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Establishing a protocol to quantify leaflet fibroblast responses to physiologic flow through a viable heart valve
M W Weston1, S Goldstein, R E Epting
1School of Chemical Engineering, Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta 30332-0100, USA.
ASAIO Journal (American Society for Artificial Internal Organs : 1992)
|November 14, 1997
Summary
Dynamic mechanical stress influences heart valve fibroblast activity. This study developed a pulsatile flow system to quantify fibroblast function and collagen synthesis in intact aortic valves under physiological conditions.
Area of Science:
- Biomedical Engineering
- Cardiovascular Research
- Cell Biology
Background:
- Mechanical stresses significantly impact cell metabolism, yet quantitative data on heart valve fibroblast responses to dynamic loading are scarce.
- Understanding fibroblast function under native conditions is crucial for developing effective tissue-engineered heart valves.
Purpose of the Study:
- To investigate heart valve leaflet fibroblast function and differentiation under dynamic flow conditions using an intact valve model.
- To establish a novel, sterile pulsatile flow system that accurately replicates the native aortic valve environment.
- To develop and apply assay protocols for assessing cellular viability and protein/collagen synthesis.
Main Methods:
- Development of a sterilizable pulsatile flow system simulating aortic valve dynamics (70 bpm, 129/82 mmHg, 2.3 L/min) for 71 hours.
- Utilized viability assays to assess cell health and proline incorporation studies to quantify protein and collagen retention within leaflet tissue.
- Employed polyacrylamide gel electrophoresis to identify and differentiate collagen Types I and III.
Main Results:
- The developed flow system maintained physiologic conditions without microbial contamination.
- Fibroblast viability decreased after prolonged antibiotic exposure.
- Leaflet tissue retained 11 times more synthesized protein than was released, with 27% identified as collagen (Types I and III).
Conclusions:
- The novel pulsatile flow system effectively recreates the native heart valve environment for studying fibroblast activity.
- Quantitative data on fibroblast viability and collagen synthesis in response to dynamic flow were successfully obtained.
- These findings provide a critical baseline for evaluating future tissue-engineered heart valve substitutes.