4c8711fa98819d1555d80cedb1b9fb80 biomedicines_a2022m2v10a465.pdf 0d1c205c6c20b0f9d32460ab1641eb833d696c93 biomedicines_a2022m2v10a465.pdf c93f5771871e94da568a177185c57a2d7eb6d19753d5b5995b01f1054695907c biomedicines_a2022m2v10a465.pdf Title: Minimum Effective Dose of DPP-4 Inhibitors for Treating Early Stages of Diabetic Retinopathy in an Experimental Model Subject: The neurovascular unit (NVU) plays an essential role in the development of diabetic retinopathy (DR). We previously reported that the topical administration (eye drops) of sitagliptin and saxagliptin, two dipeptidyl peptidase-4 inhibitors (DPP-4i), prevents retinal neurodegeneration and vascular leakage in db/db mice. The aim of the present study is to evaluate the minimum effective dose of the topical administration of these DPP-4i. For this purpose, sitagliptin and saxagliptin were tested at different concentrations (sitagliptin: 1 mg/mL, 5 and 10 mg/mL, twice per day; saxagliptin: 1 and 10 mg/mL, once or twice per day) in db/db mice. As end points of efficacy, the hallmarks of NVU impairment were evaluated: reactive gliosis, neural apoptosis, and vascular leakage. These parameters were assessed by immunohistochemistry, cell counting, and the Evans blue method, respectively. Our results demonstrated that the minimum effective dose is 5 mg/mL twice per day for sitagliptin, and 10 mg/mL twice per day for saxagliptin. In conclusion, this study provides useful results for the design of future preclinical regulatory studies and for planning clinical trials. Keywords: diabetic retinopathy; dipeptidyl peptidase-4 inhibitors; sitagliptin; saxagliptin; neurovascular unit; retinal neurodegeneration; experimental diabetes; db/db mice Author: Patricia Bogdanov, Hugo Ramos, Marta Valeri, Anna Deàs-Just, Jordi Huerta, Rafael Simó and Cristina Hernández Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Feb 16 13:41:06 2022 CET ModDate: Wed Feb 16 13:48:05 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3862723 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NEBEKD+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 GFKJOY+VnURWPalladioL Type 1 Custom yes yes no 15 0 YESBEV+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 GABUCT+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 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HCIJKC+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 253 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-04-28 02:27:39 CEST RepresentationInformation: biomedicines_a2022m2v10a465.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-04-26 17:58:48 CEST Size: 3862723 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 369 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Experimental Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Interventional Study Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Retinal Tissue Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Retinal Vascular Permeability by Evans Blue Ex Vivo Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Glial Fibrillary Acidic Protein (GFAP) and DPP-4 Immunofluorescence Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Measurements of Glial Activation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Histological Assessment with Hematoxylin-Eosin (H&E) Staining Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Cell Counting in GCL and INL Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Analysis of Retinal Thickness Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Children: Item: Title: Dose Response Effect of Topical Administration of Sitagliptin and Saxagliptin on DPP-4 Content Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Dose Response Effect of Topical Administration of Sitagliptin and Saxagliptin on Glial Activation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Dose Response Effect of Sitagliptin and Saxagliptin Eye Drops on Cell Death and Retinal Thinning Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Dose Response Effect of Topical Administration of Sitagliptin and Saxagliptin on Vascular Leackage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Info: Title: Minimum Effective Dose of DPP-4 Inhibitors for Treating Early Stages of Diabetic Retinopathy in an Experimental Model Author: Patricia Bogdanov, Hugo Ramos, Marta Valeri, Anna Deàs-Just, Jordi Huerta, Rafael Simó and Cristina Hernández Subject: The neurovascular unit (NVU) plays an essential role in the development of diabetic retinopathy (DR). We previously reported that the topical administration (eye drops) of sitagliptin and saxagliptin, two dipeptidyl peptidase-4 inhibitors (DPP-4i), prevents retinal neurodegeneration and vascular leakage in db/db mice. The aim of the present study is to evaluate the minimum effective dose of the topical administration of these DPP-4i. For this purpose, sitagliptin and saxagliptin were tested at different concentrations (sitagliptin: 1 mg/mL, 5 and 10 mg/mL, twice per day; saxagliptin: 1 and 10 mg/mL, once or twice per day) in db/db mice. As end points of efficacy, the hallmarks of NVU impairment were evaluated: reactive gliosis, neural apoptosis, and vascular leakage. These parameters were assessed by immunohistochemistry, cell counting, and the Evans blue method, respectively. Our results demonstrated that the minimum effective dose is 5 mg/mL twice per day for sitagliptin, and 10 mg/mL twice per day for saxagliptin. In conclusion, this study provides useful results for the design of future preclinical regulatory studies and for planning clinical trials. 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