Comparative efficacy of intact and apoptotic bone marrow cells at different stages of type 2 diabetes progression

Objective: to compare the therapeutic efficacy of intact and apoptotic bone marrow mononuclear cells (BMMCs) at different stages of type 2 diabetes mellitus (T2D) progression. Materials and methods. The study was conducted using a genetic model of T2D in db/db mice (n = 52; age 0.5–1.0 months). Animals were divided into four groups: control (n = 10) and three experimental groups (n = 14 each), which received allogeneic BMMCs from healthy donors at a dose of 40–45 × 106 cells at different stages of disease progression (1, 3, and 7 months of age). Each experimental group was further subdivided into two subgroups: one receiving freshly isolated intact BMMCs (iBMMCs) and the other apoptotic BMMCs (aBMMCs). Apoptotic cells were obtained by incubating intact cells in Bradschneider’s ion-balanced preservative solution to induce reversible apoptosis. Outcomes were assessed by measuring blood glucose levels, body weight, total oxidative metabolism index (TOM), and histological liver changes. Reliability of the results was assessed using statistical methods on a personal computer, employing the Shapiro–Wilk test and the Student’s t-test. Results. Both iBMMCs and aBMMCs demonstrated regulatory effects on metabolism at all stages of T2D progression; however, apoptotic BMMCs showed consistently greater efficacy. The therapeutic effect was directly associated with baseline tissue oxidative metabolism (TOM) levels at the time of administration. In early stages (adaptation at 1 month and progressive maladaptation at 3 months), when TOM levels remained relatively high, both cell types, particularly apoptotic BMMCs, produced pronounced corrective effects. In contrast, in the late stage (metabolic decompensation at 7 months), when TOM levels dropped below a critical threshold, iBMMCs, but especially aBMMCs, aggravated metabolic damage and increased animal mortality. Conclusion. Intact and apoptotic BMMCs may serve as effective adjunctive therapy in the early stages of T2D, with their therapeutic efficacy dependent on the body’s preserved stress-adaptive reserves in tissues.

1. Ramírez‑Alarcón K, Victoriano M, Mardones L, Villagran M, Al‑Harrasi A, Al‑Rawahi A et al. Phytochemicals as Potential Epidrugs in Type 2 Diabetes Mellitus. Front Endocrinol (Lausanne). 2021 Jun 1; 12: 656978. doi: 10.3389/fendo.2021.656978.

2. Artasensi A, Pedretti A, Vistoli G, Fumagalli L. Type 2 diabetes mellitus: A review of multi-target drugs. Molecules. 2020 Apr 23; 25 (8): 1987. doi: 10.3390/molecules25081987.

3. Hossan T, Kundu S, Alam SS, Nagarajan S. Epigenetic Modifications Associated with the Pathogenesis of Type 2 Diabetes Mellitus. Endocr Metab Immune Disord Drug Targets. 2019; 19 (6): 775–786. doi: 10.2174/1871530319666190301145545.

4. Taylor R, Ramachandran A, Yancy WS Jr, Forouhi NG. Nutritional basis of type 2 diabetes remission. BMJ. 2021 Jul 7; 374: n1449. doi: 10.1136/bmj.n1449.

5. Nishikawa H, Fukunishi S, Asai A, Yokohama K, Ohama H, Nishiguchi S, Higuchi K. Sarcopenia, frailty and type 2 diabetes mellitus (Review). Mol Med Rep. 2021 Dec; 24 (6): 854. doi: 10.3892/mmr.2021.12494.

6. Ma Q, Li Y, Li P, Wang M, Wang J, Tang Z et al. Research progress in the relationship between type 2 diabetes mellitus and intestinal flora. Biomed Pharmacother. 2019 Sep; 117: 109138. doi: 10.1016/j.biopha.2019.109138.

7. Zhou Z, Sun B, Yu D, Zhu C. Gut Microbiota: An Important Player in Type 2 Diabetes Mellitus. Front Cell Infect Microbiol. 2022 Feb 15; 12: 834485. doi: 10.3389/fcimb.2022.834485.

8. Bielka W, Przezak A, Pawlik A. The Role of the Gut Microbiota in the Pathogenesis of Diabetes. Int J Mol Sci. 2022 Jan 1; 23 (1): 480. doi: 10.3390/ijms23010480.

9. Zhai L, Wu J, Lam YY, Kwan HY, Bian ZX, Wong HLX. Gut-Microbial Metabolites, Probiotics and Their Roles in Type 2 Diabetes. Int J Mol Sci. 2021 Nov 27; 22 (23): 12846. doi: 10.3390/ijms222312846.

10. Xie D, Zhao X, Chen M. Prevention and treatment strategies for type 2 diabetes based on regulating intestinal flora. Biosci Trends. 2021 Nov 21; 15 (5): 313–320. doi: 10.5582/bst.2021.01275.

11. Wu J, Yang K, Fan H, Wei M, Xiong Q. Targeting the gut microbiota and its metabolites for type 2 diabetes mellitus. Front Endocrinol (Lausanne). 2023 May 9; 14: 1114424. doi: 10.3389/fendo.2023.1114424.

12. Ahmed SAH, Ansari SA, Mensah‑Brown EPK, Emerald BS. The role of DNA methylation in the pathogenesis of type 2 diabetes mellitus. Clin Epigenetics. 2020 Jul 11; 12 (1): 104. doi: 10.1186/s13148-020-00896-4.

13. Ibrahim HIM. Epigenetic Regulation of Obesity-Associated Type 2 Diabetes. Medicina (Kaunas). 2022 Sep 28; 58 (10): 1366. doi: 10.3390/medicina58101366.

14. Raciti GA, Desiderio A, Longo M, Leone A, Zatterale F, Prevenzano I et al. DNA Methylation and Type 2 Diabetes: Novel Biomarkers for Risk Assessment? Int J Mol Sci. 2021 Oct 28; 22 (21): 11652. doi: 10.3390/ijms222111652.

15. Borse SP, Chhipa AS, Sharma V, Singh DP, Nivsarkar M. Management of Type 2 Diabetes: current strategies, unfocussed aspects, challenges and alternatives. Med Princ Prat. 2021; 30 (2): 109–121. doi: 10.1159/000511002.

16. Salans S. Insulinnezavisimyy sakharnyy diabet: diagnostika i lecheniye. Endokrinologiya / Pod red. M. Lavina. M.: Praktika, 1999: 925–941.

17. Henning RJ. Type-2 diabetes mellitus and cardiovascular disease. Future Cardiol. 2018 Nov; 14 (6): 491–509. doi: 10.2217/fca-2018-0045.

18. Malone JI, Hansen BC. Does obesity cause type 2 diabetes mellitus (T2DM)? Or is it the opposite? Pediatr Diabetes. 2019 Feb; 20 (1): 5–9. doi: 10.1111/pedi.12787.

19. Ruze R, Liu T, Zou X, Song J, Chen Y, Xu R et al. Obesity and type 2 diabetes mellitus: connections in epidemiology, pathogenesis and treatments. Front Endocrinol (Lausanne). 2023 Apr 21; 14: 1161521. doi: 10.3389/fendo.2023.1161521.

20. Tanase DM, Gosav EM, Costea CF, Ciocoiu M, Lacatusu CM, Maranduca MA et al. The Intricate Relationship between Type 2 Diabetes Mellitus (T2DM), Insulin Resistance (IR), and Nonalcoholic Fatty Liver Disease (NAFLD). J Diabetes Res. 2020 Jul 31; 2020: 3920196. doi: 10.1155/2020/3920196.

21. De Lourdes Ochoa‑González F, González‑Curiel IE, Cervantes‑Villagrana AR, Fernández‑Ruiz JC, Castañeda‑Delgado JE. Innate Immunity Alterations in Type 2 Diabetes Mellitus: Understanding Infection Susceptibility. Curr Mol Med. 2021; 21 (4): 318–331. doi: 10.2174/1566524020999200831124534.

22. Golden TN, Simmons RA. Immune dysfunction in developmental programming of type 2 diabetes mellitus. Nat Rev Endocrinol. 2021 Apr; 17 (4): 235–245. doi: 10.1038/s41574-020-00464-z.

23. Babayeva AG, Gevorkyan NM, Zotikov YeA. Peredacha regeneratsionnoy informatsii kak chastnyy sluchay proyavleniya morfogeneticheskoy funktsii limfotsitov (glava 2, s. 30–50). Rol’ limfotsitov v operativnom izmenenii programmy razvitiya tkaney. Moskva: Izd. RAMN, 2009; 107.

24. Stepanova OI, Onishchenko NA, Karkishchenko NN, Baranova OV, Galakhova TV. Ispol’zovaniye kletok raznykh fraktsiy allogennogo kostnogo mozga dlya terapii sakharnogo diabeta 2-go tipa na geneticheskoy modeli. Biomeditsina. 2008; 2: 78–81.

25. Pires IGS, Silva E Souza JA, de Melo Bisneto AV, Passos XS, Carneiro CC. Clinical efficacy of stem cell therapy on diabetes mellitus: A systematic review and meta-analysis. Transpl Immunol. 2022 Dec; 75: 101740. doi: 10.10000016/j.trim.22022.101740.

26. Beer L, Mildner M, Gyongyosi M, Ankersmit HJ. Peripheral blood mononuclear cell secretome for tissue repair. Apoptosis. 2016; 21: 1336–1353. doi: 10.1007/s10495-016-1292-8.

27. Beer L, Zimmermann M, Mitterbauer A, Ellinger A, Gruber F, Narzt MS et al. Analysis of the secretome of apoptotic peripheral blood mononuclear cells: impact of released proteins and exosomes for tissue regeneration. Sci Rep. 2015; 5: 16662. doi: 10.1038/srep16662.

28. Stepanova OI, Alchinova IB, Cherepov AB, Metelkin AA, Karganov MYu, Nikolskaya AO et al. Dynamics of the state of blood cells and bone marrow in mice with the progressive development of type 2 diabetes mellitus. Pathological Physiology and Experimental Therapy, Russian journal. 2024; 68 (1): 98–108. https://doi.org/10.25557/0031-2991.2024.01.98-108.

29. Onishchenko NA, Nikolskaya AO, Gonikova ZZ, Kirsanova LA, Shagidulin MYu, Sevastianov VI. The role of apoptotic bone marrow cells in activation of liver regeneration. Russian Journal of Transplantology and Artificial Organs. 2021; 23 (4): 110–118. https://doi.org/10.15825/25/1995-1191-2021-4-110-118.

30. Stepanova OI, Klesov RA, Semenov KhKh, Pomytkin IA, Onishchenko NA, Karkischenko VN. A method for noninvasive studying tissue disorders in type 2 diabetes mellitus in db/db mice using laser Doppler flowmetry. Pathological Physiology and Experimental Therapy, Russian journal. 2023; 67 (2): 118–129. https://doi.org/10.25557/0031-2991.2023.02.118-129.