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IN VITRO AND IN OVO ASSESSMENT OF LABETALOL AND SILIBININ, ALONE AND IN COMBINATION, IN H9C2(2-1) CARDIOMYOCYTE CELLS

RADU PETROMAN 1#, IASMINA-ALEXANDRA PREDESCU 2,3,4#, GABRIELA-FLORENTINA ȚAPOȘ 2,5*, DANIELA SAŞCO 3, ALINA ANTON 3,4, CONSTANTIN TUDOR LUCA 6,7,8, SILVIA LUCA 6,7,8, SIMINA CRIȘAN 6,7,8, MIHAELA-CRISTINA NEGRU 9, CASIANA BORU 10

1 Department of Rehabilitation, Physical Medicine and Rheumatology, Research Center for Assessment of Human Motion, Functionality and Disability, “Victor Babes” University of Medicine and Pharmacy Timisoara, 300041 Timisoara, Romania
2 Doctoral School, “Victor Babes” University of Medicine and Pharmacy, 2nd Eftimie Murgu Square, 300041 Timisoara, Romania
3 Faculty of Pharmacy, “Victor Babeș” University of Medicine and Pharmacy Timișoara, 2nd Eftimie Murgu Square, 300041, Timișoara, Romania
4 Research Centre for Pharmaco-Toxicological Evaluations, “Victor Babeș” University of Medicine and Pharmacy Timișoara, 2nd Eftimie Murgu Square, 300041, Timișoara, Romania
5 Arad County Clinical Emergency Hospital, 310037, Arad, Romania
6 Cardiology Department, “Victor Babes” University of Medicine and Pharmacy, 2nd Eftimie Murgu Square, 300041 Timisoara, Romania
7 Research Center of the Institute of Cardiovascular Diseases Timisoara, 300310, Timisoara, Romania
8 Institute of Cardiovascular Diseases Timisoara, 300310, Timisoara, Romania
9 Department of ENT, "Victor Babeş" University of Medicine and Pharmacy Timisoara, 2nd Eftimie Murgu Square, 300041, Timișoara, Romania
10 Faculty of Medicine, “Vasile Goldiș” Western University of Arad, 94 Revoluției Blvd., 310130, Arad, Romania

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Cardiovascular diseases, particularly ischemic heart disease and angina pectoris, remain a major health problem worldwide. Combining conventional cardiovascular drugs with antioxidant compounds may offer complementary benefits, but biosafety must be carefully evaluated. This study aimed to assess the in vitro and in ovo biosafety of labetalol (LB; 20-500 nM) and silibinin (SIL; 5-200 µM), administered individually and in combination, using H9c2(2-1) cells and the chorioallantoic membrane model. After 24 h, cell viability, morphology, lysosomal and membrane integrity, and mitochondrial and nuclear morphology were evaluated using MTT, Neutral Red uptake, fluorescence staining, and AO/PI assays. In addition, irritation potential was assessed using the HET-CAM assay. The results showed that cell viability remained comparable to control following all treatments. Mitochondrial and nuclear analyses indicated that organelle structure and morphology were preserved. Furthermore, AO/PI staining revealed no significant induction of apoptosis or necrosis. The HET-CAM assay confirmed the absence of vascular irritation for the tested combination. Overall, the combined administration of LB and SIL did not cause additional cytotoxicity or irritation compared with the individual treatments, supporting the biosafety of this combination.