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Abnormalities of Coagulation and Fibrinolysis Assessed by Thromboelastometry in an Endotoxic Shock Model in Piglets Treated with Nitric Oxide and Hydrocortisone

Barbara Adamik
Barbara Adamik
, 
Claes Frostell
Claes Frostell
, 
Barbara Dragan
Barbara Dragan
, 
Urszula Paslawska
Urszula Paslawska
, 
Stanislaw Zielinski
Stanislaw Zielinski
, 
Robert Paslawski
Robert Paslawski
, 
Adrian Janiszewski
Adrian Janiszewski
, 
Marzena Zielinska
Marzena Zielinska
, 
Stanislaw Ryniak
Stanislaw Ryniak
, 
Johanna Albert
Johanna Albert
 e 
Waldemar Gozdzik
Waldemar Gozdzik
   | 07 giu 2024
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Article Category: Original Article
Pubblicato online: 07 giu 2024
Pagine: -
Ricevuto: 05 set 2023
Accettato: 18 apr 2024
DOI: https://doi.org/10.2478/aite-2024-0011
Parole chiave
© 2024 Barbara Adamik et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig 1.

Study design. All animals had endotoxin infusion for a total of 10 h. Administration of iNO was started 3 h after endotoxin infusion and continued until the end of the 20-h observation period. Hydrocortisone was given 3 h after the endotoxin infusion, and the dose was repeated at 8 h and 16 h. iNO and hydrocortisone were administered in the iNO(+) group, and not in the group iNO(−). iNO, inhaled nitric oxide.
Study design. All animals had endotoxin infusion for a total of 10 h. Administration of iNO was started 3 h after endotoxin infusion and continued until the end of the 20-h observation period. Hydrocortisone was given 3 h after the endotoxin infusion, and the dose was repeated at 8 h and 16 h. iNO and hydrocortisone were administered in the iNO(+) group, and not in the group iNO(−). iNO, inhaled nitric oxide.

Fig 2.

Thromboelastometry (ROTEM®) diagram. CT – represents time from the start of the test until the initiation of clotting; CFT – represents time from the initiation of clotting until a clot firmness of 20 mm was detected; MCF – represents the overall strength of the clot by the polymerized fibrin, thrombocytes, and factor XIII. LI – represents clot stability and a reduction in clot firmness after MCF. CFT, clot formation time; CT, clotting time; LI, lysis index; MCF, maximum clot firmness.
Thromboelastometry (ROTEM®) diagram. CT – represents time from the start of the test until the initiation of clotting; CFT – represents time from the initiation of clotting until a clot firmness of 20 mm was detected; MCF – represents the overall strength of the clot by the polymerized fibrin, thrombocytes, and factor XIII. LI – represents clot stability and a reduction in clot firmness after MCF. CFT, clot formation time; CT, clotting time; LI, lysis index; MCF, maximum clot firmness.

Fig 3.

Coagulation parameters measured by thromboelastometry. The parameters of the EXTEM (a), INTEM (b), and FIBTEM (c) tests in Group 1 (survivors) were measured at baseline, and the 4th, 10th, and 20th h and in Group 2 (non-survivors), were measured at baseline, and the 4th and 10th h of the experiment. p-values represent differences between groups at each time point (p > 0.05, Mann–Whitney U test). n.s., not significant.
Coagulation parameters measured by thromboelastometry. The parameters of the EXTEM (a), INTEM (b), and FIBTEM (c) tests in Group 1 (survivors) were measured at baseline, and the 4th, 10th, and 20th h and in Group 2 (non-survivors), were measured at baseline, and the 4th and 10th h of the experiment. p-values represent differences between groups at each time point (p > 0.05, Mann–Whitney U test). n.s., not significant.

Fig 4.

Fibrinolysis results measured by thromboelastometry. LI in Group 1 (survivors) and Group 2 (non-survivors) was measured at baseline, and the 4th, 10th, and 20th h of the experiment. p-values represent differences between groups at each time point (p > 0.05, Mann–Whitney U test). LI, lysis index; n.s., not significant.
Fibrinolysis results measured by thromboelastometry. LI in Group 1 (survivors) and Group 2 (non-survivors) was measured at baseline, and the 4th, 10th, and 20th h of the experiment. p-values represent differences between groups at each time point (p > 0.05, Mann–Whitney U test). LI, lysis index; n.s., not significant.

Fig 5.

Coagulation parameters measured by thromboelastometry – the effect of iNO + hydrocortisone. The parameters of the EXTEM (a), INTEM (b), and FIBTEM (c). Tests in survivors were measured at baseline, and the 4th, 10th, and 20th h of the experiment, and in non-survivors, at baseline, and the 4th, and 10th h. iNO(+), animals were treated with iNO + hydrocortisone; iNO(−) animals were not treated with iNO + hydrocortisone; p-values refer to the comparison of iNO(+) vs. iNO(−) in each group at each time point (p > 0.05, Mann–Whitney U test). iNO, inhaled nitric oxide; n.s., not significant.
Coagulation parameters measured by thromboelastometry – the effect of iNO + hydrocortisone. The parameters of the EXTEM (a), INTEM (b), and FIBTEM (c). Tests in survivors were measured at baseline, and the 4th, 10th, and 20th h of the experiment, and in non-survivors, at baseline, and the 4th, and 10th h. iNO(+), animals were treated with iNO + hydrocortisone; iNO(−) animals were not treated with iNO + hydrocortisone; p-values refer to the comparison of iNO(+) vs. iNO(−) in each group at each time point (p > 0.05, Mann–Whitney U test). iNO, inhaled nitric oxide; n.s., not significant.

Changes in ROTEM parameters and other coagulation tests before (T0), during endotoxin infusion (T4 and T10), and after stopping of endotoxin administration (T20)

Parameter Time All (N = 34) Survivors (N = 25) Non-survivors (N = 9) p
EXTEM
CT (s) 0 34.2 ± 1.2 34.9 ± 1.3 32.5 ± 2.6 0.318
4 56.9 ± 4.4 49.1 ± 4.2 74.8 ± 9.5 0.024
10 65.1 ± 5.4 58.2 ± 5.2 96.6 ± 12.2 0.009
20 52.0 ± 4.3 52.0 ± 4.30
CFT (s) 0 35.4 ± 0.8 35.1 ± 1.1 36.3 ± 1.5 0.480
4 113.1 ± 8.9 98.5 ± 8.3 153.6 ± 19.9 0.006
10 144.1 ± 14.5 132.0 ± 14.2 202.2 ± 43.3 0.050
20 134.3 ± 17.6 134.3 ± 17.6
MCF (mm) 0 75.4 ± 0.6 75.8 ± 0.6 74.5 ± 1.9 0.623
4 59.3 ± 1.5 61.0 ± 1.6 54.4 ± 2.8 0.077
10 57.1 ± 1.9 58.7 ± 2.1 49.6 ± 3.7 0.130
20 57.9 ± 2.3 57.9 ± 2.3
LI (%) 0 91.7 ± 0.4 91.4 ± 0.3 92.5 ± 1.1 0.315
4 97.0 ± 0.4 96.3 ± 0.5 99.1 ± 0.5 0.004
10 98.5 ± 0.4 98.2 ± 0.5 99.8 ± 0.2 0.237
20 97.7 ± 0.6 97.7 ± 0.6
INTEM
CT (s) 0 125.9 ± 6.1 128.1 ± 6.8 119.6 ± 13.5 0.570
4 168.8 ± 7.3 162.3 ± 9.2 186.1 ± 8.8 0.114
10 173.4 ± 10.2 163.5 ± 8.8 223.0 ± 37,7 0.224
20 170.2 ± 15.5 170.2 ± 15.5
CFT (s) 0 37.8 ± 1.1 37.6 ± 1.4 38.5 ± 1.7 0.353
4 98.7 ± 9.1 84.5 ± 8.3 138.4 ± 21.3 0.004
10 133.1 ± 17.4 116.0 ± 14.5 215.2 ± 67.5 0.043
20 135.7 ± 20.6 135.7 ± 20.6
MCF (mm) 0 71.4 ± 0.5 71.5 ± 0.6 71.0 ± 1.2 0.433
4 54.1 ± 1.4 56.1 ± 1.6 48.6 ± 2.3 0.057
10 52.4 ± 1.9 54.2 ± 2.1 44.2 ± 3.4 0.872
20 54.0 ± 2.3 54.0 ± 2.3
LI (%) 0 84.8 ± 0.4 84.8 ± 0.5 85.1 ± 0.9 0.734
4 93.4 ± 0.8 92.0 ± 0.8 97.1 ± 1.1 0.010
10 96.4 ± 0.9 95.8 ± 1.0 99.0 ± 0.4 0.360
20 96.4 ± 1.1 96.4 ± 1.1
FIBTEM
MCF (mm) 0 40.2 ± 1.4 40.5 ± 1.7 39.5 ± 2.7 0.930
4 13.0 ± 1.1 14.7 ± 1.2 8.4 ± 2.0 0.008
10 11.2 ± 1.1 12.1 ± 1.3 7.2 ± 0.85 0.009
20 17.7 ± 2.0 17.7 ± 2.0
Platelets (103/μL) 0 415.7 ± 16.8 431.4 ± 18.6 372.1 ± 34.4 0.097
4 206.1 ± 22.3 229.5 ± 28.8 143.7 ± 17.5 0.045
10 146.2 ± 14.5 153.9 ± 18.2 119.0 ± 8.4 0.434
20 119.1 ± 16.7 119.1 ± 16.7
Fibrinogen (mg · dL−1) 0 3.1 ± 0.1 3.1 ± 0.1 3.3 ± 0.4 0.482
4 2.0 ± 0.1 2.1 ± 0.1 1.7 ± 0.2 0.024
10 1.6 ± 0.1 1.7 ± 0.1 1.2 ± 0.2 0.059
20 2.2 ± 0.1 2.2 ± 0.1

The basic variables of the pigs before endotoxin infusion

Parameter All (N = 34) Survivors (N = 25) Non-survivors (N = 9) p
EXTEM
CT (s) 34.2 ± 1.2 34.8 ± 1.3 32.5 ± 2.6 0.318
CFT (s) 35.4 ± 0.8 35.1 ± 1.1 36.3 ± 1.5 0.480
MCF (mm) 75.4 ± 0.6 75.8 ± 0.6 74.5 ± 1.9 0.623
LI (%) 91.7 ± 0.4 91.4 ± 0.3 92.5 ± 1.1 0.315
INTEM
CT (s) 125.9 ± 6.1 128.1 ± 6.8 119.6 ± 13.5 0.570
CFT (s) 37.8 ± 1.1 37.6 ± 1.4 38.5 ± 1.7 0.353
MCF (mm) 71.4 ± 0.5 71.5 ± 0.6 71.0 ± 1.2 0.433
LI (%) 84.8 ± 0.4 84.8 ± 0.5 85.1 ± 0.9 0.734
FIBTEM
MCF (mm) 40.2 ± 1.4 40.4 ± 1.7 39.5 ± 2.7 0.930
Platelets (103 • μL–1) 415.7 ± 16.8 431.4 ± 18.6 372.1 ± 34.4 0.097
Fibrinogen (mg • dL–1) 3.1 ± 0.1 3.1 ± 0.1 3.3 ± 0.4 0.482
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Medicine, Basic Medical Science, Biochemistry, Immunology, Clinical Medicine, other, Clinical Chemistry
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