Cell and Organ Transplantology. 2023; 11(2):104-113.
DOI: 10.22494/cot.v11i2.156

Comparison of the therapeutic effect of native and preconditioned human umbilical cord-derived multipotent mesenchymal stromal cells on a rat model of acute pancreatitis

Pikus P.^1,2, Rymar S.^1,2, Pustovalov A.³, Shuvalova N.², Reshetnyk Ye.³, Kordium V.^1,2

• ¹Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv, Ukraine
• ²Institute of Genetic and Regenerative Medicine, M. D. Strazhesko National Scientific Center of Cardiology, Clinical and Regenerative Medicine, National Academy of Medical Sciences of Ukraine, Kyiv, Ukraine
• ³Educational and Scientific Center “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine

Abstract

Pancreatitis is a disease defined by an acute or chronic inflammatory process of the pancreas, which is induced by the activation of trypsin inside acinocytes. The consequences of these processes are necrosis of the parenchyma of the organ, abscess and systemic complications, which lead to patient mortality, which reaches 30-47 %. To date, there is no effective treatment for acute pancreatitis in the world. Potential candidates for the treatment of this disease can be multipotent mesenchymal stromal/stem cells (MMSCs) due to their immunomodulatory properties.
The purpose of the study was to compare the therapeutic effect of transplantation of native human umbilical cord-derived MMSCs (hUC-MMSCs) and hUC-MMSCs preconditioned with H₂O₂ in a rat model of acute pancreatitis.
Methods. Acute pancreatitis in rats was induced by intraperitoneal injection of L-arginine at a dose of 3.5 g/kg body weight twice at an interval of 1 hour. Rats were divided into four groups: group 1 – negative control, animals received 0.9 % saline, group 2 – positive control (pancreatitis). 3 days after the emergence of the signs of developed acute pancreatitis, part of group 2 was divided into groups 3 and 4. The group 3 rats were intraperitoneally transplanted with native hUC-MMSCs at a dose of 1.5×10⁶ cells/kg, and the group 4 were injected with the same dose of hUC-MMSCs, preconditioned with H₂O₂ at a concentration of 30 µmol for 18 hours. The effect of native or preconditioned hUC-MMSCs was evaluated 3, 7 and 14 days after their administration. The results of the study were evaluated according to the α-amylase index, the identification of insulin in the pancreatic islets, and the histological study.
Results. It was shown that 3 days after the administration of L-arginine, the level of α-amylase increased 5 times compared to the negative control, the level of glucose in the blood increased 2.5 times, which indicates damage to both the exocrine and endocrine parts of the pancreas. On microscopic examination, fibrosis of the pancreatic parenchyma was observed, which increased 3.5 times compared to the negative control. 3 days after the transplantation of native hUC-MMSCs, the level of α-amylase in the blood decreased by 1.7 times, and in the variant with preconditioned hUC-MMSCs – by 2 times compared to the group 2. After 7 days, the α-amylase index in the group with native hUC-MMSCs decreased by 2.4 times compared to the positive control in the group with preconditioned hUC-MMSCs and was almost within the norm. After 7 days, a gradual recovery of the architecture of the pancreas was observed, but in the group with preconditioned hUC-MMSCs, the recovery of the organ occurred faster. Histological sections revealed the replacement of anucleated areas with acinocytes, which was evidenced by a 3-fold decrease in the number of anucleated cells in the variant with native hUC-MMSCs, while in the variant with preconditioned hUC-MMSCs the indicator was almost normal. In both variants, a positive histochemical PAS-reaction was observed for the identification of insulin in the pancreatic islets, and blood glucose levels corresponded to the norm. After 14 days, complete recovery of the pancreas was in both variants.
Conclusion. The results of the study showed that the transplantation of hUC-MMSCs in rats with acute pancreatitis leads to the restoration of the architecture of the pancreas parenchyma and its functioning, and the administration of hUC-MMSCs preconditioned with H₂O₂ significantly improves the therapeutic effect and accelerates the recovery of the pancreas.

Keywords: acute pancreatitis; human umbilical cord-derived multipotent mesenchymal stromal cells; stem cell preconditioning; cell transplantation

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