Sucralfate, lipopolysaccharide (LPS), ferric chloride, trichloracetic acid (TCA), sodium nitrite, potassium ferricyanide, dimethyl sulfoxide (DMSO), N-(1-naphthyl) ethylenediamine dihydrochloride, phosphoric acid, and sulfanilamide were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), the antibiotic mixture (penicillin-streptomycin), and fetal bovine serum (FBS) were purchased from Hyclone (South Logan, UT, USA). All chemicals not listed were grade of reagent.
Sucralfate
Manufactured by Merck Group
Sourced in United States
Sucralfate is a laboratory product manufactured by Merck Group. It is a complex of sucrose, sulfate, and aluminum hydroxide that serves as a protective coating agent. The core function of Sucralfate is to form a protective barrier on surfaces, though its specific intended uses are not provided in this factual description.
Automatically generated - may contain errors
Lab products found in correlation
Butyric acid, by Merck Group (3 mentions)
Curcumin, by Merck Group (3 mentions)
Hydron polymer, by Merck Group (2 mentions)
Poly hema, by Merck Group (2 mentions)
Trichloracetic acid, by Merck Group (1 mentions)
Sulfanilamide, by Merck Group (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Ferric chloride, by Merck Group (1 mentions)
N 1 naphthyl ethylenediamine dihydrochloride, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Potassium ferricyanide, by Merck Group (1 mentions)
Lipopolysaccharide, by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Bio plex 200, by Bio-Rad (1 mentions)
Bio plex multiplex system, by Bio-Rad (1 mentions)
Matrigel, by Corning (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Vegf c, by R&D Systems (1 mentions)
8 protocols using sucralfate
1
Reagents for Biochemical Assays
2
Corneal Angiogenesis Assay in Mice
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The corneal micropocket assay was performed as described37 (link),38 (link). Seven-week-old Akt1WT and Akt1∆SMC mice were anesthetized with chloral hydrate (450 mg/kg, i.p.). After 10 min, alcaine was dropped into the eye. A corneal micropocket was created with a modified von Graef knife and MVR knife in both eyes. A micropellet of sucralfate (Sigma-Aldrich) coated with hydron polymer (Sigma–Aldrich) containing 200 ng of VEGF was implanted into each corneal pocket. The pellet was positioned approximately 1 mm from the corneal lymbus. Seven days later, the mice were anesthetized with 1–2% inhaled isoflurane, and the eyes were captured using a digital camera. For staining, eyes were fixed with 4% paraformaldehyde for 12 h at 4 °C. The primary antibody was incubated in blocking buffer (3% BSA in PBS-Tween-20) overnight at 4 °C. The secondary antibody was diluted in blocking buffer and incubated for 2 h at room temperature. Corneas were flat-mounted using an anti-fading reagent, and images were obtained using a confocal microscope (K1-Fluo, Nanoscope Systems, Daejeon, Korea). Sprouting was quantified by measuring the VEGF-induced vessel sprouting length using ImageJ (National Institutes of Health).
3
Corneal Neovascularization Induction and Treatment
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Pellets were prepared using poly(hydroxyethyl methacrylate) (poly-HEMA; Sigma-Aldrich, St. Louis, MO, USA), sucralfate (20 ng/pellet; Sigma-Aldrich), and mouse bFGF (80 ng/pellet; MedChemExpress, Monmouth, NJ, USA). A 12% (w/v) poly-HEMA was prepared with absolute ethanol, and a 10% (w/v) sucralfate solution was prepared with PBS. For 50 bFGF pellets, 5 µL 12% (w/v) poly-HEMA, 1 µL 10% (w/v) sucralfate solution, and 4 µL 1 µg/µL bFGF solution were mixed and vortexed thoroughly. For each pellet, 0.2 µL of the mixture was dropped onto the parafilm and dried at room temperature for 1 to 2 hours to form a pellet.40
The mice were randomly assigned to either the control group or the experimental group. Six hours before CNV induction, the experimental group received 1 mg/mL NE via subconjunctival injection, and the control group was injected with an equal volume of sterile water. Corneal micropockets were created with a Von Graefe cataract knife on the cornea of the right eye, and the bFGF pellet was implanted into the corneal micropocket. Then, 1 mg/mL NE or infection water was applied as eye drops six times a day beginning on the second day after pocket formation. Eye phenotypes were recorded with a slit lamp on day 1, 3, 5, and 7 after the operation.
The mice were randomly assigned to either the control group or the experimental group. Six hours before CNV induction, the experimental group received 1 mg/mL NE via subconjunctival injection, and the control group was injected with an equal volume of sterile water. Corneal micropockets were created with a Von Graefe cataract knife on the cornea of the right eye, and the bFGF pellet was implanted into the corneal micropocket. Then, 1 mg/mL NE or infection water was applied as eye drops six times a day beginning on the second day after pocket formation. Eye phenotypes were recorded with a slit lamp on day 1, 3, 5, and 7 after the operation.
4
Uniform Preparation of Angiogenic and Lymphangiogenic Micropellets
5
Inflammatory Cytokine Profiling of Macrophages
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Human monocytes (THP-1 cell line) were seeded onto a 6-well plate with 3 × 105 cells in 2 mL medium and then stabilized for 24 h. The stabilized cells were differentiated into macrophages by treatment with 10 ng/mL phorbol 12-myristate 13-acetate (Sigma-Aldrich, MO, USA) for 24 h. The macrophages were then exposed to 10 μmol/L of Rh1, butyric acid, curcumin, and sucralfate (Sigma-Aldrich, MO, USA) and incubated for 48 h. The cell supernatant was collected and centrifuged to remove the debris. Inflammatory cytokines, including CXCL10, CCL2, CCL8, IFN-γ-, IL-4, IL-12, and IL-21, were examined using a Bio-plex multiplex system and Bio-plex 200 (Bio-Rad, CA, USA).
6
Angiogenesis Tube Formation Assay
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The μ-slides for angiogenesis were obtained from Ibidi (Ibidi USA Inc., WI, USA). Ten microliters of Matrigel (Corning, NY, USA) were added to the inner wells of slides and incubated at 37°C for 30 min to 1 h. HUVECs were irradiated with a 10 Gy dose of radiation and were then harvested, and 1 × 104 cells were resuspended in 50 μL media containing 10 μmol/L of Rh1, butyric acid, curcumin, and sucralfate (Sigma-Aldrich, MO, USA). The cells were then seeded on Matrigel-coated slides, and their presence on the gel surface was confirmed. The cells were incubated at 37°C in 5% CO2 for 3 h. Thereafter, randomly selected fields in each group were observed under an inverted microscope, and the lengths of the tubes were measured.
7
Transwell Migration Assay with Ginsenosides
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The 6.5-mm Transwell inserts with 8.0-μm pores were obtained from Corning (Corning, NY, USA) for the migration assay. The cells were exposed to a single dose of 10 Gy and stabilized for 24 h. The cells were detached and resuspended in 200 μL of opti-MEM (Gibco, NY, USA) supplemented with 10 μmol/L of ginsenosides (Rh1, Rh2, Rb1, or Ro), butyric acid, curcumin, and sucralfate (Sigma-Aldrich, MO, USA) and then seeded in the upper chamber of transwell inserts. The lower chamber of the 24-well plate was filled with a medium containing 10% fetal bovine serum (Millipore, MA, USA) as a chemoattractant. After 24 h of incubation, the cells that migrated to the bottom of the transwell membrane were stained with crystal violet solution, and the stained cells in randomly chosen fields were counted, per group, using an inverted microscope.
8
Corneal Micropocket Angiogenesis Assay
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The procedure was performed as previously reported22 (link). Corneal micropocket was created 1.0 mm apart from limbal vascular arcade using a modified von Graefe Knife and slow-release pellet was implanted into the pocket. The pellet was made of sucralfate (Sigma Aldrich, St. Louis, MO) and hydron polymer (Sigma Aldrich) containing 400 ng VEGF-C (R&D Systems, Minneapolis, MN) and left in place for 9 days.
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