Insulin Release In Response Research Articles

Sex-specific differences in insulin response and substrate oxidation after repeated, brief whole-body immersion in 45°C water: A prospective, interventional study.

Prolonged heat exposure is suggested to improve glucose metabolism and fat oxidation, but no studies have addressed whether brief heat stimuli represent a viable, time-efficient, alternative approach. Consequently, we examined the ability of brief stimuli evoked by 45°C water to improve glucose tolerance, insulin sensitivity, and fat oxidation in young, non-obese, males and females. Twenty-four participants completed fourteen 5-min sessions involving whole body passive heating in 45°C water. Changes in resting catecholamines, cytokines, substrate oxidation, resting energy expenditure, glucose tolerance, and insulin release in response to an oral glucose tolerance test, were assessed before and 24-h after intervention, and 1 month after the end of the intervention. The results showed that repeated short-duration heat intervention had no significant effects on epinephrine, norepinephrine, interleukin-6, and tumor necrosis factor alpha production in both sexes. Glucose area under the curve (AUC) was not affected. However, females had a lower insulin AUC and improved insulin sensitivity as indicated by a decrease in homeostatic model assessment for insulin resistance, and an increase in the quantitative insulin sensitivity check index and the Matsuda insulin sensitivity index values one month after the end of the heat intervention. No effect was observed in resting energy expenditure, but carbohydrate oxidation per kilogram increased in females, and this substrate oxidation change was maintained after one month. In conclusion, fourteen sessions of brief 5-min whole-body immersion in 45°C water produced an improvement in insulin sensitivity and increased reliance on carbohydrate oxidation in females.

Iontophoresis-Integrated Smart Microneedle Delivery Platform for Efficient Transdermal Delivery and On-Demand Insulin Release.

Transdermal insulin delivery in a painless, convenient, and on-demand way remains a long-standing challenge. A variety of smart microneedles (MNs) fabricated by glucose-responsive phenylboronic acid hydrogels have been previously developed to provide painless and autonomous insulin release in response to a glucose level change. However, like the majority of MNs, their transdermal delivery efficiency was still relatively low compared to that with subcutaneous injection. Herein, we report an iontophoresis (ITP)-integrated smart MNs delivery platform with enhanced transdermal delivery efficiency and delivery depth. Carbon nanotubes (CNTs) were induced in the boronate-containing hydrogel to develop a semi-interpenetrating network hydrogel with enhanced stiffness and conductivity. Remarkably, ITP not only facilitated efficient and deeper transdermal delivery of insulin via electroosmosis and electrophoresis but also well-maintained glucose responsiveness. This ITP-combined smart MNs delivery platform, which could provide on-demand insulin delivery in a painless, convenient, and safe way, is promising to achieve persistent glycemic control. Furthermore, transdermal delivery of payloads with a wide size range was achieved by this delivery platform and thus shed light on the development of an efficient transdermal delivery platform with deep skin penetration in a minimally invasive way.

Incretin mimetics for the management of diabetes and associated comorbidities: An overview

Type 2 diabetes mellitus is commonly associated with various comorbidities that aggravate the disease’s overall impact on health. The most prevalent comorbidities of diabetes include obesity, dyslipidemia, hypertension, cardiovascular conditions, and kidney diseases. Incretin mimetics, also known as glucagon-like peptide-1 receptor agonists, mimic incretin hormones to stimulate insulin release in response to food intake. These medications help lower blood glucose by increasing insulin production, reducing glucagon secretion, slowing stomach emptying, and promoting satiety. A key advantage of incretin mimetics is their ability to reduce blood glucose levels without causing hypoglycemia, making them a safer option for many patients. They also promote weight loss, which is particularly beneficial for patients with both obesity and diabetes. Incretin mimetics are typically administered once or twice daily and are often used in combination with other treatments such as metformin or insulin. Evidence suggests that these drugs may reduce the risk of heart attack and stroke, an important consideration given the heightened cardiovascular risk in patients with diabetes. Additionally, incretin mimetics may help preserve pancreatic beta-cell function, potentially slowing the progression of diabetes. However, these drugs are costly and may be unaffordable for low-income individuals. Commonly reported side effects include nausea, vomiting, and diarrhea, which tend to decrease over time. While there have been reports of pancreatitis, current research indicates that incretin mimetics do not increase the risk of pancreatic cancer. Educating patients on proper use and potential side effects is crucial to ensure safe and effective treatment with incretin mimetics.

Acquisition of durable insulin-producing cells from human adipose tissue-derived mesenchymal stem cells as a foundation for cell- based therapy of diabetes mellitus

This study aimed to identify the suitable induction protocol to produce highly qualified insulin producing cells (IPCs) from human adipose tissue derived stem cells (ADSCs) and evaluate the efficacy of the most functionally IPCs in management of diabetes mellitus (DM) in rats. The ADSCs were isolated and characterized according to the standard guidelines. ADSCs were further induced to be IPCs in vitro using three different protocols. The success of trans-differentiation was assessed in vitro through analysis of pancreatic endocrine genes expression, and insulin release in response to glucose stimulation. Then, the functionalization of the generated IPCs was evaluated in vivo. The in vitro findings revealed that the laminin-coated plates in combination with insulin-transferrin-selenium, B27, N2, and nicotinamide could efficiently up-regulate the expression of pancreatic endocrine genes. The in vivo study indicated effectual homing of the PKH-26-labelled IPCs in the pancreas of treated animals. Moreover, IPCs infusion in diabetic rats induced significant improvement in the metabolic parameters and prompted considerable up-regulation in the expression of the pancreatic related genes. The regenerative effect of infused IPCs was determined through histological examination of pancreatic tissue. Conclusively, the utilization of laminin–coated plates in concomitant with extrinsic factors promoting proliferation and differentiation of ADSCs could efficiently generate functional IPCs.

Glucagon -like Peptide 1 Receptor Agonist Use and the Effect on Diabetic Retinopathy: An Uncertain Relationship

Glucagon-like Peptide 1 Receptor Agonists (GLP-1 RAs) are a group of relatively novel medications for the treatment of diabetes mellitus. These medications can mimic the naturally occurring incretins of the body, which promote the release of insulin in response to hyperglycaemia. The anti-glycaemic effects of these medications can be profound and carry other metabolic benefits such as promoting weight loss. Clinical trials have shown GLP-1 RAs are safe to use from a cardiovascular perspective. However, some trials have suggested a link between GLP-1 RA use and worsening diabetic retinopathy. The conclusions surrounding this link are poorly established as data is drawn primarily from cardiovascular outcome trials. If an association does exist, a possible explanation might be the observed phenomenon of early worsening diabetic retinopathy with rapid correction of hyperglycaemic states. Trials which look at diabetic retinopathy as a primary outcome in relation to use of GLP-1 RAs are sparce and warrant investigation given the growing use of this group of medications. Therefore currently, it is uncertain what effect, beneficial or adverse, GLP-1 RA use has on diabetic retinopathy. This article provides an overview of GLP-1 RA use as a treatment for diabetes mellitus and the current understanding of their relationship with diabetic retinopathy.

Could Insulin Be a Better Regulator of Appetite/Satiety Balance and Body Weight Maintenance in Response to Glucose Exposure Compared to Sucrose Substitutes? Unraveling Current Knowledge and Searching for More Appropriate Choices.

Insulin exerts a crucial impact on glucose control, cellular growing, function, and metabolism. It is partially modulated by nutrients, especially as a response to the intake of foods, including carbohydrates. Moreover, insulin can exert an anorexigenic effect when inserted into the hypothalamus of the brain, in which a complex network of an appetite/hunger control system occurs. The current literature review aims at thoroughly summarizing and scrutinizing whether insulin release in response to glucose exposure may be a better choice to control body weight gain and related diseases compared to the use of sucrose substitutes (SSs) in combination with a long-term, well-balanced diet. This is a comprehensive literature review, which was performed through searching in-depth for the most accurate scientific databases and applying effective and relevant keywords. The insulin action can be inserted into the hypothalamic orexigenic/anorexigenic complex system, activating several anorexigenic peptides, increasing the hedonic aspect of food intake, and effectively controlling the human body weight. In contrast, SSs appear not to affect the orexigenic/anorexigenic complex system, resulting in more cases of uncontrolled body weight maintenance while also increasing the risk of developing related diseases. Most evidence, mainly derived from in vitro and in vivo animal studies, has reinforced the insulin anorexigenic action in the hypothalamus of the brain. Simultaneously, most available clinical studies showed that SSs during a well-balanced diet either maintain or even increase body weight, which may indirectly be ascribed to the fact that they cannot cover the hedonic aspect of food intake. However, there is a strong demand for long-term longitudinal surveys to effectively specify the impact of SSs on human metabolic health.

Synthesis of multi-responsive poly(NIPA-co-DMAEMA)-PBA hydrogel nanoparticles in aqueous solution for application as glucose-sensitive insulin-releasing nanoparticles.

This study aimed to present an innovative method for synthesizing pH-thermo-glucose responsive poly(NIPA-co-DMAEMA)-PBA hydrogel nanoparticles via single-step aqueous free radical polymerization. The synthesis process involved free radical polymerization in an aqueous solution, and the resulting nanoparticles were characterized for their physical and chemical properties by 1H NMR, Dynamic Light Scattering (DLS) and Scanning Electron Microscopy (SEM). Insulin-loaded poly(NIPA-co-DMAEMA)-PBA hydrogel nanoparticles were prepared and evaluated for their insulin capture and release properties at different pH and temperature, in addition to different glucose concentrations, with the release profile of insulin quantitatively evaluated using the Bradford method. 1H NMR results confirmed successful PBA incorporation, and DLS outcomes consistently indicated a transition to a more hydrophobic state above the Lower Critical Solution Temperature (LCST) of NIPA and DMAEMA. While pH responsiveness exhibited variation, insulin release generally increased with rising pH from acidic to neutral conditions, aligning with the anticipated augmentation of anionic PBA moieties and increased hydrogel hydrophilicity. Increased insulin release in the presence of glucose, particularly for formulations with the lowest mol % PBA, along with a slight increase for the highest mol % PBA formulation when increasing glucose from 1 to 4mg/mL, supported the potential of this approach for nanoparticle synthesis tailored for glucose-responsive insulin release. This work successfully demonstrates a novel method for synthesizing responsive hydrogel nanoparticles and underscores their potential for controlled insulin release in response to glucose concentrations. The observed pH-dependent insulin release patterns and the influence of PBA content on responsiveness highlight the versatility and promise of this nanoparticle synthesis approach for applications in glucose-responsive drug delivery systems. Poly(NIPA) nanoparticles containing PBA moieties are normally synthesized in two or more steps in the presence of organic solvents. Here we propose a new method for the synthesis of multiresponsive hydrogel poly(NIPA-co-DMAEMA)-PBA nanoparticles in aqueous medium in a single reaction to provide a fast and effective strategy for the production of glucose-responsive multi-systems in aqueous media from free radical polymerization.

Association of glucose metabolism and insulin resistance with feed efficiency and production traits of finishing beef steers.

Increasing nutrient utilization efficiency is an important component of enhancing the sustainability of beef cattle production. The objective of this experiment was to determine the association of glucose metabolism and insulin resistance with dry matter intake (DMI), average daily gain (ADG), gain:feed ratio (G:F), and residual feed intake (RFI). Steers (n = 54; initial body weight = 518 ± 27.0kg) were subjected to an intravenous glucose tolerance test (IVGTT) where glucose was dosed through a jugular catheter and serial blood samples were collected. Three days after the last group's IVGTT, steers began a 63-d DMI and ADG test. Body weight was measured on days 0, 1, 21, 42, 62, and 63, and DMI was measured using an Insentec Roughage Intake Control system (Hokofarm Group, Emmeloord, the Netherlands). To examine relationships between DMI, ADG, G:F, and RFI with IVGTT measurements, Pearson correlations were calculated using Proc Corr of SAS 9.4 (SAS Inst. Inc., Cary, NC). Additionally, cattle were classified based on DMI, ADG, RFI, and G:F, where the medium classification was set as mean ± 0.5 SD, the low classification was < 0.5 SD from the mean, and the high classification was > 0.5 SD from the mean. No associations between DMI and IVGTT parameters were observed, and no differences were detected when classifying cattle as having low, medium, or high DMI. Peak insulin concentration in response to the IVGTT tended to be correlated with ADG (r = 0.28; P = 0.07), indicating cattle with greater ADG tend to have a greater insulin release in response to glucose. Glucose nadir concentrations tended to be positively correlated with ADG (r = 0.26; P = 0.10). Additionally, the glucose nadir was greater in high-ADG steers (P = 0.003). The association of greater glucose nadir with high-ADG could indicate that high-ADG steers do not clear glucose as efficiently as low-ADG steers, potentially indicating increased insulin resistance. Further, RFI was not correlated with IVGTT measurements, but low RFI steers had a greater peak glucose concentration (P = 0.040) and tended to have a greater glucose area under the curve (P = 0.09). G:F was correlated with glucose area under the curve (r = 0.33; P = 0.050), glucose nadir (r = 0.35; P = 0.011), and insulin time to peak (r = 0.39; P = 0.010). These results indicate that glucose metabolism and insulin signaling are associated with growth and efficiency, but the molecular mechanisms that drive these effects need to be elucidated.

Beta-cell mass adaptation to ileum nutrient flow. An experimental model.

The population with obesity has increased at an alarming rate during this century. Bariatric surgery has been demonstrated to be a good method to control weight and, most importantly, associated comorbidities, such as type 2 diabetes mellitus or high blood pressure. The reason why this happens even before losing significant weight remains unclear. Many authors believe that incretins play a main role, triggering special functions of the digestive tract. In reports, these hypotheses are known as foregut and hindgut theories. Initially, the theories were mutually exclusive; additionally, many other propositions have been analysed, according to different surgical techniques (e.g., bile acids and specific enterohormonal components). To elucidate the participation of the ileum, we developed a surgical technique to study the rapid response to nutrients in the ileum. Our goal was to study the stress functional test and histological changes in the pancreas that may explain the variations in glycaemic homeostasis in our rat model. After the oral glucose tolerance test, the experimental group presented an increased insulin release response with conserved glycaemia. We report an increasing beta-cell mass in the experimental group (+11.87 mg vs. +9.65 mg, respectively), while alpha-cell mass was not different. Based on transcription factors, the pathways that were increased were the proliferation process (as the number of PCNA-positive cells in the experimental group versus sham (+12.06 vs. +6.2 PCNA+ cells/mm²)) and transdifferentiation (ARX; +2.67 ARX+ cells/mm² in the experimental group vs. +2.04 ARX+ cells/mm² in the controls). We report the consequences of the rapid arrival of nonprocessed nutrients to the ileum on the endocrine cellular pancreas. The ileum could be a principal effector in the enterohormonal axis, which conditions endocrine pancreas cellularity.

Recent Progress in Diboronic-Acid-Based Glucose Sensors.

Non-enzymatic sensors with the capability of long-term stability and low cost are promising in glucose monitoring applications. Boronic acid (BA) derivatives offer a reversible and covalent binding mechanism for glucose recognition, which enables continuous glucose monitoring and responsive insulin release. To improve selectivity to glucose, a diboronic acid (DBA) structure design has been explored and has become a hot research topic for real-time glucose sensing in recent decades. This paper reviews the glucose recognition mechanism of boronic acids and discusses different glucose sensing strategies based on DBA-derivatives-based sensors reported in the past 10 years. The tunable pKa, electron-withdrawing properties, and modifiable group of phenylboronic acids were explored to develop various sensing strategies, including optical, electrochemical, and other methods. However, compared to the numerous monoboronic acid molecules and methods developed for glucose monitoring, the diversity of DBA molecules and applied sensing strategies remains limited. The challenges and opportunities are also highlighted for the future of glucose sensing strategies, which need to consider practicability, advanced medical equipment fitment, patient compliance, as well as better selectivity and tolerance to interferences.

Enhanced Endosomal Signaling and Desensitization of GLP-1R vs GIPR in Pancreatic Beta Cells.

The incretin receptors, glucagon-like peptide-1 receptor (GLP-1R) and glucose-dependent insulinotropic polypeptide receptor (GIPR), are prime therapeutic targets for the treatment of type 2 diabetes (T2D) and obesity. They are expressed in pancreatic beta cells where they potentiate insulin release in response to food intake. Despite GIP being the main incretin in healthy individuals, GLP-1R has been favored as a therapeutic target due to blunted GIPR responses in T2D patients and conflicting effects of GIPR agonists and antagonists in improving glucose tolerance and preventing weight gain. There is, however, a recently renewed interest in GIPR biology, following the realization that GIPR responses can be restored after an initial period of blood glucose normalization and the recent development of dual GLP-1R/GIPR agonists with superior capacity for controlling blood glucose levels and weight. The importance of GLP-1R trafficking and subcellular signaling in the control of receptor outputs is well established, but little is known about the pattern of spatiotemporal signaling from the GIPR in beta cells. Here, we have directly compared surface expression, trafficking, and signaling characteristics of both incretin receptors in pancreatic beta cells to identify potential differences that might underlie distinct pharmacological responses associated with each receptor. Our results indicate increased cell surface levels, internalization, degradation, and endosomal vs plasma membrane activity for the GLP-1R, while the GIPR is instead associated with increased plasma membrane recycling, reduced desensitization, and enhanced downstream signal amplification. These differences might have potential implications for the capacity of each incretin receptor to control beta cell function.

Glucose-Responsive Injectable Thermogels via Dynamic-Covalent Cross-Linking of Pluronic Micelles.

Stimuli-responsive hydrogels are an area of active discovery for approaches to deliver therapeutics in response to disease-specific indicators. Glucose-responsive delivery of insulin is of particular interest in better managing diabetes. Accordingly, hydrogels have been explored as platforms that enable both a rate and dose of insulin release aligning with the real-time physiological disease state; materials often include glucose sensing by dynamic-covalent cross-linking between phenylboronic acids (PBAs) and diols, with competition from ambient glucose reducing cross-link density of the material and accelerating release of encapsulated insulin. Yet, these materials historically have challenges with insulin leakage, offer limited glucose-responsive release of the insulin payload, and require unreasonably high injection pressures for syringe administration. Here, a thermogel platform prepared from temperature-induced micelles formed into a network by PBA-Diol cross-linking is optimized using a formulation-centered approach to maximize glucose-responsive insulin delivery. Importantly, the dual-responsive nature of this platform enables a low-viscosity sol at ambient temperature for facile injection, solidifying into a stable viscoelastic hydrogel network once in the body. The final optimized formulation affords acceleration in insulin release in response to glucose and enables single dose blood glucose control in diabetic rodents when subjected to multiple glucose challenges.

Oral Glucose-Responsive Nanocarrier System for Management of Diabetes

Background: The regulation of postprandial blood glucose concentration in a diabetic person is a toilsome task by the conventional approaches. The continuous monitoring and subcutaneous injection of insulin are always imperative for diabetes management. To overcome the hurdle associated with the conventional system, a glucose-responsive nanoparticle (NP)-based approach was developed for oral insulin administration. The objective of the work is the formulation and characterization of a nanocarrier-based bio-responsive, self-regulated oral insulin drug delivery system. Methods: The mannose ligand-conjugated NPs were prepared by the inotropic gelation method in which insulin and glucose oxidase enclosed alginate nanocarrier cross-linked by tripolyphosphate. The prepared NPs were filled into the enteric-coated capsule and their release profile was determined at different pH and glucose concentrations. NPs were characterized by size distribution, % drug entrapment, enzyme activity, and in vivo and in vitro drug release studies. Results: The average size, surface potential, % drug entrapment of optimized NPs were 245.52 ± 3.37 nm, 22.12 ± 2.13, and 76.15±1.3%, respectively. The release of insulin from optimized NPs (GOx-ALG-MCNPs) (98.56±0.06%) occurs only at higher sugar concentrations (? 400 mg/dL) and prevents the release at low sugar concentrations (&lt; 100 mg/dL). The enteric capsule shell protected the formulation to the gastric environment upon oral administration and directed them into the intestine. Mucoadhesion characteristics of NPs prolonged intestinal retention to make them available for cellular uptake. Glucose sensitivity and glucose-dependent effect on different pH environments study revealed that NPs released the drug only at higher concentrations of sugar solution while inhibiting the release at normal and lower sugar concentrations. Besides, compared to other prepared formulations, the GOx-ALG-MCNPs displayed controlled insulin release in response to glucose concentration. Conclusions: An important finding of GOx-ALG-MCNPs is auto-regulating, glucose-responsive behavior imparting a significantly controlled lowering of blood glucose in an animal model. The findings reveal that the developed glucose-responsive NPs may prove to be a potential strategy for oral insulin delivery. J Endocrinol Metab. 2022;12(4-5):146-160 doi: https://doi.org/10.14740/jem747

Recent Advances of Diabetic Screening and Treatment

Blood glucose lever monitoring is an essential routine work for patients who are diagnosed with diabetes including type I and II. Insulin therapy plays a crucial role in treatment for both types of diabetes, whereas insulin delivery system has limited the capacities of insulin-loading, even worse, is not able to manufacture easily. Existing machines for measuring blood glucose are cumbersome, in addition, people need an injection of insulin for blood glucose regulation. The delivery route imposes restrictions on portability, the transdermal alternative seems favorable for the drug delivery. Naked-eye glucose monitoring base on colloidal crystal (GCC)-MN, aim to enhance health, improve portability and minimize invasiveness. This review discussed the sophisticated painless device named colloidal crystal (GCC)-MN patch for diabetes treatment and the process of glucose monitoring and responsive insulin release.

PSII-B-12 Association of Glucose Metabolism with Feed Intake, Growth, and Efficiency of Beef Steers

Abstract The objective of this experiment was to determine the association of glucose metabolism and insulin sensitivity with dry matter intake (DMI), average daily gain (ADG), and feed efficiency (G:F) of steers fed a finishing diet. Steers (n = 55; initial body weight 518 ± 26.7 kg) were subjected to an intravenous glucose tolerance test (IVGTT) where glucose was dosed through a jugular catheter and serial blood samples were collected. Three days after the final IVGTT, steers began a 63-d DMI and ADG test. Body weight was measured on d 0, 1, 21, 42, 62, and 63, and DMI was measured using an Insentec Roughage Intake Control system (Insentec, Markenesse, The Netherlands). Peak insulin concentration in response to the IVGTT tended to be correlated with ADG (r = 0.25; P = 0.07) and DMI (r = 0.23; P = 0.09), indicating cattle with greater ADG and DMI tend to require a greater insulin release in response to glucose. Glucose nadir concentrations were correlated with ADG (r = 0.27; P = 0.044) and G:F (r = 0.37; P = 0.005). Additionally, when steers within 0.5 standard deviations of the mean for DMI are classed as displaying high- or low-ADG, glucose nadir was greater in high-ADG steers (P = 0.042). The association of greater glucose nadir with high-ADG could indicate that high-ADG steers do not clear glucose as efficiently as low-ADG steers, potentially indicating reduced insulin sensitivity. Carcass marbling score was negatively correlated with area under the glucose curve (r = -0.38; P = 0.006), glucose clearance rate (r = -0.33; P = 0.016), and glucose nadir concentration (r = -0.40; P = 0.004). These results indicate that glucose metabolism and insulin signaling are associated with growth, efficiency, and carcass quality, but the molecular mechanisms that drive these effects need to be elucidated.

In depth functional characterization of human induced pluripotent stem cell-derived beta cells in vitro and in vivo.

In vitro differentiation of human induced pluripotent stem cells (iPSCs) into beta cells represents an important cell source for diabetes research. Here, we fully characterized iPSC-derived beta cell function in vitro and in vivo in humanized mice. Using a 7-stage protocol, human iPSCs were differentiated into islet-like aggregates with a yield of insulin-positive beta cells comparable to that of human islets. The last three stages of differentiation were conducted with two different 3D culture systems, rotating suspension or static microwells. In the latter, homogeneously small-sized islet-like aggregates were obtained, while in rotating suspension size was heterogeneous and aggregates often clumped. In vitro function was assessed by glucose-stimulated insulin secretion, NAD(P)H and calcium fluctuations. Stage 7 aggregates slightly increased insulin release in response to glucose in vitro. Aggregates were transplanted under the kidney capsule of NOD-SCID mice to allow for further in vivo beta cell maturation. In transplanted mice, grafts showed glucose-responsiveness and maintained normoglycemia after streptozotocin injection. In situ kidney perfusion assays showed modulation of human insulin secretion in response to different secretagogues. In conclusion, iPSCs differentiated with equal efficiency into beta cells in microwells compared to rotating suspension, but the former had a higher experimental success rate. In vitro differentiation generated aggregates lacking fully mature beta cell function. In vivo, beta cells acquired the functional characteristics typical of human islets. With this technology an unlimited supply of islet-like organoids can be generated from human iPSCs that will be instrumental to study beta cell biology and dysfunction in diabetes.

Fabrication of bioartificial pancreas using decellularized rat testicular tissue

AimsDiabetes is a chronic disease that is associated with a decrease or disfunction of β-cell. In the present study, fabrication of bioartificial pancreas using MIN-6 β-cell line seeded in decellularized rat testicles was investigated. Main methodsIn this experimental study, the whole body of testes were decellularized and after characterization, were seeded by MIN-6 cell line. The expression of insulin-related genes and proteins including PDX-1, Glut2, Insulin, and Neurogenin-3 were evaluated. Insulin secretion was assessed under different concentrations of glucose. Seeded scaffolds with or without MIN-6 cells were transplanted to the rat's mesentery and their blood sugar and body weight were evaluated every three days for 28 days and analyzed with H&E staining. ResultsHistological assessments indicated the cells were completely removed after decellularization. The scaffold had no toxic impacts on the MIN-6 cells (P˂ 0.02).Insulin release in response to different concentrations of glucose in 3D culture (testis-ECM) was significantly more than the traditional 2D monolayer culture (P < 0.001). Moreover, the relative genes and proteins expression were significantly higher in the 3D culture, compared to the 2D control group. In vivo transplantation of the (testis- Extra Cellular Matrix) testis-ECM scaffolds showed appropriate positions for transplantation with angiogenesis and low infiltration of inflammatory cells. The recellularized scaffolds could drop blood sugar levels and increase the body-weight of STZ-diabetic rats (P < 0.01). SignificanceOur study clearly confirmed that ECM valuable organ scaffolds prepared by decellularization of the testicular tissue is suitable for the fabrication of bioartificial pancreas for transplantation.

Insulinotropic Effects of Neprilysin and/or Angiotensin Receptor Inhibition in Mice.

Treatment of heart failure with the angiotensin receptor-neprilysin inhibitor sacubitril/valsartan improved glycemic control in individuals with type 2 diabetes. The relative contribution of neprilysin inhibition versus angiotensin II receptor antagonism to this glycemic benefit remains unknown. Thus, we sought to determine the relative effects of the neprilysin inhibitor sacubitril versus the angiotensin II receptor blocker valsartan on beta-cell function and glucose homeostasis in a mouse model of reduced first-phase insulin secretion, and whether any beneficial effects are additive/synergistic when combined in sacubitril/valsartan. High fat-fed C57BL/6J mice treated with low-dose streptozotocin (or vehicle) were followed for eight weeks on high fat diet alone or supplemented with sacubitril, valsartan or sacubitril/valsartan. Body weight and fed glucose levels were assessed weekly. At the end of the treatment period, insulin release in response to intravenous glucose, insulin sensitivity, and beta-cell mass were determined. Sacubitril and valsartan, but not sacubitril/valsartan, lowered fasting and fed glucose levels and increased insulin release in diabetic mice. None of the drugs altered insulin sensitivity or beta-cell mass, but all reduced body weight gain. Effects of the drugs on insulin release were reproduced in angiotensin II-treated islets from lean C57BL/6J mice, suggesting the insulin response to each of the drugs is due to a direct effect on islets and mechanisms therein. In summary, sacubitril and valsartan each exert beneficial insulinotropic, glycemic and weight-reducing effects in obese and/or diabetic mice when administered alone; however, when combined, mechanisms within the islet contribute to their inability to enhance insulin release.

Deletion of the Vitamin D Receptor in Skeletal Muscle is Associated with Improved Glucose Tolerance and Reduced Muscle Function

Skeletal muscle insulin resistance contributes to the pathogenesis of type 2 diabetes mellitus. Epidemiological studies have demonstrated that vitamin D deficiency is associated with the presence of type 2 diabetes in humans. To probe the role of the vitamin D receptor (VDR), an intracellular and nuclear protein that mediates the bioactivity of the active vitamin D ligand, 1,25‐dihydroxyvitamin D, in skeletal muscle mediated glucose disposal, we deleted the skeletal muscle VDR in adult mice. We assessed glucose disposal, insulin concentrations and sensitivity, and skeletal muscle gene expression in Vdr knockout and wild‐type Vdr mice. Following the induction of a skeletal muscle‐specific Crerecombinase with tamoxifen in adult male Vdrfl/flmice, efficient deletion of the Vdrwas demonstrated. In skeletal muscle‐specific Vdrknockout mice, basal glucose concentrations were similar to those in wild‐type Vdrmice. In Vdrknockout mice, blood glucose concentrations decreased more rapidly following the administration of intraperitoneal glucose than in control mice expressing wild‐type Vdr (Figure 1A and Figure 1B). The AUC for glucose concentrations in Vdrfl/fl.actin iCre mice treated with tamoxifen was reduced by 20% (p=0.015) compared to the AUC of blood glucose concentrations prior to tamoxifen treatment. Pre‐and post‐treatment AUC’s for blood glucose in Vdrfl/fl.actin iCre mice treated with vehicle and in Vdrfl/flmice treated with tamoxifen were unchanged (p=0.97,0.23). Plasma insulin concentrations were identical in knockout and wild‐type Vdr mice following a bolus of glucose, demonstrating that deletion of the Vdr in skeletal muscle did not influence the release of insulin in response to glucose (one‐way ANOVA, p=0.45). Insulin sensitivity was similar in Vdrknockout and wild‐type Vdrmice. Gene expression analysis of skeletal muscle mRNA derived from Vdrknockout mice and wild‐type mice demonstrated an increase in the expression of genes associated with oxidative phosphorylation, fatty acid oxidation and adipogenesis pathways that could potentially alter glucose utilization. We conclude that selective deletion of the Vdr in the skeletal muscle of adult mice results in more efficient glucose disposal than in wild‐type Vdrmice; basal and glucose‐stimulated insulin serum concentrations are identical in knock‐out and wild type VDR mice with no changes in insulin sensitivity. The data suggest that modulation of the vitamin‐D endocrine system is unlikely to improve glucose utilization.

Elevated Glucose Negatively Regulates Nkx6.1 Protein Level in the Pancreatic Beta Cell

Millions of people suffer from T2D. A key characteristic of T2D is damage to the pancreatic beta cell. This damage affects the beta cell’s ability to sense glucose, produce insulin, and release insulin in response to elevated blood glucose levels. Homeobox protein Nkx6.1 is a beta cell transcription factor essential for beta cell differentiation, proliferation, and insulin secretion. To test the effect of hyperglycemia on the beta cell, INS‐1 832/13 beta cells were cultured under hyperglycemic conditions (20mM glucose) for 24 and 48 hours. This treatment significantly increased cellular ROS levels and impairs beta cell insulin secretion. Similarly, 20mM culture resulted in decreased nuclear localization of Nkx6.1, decreased overall Nkx6.1 protein levels, and decreased Nkx6.1 mRNA levels. We demonstrate that culturing beta cells in 20mM glucose increase Nkx6.1 ubiquitination and appears to impair new Nkx6.1 translation. Our data suggest that beta cells cultured in 20mM glucose result in increased ROS levels that result in Nkx6.1 relocalization and degradation, thus changing the ability of the beta cell to properly secrete insulin.