Glycemic Control In Diabetic Rats Research Articles

Differentiation of Mesenchymal Stem Cells towards an insulin-releasing phenotype after co-culture with Pancreatic Islets

Transplantation of pancreatic islets has become a promising clinical option to treat patients with type 1 diabetes, alternative to the standard therapy with insulin injections. Islet transplantation is a minimally invasive therapeutic approach, and it allows a better metabolic control and a long-term insulin independence in more than 80% of patients (Ryan et al., 2002). However this therapeutic treatment has some side effects, such as the poor yield of pancreatic islet explants and even more the immune graft rejection, which have as a consequence the very limited lifespan of transplanted pancreatic islets. To avoid these side effects several strategies have been proposed and, besides the treatment with immunosuppressive drugs, promising results have been obtained with the use of Mesenchymal Stem cells (MSCs), already known in literature to be able to support the survival of many cell types (Scuteri et al., 2006). Several in vivo studies have demonstrated that the concurrent transplantation of pancreatic islets with MSCs reduces the number of islets required to achieve glycemic control in diabetic rats, but the mechanisms of these encouraging results are still unknown (Figliuzzi et al., 2009). For these reasons in this in vitro study we characterized the effect of co-culture of rat MSC on survival and functioning of rat pancreatic islets, by evaluating for 4 weeks: i) MSC adhesion to pancreatic islets; ii) viability of pancreatic islets co-cultured with MSCs; iii) the expression of insulin after co-culture; iv) the ability of co-cultured pancreatic islets to correctly adjust insulin release after variation of glucose concentration. Our results demonstrated that MSCs are able to adhere to pancreatic islets, but to increase only partly the pancreatic islet survival, which retain the ability to express and correctly release insulin after glucose variation in medium culture. Noteworthy that the insulin level in the medium of co-cultured pancreatic islets is always higher with respect to medium of pancreatic islets alone. The immunofluorescence analysis reveals that also MSCs (and not only pancreatic islets) are able to express insulin, but only in co-culture. These results, which justify the in vivo observation reported above, suggest that MSCs undergo to differentiation into a insulin-releasing phenotype after co-culture with pancreatic islets. We are now evaluating the molecular mechanisms which drive this effect, by analyzing the role of soluble factors and of proteins able to induce insulin expression. This study was granted by MIUR – FIRB Futuro in Ricerca 2008 RBFR08VSVI_001.

Bone Marrow–Derived Mesenchymal Stem Cells Improve Islet Graft Function in Diabetic Rats

Type 1 diabetes is associated with a progressive loss of β cells and pancreatic islet transplantation could represent a cure for this disease. Herein we explored whether transplantation of bone marrow–derived mesenchymal stem cells (MSCs) allowed a reduced number of pancreatic islets to improve glycemic control in diabetic rats, by promoting islet vascularization. We transplanted 2000 syngenic islets alone or in combination with MSCs (10 6 cells) under the kidney capsules of diabetic Lewis rats. Animals transplanted with 2000 islets never reached normoglycemia. In contrast, rats transplanted with 2000 islets plus MSCs, showed a gradual fall in glycemia after transplantation, with normoglycemia maintained until killing. Comparable glycemic control was obtained with transplantation of 3000 islets alone. The MSC preparation used for in vivo experiments expressed high levels of vascular endothelial growth factor (VEGF 165) and, at less extent, VEGF 189, as evaluated by reverse transcriptase polymerase chain reaction (RT-PCR). In transplanted animals, vascularization was quantified by morphometric analysis of islet grafts with anti–RECA and anti-insulin antibodies. MSCs were stained with PKH-26. Mean capillary density was 1002 ± 55 capillaries/mm 2 in islets transplanted alone. Co-infusion of MSCs with islets significantly increased the number of capillaries to 1459 ± 66 capillaries/mm 2. In conclusion, our study indicated that co-transplantation of MSCs with pancreatic islets improved islet graft function by promoting graft vascularization.

Effects of garlic oil and diallyl trisulfide on glycemic control in diabetic rats

We investigated the effects of garlic oil and diallyl trisulfide on glycemic control in rats with streptozotocin-induced diabetes. Diabetic rats received by gavage garlic oil (100 mg/kg body weight), diallyl trisulfide (40 mg/kg body weight), or corn oil every other day for 3 weeks. Control rats received corn oil only. Both garlic compounds significantly raised the basal insulin concentration. The insulin resistance index as assessed by homeostasis model assessment and the first-order rate constant for glucose disappearance were significantly improved by both garlic compounds ( P < 0.05). Oral glucose tolerance was also improved by both garlic compounds and was accompanied by a significantly increased rate of insulin secretion ( P < 0.05). Glycogen formation (but not that of lactate or carbon dioxide) from glucose by the soleus muscle in the presence of 10 or 100 μU/ml of insulin was significantly better after treatment with both garlic compounds. Both garlic oil and diallyl trisulfide improve glycemic control in diabetic rats through increased insulin secretion and increased insulin sensitivity.

Quantification of Insulin Release from Implantable Polymer-Based Delivery Systems and Augmentation of Therapeutic Effect with Simultaneous Release of Somatostatin

Insulin injections control diabetes mellitus but do not reproduce physiologic regulation. Polymer-based controlled-release technology has enabled us to demonstrate: that the controlled release of insulin from polymer matrices can indeed be used to control diabetes mellitus but does so at the expense of hyperinsulinemia and hypoglycemia; and that somatostatin can be delivered in similar fashion, so as to provide glucose homeostasis in a more physiologic range, at lower insulin levels and at somatostatin doses below those used in intermittent infusion studies; and, that microgram quantities of a drug can be delivered successfully in vivo with intact biological function and in a manner that can be monitored continuously. In the present study the simultaneous polymer-matrix-controlled release of insulin with somatostatin extended glycemic control in diabetic rats. Eleven rats received subcutaneous polymer matrix implants containing insulin alone and 11 rats received implants containing insulin and somatostatin. Plasma and urinary glucose control were improved in both groups. Glucose concentrations in the insulin alone group remained depressed for 5days until insulin release from the matrices declined below 11.6 units/kg/day. When somatostatin was delivered at 0.75–1.1 µg/kg/day together with insulin, plasma glucose control persisted for 12days until insulin release decreased below 3.6 units/kg/day. It is our hope that further experiments regarding the potential role of both controlled-release devices and somatostatin will be performed to provide continuing therapeutic alternatives to the insulin-dependent diabetic. This is also the first in vivo demonstration of the simultaneous release of two biologically active peptide hormones from polymer matrices. The use of the polymer matrix systems may not only have profound effects on the ambulatory care of diabetes but might also permit the investigation of the synergistic effects of other families of compounds.