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The influence of sodium hypochlorite concentration on the fibrin structure of human blood clots and transforming growth factor-beta 1 release: an ex vivo study

  • Anisha Mishra (Department of Conservative Dentistry and Endodontics, Faculty of Dentistry, Meenakshi Ammal Dental College & Hospital, Meenakshi Academy of Higher Education and Research (MAHER)) ;
  • Velmurugan Natanasabapathy (Department of Conservative Dentistry and Endodontics, Faculty of Dentistry, Meenakshi Ammal Dental College & Hospital, Meenakshi Academy of Higher Education and Research (MAHER)) ;
  • Nandini Suresh (Department of Conservative Dentistry and Endodontics, Faculty of Dentistry, Meenakshi Ammal Dental College & Hospital, Meenakshi Academy of Higher Education and Research (MAHER))
  • Received : 2022.05.26
  • Accepted : 2022.08.10
  • Published : 2022.11.30

Abstract

Objective: This study investigated the effects of various concentrations of sodium hypochlorite (NaOCl) on human whole-blood clotting kinetics, the structure of the blood clots formed, and transforming growth factor (TGF)-β1 release. Materials and Methods: Human whole blood was collected from 5 healthy volunteers and divided into 4 groups: CG (control, 0.5 mL of blood), BN0.5 (0.5 mL of blood with 0.5 mL of 0.5% NaOCl), BN3 (0.5 mL of blood with 0.5 mL of 3% NaOCl), and BN5.25 (0.5 mL of blood with 0.5 mL of 5.25% NaOCl). The effects of NaOCl on clotting kinetics, structure of fibrin and cells, and release of TGF-β1 were assessed using thromboelastography (TEG), scanning electron microscopy (SEM), and enzyme-linked immunosobent assay, respectively. Statistical analysis was conducted using the Kruskal Wallis and Mann-Whitney U tests, followed by the post hoc Dunn test. A p value < 0.05 indicated statistical significance. Results: The blood samples in BN0.5 and BN3 did not clot, whereas the TEG of BN5.25 showed altered clot formation. Samples from the CG and BN3 groups could only be processed with SEM, which showed that the latter lacked fibrin formation and branching of fibers, as well as clumping of red blood cells with surface roughening and distortion. TGF-β1 release was significantly highest in BN3 when all groups were compared to CG (p < 0.05). Conclusions: Each concentration of NaOCl affected the release of TGF-β1 from blood clots and altered the clotting mechanism of blood by affecting clotting kinetics and cell structure.

Keywords

Acknowledgement

The authors would like to acknowledge: the guidance of Dr. Ganesh Venkatraman, Professor and Ms. Vaishnavi Balasubramanian, Research Scholar, Department of Human Genetics, Sri Ramachandra Institute of Higher Education and Research in procuring and aiding in the analysis of TGF-β1 growth factor release using enzyme-linked immunosorbent assay; the Department of Biomedical Sciences, Apollo Hospitals, Greams Road, Chennai for permitting the use of the Thromboelastograph 5000 Hemostasis Analyzer; Prof. AK Mishra, Dean, IIT-Madras; Mr. Sundarraman and Ms. Kalpana, Sophisticated Analytical Instrument Facility, IIT-Madras; Dr. Thangavel, Vice Principal, Allied Health Sciences, Meenakshi Academy of higher Education and research; Dr. Lokeshwari, Department of Biotechnology, Sri Ramachandra Institute of Higher Education and Research for aiding in the analysis of samples under a scanning electron microscope.

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