The effect of mesenchymal stromal cells, their microvesicles and platelet-rich plasma on pathological changes in pancreatic tissue in acute necrotizing pancreatitis in rats

Background. Improving treatment outcomes for acute necrotizing pancreatitis (ANP) remains the most difficult and unresolved problem for surgeons and intensive care specialists.

Objective. To evaluate the effect of mesenchymal stromal cells (MSCs), MSC-derived microvesicles (MSC MVs) and platelet-rich plasma (PRP) on morphological and immunohistochemical characteristics of the pancreas in ANP in rats with regard to the route and timing of PRP administration, as well as separate and combined application.

Material and methods. The effects of MSCs, MSC MVs and PRP on the morphological and immunohistochemical characteristics of the pancreas in ANP in rats were studied, taking into the account the route (intraperitoneal/ intravenous) and time of administration (6 and 24 hours from the start of disease modeling), as well as their separate or combined use (PRP and MSCs, PRP and MSC MVs). The study was conducted on 72 adults Wistar rats. Acute pancreatitis was induced by the administration of a 0.3 ml of 5% solution of non-ionic polyethylene glycol octylphenol ether detergent into the caudal part of the rat pancreas. Organ and tissue sampling was performed on the 3^rd day from the beginning of the disease modeling. Histological changes in pancreatic tissues were studied by hematoxylin and eosin staining the preparations. Immunohistochemical staining of pancreatic tissue with anti-TGF- â1 and anti-SCARD1 primary monoclonal antibodies was performed followed by studying the nature of their expression.

Results. A comparative assessment of the effects of MSCs, MSC MVs and PRP on the histomorphological and immunohistochemical changes in pancreatic tissue in ANP in rats, found that more pronounced regeneration and neoangiogenesis processes were observed with the intravenous delivery of MSCs and MSC MVs, regardless of the timing of administration (6 or 24 hours from the onset of the disease). The obtained data suggested the immunomodulatory and pro-regenerative effects of MSC and MSC MVs produced by potentiating the polarization from inflammatory M1 macrophages to anti-inflammatory M2, as evidenced by a pronounced immunohistochemical reaction to TGF- â1, which is secreted predominantly by macrophages of the M2 phenotype.

Conclusion. The experimental use of MSCs and MSC MVs to treat АNP at early stages of the disease provides more pronounced processes of repair and neoangiogenesis in pathologically altered pancreatic tissue.

1. Goodman RR, Jong MK, Davies JE. Concise review: the challenges and opportunities of employing mesenchymal stromal cells in the treatment of acute pancreatitis. Biotechnol Adv. 2020;42:107338. PMID: 30639517 https://doi.org/10.1016/j.biotechadv.2019.01.005

2. Kudelich OA, Kondratenko GG, Potapnev MP. Stem cell technologies in the treatment of acute experimental pancreatitis. Voennaya medicina. 2022;3(64):90–99. (In Russ.). https://doi.org/10.51922/2074-5044.2022.3.90

3. Ahmed SM, Morsi M, Ghoneim NI, Abdel-Daim MM, El-Badri N. Mesenchymal stromal cell therapy for pancreatitis: a systematic review. Oxid Med Cell Longev. 2018;2018:3250864. PMID: 29743979 https://doi.org/10.1155/2018/3250864

4. Hu F, Lou N, Jiao J, Guo F, Xiang H, Shang D. Macrophages in pancreatitis: mechanisms and therapeutic potential. Biomed Pharmacother. 2020;131:110693. PMID: 32882586 https://doi.org/10.1016/j.biopha.2020.110693

5. Rehg JE, Bush D, Ward JM. The utility of immunohistochemistry for the identification of hematopoietic and lymphoid cells in normal tissues and interpretation of proliferative and inflammatory lesions of mice and rats. Toxicol Pathol. 2012;40(2):345– 374. PMID: 22434870 https://doi.org/10.1177/0192623311430695

6. Saito N, Pulford KA, Breton-Gorius J, Massé JM, Mason DY, Cramer EM. Ultrastructural localization of the CD68 macrophage-associated antigen in human blood neutrophils and monocytes. Am J Pathol. 1991;139(5):1053–1059. PMID: 1719819

7. Deng Z, Fan T, Xiao C, Tian H, Zheng Y, Li C, et al. TGF- β signaling in health, disease, and therapeutics. Signal Transduct Target Ther. 2024;9(1):61. PMID: 38514615 https://doi.org/10.1038/s41392-024-01764-w

8. Peng C, Li Z, Yu X. The role of pancreatic infiltrating innate immune cells in acute pancreatitis. Int J Med Sci. 2021;18(2):534–545. PMID: 33390823 https://doi.org/10.7150/ijms.51618

9. Nishikawa Y, Wang M, Carr BI. Changes in TGF-beta receptors of rat hepatocytes during primary culture and liver regeneration: increased expression of TGF-beta receptors associated with increased sensitivity to TGF-beta-mediated growth inhibition. J Cell Physiol. 1998;176(3):612–623. PMID: 9699514 https://doi.org/10.1002/(SICI)1097-4652(199809)176:33.0.CO;2-0

10. Riesle E, Friess H, Zhao L, Wagner M, Uhl W, Baczako K, et al. Increased expression of transforming growth factor beta s after acute oedematous pancreatitis in rats suggests a role in pancreatic repair. Gut. 1997;40(1):73–79. PMID: 9155579 https://doi.org/10.1136/gut.40.1.73

11. Friess H, Lu Z, Riesle E, Uhl W, Bründler AM, Horvath L, et al. Enhanced expression of TGF-betas and their receptors in human acute pancreatitis. Ann Surg. 1998;227(1):95–104. PMID: 9445116 https://doi.org/10.1097/00000658-199801000-00014

12. Gress T, Müller-Pillasch F, Elsässer HP, Bachem M, Ferrara C, Weidenbach H, et al. Enhancement of transforming growth factor beta 1 expression in the rat pancreas during regeneration from caerulein-induced pancreatitis. Eur J Clin Invest. 1994;24(10):679– 685. PMID: 7851468 https://doi.org/10.1111/j.1365-2362.1994.tb01060.x

13. Wan M, Li C, Zhen G, Jiao K, He W, Jia X, et al. Injury-activated transforming growth factor β controls mobilization of mesenchymal stem cells for tissue remodeling. Stem Cells. 2012;30(11):2498–2511. PMID: 22911900 https://doi.org/10.1002/stem.1208

14. Bax NA, van Oorschot AA, Maas S, Braun J, van Tuyn J, de Vries AA, et al. In vitro epithelial-to-mesenchymal transformation in human adult epicardial cells is regulated by TGF β -signaling and WT1. Basic Res Cardiol. 2011;106(5):829– 847. PMID: 21516490 https://doi.org/10.1007/s00395-011-0181-0

15. Redini F, Galera P, Mauviel A, Loyau G, Pujol JP. Transforming growth factor beta stimulates collagen and glycosaminoglycan biosynthesis in cultured rabbit articular chondrocytes. FEBS Lett. 1988;234(1):172–176. PMID: 3164687 https://doi.org/10.1016/0014-5793(88)81327-9

16. Buss A, Pech K, Kakulas BA, Martin D, Schoenen J, Noth J, et al. TGF-beta1 and TGF-beta2 expression after traumatic human spinal cord injury. Spinal Cord. 2008;46(5):364–371. PMID: 18040277 https://doi.org/10.1038/sj.sc.3102148

17. Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2018;7(1):1535750. PMID: 30637094 https://doi.org/10.1080/20013078.2018.1535750

18. Yamaguchi R, Terashima H, Yoneyama S, Tadano S, Ohkohchi N. Effects of platelet-rich plasma on intestinal anastomotic healing in rats: PRP concentration is a key factor. J Surg Res. 2012;173(2):258–266. PMID: 21074782 https://doi.org/10.1016/j.jss.2010.10.001

19. Kudelich OA, Kondratenko GG, Potapnev MP, Kolesnikova T, Klimenkova OV, Goncharova NV. Comparative evaluation of cellular origin bioproducts effects on the course of аcute necrotizing pancreatitis in experiment. Surgery. Eastern Europe. 2024;13(4):585–601. (In Russ.). https://doi.org/10.34883/PI.2024.13.4.024

20. Kang R, Lotze MT, Zeh HJ, Billiar TR, Tang D. Cell death and DAMPs in acute pancreatitis. Mol Med. 2014;20(1):466– 477. PMID: 25105302 https://doi.org/10.2119/molmed.2014.00117

21. Fedorov AA, Ermak NA, Gerashchenko TS, Topolnitskii EB, Shefer NA, Rodionov EO, et al. Polarization of macrophages: mechanisms, markers and factors of induction. Siberian journal of oncology. 2022;21(4):124–136. (In Russ.). https://doi.org/10.21294/1814-4861-2022-21-4-124-136

22. Bulava GV. Immunopathogenesis of acute pancreatitis. Russian Sklifosovsky Journal "Emergency Medical Care". 2022;11(3):484–492. (In Russ.). https://doi.org/10.23934/2223-9022-2022-11-3-484-492

23. Roch AM, Maatman TK, Cook TG, Wu HH, Merfeld-Clauss S, Traktuev DO, et al. Therapeutic use of adipose-derived stromal cells in a murine model of acute pancreatitis. J Gastrointest Surg. 2020;24(1):67–75. PMID: 31745900 https://doi.org/10.1007/s11605-019-04411-w

24. Bernardo ME, Fibbe WE. Mesenchymal stromal cells: sensors and switchers of inflammation. Cell Stem Cell. 2013;13(4):392–402. PMID: 24094322 https://doi.org/10.1016/j.stem.2013.09.006

25. Jung KH, Song SU, Yi T, Jeon MS, Hong SW, Zheng HM, et al. Human bone marrow-derived clonal mesenchymal stem cells inhibit inflammation and reduce acute pancreatitis in rats. Gastroenterology. 2011;140(3):998–1008. PMID: 21130088 https://doi.org/10.1053/j.gastro.2010.11.047

26. Goswami TK, Singh M, Dhawan M, Mitra S, Emran TB, Rabaan AA, et al. Regulatory T cells (Tregs) and their therapeutic potential against autoimmune disorders - advances and challenges. Hum Vaccin Immunother. 2022;18(1):2035117. PMID: 35240914 https://doi.org/10.1080/21645515.2022.2035117