Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells
Chitosan and heparin-based hydrogels possess potent properties such as air permeability, absorption and water retention. Our group has previously reported heparin-loaded, freeze-dried porous hydrogels, using chitosan/PVA and variable concentrations of PCL. Significant angiogenic effect of these hydr...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-09-01
|
| Series: | Journal of King Saud University: Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1018364721001956 |
| _version_ | 1818715796013056000 |
|---|---|
| author | Sadia Fazil Humaira Shah Mamoona Noreen Muhammad Yar Ather Farooq Khan Sher Zaman Safi Suliman Yousef Alomar Afrah Fahad Alkhuriji Hanan Mualla Alharbi Muhammad Sohail Afzal |
| author_facet | Sadia Fazil Humaira Shah Mamoona Noreen Muhammad Yar Ather Farooq Khan Sher Zaman Safi Suliman Yousef Alomar Afrah Fahad Alkhuriji Hanan Mualla Alharbi Muhammad Sohail Afzal |
| author_sort | Sadia Fazil |
| collection | DOAJ |
| description | Chitosan and heparin-based hydrogels possess potent properties such as air permeability, absorption and water retention. Our group has previously reported heparin-loaded, freeze-dried porous hydrogels, using chitosan/PVA and variable concentrations of PCL. Significant angiogenic effect of these hydrogels was observed. In this follow-up study, we have investigated the underlying molecular mechanisms of increased angiogenesis due to these heparin-releasing chitosan-based hydrogels, in human umbilical vein endothelial cells (HUVECs). Cells were cultured in complete ECM media and supplemented with 10% FBS and 1% Penicillin-Streptomycin. HUVECs were treated with heparin-releasing chitosan-based hydrogels in different concentrations (H4, H8 and H16) for 1, 3 and 7 days. RNA was extracted using trizol and quantified by nanodrop. Primers were designed for the target genes including VEGF, VEGFR, Ap-1, BCL-2, HIF-1, BAX, IL-1 and TNF-α. Gene expression analysis were performed using PCR. Gel electrophoresis was used to visualize results. MTT and DCFH-DA assays were performed to check cytotoxicity and reactive oxygen species (ROS) respectively. Hydrogels H8, had significantly high effect on the expression of VEGF and VEGFR, as compared to H4 and H16. At day 1, most of the target genes showed higher expression. HIF-1 showed high expression at day one, with a gradual decrease at day 3 and 4. We observed no expression of AP-1 with hydrogels H4, H16 and control, but a high expression with hydrogels H8, at day 1 and 3. BCL-2, IL-1and TNF-α also revealed high expression with hydrogels H8. A low expression of BAX was observed in all cases. Our study demonstrates high expression of genes with H4 probably due to high concentration of heparin. Similarly, at day 1, the high expression is attributed to the fact that these hydrogels release maximum heparin in the first 24 h. |
| first_indexed | 2024-12-17T19:09:03Z |
| format | Article |
| id | doaj.art-f790bea6114c47ce99e447d58cd29e2c |
| institution | Directory Open Access Journal |
| issn | 1018-3647 |
| language | English |
| last_indexed | 2024-12-17T19:09:03Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of King Saud University: Science |
| spelling | doaj.art-f790bea6114c47ce99e447d58cd29e2c2022-12-21T21:35:54ZengElsevierJournal of King Saud University: Science1018-36472021-09-01336101534Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cellsSadia Fazil0Humaira Shah1Mamoona Noreen2Muhammad Yar3Ather Farooq Khan4Sher Zaman Safi5Suliman Yousef Alomar6Afrah Fahad Alkhuriji7Hanan Mualla Alharbi8Muhammad Sohail Afzal9School of Science, University of Management and Technology, Lahore, PakistanSchool of Science, University of Management and Technology, Lahore, PakistanDepartment of Zoology, Women University Multan, PakistanInterdisciplinary Research Center in Biomedical Materials (IRCBM), COMSATS University Islamabad, Lahore Campus, PakistanInterdisciplinary Research Center in Biomedical Materials (IRCBM), COMSATS University Islamabad, Lahore Campus, PakistanInterdisciplinary Research Center in Biomedical Materials (IRCBM), COMSATS University Islamabad, Lahore Campus, Pakistan; Corresponding author.Department of Zoology, College of Science, King Saud University, Riyadh, Saudi ArabiaDepartment of Zoology, College of Science, King Saud University, Riyadh, Saudi ArabiaDepartment of Zoology, College of Science, King Saud University, Riyadh, Saudi ArabiaSchool of Science, University of Management and Technology, Lahore, PakistanChitosan and heparin-based hydrogels possess potent properties such as air permeability, absorption and water retention. Our group has previously reported heparin-loaded, freeze-dried porous hydrogels, using chitosan/PVA and variable concentrations of PCL. Significant angiogenic effect of these hydrogels was observed. In this follow-up study, we have investigated the underlying molecular mechanisms of increased angiogenesis due to these heparin-releasing chitosan-based hydrogels, in human umbilical vein endothelial cells (HUVECs). Cells were cultured in complete ECM media and supplemented with 10% FBS and 1% Penicillin-Streptomycin. HUVECs were treated with heparin-releasing chitosan-based hydrogels in different concentrations (H4, H8 and H16) for 1, 3 and 7 days. RNA was extracted using trizol and quantified by nanodrop. Primers were designed for the target genes including VEGF, VEGFR, Ap-1, BCL-2, HIF-1, BAX, IL-1 and TNF-α. Gene expression analysis were performed using PCR. Gel electrophoresis was used to visualize results. MTT and DCFH-DA assays were performed to check cytotoxicity and reactive oxygen species (ROS) respectively. Hydrogels H8, had significantly high effect on the expression of VEGF and VEGFR, as compared to H4 and H16. At day 1, most of the target genes showed higher expression. HIF-1 showed high expression at day one, with a gradual decrease at day 3 and 4. We observed no expression of AP-1 with hydrogels H4, H16 and control, but a high expression with hydrogels H8, at day 1 and 3. BCL-2, IL-1and TNF-α also revealed high expression with hydrogels H8. A low expression of BAX was observed in all cases. Our study demonstrates high expression of genes with H4 probably due to high concentration of heparin. Similarly, at day 1, the high expression is attributed to the fact that these hydrogels release maximum heparin in the first 24 h.http://www.sciencedirect.com/science/article/pii/S1018364721001956Molecular mechanismsCell biologyBiomaterialsChitosanHeparinAngiogenesis |
| spellingShingle | Sadia Fazil Humaira Shah Mamoona Noreen Muhammad Yar Ather Farooq Khan Sher Zaman Safi Suliman Yousef Alomar Afrah Fahad Alkhuriji Hanan Mualla Alharbi Muhammad Sohail Afzal Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells Journal of King Saud University: Science Molecular mechanisms Cell biology Biomaterials Chitosan Heparin Angiogenesis |
| title | Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells |
| title_full | Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells |
| title_fullStr | Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells |
| title_full_unstemmed | Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells |
| title_short | Evaluation of molecular mechanisms of heparin-induced angiogenesis, in human umbilical vein endothelial cells |
| title_sort | evaluation of molecular mechanisms of heparin induced angiogenesis in human umbilical vein endothelial cells |
| topic | Molecular mechanisms Cell biology Biomaterials Chitosan Heparin Angiogenesis |
| url | http://www.sciencedirect.com/science/article/pii/S1018364721001956 |
| work_keys_str_mv | AT sadiafazil evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT humairashah evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT mamoonanoreen evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT muhammadyar evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT atherfarooqkhan evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT sherzamansafi evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT sulimanyousefalomar evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT afrahfahadalkhuriji evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT hananmuallaalharbi evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells AT muhammadsohailafzal evaluationofmolecularmechanismsofheparininducedangiogenesisinhumanumbilicalveinendothelialcells |