Tooth micro-hardness changes after applying bioactive glass-containing, anti-microbial sealants

The AAPD recommends placement of dental pit and fissure sealants on surfaces that are high risk or that already exhibit incipient carious lesions. Protection provided by sealants may be enhanced by the addition of ion-releasing, anti-microbial filler particles of bioactive glass. Objective: We prepared novel dental pit and fissure sealant materials containing bioactive glass (BAG) fillers and tested their ability to prevent tooth demineralization in a bacterial broth. Methods: Two types of BAG were synthesized in our lab: BAG1 (61 wt% silica - 31 wt% calcia - 4 wt% phosphate - 4 wt% flouride); and BAG2 (81 wt% silica - 11 wt% calcia - 4 wt% phosphate - 4 wt% flouride). Ultraseal XT (USXT) resin without filler was supplied by the manufacturer (Ultradent Products, Inc. South Jordan, UT). BAGs were individually incorporated into the resin (25 wt%) and provided handling properties similar to USXT. Caries-free teeth (n=5 each) were randomly assigned to three groups (BAG1-sealant, BAG2-sealant,or USXT) and sealants were placed by the same practitioner. Acid-resistant nail polish was used to cover half of the tooth surface. Teeth were immersed in a bacterial culture system of sucrose-rich brain-heart infusion (BHI) media containing Streptococcus mutans strain #25175, an acid-producing microbe and incubated at 37˚C, 5%CO2; media was changed every other day. Bacterial growth was confirmed throughout the test period. At two weeks, teeth were sectioned sagitally and microhardness testing compared changes in hardness as a function of location on the tooth. Results : Overall, the BAG2-sealant samples were significantly harder than the BAG1-sealant or USXT samples. Areas adjacent to the BAG2-sealant were harder than the original tooth surface. (ANOVA/Tukey’s; α=0.05). Conclusion: The inclusion of anti-bacterial, ion-releasing BAG as a filler component results in a harder tooth that may be better able to resist demineralization.This presentation was presented at the American Association for the Advancement of Science (AAAS) Pacific Division 93rd Annual Meeting in Boise, Idaho. It was also presented at the American Association for Dental Research Annual Conference in 2012 (AADR)

A KATP Channel-Dependent Pathway within α Cells Regulates Glucagon Release from Both Rodent and Human Islets of Langerhans

Glucagon, secreted from pancreatic islet α cells, stimulates gluconeogenesis and liver glycogen breakdown. The mechanism regulating glucagon release is debated, and variously attributed to neuronal control, paracrine control by neighbouring β cells, or to an intrinsic glucose sensing by the α cells themselves. We examined hormone secretion and Ca2+ responses of α and β cells within intact rodent and human islets. Glucose-dependent suppression of glucagon release persisted when paracrine GABA or Zn2+ signalling was blocked, but was reversed by low concentrations (1–20 μM) of the ATP-sensitive K+ (KATP) channel opener diazoxide, which had no effect on insulin release or β cell responses. This effect was prevented by the KATP channel blocker tolbutamide (100 μM). Higher diazoxide concentrations (≥30 μM) decreased glucagon and insulin secretion, and α- and β-cell Ca2+ responses, in parallel. In the absence of glucose, tolbutamide at low concentrations (<1 μM) stimulated glucagon secretion, whereas high concentrations (>10 μM) were inhibitory. In the presence of a maximally inhibitory concentration of tolbutamide (0.5 mM), glucose had no additional suppressive effect. Downstream of the KATP channel, inhibition of voltage-gated Na+ (TTX) and N-type Ca2+ channels (ω-conotoxin), but not L-type Ca2+ channels (nifedipine), prevented glucagon secretion. Both the N-type Ca2+ channels and α-cell exocytosis were inactivated at depolarised membrane potentials. Rodent and human glucagon secretion is regulated by an α-cell KATP channel-dependent mechanism. We propose that elevated glucose reduces electrical activity and exocytosis via depolarisation-induced inactivation of ion channels involved in action potential firing and secretion

Pulsatile insulin secretion, impaired glucose tolerance and type 2 diabetes

Type 2 diabetes (T2DM) results when increases in beta cell function and/or mass cannot compensate for rising insulin resistance. Numerous studies have documented the longitudinal changes in metabolism that occur during the development of glucose intolerance and lead to T2DM. However, the role of changes in insulin secretion, both amount and temporal pattern has been understudied. Most of the insulin secreted from pancreatic beta cells of the pancreas is released in a pulsatile pattern, which is disrupted in T2DM. Here we review the evidence that changes in beta cell pulsatility occur during the progression from glucose intolerance to T2DM in humans, and contribute significantly to the etiology of the disease. We review the evidence that insulin pulsatility improves the efficacy of secreted insulin on its targets, particularly hepatic glucose production, but also examine evidence that pulsatility alters or is altered by changes in peripheral glucose uptake. Finally, we summarize our current understanding of the biophysical mechanisms responsible for oscillatory insulin secretion. Understanding how insulin pulsatility contributes to normal glucose homeostasis and is altered in metabolic disease states may help improve the treatment of T2DM

Long Lasting Synchronization of Calcium Oscillations by Cholinergic Stimulation in Isolated Pancreatic Islets

Individual mouse pancreatic islets exhibit oscillations in [Ca2+]i and insulin secretion in response to glucose in vitro, but how the oscillations of a million islets are coordinated within the human pancreas in vivo is unclear. Islet to islet synchronization is necessary, however, for the pancreas to produce regular pulses of insulin. To determine whether neurohormone release within the pancreas might play a role in coordinating islet activity, [Ca2+]i changes in 4–6 isolated mouse islets were simultaneously monitored before and after a transient pulse of a putative synchronizing agent. The degree of synchronicity was quantified using a novel analytical approach that yields a parameter that we call the “Synchronization Index”. Individual islets exhibited [Ca2+]i oscillations with periods of 3–6 min, but were not synchronized under control conditions. However, raising islet [Ca2+]i with a brief application of the cholinergic agonist carbachol (25 μM) or elevated KCl in glucose-containing saline rapidly synchronized islet [Ca2+]i oscillations for ≥30 min, long after the synchronizing agent was removed. In contrast, the adrenergic agonists clonidine or norepinephrine, and the KATP channel inhibitor tolbutamide, failed to synchronize islets. Partial synchronization was observed, however, with the KATP channel opener diazoxide. The synchronizing action of carbachol depended on the glucose concentration used, suggesting that glucose metabolism was necessary for synchronization to occur. To understand how transiently perturbing islet [Ca2+]i produced sustained synchronization, we used a mathematical model of islet oscillations in which complex oscillatory behavior results from the interaction between a fast electrical subsystem and a slower metabolic oscillator. Transient synchronization simulated by the model was mediated by resetting of the islet oscillators to a similar initial phase followed by transient “ringing” behavior, during which the model islets oscillated with a similar frequency. These results suggest that neurohormone release from intrapancreatic neurons could help synchronize islets in situ. Defects in this coordinating mechanism could contribute to the disrupted insulin secretion observed in Type 2 diabetes

Synchronization of Pancreatic Islet Oscillations by Intrapancreatic Ganglia: A Modeling Study

Plasma insulin measurements from mice, rats, dogs, and humans indicate that insulin levels are oscillatory, reflecting pulsatile insulin secretion from individual islets. An unanswered question, however, is how the activity of a population of islets is coordinated to yield coherent oscillations in plasma insulin. Here, using mathematical modeling, we investigate the feasibility of a potential islet synchronization mechanism, cholinergic signaling. This hypothesis is based on well-established experimental evidence demonstrating intrapancreatic parasympathetic (cholinergic) ganglia and recent in vitro evidence that a brief application of a muscarinic agonist can transiently synchronize islets. We demonstrate using mathematical modeling that periodic pulses of acetylcholine released from cholinergic neurons is indeed able to coordinate the activity of a population of simulated islets, even if only a fraction of these are innervated. The role of islet-to-islet heterogeneity is also considered. The results suggest that the existence of cholinergic input to the pancreas may serve as a regulator of endogenous insulin pulsatility in vivo

Characterization of Erg K+ Channels in α- and β-Cells of Mouse and Human Islets*

Voltage-gated eag-related gene (Erg) K+ channels regulate the electrical activity of many cell types. Data regarding Erg channel expression and function in electrically excitable glucagon and insulin producing cells of the pancreas is limited. In the present study Erg1 mRNA and protein were shown to be highly expressed in human and mouse islets and in α-TC6 and Min6 cells α- and β-cell lines, respectively. Whole cell patch clamp recordings demonstrated the functional expression of Erg1 in α- and β-cells, with rBeKm1, an Erg1 antagonist, blocking inward tail currents elicited by a double pulse protocol. Additionally, a small interference RNA approach targeting the kcnh2 gene (Erg1) induced a significant decrease of Erg1 inward tail current in Min6 cells. To investigate further the role of Erg channels in mouse and human islets, ratiometric Fura-2 AM Ca2+-imaging experiments were performed on isolated α- and β-cells. Blocking Erg channels with rBeKm1 induced a transient cytoplasmic Ca2+ increase in both α- and β-cells. This resulted in an increased glucose-dependent insulin secretion, but conversely impaired glucagon secretion under low glucose conditions. Together, these data present Erg1 channels as new mediators of α- and β-cell repolarization. However, antagonism of Erg1 has divergent effects in these cells; to augment glucose-dependent insulin secretion and inhibit low glucose stimulated glucagon secretion