Certains neuropeptides, comme les orexines, pourraient être impliqués dans les mécanismes neurobiologiques qui sous-tendent la dépression majeure. Nous avons utilisé un modèle murin de dépression, le stress chronique imprédictible modéré, pour étudier précisément le rôle du système orexinergique dans la dépression. Les souris stressées présentaient une augmentation de l’activation des neurones orexinergiques dans la partie dorsomédiale et périfornicale de l’hypothalamus, ainsi que des changements de l’expression des récepteurs orexine-2, contrecarrés par un traitement chronique avec un antidépresseur. De plus, le blocage pharmacologique du système orexinergique chez des souris stressées a engendré des effets antidépresseurs sous-tendus par le rétablissement du rétrocontrôle négatif de l’axe HPA indépendamment d’une augmentation de la neurogenèse hippocampique. Ces résultats montrent que le système orexinergique pourrait être impliqué dans la physiopathogenèse des troubles dépressifs. / Neuropeptides, especially orexins, could be involved in the neurobiological mechanisms underlying maj or depression. We used a mouse mode! of depression, the unpredictable chronic mild stress, to specifically study the role ofthe orexinergic system in depression. Stressed mice showed an increase of orexinergic neuronal activation in the dorsomedial and perifornical hypothalamus, as well as change in orexin receptor-2 expression, reversed by chronic treatment with an antidepressant. In addition, pharmacological blockade ofthe orexinergic system in stressed mice induced antidepressant-like effects, underpinned by the restoration ofthe HPA axis negative feedback, independently of an increase of hippocampal neurogenesis. These results show that the orexinergic system could be involved in the pathophysiology of depressive disorders.

A hipertensão arterial sistêmica (HAS) é um dos principais fatores de risco para a instalação e progressão das complicações crônicas do diabetes melito (DM) tipo 2. A medida da pressão arterial (PA) através da monitorização ambulatorial da PA (MAPA) apresenta melhor correlação com o desenvolvimento de lesões em órgãos-alvo do que a medida no consultório. Além disso, permite a avaliação de parâmetros pressóricos distintos como as médias das PAs sistólica e diastólica das 24 h, do dia e da noite, cargas pressóricas e ausência do descenso noturno, além da identificação de pacientes com HAS do avental branco e mascarada. Os pacientes com DM apresentam maiores médias de PA diurna e noturna do que os sem DM. Além disso, um terço do pacientes normotensos com DM tipo 2 apresentam HAS mascarada, que está associada a um aumento da albuminúria e da espessura das paredes do ventrículo esquerdo. Por outro lado, a prevalência e o efeito da HAS do avental branco nos pacientes com DM ainda não foram adequadamente avaliados. A determinação da ausência do descenso noturno da PA não acrescenta informação às medidas da PA nas 24 h, no dia ou na noite, mas a medida da PA noturna parece ser relevante na retinopatia do DM. Em conclusão, a determinação da PA através da MAPA é capaz de estratificar de forma mais adequada os pacientes em risco para o desenvolvimento das complicações crônicas do DM e tornou-se um instrumento indispensável para o controle efetivo da PA nestes pacientes. / Hypertension is one of the main risk factors for the onset and progression of chronic complications in type 2 diabetes mellitus (DM). Ambulatory blood pressure (BP) monitoring (ABPM) provides a better correlation with target organ lesions than BP obtained in the office. Furthermore, it allows the evaluation of distinct BP parameters such as the 24-h, daytime and nighttime systolic and diastolic BP means, BP loads and the absence of nocturnal drop of BP, as well as the identification of white-coat and masked hypertension. DM patients have higher daytime and nighttime BP means than non-DM patients. In addition, one third of normotensive type 2 DM patients have masked hypertension, which is associated with an increase in albuminuria and in left ventricle wall thickness. On the other hand, the prevalence and effect of white-coat hypertension in type 2 DM patients have not yet been properly evaluated. The absence of nocturnal drop of BP does not add information to the 24 h, daytime or nighttime BP measurements, but the nighttime BP means seem to be relevant in DM retinopathy. In conclusion, BP determination by ABPM allows better patient risk stratification for the development of DM chronic complications and is an essential instrument for effective BP control in these patients.

Objetivos: Avaliar a variabilidade glicêmica, respostas metabólicas e cardiovasculares após uma sessão de exercício aeróbico em pacientes com diabetes em tratamento com vildagliptina ou glibenclamida. Métodos: Foi realizado ensaio clínico aberto e paralelo que incluiu 13 pacientes com diabetes tipo 2 tratados com vildagliptina (50mg bid) ou glibenclamida (5mg bid) por 12 semanas. Antes e após a intervenção, a variabilidade glicêmica (glicose média, variância da glicose, coeficiente de variação e média da amplitude das oscilações glicêmicas), respostas metabólicas (HbA1c, glicose, insulina e 8-iso prostaglandina F2α) e cardiovasculares (débito cardíaco, variabilidade da frequência cardíaca e componentes do controle autonômico) foram avaliadas no repouso, durante e após uma sessão de exercício aeróbico de 30 minutos. A variabilidade da pressão arterial foi aferida nas 24 horas após o exercício. Resultados: Doze semanas de tratamento resultou em redução da glicemia de 18% com vildagliptina e 35% com glibenclamida (p grupo=0.007). A HbA1c reduziu significativamente (1.24 % e 1.52%) nos grupos vildagliptina e glibenclamida, respectivamente.A variabilidade glicêmica não se alterou após o tratamento com glibenclamida ou vildagliptina (MAGE=55.8 ±5.3 mg/dL e 69.9 ± 13.3 mg/dL, respectivamente; p grupo=0.091; p tempo=0.234). Foi encontrada uma diminuição da glicose avaliada por monitoramento contínuo durante as 3 horas de recuperação do exercício, com AUC (de 6h) menor no grupo glibenclamida vs vildagliptina (p=0.04). A glibenclamida induziu maiores concentrações de insulina na recuperação do exercício. O grupo tratado com vildagliptina obteve 6.3mmHg de redução da pressão arterial sistólica, enquanto a glibenclamide reduziu 3.6mmHg, sem diferença entre os grupos. Os pacientes tratados com vildagliptina apresentaram menor variabilidade da pressão arterial sistólica (0.0445 ±0.05 mm/Hg), medida por time rate, comparados aos tratados com glibenclamida (0.601 ±0.12 mm/Hg), p=0.012. Conclusão: Este é o primeiro estudo conduzido em pacientes com DM2 que fornece dados sobre a influência da terapia medicamentosa padrão (metformina e glibenclamida) vs outra classe disponível (metformina e vildagliptina) em respostas a uma sessão de exercício aeróbico, um dos tratamentos recomendados para pacientes com DM2. Além de melhora no controle glicêmico e redução da pressão arterial sistólica obtidas por ambos tratamentos, foi observada menor variabilidade da pressão arterial nos 9 pacientes submetidos ao tratamento com vildagliptina. Não foi encontrada menor variabilidade da glicemia após o exercício nos pacientes tratados com vildagliptina comparados aos tratados com glibenclamida, refutando a hipótese de estudo.

Ces travaux concernent le développement d’un amplificateur à fibre optique souple, microstructurée, double-gaine, dopée ytterbium (Yb), et monomode à large coeur, dans la gamme d’impulsion nanoseconde, multi-kiloHertz et milliJoule, pour l’injection de chaînes lasers de puissance. L’architecture amplificatrice est mise en oeuvre dans une configuration MOPA (Master Oscillator Power Amplifier) à plusieurs étages. Un modèle numérique de l’amplification sur fibre double-gaine dopée Yb, incluant l’émission spontanée amplifiée, a été développé pour étudier le comportement de ce type d’amplificateur fibré et procéder au dimensionnement du dispositif expérimental. Afin de s’affranchir du processus de saturation par le gain, un algorithme de contre-réaction permettant de déterminer numériquement la forme temporelle optimale a été associé au modèle. Nous avons obtenu des résultats expérimentaux en bon accord avec les simulations numériques, et avec les performances suivantes : une énergie de 0.5 mJ par impulsion à une fréquence de répétition dans la gamme de 1 kHz à 10 kHz, sur des impulsions à spectre étroit centré à la longueur d’onde 1053 nm, à profil temporel super-gaussien d’ordre 20 de durée 10 ns, avec un rapport signal-sur-bruit optique supérieur à 50 dB et un taux de maintien de la polarisation à 20 dB. Le profil spatial en sortie de système est monomode (M²=1.1). Ce dispositif peut également délivrer des énergies jusqu’à 1.5 mJ. Nous avons ensuite mis à profit ces performances pour l’amplification d’impulsions à dérive de fréquence, et avons obtenu une énergie par impulsion de 0.7 mJ sur une durée de 570 fs, à une fréquence de répétition de 10 kHz. / This work concerns the development of a double-clad ytterbium-doped single-mode microstructured flexible fiber-based amplifier, in the nanosecond, multi-kiloHertz and milliJoule regime, for large-scale laser facilities seeding. We have used a multi-stage master oscillator power amplifier fibered architecture. A numerical model of ytterbium-doped double-clad fiber-based amplification, including amplified spontaneous emission, was developed in order to study the behaviour of such amplifier and to correctly design the experimental set-up. This model was completed by a feed-back algorithm to numerically predict the optimal temporal shape to compensate the gain saturation process. We demonstrated experimental results in good agreement with numerical simulations, with the following performances: 0.5 mJ pulse energy, at a frequency repetition from 1 kHz to 10 kHz, with a narrow bandwidth spectrum centred at 1053 nm wavelength, with 10 ns pulse duration on a perfect super-Gaussian temporal profile, an optical signal-to-noise ratio better than 50 dB and a polarization extinction ratio of 20 dB. We checked that the beam quality was diffraction limited, with an M² measurement of 1.1. Moreover, the system can deliver energies up to 1.5 mJ. Then, we took the advantage of such results to amplify chirped pulses. We demonstrated 0.7 mJ pulse energy, with 570 fs duration at 10 kHz repetition frequency.

Angiogenesis supports tumor growth and facilitates metastasis, the leading cause of patient mortality in breast and other types of solid tumors. Angiopoietin-2 (Ang-2) is an angiogenic factor whose overexpression is associated with increased tumor vascularity, metastasis, and decreased patient survival. We assessed the effects of Ang-2 on breast tumor vasculature by comparing vascular morphology and metastasis of orthotopically implanted metastatic breast carcinoma line MDA-MB-231 that either lacked or overexpressed Ang-2. Methods: Luciferase-tagged MDA-MB-231 breast carcinoma cells designated hAng2 were engineered to overexpress human Ang-2. The control line expressed hAng-2 in reverse orientation. Stable production of hAng-2 was confirmed by RT-PCR, qRT-PCR, Western blot, and ELISA. Functionality of recombinant hAng-2 was assessed by migration and proliferation assays. MDA-MB-231 tumors, hAng2 and control, were orthotopically implanted into female Nu/Nu and SCID mice. Metastasis to lymph node and lung were determined by measuring luciferase activity in tissue extracts. Tumor blood vessel morphology was analyzed by immunohistochemistry (IHC) using antibodies against MECA-32, smooth muscle actin (SMA-alpha), VEGFR-3 and Notch-1. Results: MDA-MB-231 hAng2 lines were stably generated to produce 0-370 ng/ml of hAng-2 while expression in control line was undetectable. Tumor-produced hAng-2 was functional as demonstrated by a dose-dependent induction of migration of lymphatic endothelial cells and mouse mesenchymal stem cells with a maximum increase of 3-fold. Overexpression of Ang-2 had no significant effect on proliferation of cultured MDA-MB-231 cells, growth rate of hAng2 tumors or blood vessel density. In contrast, we detected substantial differences in vascular morphology of Ang-2 overexpressing tumors including 1.7-fold increase in blood vascular area, 2.8-fold increase in number of vessels with open lumen, and 6-fold increase in lumens' cross-sectional area. Blood vascular pericyte coverage shown by SMA-α staining decreased (p>0.001) in hAng2 tumors, demonstrating vessel destabilization. Blood vascular invasion by tumor cells and pulmonary metastasis increased in Ang-2 overexpressing tumors by 500% and 1100%, as compared with control tumors. Blood vessels in hAng2 tumors, but not lymphatic vessels, displayed significantly upregulated Notch-1 and VEGFR-3 expression amplified by a 2.2-fold. Conclusions: These data suggest that Ang-2 increases metastasis because of suppression of pericytes' recruitment that leads to destabilization of the tumor vessels, which facilitates the entry of tumor cells into the vessels and increases metastatic spread. These effects are associated with up-regulation of Notch-1 and VEGFR-3 on tumor vasculature suggesting that signaling of these proteins underlie the morphologic changes in Ang-2 overexpressing tumors. This is consistent with prior data demonstrating up-regulation of VEGFR-3 on tumor blood vessels and association of both proteins with increased metastasis. This study demonstrates that Ang-2 plays a key role in hematogenous metastasis and suggests that Ang-2 might represent novel a target for inhibition of breast cancer metastasis.

HIV and TB are among top causes of mortality and morbidity globally. Incident TB cases in 2015 were approximately 10.5 million, with 11% having HIV. Having HIV is also the biggest risk factor for developing TB disease. Clinical studies show HIV-2/TB coinfection cases progress to disease slower than HIV-1/TB. However, the underlying cellular, immunological, and molecular host-pathogen interactions accounting for these observations remain unclear. The host immune response to both viral and mycobacterial infection of macrophages is partly dependent on type I interferon, which in turn induces expression of various interferon-stimulated genes (ISGs) to control the pathogens. GBP5, a GTPase with a role in activating the inflammasome; and Sp110, a nuclear body protein, are among various ISGs whose expression levels are elevated during individual viral and bacterial infections in macrophages. GBP5 seems to restrict HIV-1 replication in immune cells, while Sp110 is known to restrict MTB proliferation in macrophages, but is now thought to also promote HIV-1 replication. We hypothesized differences in the effect of these proteins on the infectivity of the two HIV subtypes in macrophages. We also hypothesized that the viral protein Vpr is associated with the host expression levels of GBP5 and Sp110. We used small hairpin RNA to knock out GBP5 and Sp110 from THP-1 human monocytoidcell line, differentiated using PMA into a macrophage phenotype;and infected these cells with GFP-expressing HIV-2 ROD 9ΔenvΔnefstrain pseudotyped with VSV-G envelope for single cycle infection. Percentages of macrophages infected with GFP-expressing viruses were measured by Flow cytometry. To determine effect of GBP5 and Sp110 onreplication of the two HIV subtypes, HIV-1 and HIV-2, we measured virus production in supernatants of productively infected THP-1 macrophages by titering cell-free supernatants on TZMBl-reporter cells at day 3 and 6 post infection. There was a marginal increase, and a dramatic decrease, of HIV-2ΔenvΔnef GFP+ virus infectivity inall THP-1 macrophage conditions in the absence of viral Vpr, and Vpx, respectively. Knocking downmacrophage Sp110 gene expression reduced single cycle (VSV-G pseudotyped) HIV-2 ROD 9ΔenvΔnefGFP+wild type and Vpr mutant virus infectivity but promoted Vpx mutant.There was a reduction in number of infectious HIV-1 particles released from Sp110 knockdown compared to normal macrophages over 6 days post infection; no difference in HIV-1 virus production between GBP5 knock down and normal macrophages in one experiment; and a decrease in HIV-1 virus production from GBP5 knockout compared to normal macrophages 3 and 6 days post infection in a follow up experiment.Interestingly, reduced Sp110 expression corresponded to increased HIV-2 production from macrophages in one experiment but no difference in viral production between normal and Sp110 knock down macrophages by day 3 post infection in the follow up experiment. There was no HIV-2 virus production from the Sp110 knock down cells on day 6 in the follow up experiment and virus spread was reduced in GBP5 knock down, relative to normal macrophages. Interestingly, reduced Sp110 expression corresponded to increased HIV-2 production from macrophages in one experiment, but there was no difference in viral production between normal and Sp110 knock down macrophages by day 3 post infection in the follow up experiment. There was no virus production from the Sp110 knock down cells on day 6 in the follow up experiment. HIV-2 virus production was also reduced in GBP5 knock down, relative to normal, productively infected THP1/PMA macrophages. Our results support the argument that HIV-2, just like HIV-1, possibly utilizes Vpr to prevent sensing of virus infection in macrophages, thus allowing viral replication without inducing interferon stimulation, and subsequent viral restriction. Our results also suggest that the host macrophage restriction factor Sp110 may indeed enhance VSV-G pseudotyped GFP+ HIV-2 and HIV-2delVpr infectivity but restrict HIV-2 Vpx mutant. This could imply presence of a yet unknown association between another HIV-2 viral accessory protein, Vpx, and the macrophage restriction factor, Sp110 known to counter invading intracellular bacteria. We report much lower productive infectious virus release of HIV-2, compared to HIV-1, from macrophages. Overall, our results suggest that Sp110 may transiently restrict HIV-2 infection of macrophages, but is eventually manipulated by the virus to promote viral replication.

Interest in the ketogenic diet and its potential to treat obesity and type 2 diabetes has been steadily growing in recent years. With a very limited amount of calories coming from carbohydrates (typically < 50 gm/day), and the majority of calories coming from fat, this diet leads to a states of physiological ketosis, in which ketone bodies replace glucose as the primary source of energy. Early clinical trials found this to lead to a spontaneous reduction in calories consumed, and it has been suggested that a state of ketosis has appetite suppressing properties. There are a few studies evaluating self-reported decrease in hunger while consuming a ketogenic diet, as well as the changes in hormone levels associated with appetite, but this evidence is limited. The primary determinant of weight change is the difference between calories consumed and calories burned, however there is some suggestion that the macronutrient composition of the ketogenic diet may have a specific metabolic advantage for weight loss separate from the total number of calories in deficit. Multiple diet comparison studies have found the ketogenic diet to be effective for weight loss in the short term, particularly when compared to low fat diets. It is questionable whether the ketogenic diet is sustainable in the long term, particularly without frequent nutritional counseling and monitoring. Still, there is preliminary evidence that the ketogenic diet can lead not only to a large amount of weight loss, but may also be effective in treating and reversing type 2 diabetes. Clinical trials have shown decreases in HbA1c, fasting glucose, and reduction of antiglycemic medication requirements, though it is unclear if these effects are primarily due to weight loss itself, or the specific composition of the ketogenic diet. These benefits need to be weighed against the risks, and a common criticism is its high fat content which can adversely affect serum lipid levels and thus risk of cardiovascular disease. Consuming a ketogenic diet with a high intake of saturated fat has been found to increase LDL cholesterol, however this effect can be mitigated by favoring polyunsaturated or monounsaturated fats instead. While the above findings provide a preliminary understanding of the effects of the ketogenic diet, more research is needed to further elucidate the effectiveness and safety of the diet for treating obesity and type 2 diabetes.

Graphene and structurally similar 2-dimensional (2d) materials such as transition metal dichalcogenides (TMDs) and black phosphorus (BP) hold enormous potential for the next generation optoelectronics and photonics. Pairing 2d materials with printing is an emerging cost-effective large-scale device fabrication strategy. However, the current inks are far from ideal to support reproducible device fabrication. In addition, the instability of BP in ambient limits its applications. In this thesis, I present formulation of 2d material inks for inkjet printing for optoelectronic and photonic applications. To begin with, I produce mono- and few-layer 2d material flakes via ultrasonic assisted liquid phase exfoliation. This allows one-step formulation of a polymer stabilised graphene ink. For TMDs and BP, I design a binary solvent carrier for binder-free ink formulation. I show that these 2d material inks have optimal fluidic properties, drying dynamics and interaction with substrates for spatially uniform, highly controllable and print-to-print consistent large-scale printing on untreated substrates. In particular, the rapid ink drying at low temperatures leads to minimal oxidation of BP during ambient printing; the printed BP with passivation retains a stability over one month. On this basis, the printed graphene is employed as active sensing layer in CMOS integrated humidity sensors and as counter-electrodes in dye-sensitised solar cells, while the printed TMDs and BP are used to develop nonlinear photonic devices (i.e. saturable absorbers for femtosecond pulsed laser generation) and visible to near-infrared photodetectors (e.g. MoS$_2$ and BP/graphene/silicon hybrid photodetectors). Beyond inkjet printing, I present an ink formulation of commercial graphene nanoplatelets for roll-to-roll flexographic press ($\sim$100 m min$^{−1}$ printing speed). This allows hundreds of conductive electronic circuits to be printed in a minute for capacitive touchpads. Though I investigate only graphene, TMDs and BP, the ink formulation strategies can be effortlessly transferred to other 2d materials such as boron nitride, MXenes and mica. In addition to the demonstrated applications, printing of 2d materials can be potentially exploited to fabricate devices such as transistors, light emitters, energy storage conversion, and biosensors. This significantly expands the prospect of printable 2d material optoelectronics and photonics.

During the normal menstrual cycle, the human endometrium undergoes extensive tissue remodelling under the influence of ovarian-derived hormones. The endometrium has well defined stromal and epithelial compartments with the former containing both a well-developed vasculature as well as a diverse population of immune cells. Mast cells (MCs) are long-lived tissue resident immune cells characterised by the presence of granules containing proteases. Mast cells have been detected in the human uterus but little is known about their regulation or the impact of steroids on their differentiation status. Recently MCs have been implicated as key players in physiological and pathological pain pathways but little is known about their role in endometrial pathologies. Endometriosis is a chronic incurable condition characterized by the presence of endometrial tissue outside the uterine cavity: women with endometriosis can suffer from a debilitating range of symptoms including chronic pain. Whilst the aetiology of endometriosis is uncertain, close proximity between MCs and nerves has implicated them in aberrant activation of pain pathways. The aims of the current project were: 1. To determine the spatial and temporal location of uterine MCs and to explore their phenotype including expression of steroid receptors. 2. To explore the activation status of MCs in women with endometriosis and/or pain, 3. To explore the use of cells and mice as models to investigate the phenotype of mast cells and their regulation by steroids. Mast cell proteases tryptase and chymase were detected by RTPCR and immunohistochemistry in “full thickness” (uterine lumen to endometrial-myometrial junction) biopsies from women undergoing hysterectomy. In agreement with previous findings MCs were most abundant in the myometrium. Uterine MCs were predominantly of the classical MC subtypes: tryptasepos/chymaseneg and tryptasepos/chymasepos but a rare third subtype was also identified as tryptaseneg/chymasepos. Mast cell activation/degranulation was cycle stage dependent and for the first time their steroid receptor phenotype was identified as ERαneg/ERβpos/GRpos, suggesting potential regulation by the uterine steroid microenvironment. Studies on tissue samples from women with endometriosis revealed MCs with an altered activation status in the pelvic peritoneal wall, compared to controls, which showed an intense diffuse immunoexpression of chymase suggestive of MC activation and release of this protease during normal physiology of the peritoneum. Surprisingly, analysis of peritoneal fluids from controls, women with pain but no endometriosis, and pain with endometriosis did not detect differences in numbers of MCs or concentrations of tryptase or chymase. Analysis of peritoneal biopsies also provided the first evidence for a striking increase in immunoexpression of PAR-2, a protease-activated receptor, in women suffering from chronic pelvic pain and/or endometriosis which may provide a mechanism by which mast cell derived factors may alter pain pathways. Studies in a mouse model of endometriosis identified MCs within endometria-llike lesions and offer a platform for future studies. In vitro explorations using MCs derived from peripheral blood precursors and HMC-1, a cell line derived from a patient with MC leukaemia confirmed expression of ERβ but did not support previous studies claiming cells were ERαpos. In summary, this study has provided novel insights into the phenotype of endometrial mast cells in the normal cycling endometrium and contrasted them with those in women with endometriosis and pelvic pain. This is the first study to identify MCs as ERβpos. Further studies are required to determine whether inhibition of PAR- 2 might offer a therapeutic target in women with chronic pelvic pain.

Trabajo de Investigación Requisito para optar al Título de Cirujano Dentista / Autor no autoriza el acceso a texto completo de su documento / La diabetes mellitus es una patología que se caracteriza por desregulación metabólica de los hidratos de carbono en que se puede producir neuropatía diabética, complicación que se manifiesta con la alteración de la sensibilidad en una zona del cuerpo. El objetivo de este trabajo fue determinar la respuesta pulpar a las pruebas de vitalidad térmicas en enfermos con diabetes mellitus tipo II y describir la distribución de las terminaciones nerviosas en la pulpa dentaria, en dientes extraídos por enfermedad periodontal. Se evaluaron 26 casos, 9 diabéticos y 17 no diabéticos, que tenían indicación de exodoncia por enfermedad periodontal avanzada. En estos pacientes se realizaron pruebas de vitalidad pulpar térmicas y a continuación se realizaron las exodoncias obteniéndose 23 dientes de pacientes diabéticos y 27 dientes de pacientes sin esta patología. A continuación los dientes se fijaron en formalina al 10 % y luego se descalcificaron con E.D.T.A. al 4.13% y pH 7.4. Se procesaron con técnica histológica corriente, se tiñeron con hematoxilina eosina y se realizó inmunomarcación con S-100. Los enfermos diabéticos presentaron una menor respuesta pulpar frente a las pruebas de vitalidad (p<0,001). También se encontró que los diabéticos tienen una menor respuesta al calor en sus dientes (p<0,001). Los enfermos diabéticos presentaron un mayor porcentaje de calcificaciones intrapulpares. Con técnica inmunohistoquímica se observó que la extensión de las fibras nerviosas abarcó hasta los túbulos dentinarios en ambos grupos, no mostrando diferencias entre ellos.