-26 NPs 1µg), N=22 (P-C-Col IV-Ac2-26 NPs 10µg). Three animals were excluded from analyses for reaching abort criteria (1 per each treatment group), 3 animals were excluded for intraoperative technical problems (2 in P-C-Col IV-Ac2-26 NPs 1 µg, 1 in P-C-Col IV-Ac2-26 NPs 10µg).

Article Snippet: The Collagen IV targeting peptide (KLWVLPKGGGC) and the scrambled Ac2-26 peptide (WLKQKFQESVEQIAYVMENATEFEV) were purchased from Mimotopes.

Techniques: Staining, MANN-WHITNEY