Poster 90 - Impact of TNF-α on Endothelial Cell Survival Following Laser-Induced Nuclear Damage

Saturday, October 26, 2024

10:00 AM - 12:00 PM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Alex Sur

Intern
UCSD Institute of Engineering in Medicine
San Diego, California, United States

Co-Author(s)

Grace He

Student Mentor
UCSD Institute of Engineering in Medicine, United States

Co-Author(s)

Evan Lin (he/him/his)

Student
University of California San Diego
Palos Verdes Peninsula, California, United States

Co-Author(s)

Sammy H. Komen (he/him/his)

High School Student
UCSD Institute of Engineering in Medicine
Cardiff, California, United States
Alex P. Chung

High School Student
UCSD Institute of Engineering in Medicine
Vista, United States

Co-Author(s)

ED

Eesha A. Dumbre (she/her/hers)

Student
UCSD Institute of Engineering in Medicine, California, United States

Co-Author(s)

Henry F. Hou (he/him/his)

High School Student
UCSD Institute of Engineering in Medicine
Rancho Santa Fe, California, United States
Felix E. Yang (he/him/his)

High School Student
UCSD Institute of Engineering in Medicine
San Diego, California, United States
Sarah Kocian-Rudenberg

Student Mentor
University of California, San Diego: Institute of Engineering in Medicine, California, United States
Lijing Chen, PhD

Assistant Project Scientist
UCSD Institute of Engineering in Medicine, United States
SC

Shu Chien

Professor Emeritus
UCSD Institute of Engineering in Medicine, United States

Primary Investigator(s)

Linda Shi, PhD (she/her/hers)

OPALS Program Director, Project Scientist, Co-director IEM
University of California, San Diego: Institute of Engineering in Medicine
La Jolla, California, United States

Introduction: Cellular responses to stress and damage is a crucial aspect of understanding disease mechanisms and developing effective treatments. Tumor Necrosis Factor Alpha (TNF-α) is a key inflammatory cytokine that significantly impacts cellular behavior and stress responses, particularly in endothelial cells, which are crucial for vascular health and disease progression. This experiment aims to compare the survivability of wild-type (WT) cells and Inflamed TNF-α (Tumor Necrosis Factor Alpha) endothelial cells following nuclear damage induced by laser ablation. By assessing if these two cell types withstand and recover from targeted nuclear injury, we aim to reveal differences in their stress response mechanisms and repair capabilities. The findings from this study are expected to deepen the scientific understanding of how inflammatory conditions affect cellular resilience and contribute to strategies for managing diseases related to endothelial dysfunction and inflammation.

Materials and

Methods: The experiment involved four endothelial cell dishes: two containing inflamed cells and two containing wild-type cells. Next, the Zeiss Axiovert 200M Motorized Microscope and Photometrics Quantem 512SC EMCCD Camera were used to locate optimal cells for laser ablation and observe subsequent changes. Laser ablation and image collection were managed with LabView and Robolase III over a 12-minute period, while the MaiTai application adjusted the laser settings. Images were obtained for 30 wild-type and 30 TNF-α inflamed cells over a 12-minute period with 3-minute intervals after the nuclear envelope was cut using laser ablation. The images were then sorted and analyzed using ImageJ and using NIH Classification of Cell Death criteria the cells were determined to be alive or dead in a given image taken a specific amount of time after being initially ablated.

Results, Conclusions, and Discussions: RESULTS AND DISCUSSIONS
Cutting the nuclear envelope using laser ablation led to cell death in some instances. In addition, TNF-α inflamed cells were less likely to die than the WT cells as seen in Figure 1. 33.33% of the TNF-α cells were dead twelve minutes after their nuclear envelope was laser ablated while this happened to 76.67% of WT cells.
CONCLUSION
To conclude, this experiment successfully illustrates that TNF-α affected endothelial cells have higher survival rates compared to their wild-type counterparts following laser ablation. This increased survivability may be attributed to the heightened stiffness of TNF-α induced cells, which leads to a greater mechanical stability and resilience in the face of nuclear injury. These results could inform the development of targeted interventions to enhance cell survival and repair in inflammatory diseases, potentially improving therapeutic strategies for managing vascular and inflammatory disorders. Future studies could include studying the signal pathways that lead to inflamed cells having a higher survival rate or testing anti-inflammatory agents on the survival of endothelial cells.

Acknowledgements (Optional): This work was supported by a generous gift from Beckman Laser Institute Inc. to LS. The high school students (*) participated in the UCSD IEM OPALS internship. Special thanks to Dr. Shu Chien from UCSD Bioengineering and Dr. Lizhu Chen from CorDx Inc. for their donations.

References (Optional): Kroemer, G., Nomenclature Committee on Cell Death 2009. Classification of Cell Death: Recommendations of the Nomenclature Committee on Cell Death 2009. (2009, January). https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2744427/
Lee SY. Probing the mechanical properties of TNF-α stimulated endothelial cell with atomic force microscopy. International Journal of Nanomedicine. 2011 Jan;179.
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