Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity
Background: Vascular calcification is an active process that increases cardiovascular disease (CVD) risk. There is still no consensus on an appropriate biomarker for vascular calcification. We reasoned that the biomarker for vascular calcification is the collection of all blood components that can b...
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MDPI AG
2021-08-01
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| author | Armand M. G. Jaminon Asim C. Akbulut Niko Rapp Rafael Kramann Erik A. L. Biessen Lieve Temmerman Barend Mees Vincent Brandenburg Robert Dzhanaev Willi Jahnen-Dechent Juergen Floege Jouni Uitto Chris P. Reutelingsperger Leon J. Schurgers |
| author_facet | Armand M. G. Jaminon Asim C. Akbulut Niko Rapp Rafael Kramann Erik A. L. Biessen Lieve Temmerman Barend Mees Vincent Brandenburg Robert Dzhanaev Willi Jahnen-Dechent Juergen Floege Jouni Uitto Chris P. Reutelingsperger Leon J. Schurgers |
| author_sort | Armand M. G. Jaminon |
| collection | DOAJ |
| description | Background: Vascular calcification is an active process that increases cardiovascular disease (CVD) risk. There is still no consensus on an appropriate biomarker for vascular calcification. We reasoned that the biomarker for vascular calcification is the collection of all blood components that can be sensed and integrated into a calcification response by human vascular smooth muscle cells (hVSMCs). Methods: We developed a new cell-based high-content assay, the BioHybrid assay, to measure in vitro calcification. The BioHybrid assay was compared with the o-Cresolphthalein assay and the T50 assay. Serum and plasma were derived from different cohort studies including chronic kidney disease (CKD) stages III, IV, V and VD (on dialysis), pseudoxanthoma elasticum (PXE) and other cardiovascular diseases including serum from participants with mild and extensive coronary artery calcification (CAC). hVSMCs were exposed to serum and plasma samples, and in vitro calcification was measured using AlexaFluor<sup>®</sup>-546 tagged fetuin-A as calcification sensor. Results: The BioHybrid assay measured the kinetics of calcification in contrast to the endpoint o-Cresolphthalein assay. The BioHybrid assay was more sensitive to pick up differences in calcification propensity than the T50 assay as determined by measuring control as well as pre- and post-dialysis serum samples of CKD patients. The BioHybrid response increased with CKD severity. Further, the BioHybrid assay discriminated between calcification propensity of individuals with a high CAC index and individuals with a low CAC index. Patients with PXE had an increased calcification response in the BioHybrid assay as compared to both spouse and control plasma samples. Finally, vitamin K1 supplementation showed lower in vitro calcification, reflecting changes in delta Agatston scores. Lower progression within the BioHybrid and on Agatston scores was accompanied by lower dephosphorylated-uncarboxylated matrix Gla protein levels. Conclusion: The BioHybrid assay is a novel approach to determine the vascular calcification propensity of an individual and thus may add to personalised risk assessment for CVD. |
| first_indexed | 2024-03-10T08:55:12Z |
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| last_indexed | 2024-03-10T08:55:12Z |
| publishDate | 2021-08-01 |
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| spelling | doaj.art-12529c17190443269e9d4a251ecbaf1e2023-11-22T07:11:43ZengMDPI AGCells2073-44092021-08-01108209710.3390/cells10082097Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification PropensityArmand M. G. Jaminon0Asim C. Akbulut1Niko Rapp2Rafael Kramann3Erik A. L. Biessen4Lieve Temmerman5Barend Mees6Vincent Brandenburg7Robert Dzhanaev8Willi Jahnen-Dechent9Juergen Floege10Jouni Uitto11Chris P. Reutelingsperger12Leon J. Schurgers13Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, 6200 MD Maastricht, The NetherlandsDepartment of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, 6200 MD Maastricht, The NetherlandsDepartment of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, 6200 MD Maastricht, The NetherlandsInstitute of Experimental Medicine and Systems Biology, RWTH Aachen University, 52074 Aachen, GermanyDepartment of Pathology, Cardiovascular Research Institute Maastricht, Maastricht University, 6229 HX Maastricht, The NetherlandsDepartment of Pathology, Cardiovascular Research Institute Maastricht, Maastricht University, 6229 HX Maastricht, The NetherlandsDepartment of Vascular Surgery, Maastricht University Medical Centre, 6229 HX Maastricht, The NetherlandsDepartment of Cardiology, Rhein-Maas-Klinikum Würselen, 52146 Würselen, GermanyHelmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, 52074 Aachen, GermanyHelmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, 52074 Aachen, GermanyDepartment of Nephrology and Clinical Immunology, RWTH Aachen University Hospital, 52074 Aachen, GermanyDepartment of Dermatology and Cutaneous Biology, Sidney Kimmel Medical College, Jefferson Institute of Molecular Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, 6200 MD Maastricht, The NetherlandsDepartment of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University Medical Centre, 6200 MD Maastricht, The NetherlandsBackground: Vascular calcification is an active process that increases cardiovascular disease (CVD) risk. There is still no consensus on an appropriate biomarker for vascular calcification. We reasoned that the biomarker for vascular calcification is the collection of all blood components that can be sensed and integrated into a calcification response by human vascular smooth muscle cells (hVSMCs). Methods: We developed a new cell-based high-content assay, the BioHybrid assay, to measure in vitro calcification. The BioHybrid assay was compared with the o-Cresolphthalein assay and the T50 assay. Serum and plasma were derived from different cohort studies including chronic kidney disease (CKD) stages III, IV, V and VD (on dialysis), pseudoxanthoma elasticum (PXE) and other cardiovascular diseases including serum from participants with mild and extensive coronary artery calcification (CAC). hVSMCs were exposed to serum and plasma samples, and in vitro calcification was measured using AlexaFluor<sup>®</sup>-546 tagged fetuin-A as calcification sensor. Results: The BioHybrid assay measured the kinetics of calcification in contrast to the endpoint o-Cresolphthalein assay. The BioHybrid assay was more sensitive to pick up differences in calcification propensity than the T50 assay as determined by measuring control as well as pre- and post-dialysis serum samples of CKD patients. The BioHybrid response increased with CKD severity. Further, the BioHybrid assay discriminated between calcification propensity of individuals with a high CAC index and individuals with a low CAC index. Patients with PXE had an increased calcification response in the BioHybrid assay as compared to both spouse and control plasma samples. Finally, vitamin K1 supplementation showed lower in vitro calcification, reflecting changes in delta Agatston scores. Lower progression within the BioHybrid and on Agatston scores was accompanied by lower dephosphorylated-uncarboxylated matrix Gla protein levels. Conclusion: The BioHybrid assay is a novel approach to determine the vascular calcification propensity of an individual and thus may add to personalised risk assessment for CVD.https://www.mdpi.com/2073-4409/10/8/2097vascular calcificationvascular smooth muscle cellsBioHybridfetuin-Amatrix Gla proteinvitamin K |
| spellingShingle | Armand M. G. Jaminon Asim C. Akbulut Niko Rapp Rafael Kramann Erik A. L. Biessen Lieve Temmerman Barend Mees Vincent Brandenburg Robert Dzhanaev Willi Jahnen-Dechent Juergen Floege Jouni Uitto Chris P. Reutelingsperger Leon J. Schurgers Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity Cells vascular calcification vascular smooth muscle cells BioHybrid fetuin-A matrix Gla protein vitamin K |
| title | Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity |
| title_full | Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity |
| title_fullStr | Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity |
| title_full_unstemmed | Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity |
| title_short | Development of the BioHybrid Assay: Combining Primary Human Vascular Smooth Muscle Cells and Blood to Measure Vascular Calcification Propensity |
| title_sort | development of the biohybrid assay combining primary human vascular smooth muscle cells and blood to measure vascular calcification propensity |
| topic | vascular calcification vascular smooth muscle cells BioHybrid fetuin-A matrix Gla protein vitamin K |
| url | https://www.mdpi.com/2073-4409/10/8/2097 |
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