An update on periodontal inflammation and bone loss.

Periodontal disease is a chronic inflammatory condition that affects the supporting structures of the teeth, including the periodontal ligament and alveolar bone. Periodontal disease is due to an immune response that stimulates gingivitis and periodontitis, and its systemic consequences. This immune response is triggered by bacteria and may be modulated by environmental conditions such as smoking or systemic disease. Recent advances in single cell RNA-seq (scRNA-seq) and in vivo animal studies have provided new insight into the immune response triggered by bacteria that causes periodontitis and gingivitis. Dysbiosis, which constitutes a change in the bacterial composition of the microbiome, is a key factor in the initiation and progression of periodontitis. The host immune response to dysbiosis involves the activation of various cell types, including keratinocytes, stromal cells, neutrophils, monocytes/macrophages, dendritic cells and several lymphocyte subsets, which release pro-inflammatory cytokines and chemokines. Periodontal disease has been implicated in contributing to the pathogenesis of several systemic conditions, including diabetes, rheumatoid arthritis, cardiovascular disease and Alzheimer's disease. Understanding the complex interplay between the oral microbiome and the host immune response is critical for the development of new therapeutic strategies for the prevention and treatment of periodontitis and its systemic consequences.

Promotion of cutaneous diabetic wound healing by subcutaneous administration of Wharton's jelly mesenchymal stem cells derived from umbilical cord.

Wound healing is a major problem in diabetic patients, and current treatments have been confronted with limited success. The present study examined the benefit of Wharton's jelly mesenchymal stem cells (WJ-MSCs) derived from the human umbilical cord (UC) in wound healing in diabetic rats. Thirty days after inducing diabetes, a circular excision was created in the skin of rats, and the treatments were performed for 21 days. Two groups were studied, which included the Control group and WJ-MSCs group. The studied groups were sampled on the 7th, 14th, and 21st days after wounding. Histological ultrasound imaging of dermis and epidermis in the wound area for thickness and density measurement and skin elasticity were evaluated. Our results on post-wounding days 7, 14, and 21 showed that the wound closure, thickness, and density of new epidermis and dermis, as well as skin elasticity in the healed wound, were significantly higher in the WJ-MSCs group compared to the Control group. Subcutaneous administration of WJ-MSCs in diabetic wounds can effectively accelerate healing. Based on this, these cells can be used along with other treatment methods in the healing of different types of chronic wounds.

Chronic nitrate administration increases the expression the genes involved in the browning of white adipose tissue in female rats.

Nitrate, a nitric oxide (NO) donor, has antiobesity effect in female rats. This study hypothesized that the antiobesity effect of nitrate in female rats is due to the browning of white adipose tissue (WAT). Female Wistar rats (aged 8 months) were divided into two groups (n = 10/group): the control group received tap water and the nitrate group received water containing 100 mg/L of sodium nitrate for 9 months. At months 0, 3, 6, and 9, obesity indices were measured. At month 9, gonadal adipose tissue was used to measure messenger RNA (mRNA) and protein levels of peroxisome proliferator-activated receptor-γ (PPAR-γ), PPAR-γ coactivator 1-α (PGC1-α), uncoupling protein 1 (UCP1), and adipocyte density and area. After the 9-month intervention, nitrate-treated rats had lower body weight, body mass index, thoracic circumference, and abdominal circumference by 6.4% (p = .012), 9.1% (p = .029), 6.0% (p = .056), and 5.7% (p = .098), respectively. In addition, nitrate-treated rats had higher PPAR-γ (mRNA: 1.78-fold, p = .016 and protein: 19%, p = .076), PGC1-α (mRNA: 1.69-fold, p = .012 and protein: 68%, p = .001), and UCP1 (mRNA: 2.50-fold, p = .001 and protein: 81%, p = .001) in gonadal adipose tissue. Nitrate also reduced adipocyte area by 35% (p = .054) and increased adipocyte density by 31% (p = .086). In conclusion, antiobesity effect of nitrate in female rats is associated with increased browning of gonadal adipose tissue as indicated by higher expression of PPAR-γ, PGC1-α, and UCP1 and reduced adipocyte area and increased adipocyte density.

Role of Cultured Skin Fibroblasts in Regenerative Dermatology.

The skin, as the largest organ, covers the entire outer part of the body, and since this organ is directly exposed to microbial, thermal, mechanical and chemical damage, it may be destroyed by factors such as acute trauma, chronic wounds or even surgical interventions. Cell therapy is one of the most important procedures to treat skin lesions. Fibroblasts are cells that are responsible for the synthesis of collagen, elastin, and the organization of extracellular matrix (ECM) components and have many vital functions in wound healing processes. Today, cultured autologous fibroblasts are used to treat wrinkles, scars, wounds and subcutaneous atrophy. The results of many studies have shown that fibroblasts can be effective and beneficial in the treatment of skin lesions. On the other hand, skin substitutes are used as a regenerative model to improve and regenerate the skin. The use of these alternatives, restorative medicine and therapeutic cells such as fibroblasts has tremendous potential in the treatment of skin diseases and can be a new window for the treatment of diseases with no definitive treatment. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description ofthese Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Hydrogen sulfide potentiates the protective effects of nitrite against myocardial ischemia-reperfusion injury in type 2 diabetic rats.

Decreased heart levels of nitric oxide (NO) and hydrogen sulfide (H2S) in type 2 diabetes (T2D) are associated with a higher risk of mortality following ischemia-reperfusion (IR) injury. This study aimed to determine the effects of co-administration of sodium nitrite and sodium hydrosulfide (NaSH) on IR injury in the isolated heart from rats with T2D. Two-month-old male rats were divided into 5 groups (n = 7/group): Control, T2D, T2D + nitrite, T2D + NaSH, and T2D + nitrite + NaSH. T2D was induced using a high-fat diet and a single low dose streptozotocin (30 mg/kg) in intraperitoneal injection. Nitrite (50 mg/L in drinking water) and NaSH (0.28 mg/kg, daily intraperitoneal injection) were administrated for 9 weeks. At the end of the study, hemodynamic parameters were recorded, and infarct size and mRNA expression of H2S- and NO-producing enzymes were measured in the isolated hearts. Nitrite administration to rats with T2D improved recovery of left ventricular developed pressure (LVDP) and the peak rates of positive and negative changes in LV pressure (±dp/dt) by 30%, 17%, and 7.9%, respectively, and decreased infarct size by 18.4%. Co-administration of nitrite and NaSH resulted in further improve in recovery of LVDP, +dp/dt, and -dp/dt by 8.3% (P = 0.0478), 8.4% (P = 0.0085), and 9.0% (P = 0.0004), respectively, and also further decrease in infarct size by 24% (P = 0.0473). Nitrite treatment decreased inducible and neuronal NO synthases (iNOS, 0.4-fold; nNOS, 0.4-fold) and cystathionine ß-synthase (CBS, 0.1-fold) expression in the isolated heart from rats with T2D. Co-administration of nitrite and NaSH further increased cystathionine γ-lyase (CSE, 2.8-fold) and endothelial NOS (eNOS, 2.0-fold) expression and further decreased iNOS (0.4-fold) expression. In conclusion, NaSH at a low dose potentiates the favorable effects of inorganic nitrite against myocardial IR injury in a rat model of T2D. These anti-ischemic effects, following co-administration of nitrite and NaSH, were associated with higher CSE-derived H2S and eNOS-derived NO as well as lower iNOS-derived NO in the diabetic hearts.

Topical spray of Wharton's jelly mesenchymal stem cells derived from umbilical cord accelerates diabetic wound healing.

BACKGROUND AND AIM: Cell-based therapy utilizing mesenchymal stem cells (MSCs) is currently being investigated as a therapeutic agent for chronic wounds. There is no evidence regarding effectiveness of the spray and local transfer of this cellular product in diabetic wound healing. Accordingly, the present study, using clinical, pathological and biometric parameters, investigated the effectiveness of the spray of these cells in the healing of diabetic wounds in rats. METHODS: Three days after the induction of diabetes (50 mg/kg single dose of streptozotocin) a circular excision was created on the back of rats. Diabetic rats were divided into two groups (n = 21): Control and WJ-MSCs group. Sampling of the studied groups was performed on Days 7, 14, and 21 after wounding. Histological, ultrasound imaging of dermis and epidermis in the wound area for thickness and density measurement and skin elasticity were evaluated. RESULTS: Our results on Days 7, 14, and 21 after wounding showed that the wound closure, thickness, and density of new epidermis and dermis, as well as skin elasticity in healed wound were significantly higher in WJ-MSCs group compared with the Control group. CONCLUSION: Application of WJ-MSCs suspension spray on the wound area can accelerate healing in diabetic wounds. Our findings may potentially provide a helpful therapeutic strategy for patients with a diabetic wound.

Combination of epidermal keratinocyte-melanocyte cells suspension and microneedling: Safe surgical approach in vitiligo.

INTRODUCTION: Vitiligo is a skin disease that is associated with impaired skin immune systems and pigment degradation in skin melanocyte cells. Despite the significant impact of the disease on the quality of life of patients, treatment of the disease using an effective method such as the transplantation of uncultivated melanocytes was considered by researchers around the world. The goal of this research was to use microneedling to transplant epidermal keratinocyte-melanocyte cells suspension for the treatment of vitiligo patients. METHODS: In this study, 15 male and female vitiligo in face region patients aged 18-45 years were studied. In this study, melanocyte-keratinocyte cells suspension was sprayed. Patients underwent microneedling treatment after spraying the cells. Before and after transplantation, patients were biometrically examined, and the quantity of pigmentation and changes in the transplanted region were documented. Statistical software was used to examine the results. RESULTS: The color difference between the lesion area and normal skin in one, two, and six months after treatment with cell suspension was significantly reduced compared with before treatment (by 48.95%). Moreover, the amount of melanin was significantly increased 6 months after treatment compared with before treatment (129.8 ± 4.16 vs. 195.2 ± 3.54, p = 0.000). A significant decrease in skin brightness in the skin of the lesion area was observed compared with normal skin, 6 months after treatment compared with before treatment (43.7 ± 1.44 vs. 27.9 ± 1.24, p = 0.000). CONCLUSION: Epidermal keratinocyte-melanocyte cells suspension in combination with microneedling could be considered as an effective treatment of vitiligo.

Tissue engineering in dermatology - from lab to market.

Tissue Engineering is a branch of regenerative medical technology which helps replace damaged tissue using appropriate scaffolding, living cells, and growth factors. Using tissue engineering products can be a promising method for treating skin lesions such as wounds and deep burns. The interaction and interconnection of cells within the bio-culture medium or within a three-dimensional scaffold provides the conditions for tissue regeneration and subsequent healing of skin wounds. Tissue engineering in the field of dermatology has evolved over time from a single application of skin cells or biopolymer scaffolds to the use of cell and scaffold combinations for the treatment, repair, and closure of acute and chronic skin wounds. It has evolved. This technology has reached a point where most products are accepted, and the body rejects a small number, which strengthens the tissue engineering market. In this article, we aimed to review and study the market of this field by reviewing various articles on tissue engineering in the field of dermatology. Tissue-engineered skin substitutes are future options for wound healing and tissue regeneration strategies.

Long Term Sodium Nitrate Administration Positively Impacts Metabolic and Obesity Indices in Ovariectomized Rats.

BACKGROUND: In postmenopausal women, nitric oxide (NO) deficiency is associated with obesity and type 2 diabetes (T2D). This study aims at determining the long-term effects of low-dose nitrate administration on metabolic and obesity indices in ovariectomized (OVX) rats. METHODS: OVX rat model was induced using the two dorsolateral skin incision method. Two months after ovariectomy, rats were divided into three groups (n = 10/group): Control, OVX, and OVX+nitrate, and the latter received sodium nitrate at a dose of 100 mg/L in their drinking water for nine months. Fasting serum glucose and lipid profile were measured every month. A glucose tolerance test was performed at months 1, 3, and 9 (the end of the study). Obesity indices were calculated, and histological analyses were performed on the gonadal adipose tissues at month 9. RESULTS: OVX rats had impaired fasting glucose, glucose intolerance, and dyslipidemia with higher obesity indices at month 9. Nitrate improved glucose and lipid metabolism in OVX rats and decreased body weight (6.9%), body mass index (12.5%), Lee index (5.4%), adiposity index (23.9%), abdominal circumference (10.5%), and thoracic circumference (17.1%). Also, nitrate decreased adipocyte area by 49% and increased adipocyte density by 193% in gonadal adipose tissue. CONCLUSION: Long-term low-dose nitrate administration improves glucose and lipid metabolism in OVX rats in association with decreasing OVX-induced adiposity, increasing adipocyte density, and decreasing adipocyte area. These findings provide support for a potential therapeutic role of nitrate in postmenopausal women with some features of metabolic syndrome.

Effect of oral nitrite administration on gene expression of SNARE proteins involved in insulin secretion from pancreatic islets of male type 2 diabetic rats.

BACKGROUND: Nitrite stimulates insulin secretion from pancreatic ß-cells; however, the underlying mechanisms have not been completely addressed. The aim of this study is to determine effect of nitrite on gene expression of SNARE proteins involved in insulin secretion from isolated pancreatic islets in Type 2 diabetic Wistar rats. METHODS: Three groups of rats were studied (n = 10/group): Control, diabetes, and diabetes + nitrite, which treated with sodium nitrite (50 mg/L) for 8 weeks. Type 2 diabetes was induced using a low-dose of streptozotocin (25 mg/kg) combined with high-fat diet. At the end of the study, pancreatic islets were isolated and mRNA expressions of interested genes were measured; in addition, protein expression of proinsulin and C-peptide in pancreatic tissue was assessed using immunofluorescence staining. RESULTS: Compared with controls, in the isolated pancreatic islets of Type 2 diabetic rats, mRNA expression of glucokinase (59%), syntaxin1A (49%), SNAP25 (70%), Munc18b (48%), insulin1 (56%), and insulin2 (52%) as well as protein expression of proinsulin and C-peptide were lower. In diabetic rats, nitrite administration significantly increased gene expression of glucokinase, synaptotagmin III, syntaxin1A, SNAP25, Munc18b, and insulin genes as well as increased protein expression of proinsulin and C-peptide. CONCLUSION: Stimulatory effect of nitrite on insulin secretion in Type 2 diabetic rats is at least in part due to increased gene expression of molecules involved in glucose sensing (glucokinase), calcium sensing (synaptotagmin III), and exocytosis of insulin vesicles (syntaxin1A, SNAP25, and Munc18b) as well as increased expression of insulin genes.

Acidified Nitrite Accelerates Wound Healing in Type 2 Diabetic Male Rats: A Histological and Stereological Evaluation.

Impaired skin nitric oxide production contributes to delayed wound healing in type 2 diabetes (T2D). This study aims to determine improved wound healing mechanisms by acidified nitrite (AN) in rats with T2D. Wistar rats were assigned to four subgroups: Untreated control, AN-treated control, untreated diabetes, and AN-treated diabetes. AN was applied daily from day 3 to day 28 after wounding. On days 3, 7, 14, 21, and 28, the wound levels of vascular endothelial growth factor (VEGF) were measured, and histological and stereological evaluations were performed. AN in diabetic rats increased the numerical density of basal cells (1070 ± 15.2 vs. 936.6 ± 37.5/mm3) and epidermal thickness (58.5 ± 3.5 vs. 44.3 ± 3.4 µm) (all p < 0.05); The dermis total volume and numerical density of fibroblasts at days 14, 21, and 28 were also higher (all p < 0.05). The VEGF levels were increased in the treated diabetic wounds at days 7 and 14, as was the total volume of fibrous tissue and hydroxyproline content at days 14 and 21 (all p < 0.05). AN improved diabetic wound healing by accelerating the dermis reconstruction, neovascularization, and collagen deposition.

Long-term nitrate administration increases expression of browning genes in epididymal adipose tissue of male type 2 diabetic rats.

Expression of browning genes are lower in both humans and animals with type 2 diabetes (T2D). This study aims at determining effects of long-term nitrate administration on protein and mRNA levels of uncoupling protein 1 (UCP1), peroxisome proliferator activated receptor gamma (PPAR-γ), and PPAR-γ coactivator 1 alpha (PGC1-α) in epididymal adipose tissue (eAT) of rats with T2D. Male Wistar rats were divided into 4 groups (n = 6/group): Control, diabetes, control + nitrate (CN), and diabetes + nitrate (DN). T2D was induced using high fat diet combined with a low-dose of streptozotocin (30 mg/kg body weight). Sodium nitrate was administrated at a dose of 100 mg/L for 6 months in nitrate-treated rats. Fasting serum glucose and insulin concentrations were measured at months 0 (i.e. at start of the protocol), 3, and 6. At month 6, protein and mRNA levels of UCP1, PPAR-γ, and PGC1-α were measured in eAT samples. In addition, tissue concentration of cyclic guanosine monophosphate (cGMP) was measured and histological analyses were done at month 6. In rats with T2D, 6-month administration of nitrate decreased serum glucose and insulin concentrations by 13% and 23%, respectively and increased cGMP level by 85%. Rats with T2D had lower mRNA and protein levels of PPAR-γ (62%, P < 0.0001 and 18%, P = 0.0472), PGC1-α (49%, P = 0.0019 and 21%, P = 0.0482), and UCP1 (35%, P = 0.0613 and 30%, P = 0.0031) in eAT; 6-month nitrate administration restored these decreased levels to near control values. In addition, nitrate increased adipocyte density by 193% and decreased adipocyte area by 53% in rats with T2D. In conclusion, long-term low-dose nitrate administration increased mRNA and protein expressions of browning genes in white adipose tissue of male rats with T2D; these findings partly explain favorable metabolic effects of nitrate administration in diabetes.

Acidified nitrite improves wound healing in type 2 diabetic rats: Role of oxidative stress and inflammation.

PURPOSE: Decreased nitric oxide bioavailability in skin contributes to impaired wound healing in type 2 diabetes (T2D). This study aims at determining effects of acidified nitrite on wound closure as well as inflammatory and antioxidants markers in wound tissue of rats with T2D. MAIN METHODS: Skin wound was made on the back of rats 28 days after the induction of T2D (high-fat diet/low-dose of streptozotocin). Control and diabetic rats were subdivided into two subgroups: Untreated control (C), acidified nitrite-treated control (CN), untreated diabetes (D), and acidified nitrite-treated diabetes (DN). Acidified nitrite was applied once daily from day 3 to day 28 and the wounds were photographed for macroscopic changes. On days 3, 7, 14, 21, and 28 after wounding, wound levels of inflammatory and antioxidant markers were measured. RESULTS: Half closure time (CT50%) was significantly lower in acidified nitrite-treated diabetic rats compared to untreated ones (5.1 vs. 8.0 days, P < 0.001). Inflammatory response was delayed in diabetic rats and persistent inflammatory response was observed at day 14 after wounding. Acidified nitrite application restored the inflammatory response and antioxidant levels to control values. CONCLUSIONS: Acidified nitrite accelerated wound healing in rats with T2D by restoring delayed inflammatory response and augmentation of antioxidant defense.

Acute physical and psychological stress effects on visceral hypersensitivity in male rat: role of central nucleus of the amygdala / Efeitos agudos do estresse físico e psicológico na hipersensibilidade visceral em rato macho: papel do núcleo central da amídala

ABSTRACT Objective: The aim of this study was to investigate the effects of acute physical and psychological stress and temporary central nucleus of the amygdala (CeA) block on stress-induced visceral hypersensitivity. Methods: Forty two male Wistar rats were used in this study. Animals were divided into 7 groups (n = 6); 1 - Control, 2 - physical stress, 3 - psychological stress, 4 - sham, 5 - lidocaine, 6 - lidocaine + physical stress and 7 - lidocaine + psychological stress. Stress induction was done using a communication box. Results: Abdominal withdrawal reflex (AWR) score was monitored one hour after stress exposure. AWR score significantly heightened at 20, 40 and 60 mmHg in the psychological stress group compared with control (p < 0.05), while, it was almost unchanged in other groups. This score was strikingly decreased at 20, 40 and 60 mmHg in lidocaine + psychological stress group compared with psychological stress with no tangible response on physical stress. Total stool weight was significantly increased in psychological stress group compared with control (0.72 ± 0.15, 0.1 ± 0.06 g) (p < 0.05), but it did not change in physical stress compared to control group (0.16 ± 0.12, 0.1 ± 0.06 g) (p < 0.05). Concomitant use of lidocaine with stress followed the same results in psychological groups (0.18 ± 0.2, 0.72 ± 0.15 g) (p < 0.05), while it did not have any effect on physical stress group (0.25 ± 0.1, 0.16 ± 0.12 g) (p < 0.05). Conclusions: Psychological stress could strongly affect visceral hypersensitivity. This effect is statistically comparable with physical stress. Temporary CeA block could also reduce visceral hypersensitivity post-acute psychological stress.

RESUMEN Objetivo: O objetivo desse estudo foi investigar os efeitos do estresse físico e psicológico agudo e bloqueio temporário do núcleo central da amídala (CeA) na hipersensibilidade visceral induzida por estresse. Métodos: Quarenta e dois ratos Wistar machos foram empregados nesse estudo. Os animais foram divididos em 7 grupos (n = 6): 1 - Controle, 2 - estresse físico, 3 - estresse psicológico, 4 - simulacro, 5 - lidocaína, 6 - lidocaína + estresse físico e 7 - lidocaína + estresse psicológico. A indução do estresse foi feita com o uso de uma caixa de comunicação. Resultados: O escore do reflexo de retirada abdominal (RRA) foi monitorado uma hora depois da exposição ao estresse. O escore RRA aumentou significativamente a 20, 40 e 60 mmHg no grupo de estresse psicológico versus controle (p < 0,05), enquanto que praticamente permaneceu inalterado nos demais grupos. Esse escore diminuiu drasticamente a 20, 40 e 60 mmHg no grupo de lidocaína + estresse psicológico versus estresse psicológico, sem resposta tangível no estresse físico. O peso total das fezes aumentou significativamente no grupo de estresse psicológico versus controle (0,72 ± 0,15, 0,1 ± 0,06 g) (p < 0,05), mas não houve mudança no grupo de estresse físico versus controle (0,16 ± 0,12, 0,1 ± 0,06 g) (p < 0,05). O uso simultâneo da lidocaína com o estresse acompanhou os mesmos resultados nos grupos psicológicos (0,18 ± 0,2, 0,72 ± 0,15 g) (p < 0,05), enquanto que não foi observado qualquer efeito no grupo de estresse físico (0,25 ± 0,1, 0,16 ± 0,12 g) (p < 0,05). Conclusões: O estresse psicológico pode afetar fortemente a hipersensibilidade visceral. Esse efeito é estatisticamente comparável com o estresse físico. Um bloqueio temporário do CeA também pode reduzir a hipersensibilidade visceral pós-estresse psicológico agudo.