Clinical Study on the Treatment of Type 2 Diabetes with Bone Marrow Platelet-Rich Plasma (BMPRP)

Objective: To retrospectively analyze the effects of (bone marrow platelet-rich plasma BMPRP) precise pancreatic infusion therapy versus conventional treatment.

Methods: Bone marrow was collected from the iliac crest anterior superior spine, and platelet-rich plasma was separated by centrifugation. BMPRP was infused into the pancreas under ultrasound guidance. It was compared with conventional hypoglycemic drugs and insulin therapy.

Results: In the BMPRP treatment group of 32 cases, the fasting blood sugar and hemoglobin A1c were significantly lower than pre-treatment levels, while the C-peptide level did not change significantly. The insulin dose was reduced. In the conventional treatment group of 28 cases, the fasting blood sugar, hemoglobin A1c, and C-peptide levels did not change significantly after continuous treatment for one year, and the insulin dose was not reduced.

Conclusion: BMPRP precise pancreatic infusion therapy can improve pancreatic function, reduce insulin resistance, lower blood sugar, and reduce the required insulin dosage.

Type 2 diabetes is a metabolic disease caused by insulin resistance and β cell damage caused by over-nutrition and insufficient activity. Conventional treatments for diabetes include hypoglycemic agents and insulin, which still cannot prevent diabetic patients from developing microvascular and macrovascular complications. Stem cells are a kind of cells with multiple differentiation potential, which can differentiate into different cells in different organ microenvironment, or secrete some factors to repair damaged cells and improve organ function. Bone marrow contains multipotent stem cells. Bone marrow platelet-enriched plasma (BMPRP) containing bone marrow stem cells can be separated by density gradient centrifugation. Injecting BMPRP into pancreatic tissue of diabetic patients can improve islet function. This study retrospectively analyzed the clinical data of patients treated with pancreatic infusion of BMPRP and routine treatment, and compared the therapeutic effects to explore the therapeutic effect and principle of BMPRP in treating diabetes.

From January 2022 to December 2023, the medical records of diabetic patients treated with BMPRP in Zigong Hospital affiliated to Southwest Medical University, Renji Hospital of Zhengzhou and Shandong Public Health Clinical Center were analyzed retrospectively. Inclusion criteria: male or female, aged 20-70. Fasting blood sugar is more than 7 mmol, and blood sugar is more than 12 mmol 2 hours after meals. It is necessary to use hypoglycemic agents or insulin to control blood sugar close to normal or still significantly higher than normal, and to exclude patients with evident coagulation disorders.

Treatment

32 cases in BMPRP treatment group, including 17 males and 15 females, aged 31-64 years, with an average age of 54 years. Bone marrow was punctured in the anterior superior iliac crest, and 4 ml heparin saline was pre-filled with 20 mL syringes. Bone marrow was continuously aspirated and collected in five 20 mL syringes. The collected bone marrow was injected into eight 15 ml centrifuge tubes and placed in a centrifuge for density gradient centrifugation. After centrifugation, red blood cells settled at the bottom of the centrifuge tube, plasma and fat cells are located in the upper layer, and nucleated cells and platelets in bone marrow are located in the white membrane layer close to the upper part of the red blood cell layer. First, the plasma from the upper layer of the centrifuge tube was removed, and then 6 ml of bone marrow plasma including bone marrow nucleated cells and platelets was aspirated. Under B-ultrasound guidance, a fine-needle puncture was performed to infuse BMPRP into the pancreatic parenchyma. Then, the bone marrow plasma separated from BMPRP was mixed with red blood cells and reinfused from peripheral veins. Following the initial treatment, a one-month interval was maintained, and then the second and third treatments were done two months. Insulin and hypoglycemic agents were maintained post-treatment, and when the fasting blood sugar drops below 6 mmol, the amount of insulin and hypoglycemic agents should be gradually reduced. 28 patients in the conventional treatment group, including 15 males and 13 females, aged 32-69 years, with an average age of 55 years. Continuous treatment with hypoglycemic agents and insulin.

Statistical methods

SPSS 20.0 statistical software was used for analysis. The chi-square test was used for counting data, and mean ± standard deviation (SD) was used for measurement data. A p - value < 0.05 was considered statistically significant.

Diabetes is a major disease that threatens human health. At present, the commonly used treatment methods are difficult to achieve accurate regulation of blood sugar, which leads to a variety of complications, seriously affecting the quality of life of patients and even endangering their lives. Traditional medical treatment can’t solve the problem of insulin resistance and islet β cell dysfunction from the source. In order to overcome this situation, the current research focus has shifted to the field of stem cell therapy for diabetes [1-5].

There are stem cells in bone marrow [6-9]. Transplantation of bone marrow stem cells into pancreatic microenvironment with islet cell injury may transform bone marrow stem cells into islet β cells, or secrete some cytokines to promote the repair and reconstruction of islet damaged cells, and the regulation function of blood sugar will improve. If autologous bone marrow cells are infused through peripheral veins, most stem cells may stay in the lungs after pulmonary circulation, and a small amount of stem cells may enter the pancreas through systemic circulation. Due to the limited number of stem cells, the improvement of islet function may be minimal. Studies have reported that agents such as granulocyte colony-stimulating factor are used to mobilize bone marrow stem cells into peripheral blood, and then bone marrow stem cells are collected from peripheral blood. Through interventional radiology, bone marrow stem cells can be infused from femoral artery cannula into pancreaticoduodenal artery, which can improve islet function and have a good effect on the treatment of type 2 diabetes. This method further supports the hypothesis that bone marrow stem cells transplanted into pancreatic microenvironment can be transformed into islet β cells to improve islet function [10-13].

We can collect bone marrow from the anterior superior iliac spine under local anesthesia, and we can collect more primitive bone marrow stem cells than those from peripheral blood after applying granulocyte stimulating factor. After collecting bone marrow, density gradient centrifugation is used to stratify bone marrow components. Red blood cells in bone marrow are thrown to the lowest part of the centrifuge tube because they contain the highest specific gravity of hemoglobin. White blood cells in bone marrow include stem cells, and platelets are lighter than red blood cells and heavier than plasma. After centrifugation, it clings to the red blood cell layer. The plasma is in the upper part of the centrifuge tube. First, remove the plasma from the upper part of the centrifuge tube, and then suck out nucleated cells such as stem cells and platelets, including some plasma, which is bone marrow-derived platelet-rich plasma (BMPRP). Some researchers refer to this as bone marrow aspirate concentrate (BMAC) [14], but it should be bone marrow aspirate concentrate after removing red blood cells. BMPRP may be a more precise term in this context.

Under the guidance of B-ultrasound, the No.7 puncture needle is punctured through the upper abdomen, and the fine needle enters the pancreas through the stomach and injects BMPRP into pancreatic islet tissue. Because the density of BMPRP is different from that of pancreatic tissue, ultrasonic waves will echo at different density interfaces, and the ultrasonic probe emits ultrasonic waves and receives the reflected ultrasonic waves. After computer processing, these reflected ultrasonic waves are displayed on the screen in the form of bright spots, so B-ultrasound can show that BMPRP is infused into pancreatic tissue and the brightness of the pancreas is increased. Because the puncture needle is very thin, after infusion of bone marrow cells, Patients could resume movement shortly after needle removal, which is more convenient and less harmful than radiotherapy [15-19].

Comparing the general conditions, gender, age, body type, exercise, diet, alcohol consumption, insulin antibodies, etc. between the BMPRP treatment group and the conventional treatment group showed no significant differences (Table 1). Bone marrow stem cells may differentiate into pancreatic beta cells or secrete certain factors to promote the repair of damaged pancreatic beta cells in the pancreatic microenvironment. Clinical observations indicated a progressive decline in blood glucose levels and glycated hemoglobin levels of patients treated with BMPRP. There was a statistically significant difference in fasting blood glucose and glycated hemoglobin between the two groups (Tables 2,3), but there was no significant difference in C-peptide levels between the two groups (Table 4). This may be attributed to the average C-peptide levels in both groups of type 2 diabetes patients was not significantly low, and some patients had lower C-peptide levels, while others had higher C-peptide levels than normal. After self-bone marrow cell pancreatic infusion treatment, combined with appropriate exercise and control of carbohydrate diet, pancreatic function improved, fasting blood glucose and glycated hemoglobin levels decreased, and some patients’ C-peptide levels also decreased, which may indicate a reduction in insulin resistance, allowing blood sugar to be controlled within a normal range without the need for more insulin [20,21]. Some diabetic patients had lower C-peptide levels prior to BMPRP treatment, and their pancreatic function improved and C-peptide levels increased after treatment, suggesting an increase in insulin secretion. After BMPRP treatment, fasting blood glucose and glycated hemoglobin levels decreased, and insulin use was gradually reduced, with some patients being able to maintain normal blood sugar without insulin after stopping insulin use (Table 5). The conventional treatment group did not see a decrease in blood sugar, and insulin use could not be reduced.

This study conducted a retrospective statistical analysis of clinical data. Randomization was not applicable. The sample size was determined based on the availability of complete medical records. A formal power analysis was not feasible, but the sample was deemed sufficient to detect statistically significant differences in key outcome measures. Due to the absence of some clinical data, statistical analysis was carried out after deleting these incomplete clinical cases. Therefore, the statistical results may have some limitations. In the future, with an increase in the number of enrolled clinical cases, more comprehensive conclusions may be drawn.

After BMPRP pancreatic infusion therapy, blood sugar levels decreased. Insulin sensitivity improved, and insulin resistance decreased. It is an effective treatment for type 2 diabetes.

Authors’ contributions

Baochi Liu conceptualized and designed the study. All authors read and approved the final manuscript.

Ethics approval and consent to participate

This study received ethical approval from the Ethics Committee of the Zigong Hospital, Affiliated with Southwest Medical University. The ethical approval number ZGLCZXEC2022-0314. Written informed consent was obtained from all participants for the use of their samples for the detection and publication of their relevant data.

Patient consent for publication

All participants in this study provided written informed consent for the use of their samples and publication of their data.

1. Memon B, Abdelalim EM. Stem cell therapy for diabetes: beta cells versus pancreatic progenitors. Cells. 2020;9(2):283. Available from: https://doi.org/10.3390/cells9020283
2. Zhang S, Chen L, Zhang G, Zhang B. Umbilical cord-matrix stem cells induce the functional restoration of vascular endothelial cells and enhance skin wound healing in diabetic mice via the polarized macrophages. Stem Cell Res Ther. 2020;11(1):39. Available from: https://doi.org/10.1186/s13287-020-1561-x
3. Su G, Mi S, Tao H, Li Z, Yang H, Zheng H, et al. Association of glycemic variability and the presence and severity of coronary artery disease in patients with type 2 diabetes. Cardiovasc Diabetol. 2011;10:19. Available from: https://doi.org/10.1186/1475-2840-10-19
4. Zhang X, Yang X, Sun B, Zhu C. Perspectives of glycemic variability in diabetic neuropathy: a comprehensive review. Commun Biol. 2021;4(1):1366. Available from: https://doi.org/10.1038/s42003-021-02896-3
5. Li Y, Teng D, Shi X, Qin G, Qin Y, Quan H, et al. Prevalence of diabetes recorded in mainland China using 2018 diagnostic criteria from the American Diabetes Association: national cross sectional study. BMJ. 2020;369:m997. Available from: https://doi.org/10.1136/bmj.m997
6. Shi L, Tee BC, Sun Z. Effects of porcine bone marrow-derived platelet-rich plasma on bone marrow-derived mesenchymal stem cells and endothelial progenitor cells. Tissue Cell. 2021;71:101587. Available from: https://doi.org/10.1016/j.tice.2021.101587
7. Murphy MB, Blashki D, Buchanan RM, Yazdi IK, Ferrari M, Simmons PJ, et al. Adult and umbilical cord blood-derived platelet-rich plasma for mesenchymal stem cell proliferation, chemotaxis, and cryo-preservation. Biomaterials. 2012;33(21):5308-16. Available from: https://doi.org/10.1016/j.biomaterials.2012.04.007
8. Park G, Hwang DY, Kim DY, Han JY, Lee E, Hwang H, et al. Identification of CD141+ vasculogenic precursor cells from human bone marrow and their endothelial engagement in the arteriogenesis by co-transplantation with mesenchymal stem cells. Stem Cell Res Ther. 2024;15(1):388. Available from: https://doi.org/10.1186/s13287-024-03994-9
9. Nammian P, Asadi-Yousefabad SL, Daneshi S, Sheikhha MH, Tabei SMB, Razban V. Comparative analysis of mouse bone marrow and adipose tissue mesenchymal stem cells for critical limb ischemia cell therapy. Stem Cell Res Ther. 2021;12(1):58. Available from: https://doi.org/10.1186/s13287-020-02110-x
10. Peng X, Liang B, Wang H, Hou J, Yuan Q. Hypoxia pretreatment improves the therapeutic potential of bone marrow mesenchymal stem cells in hindlimb ischemia via upregulation of NRG-1. Cell Tissue Res. 2022;388(1):105-116. Available from: https://doi.org/10.1007/s00441-021-03562-0
11. Battelino T, Danne T, Bergenstal RM, Amiel SA, Beck R, Biester T, Bosi E, et al. Clinical targets for continuous glucose monitoring data interpretation: recommendations from the international consensus on time in range. Diabetes Care. 2019;42(8):1593-1603. Available from: https://doi.org/10.2337/dci19-0028
12. Sassoli C, Vallone L, Tani A, Chellini F, Nosi D, Zecchi-Orlandini S. Combined use of bone marrow-derived mesenchymal stromal cells (BM-MSCs) and platelet rich plasma (PRP) stimulates proliferation and differentiation of myoblasts in vitro: new therapeutic perspectives for skeletal muscle repair/regeneration. Cell Tissue Res. 2018;372(3):549-570. Available from: https://doi.org/10.1007/s00441-018-2792-3
13. Lu J, Wang C, Shen Y, Chen L, Zhang L, Cai J, et al. Time in range in relation to all-cause and cardiovascular mortality in patients with type 2 diabetes: a prospective cohort study. Diabetes Care. 2021;44(2):549-555. Available from: https://doi.org/10.2337/dc20-1862
14. Lee JS, Gillinov SM, Siddiq BS, Dowley KS, Martin SD. Surgical applications for bone marrow aspirate concentrate. Arthroscopy. 2024;40(9):2350-2352. Available from: https://doi.org/10.1016/j.arthro.2024.05.002
15. Li L, Si Y, Cheng M, Lang L, Li A, Liu B. Therapeutic effect of autologous bone marrow cells injected into the liver under the guidance of B ultrasound in the treatment of HBV-related decompensated liver cirrhosis. Exp Ther Med. 2022;24:633. Available from: https://doi.org/10.3892/etm.2022.11570
16. Liu BC, Cheng MR, Lang L, Li L, Si YH, Li AJ, et al. Autologous bone marrow infusion via portal vein combined with splenectomy for decompensated liver cirrhosis: A retrospective study. World J Gastrointest Surg. 2023;15(9):1919-1931. Available from: https://doi.org/10.4240/wjgs.v15.i9.1919
17. Liu B, Gao X, Chen Y, Dong Q, Wang J, Zhao B. B-ultrasound-guided intrahepatic infusion of autologous bone marrow cells for decompensated cirrhosis. Int J Bone Marrow Res. 2024;7(1):001-006. Available from: https://dx.doi.org/10.29328/journal.jbmr.1001017
18. Liu B, Gao X, Chen Y, Zheng R, Dong Q, Wang J. Analysis of clinical data on the treatment of type 2 diabetes with BMPRP. Am J Biosci Bioeng. 2024;12(6):128-134. Available from: https://doi.org/10.11648/j.bio.20241206.14
19. Wang X, Zhao X, Dorje T, Yan H, Qian J, Ge J. Glycemic variability predicts cardiovascular complications in acute myocardial infarction patients with type 2 diabetes mellitus. Int J Cardiol. 2014;172(2):498-500. Available from: https://doi.org/10.1016/j.ijcard.2014.01.015
20. Ceriello A, Esposito K, Piconi L, Ihnat MA, Thorpe JE, Testa R, et al. Oscillating glucose is more deleterious to endothelial function and oxidative stress than mean glucose in normal and type 2 diabetic patients. Diabetes. 2008;57(5):1349-1354. Available from: https://doi.org/10.2337/db08-0063
21. Saboo B, Kesavadev J, Shankar A, Krishna MB, Sheth S, Patel V, et al. Time-in-range as a target in type 2 diabetes: an urgent need. Heliyon. 2021;7(1):e05967. Available from: https://doi.org/10.1016/j.heliyon.2021.e05967