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St. Thomas Modified Cardioplegia Effects on Myoblasts' Viability and Morphology
Rafał Nowicki1, Katarzyna Bieżuńska-Kusiak2, Julita Kulbacka2
1Department of Cardiac Surgery, Wroclaw Medical University, Curie-Skłodowskiej, 50-369 Wrocław, Poland.
Insights
St. Thomas cardioplegia (ST2C) protects heart cells in vitro for up to 4 hours. Longer exposure showed cell vacuolization, indicating optimal use duration for cardioprotection during cardiac surgery.
Area of Science:
- Cardiology
- Cell Biology
- Biochemistry
Background:
- Cardioprotection is vital in cardiac surgery, often involving cardioplegic arrest.
- In vitro studies using cultured cells are less common than clinical trials for evaluating cardioprotective agents.
- St. Thomas cardioplegia (ST2C) is a widely used cardioplegic solution.
Purpose of the Study:
- To assess the suitability of rat heart H9C2 cells as an in vitro model for studying ST2C.
- To determine the protective effects and safe exposure limits of ST2C on cardiac cells in vitro.
- To investigate the impact of ST2C on cell viability, proliferation, and morphology.
Main Methods:
- H9C2 cells were incubated with ST2C for up to 24 hours.
- Assessed cell viability, confluency, and size.
- Measured thiol group levels, Ki67, and PCNA expression.
- Utilized Transmission Electron Microscopy (TEM) and 3D holographic imaging for morphological analysis.
Main Results:
- Optimal ST2C exposure for cell viability and confluency was determined to be no more than 4 hours.
- Increased expression of Ki67 and PCNA indicated a proliferative response.
- TEM and 3D imaging revealed cellular vacuolization after 24 hours of ST2C exposure.
Conclusions:
- Rat heart H9C2 cells serve as a viable in vitro model for ST2C research.
- ST2C demonstrates protective potential in cardiac surgery, with a recommended exposure limit of 4 hours in this model.
- Understanding ST2C's effects on cellular level aids in optimizing cardioprotection strategies.
Abstract:
Background and Objectives: The cardioplegic arrest of the heart during cardiosurgical procedures is the crucial element of a cardioprotection strategy. Numerous clinical trials compare different cardioplegic solutions and cardioprotective protocols, but a relatively small number of papers apply to in vitro conditions using cultured cells. This work aimed to analyze whether it is possible to use the rat heart myocardium cells as an in vitro model to study the protective properties of St. Thomas cardioplegia (ST2C). Methods: The rat heart myocardium cells-H9C2 were incubated with cold cardioplegia for up to 24 h. After incubation, we determined: viability, confluency, and cell size, the thiol groups' level by modifying Ellman's method, Ki67, and Proliferating Cell Nuclear Antigen expression (PCNA). The impact on cells' morphology was visualized by the ultrastructural (TEM) study and holotomograpic 3D imaging. Results: The viability and confluency analysis demonstrated that the safest exposure to ST2C, should not exceed 4h. An increased expression of Ki67 antigen and PCNA was observed. TEM and 3D imaging studies revealed vacuolization after the longest period of exposure (24). Conclusions: According to obtained results, we conclude that STC can play a protective role in cardiac surgery during heart arrest.
