Effect of estrogen depression on alveolar bone microarchitecture and periodontal ligament cells during orthodontic movement.

This study aimed to evaluate the effects of the estrogen depression during orthodontic tooth movement on alveolar bone microarchitecture and periodontal ligament. Female Wistar rats were divided into two groups, one consisting of non-ovariectomized animals subjected to orthodontic tooth movement, and one comprising ovariectomized animals subjected to orthodontic tooth movement. Micro-CT assessment of bone volume to total volume (BV/TV), total porosity, trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) in the alveolar bone of the orthodontically moved tooth was performed. Histomorphometric analyses were made in the periodontal ligament, and immunoexpression of RANK, RANKL, OPG, and TUNEL were quantified. Orthodontic tooth movement in the group of ovariectomized rats was faster than in non-ovariectomized animals. The alveolar bone area showed lower values of BV/TV and trabecular thickness, and higher bone porosity and trabeculae numbers in the ovariectomized rats. Histological analyses in the ovariectomized group revealed an increase in collagen fibers in the periodontal ligament. The apoptotic cell counts in the periodontal ligament were higher in the group of ovariectomized rats than in the sham-operated rats. Ovariectomy resulted in an increase in tooth movement and alteration of the alveolar bone microstructure in the first 7 day of orthodontic tooth movement, and in the presence of apoptotic cells in the periodontal ligament.

Mandible development under gestational protein restriction: cellular and molecular mechanisms.

Insufficient evidence regarding how maternal undernutrition affects craniofacial bone development persists. With its unique focus on the impact of gestational protein restriction on calvaria and mandible osteogenesis, this study aims to fill, at least in part, this gap. Female mice were mated and randomized into NP (normal protein) or LP (low protein) groups. On the 18th gestational day (GD), male embryos were collected and submitted to microtomography (µCT), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), PCR, and autophagy dynamic analyses. The study shows that the LP offspring exhibited lower body mass than the NP group, with µCT analysis revealing no volumetric differences in fetus's head. EDS analysis showed lower calcium and higher phosphorus percentages in mandibles and calvaria. SEM assessment evidenced higher hydroxyapatite crystal-like (HC) deposition on the calvaria surface in LP fetus. Conversely, lower HC deposition was observed on the mandible surface, suggesting delayed matrix mineralization in LP fetuses with a higher percentage of collagen fibers in the mandible bone. The autophagy process was reduced in the mesenchyme of LP fetuses. PCR array analysis of 84 genes revealed 27 genes with differential expression in the LP progeny-moreover, increased mRNA levels of Akt1, Mtor, Nfkb, and Smad1 in the LP offspring. In conclusion, the results suggest that gestational protein restriction anticipated bone differentiation in utero, before 18GD, where this process is reduced compared to the control, leading to the reduction in bone area at 15 postnatal day previously observed. These findings provide insights into the molecular and cellular mechanisms of mandible development and suggest potential implications for the Developmental Origins of Health and Disease (DOHaD).

Absence of melatonin during development impairs craniofacial and dental onset in rats.

OBJECTIVE: Herein, we evaluated pinealectomy-induced melatonin absence to determine its effects on craniofacial and dental development in the offspring. DESIGN: Female Wistar rats in three groups, i.e., intact pregnant rats, pinealectomized pregnant rats (PINX), and pinealectomized pregnant rats subjected to oral melatonin replacement therapy, were crossed 30 days after surgery. The heads of 7-day-old pups were harvested for cephalometric and histological analyses, and maxillae and incisors were collected for mRNA expression analysis. RESULTS: The PINX pups exhibited a reduction in neurocranial and facial parameters such as a decrease in alveolar bone area, incisor size and proliferation, and an increase in odontoblasts and the dentin layer. Based on incisor mRNA expression analysis, we found that Dmp1 expression was upregulated, whereas Col1a1 expression was downregulated. Maxillary mRNA expression revealed that Rankl expression was upregulated, whereas that of Opn and Osx was downregulated. CONCLUSION: Our results demonstrated that the absence of maternal melatonin during early life could affect dental and maxillary development in offspring, as well as delay odontogenesis and osteogenesis in maxillary tissues. CLINICAL RELEVANCE: Our findings suggest that disruptions or a lack of melatonin during pregnancy may cause changes in craniofacial and dental development, at least in animal experiments; however, in humans, these feedings are still poorly understood, and thus careful evaluations of melatonin levels in humans need to be investigated in craniofacial alterations.

Caloric restriction prevents inflammation and insulin dysfunction in middle-aged ovariectomized mice.

BACKGROUND: Loss of ovarian function is associated with increased visceral fat. In this study, we aimed to study the effects of caloric restriction (CR) on metabolism in ovariectomized mice. METHODS AND RESULTS: Female, 8-12-month-old mice were divided into three groups: OVX (ovariectomized mice), OVXR (40% CR) and Sham. CR increased insulin sensitivity and glucose tolerance. AMPK phosphorylation was observed in the liver of OVXR mice. CR also increased hepatic cholesterol and triglyceride levels. The reductions in the level of TBARS in the serum and liver and of H2O2 in the liver of OVXR mice suggested alterations in the redox state of the liver. Although expression of catalase protein was reduced by CR, expression of superoxide dismutase was not altered by CR. Although interleukin IL-6 and IL-10 levels in OVXR mice were similar to those in Sham mice, macrophage infiltration was reduced in OVXR mice. OVXR mice had increased sirtuin1 levels and decreased sirtuin3 levels in the liver. CONCLUSIONS: In conclusion, CR improved the condition of ovariectomized mice by reducing adiposity and increasing insulin sensitivity and glucose tolerance through a mechanism that may involve AMPK.

Maxillary and dental development in the offspring of protein-restricted female rats.

Nutritional restriction during developmental periods impairs organ physiology. Female rats were subjected to protein restriction during pregnancy and lactation to analyze dental and maxillary development. Four exposure groups were considered: normal-protein diet during pregnancy and lactation (NP, 17% casein), low-protein diet during lactation (LP-L, 6% casein), low-protein diet during pregnancy and lactation (LP), and low-protein diet during pregnancy (LP-G). Maxillae from 15-day-old male pups were collected. All protein-restricted groups presented increased dentin thickness and reduced alveolar bone area. When protein restriction was applied during both gestation and lactation (LP), harmful effects were observed in the form of loss of protective OPG (osteoprotegerin) in tooth epithelium-mesenchyme, due to higher RANKL expression, delay in odontoblast maturation, less dental pulp vascularity, reduction in amount of alveolar bone, and less matrix mineralization. In the LP-L group, effects of protein restriction seemed less harmful, and despite less alveolar bone, the enhancement in BMP-7, VEGF, and RANKL seems a compensatory signal to maintain maxillary osteogenesis. In LP-G animals, Dspp expression was higher, suggesting a delay in odontoblast maturation or expression recuperation. In conclusion, maternal protein restriction affects dental and maxillary development. A low-protein diet only in gestation allows for normal development. A low-protein diet during gestation-lactation results in impaired odontogenesis that may increase susceptibility of dental anomalies.

Quercetin prevents insulin dysfunction in hypertensive animals.

Angiotensin II induced increase in hypertension enhances oxidative stress and compromises insulin action and pancreatic function. Quercetin-rich foods are beneficial for hypertensive and diabetic animals owing to their antioxidant function. The aim of this study was to evaluate the antioxidant effects of quercetin in hypertensive rats on insulin action, signaling, and secretion. Wistar rats were randomly divided into three groups: sham, hypertensive rats (H), and hypertensive rats supplemented with quercetin (HQ). After three months of initial hypertension, quercetin was administered at 50 mg/kg/day for 30 days. Our results indicate that hypertension and serum lipid peroxidation levels were reduced by quercetin supplementation. We observed increased insulin sensitivity in adipose tissue, corroborating the insulin tolerance test, HOMA index, and improvements in lipid profile. Despite normal insulin secretion at 2.8 and 20 mM of glucose, animals treated with quercetin exhibited increased number of islets per section; increased protein expression of muscarinic receptor type 3, VEGF, and catalase in islets; and hepatic mRNA levels of Ide were normalized. In conclusion, supplementation with quercetin improved insulin action and prevented pancreatic and metabolic dysfunction.

Morphofunctional and immunological cardiac evaluation of protein restriction on rat offspring.

Cardiovascular diseases affect a large part of the population during adulthood. Several epidemiological and experimental studies have demonstrated an association between maternal protein restriction and a marked risk of developing heart disease during early intrauterine and postnatal life. Maternal nutritional conditions act by modulating the microenvironment, thus favoring adaptive processes of the fetal organism for development to occur. However, the physiological profile is established in this period and its effects can be observed in the long term. In the present study, the cardiac muscles of 15-day-old offspring of Wistar rats subjected to maternal protein restriction during pregnancy and/or lactation were evaluated to identify possible cardiac changes relevant for heart disease in adulthood. The offspring of restricted female rats during pregnancy had a lower birth weight. Male offspring subjected to restriction during pregnancy and lactation showed an increase in the concentration of H2O2, a reduction in the expression of the Mn-SOD enzyme, and a greater expression of ß-MHC and Connexin 43. There was also an increase in the MPO enzyme activity in the tissue. It was observed that the effects of protein restriction are sex-specific, since the cardiac muscle of male animals showed alterations suggestive of oxidative stress, hypertrophy, signs of tissue inflammation, and increased expression of important proteins in intercellular communication. These changes characterize the ongoing cardiac remodeling process. Finally, the data revealed that the lactation phase accentuated harmful effects on the cardiac tissue of the offspring.

Pulpal outcomes in orthodontic tooth movement in diabetes mellitus.

Diabetes mellitus impairs angiogenesis and tissue reorganization during orthodontic tooth movement (OTM). Thus, this study evaluated pulpal outcomes in orthodontic tooth movement through metabolic changes in diabetes. Male Wistar rats were used, and the in vivo study design consisted of four groups (n = 10/group): C-non-diabetic animals not subjected to orthodontic tooth movement; D-diabetic animals not subjected to orthodontic tooth movement; OTM-non-diabetic animals subjected to orthodontic tooth movement; and D + OTM-diabetic animals subjected to orthodontic tooth movement. In addition, the pulps of the distovestibular root (DV) and mesiovestibular root (MV) were assessed by histomorphometric analyses and immunoexpression of the RANKL/OPG system. Pulpal analysis of the MV root showed an increase in blood vessels in diabetic animals. Inflammatory infiltrate and fibroblastic cells were elevated in diabetic animals with tooth movement in the DV and MV roots. In the DV and MV roots, diabetic rats with OTM showed a reduction in birefringent collagen fibers. The immunostaining for RANKL was higher in the pulp tissue of OTM in diabetic and non-diabetic animals. It was concluded that the pulp tissue has less adaptive and repair capacity during OTM in diabetes. Orthodontic strength can alter the inflammatory processes in the pulp.

Gestational protein restriction alters the RANKL/OPG system in the dental germ of offsprings.

Evaluate the dentinogenesis in the offspring of rats submitted to gestational protein restriction (GPR). DESIGN: The offspring were evaluated at the 21st day of gestation (21 dG). Assessments were made of morphological parameters and the RANKL/OPG system - bone tissue maturation markers - in the upper incisor tooth germ. Pregnant 10-week-old female Wistar rats were divided into normal protein (NP, 17% casein, n = 5) and low protein (LP, 6% casein, n = 5) diet groups. At 21 dG, the offspring maxillae were collected for histomorphometric and immunohistochemical analyses. RESULTS: The LP group showed decreased thickness of the dentin and odontoblast cell layers on the tooth germ. GPR led to decreased OPG expression and increased RANKL expression in the incisor germ. CONCLUSION: The results suggested that gestational protein restriction altered odontoblast RANKL/OPG expression and decreased dentin matrix deposition and thickness in tooth development.