Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion
The placenta plays a critical role in nutrient and oxygen exchange during pregnancy, yet the effects of medicinal drugs on this selective barrier remain poorly understood. To overcome this, this study presents a cost-effective bioimpedance spectroscopy (BIS) system to assess tight junction integrity...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
|
| Series: | Micromachines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-666X/15/12/1506 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850239391326273536 |
|---|---|
| author | Karim Saadé Mohammed Areeb Hussain Shannon A. Bainbridge Raphael St-Gelais Fabio Variola Marianne Fenech |
| author_facet | Karim Saadé Mohammed Areeb Hussain Shannon A. Bainbridge Raphael St-Gelais Fabio Variola Marianne Fenech |
| author_sort | Karim Saadé |
| collection | DOAJ |
| description | The placenta plays a critical role in nutrient and oxygen exchange during pregnancy, yet the effects of medicinal drugs on this selective barrier remain poorly understood. To overcome this, this study presents a cost-effective bioimpedance spectroscopy (BIS) system to assess tight junction integrity and monolayer formation in BeWo b30 cells, a widely used model of the multinucleated maternal–fetal exchange surface of the placental barrier. Cells were cultured on collagen-coated porous membranes and treated with forskolin to induce controlled syncytialization. Electrical impedance was measured using an entry level impedance analyzer, while immunofluorescence staining was used to confirm monolayer formation and syncytialization. The measurements and staining confirmed the formation of a confluent monolayer on day 4. In fact, the electrical resistance tripled for treated samples indicating a more electrically restrictive barrier. This resistance remained constant for treated samples reflecting the intact barrier’s integrity over the next 3 days. The measurements show that, on day 4, the electrical capacitance of the cells decreased for the treated samples as opposed to the untreated samples. This reflects that the surface area of the BeWo b30 cells decreased when the samples were treated with forskolin. Finally, a COMSOL model was developed to explore the effects of electrode positioning, depth, and distance on TEER measurements, explaining discrepancies in the literature. In fact, there was a substantial 97% and 39.4% difference in the obtained TEER values. This study demonstrates the AD2 device’s feasibility for monitoring placental barrier integrity and emphasizes the need for standardized setups for comparable results. The system can hence be used to analyze drug effects and nutrient transfer across the placental barrier. |
| format | Article |
| id | doaj-art-0c7ad4fedba748b69f00d1f732dd56a2 |
| institution | OA Journals |
| issn | 2072-666X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj-art-0c7ad4fedba748b69f00d1f732dd56a22025-08-20T02:01:10ZengMDPI AGMicromachines2072-666X2024-12-011512150610.3390/mi15121506Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell FusionKarim Saadé0Mohammed Areeb Hussain1Shannon A. Bainbridge2Raphael St-Gelais3Fabio Variola4Marianne Fenech5Department of Mechanical Engineering, University of Ottawa, Ottawa, ON K1N 6N5, CanadaDepartment of Mechanical Engineering, University of Ottawa, Ottawa, ON K1N 6N5, CanadaFaculty of Health Sciences, University of Ottawa, Ottawa, ON K1S 5S9, CanadaDepartment of Mechanical Engineering, University of Ottawa, Ottawa, ON K1N 6N5, CanadaDepartment of Mechanical Engineering, University of Ottawa, Ottawa, ON K1N 6N5, CanadaDepartment of Mechanical Engineering, University of Ottawa, Ottawa, ON K1N 6N5, CanadaThe placenta plays a critical role in nutrient and oxygen exchange during pregnancy, yet the effects of medicinal drugs on this selective barrier remain poorly understood. To overcome this, this study presents a cost-effective bioimpedance spectroscopy (BIS) system to assess tight junction integrity and monolayer formation in BeWo b30 cells, a widely used model of the multinucleated maternal–fetal exchange surface of the placental barrier. Cells were cultured on collagen-coated porous membranes and treated with forskolin to induce controlled syncytialization. Electrical impedance was measured using an entry level impedance analyzer, while immunofluorescence staining was used to confirm monolayer formation and syncytialization. The measurements and staining confirmed the formation of a confluent monolayer on day 4. In fact, the electrical resistance tripled for treated samples indicating a more electrically restrictive barrier. This resistance remained constant for treated samples reflecting the intact barrier’s integrity over the next 3 days. The measurements show that, on day 4, the electrical capacitance of the cells decreased for the treated samples as opposed to the untreated samples. This reflects that the surface area of the BeWo b30 cells decreased when the samples were treated with forskolin. Finally, a COMSOL model was developed to explore the effects of electrode positioning, depth, and distance on TEER measurements, explaining discrepancies in the literature. In fact, there was a substantial 97% and 39.4% difference in the obtained TEER values. This study demonstrates the AD2 device’s feasibility for monitoring placental barrier integrity and emphasizes the need for standardized setups for comparable results. The system can hence be used to analyze drug effects and nutrient transfer across the placental barrier.https://www.mdpi.com/2072-666X/15/12/1506bioimpedance spectroscopyCOMSOL (Version 6.2) multiphysicselectrical capacitancesyncytiotrophoblasttransepithelial/transendothelial electrical resistance (TEER) |
| spellingShingle | Karim Saadé Mohammed Areeb Hussain Shannon A. Bainbridge Raphael St-Gelais Fabio Variola Marianne Fenech Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion Micromachines bioimpedance spectroscopy COMSOL (Version 6.2) multiphysics electrical capacitance syncytiotrophoblast transepithelial/transendothelial electrical resistance (TEER) |
| title | Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion |
| title_full | Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion |
| title_fullStr | Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion |
| title_full_unstemmed | Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion |
| title_short | Cost-Effective Bioimpedance Spectroscopy System for Monitoring Syncytialization In Vitro: Experimental and Numerical Validation of BeWo Cell Fusion |
| title_sort | cost effective bioimpedance spectroscopy system for monitoring syncytialization in vitro experimental and numerical validation of bewo cell fusion |
| topic | bioimpedance spectroscopy COMSOL (Version 6.2) multiphysics electrical capacitance syncytiotrophoblast transepithelial/transendothelial electrical resistance (TEER) |
| url | https://www.mdpi.com/2072-666X/15/12/1506 |
| work_keys_str_mv | AT karimsaade costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion AT mohammedareebhussain costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion AT shannonabainbridge costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion AT raphaelstgelais costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion AT fabiovariola costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion AT mariannefenech costeffectivebioimpedancespectroscopysystemformonitoringsyncytializationinvitroexperimentalandnumericalvalidationofbewocellfusion |