Eicosapentaenoic acid potentiates the therapeutic effects of adipose tissue-derived mesenchymal stromal cells on lung and distal organ injury in experimental sepsis

Table 2 Semiquantitative analysis of distal organ damage

Pathologic findings	Groups
Pathologic findings	C	CLP-SAL	CLP-AD-MSC	CLP-AD-MSC-EPA
Liver
Hepatocyte disarrangement	1 (0.5–1)	9 (8.5–10.5)*	8 (7–8.5)*	2 (2–3)*^,#,‡
Kupffer cell hyperplasia	1 (1–1)	9 (9–9)*	6 (6–6)*^,#	4 (3–4)*^,#,‡
Hydropic degeneration	1 (1–1)	6 (5–7.5)*	3 (3–4.5)*^,#	2 (2–3)*^,#
Kidney
Tubular acute necrosis	0 (0–0.5)	12 (10.5–12)*	6 (5–6)*^,#	2 (2–3)*^,#,‡
Heart
Disarrangement of cardiac myocyte architecture	1 (0.5–1)	6 (5–6)*	4 (3–4)*^,#	2 (2–3)*^,#,‡
Interstitial edema	1 (0.5–1)	6 (6–7.5)*	2 (2–2)*^,#	2 (1.5–2)*^,#
Microvacuolization	1 (1–1)	9 (7.5–9)*	2 (1.5–2)*^,#	2 (1.5–2)*^,#
Spleen
Increased number of megakaryocytes	1 (0.5–1)	12 (10.5–12)*	6 (5–6)*^,#	2 (2–3)*^,#,‡
Increased number of lymphocytes	1 (1–1.5)	8 (7–8)*	6 (5–6)*^,#	2 (2–2)*^,#,‡
Small bowel
Villi architecture disorganization	1 (0.5–1)	12 (10–12)*	6 (5–6)*^,#	2 (2–3)*^,#,‡
Edema of lamina propria	0 (0–0.5)	9 (8.5–9)*	4 (4–5)*^,#	4 (3–4)*^,#
Number of enteroblasts (crypt cells)	0 (0–0.5)	6 (6–7)*	4 (4–5)*^,#	4 (3–4)*^,#

Values are median (interquartile range) of eight mice in each group. A semiquantitative scoring system based on severity and extension of injury was used to assess the degree of distal organ damage. Severity was graded as follows: 1 = normal tissue, 2 = mild injury, 3 = moderate injury, and 4 = severe injury. Extension of injury was graded as follows: 0 = normal tissue, 1 = damage to 1–25% of total tissue examined, 2 = damage to 26–50% of total tissue examined, 3 = damage to 51–75% of total tissue examined, and 4 = damage to 76–100% of total tissue examined. The final score is obtained by multiplying severity and extension of injury in each tissue examined. Scoring was done independently by two different investigators (J.D.S. and V.L.C.). Control (C) mice were subjected to an abdominal incision alone, while animals were subjected to cecal ligation and puncture (CLP) to induce polymicrobial sepsis. After 24 h, the CLP group was further randomized to receive saline (0.05 mL, SAL) or adipose tissue-derived mesenchymal stromal cells maintained under regular conditions (MSC; 10⁵ cells) or preconditioned with eicosapentaenoic acid (AD-MSC-EPA; 10⁵ cells). *Significantly different from the C group (P < 0.05). ^#Significantly different from the CLP-SAL group (P < 0.05). ^‡Significantly different from CLP-AD-MSC group (P < 0.05)

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