Higher Numbers Of Inflammatory Cells Research Articles

Inflammatory Cells in Adipose Tissue and Skeletal Muscle of Patients with Peripheral Arterial Disease or Chronic Venous Disease: A Prospective, Observational, and Histological Study.

The main goal of this study was to assess whether the presence of peripheral arterial disease (PAD) correlates with increased inflammatory cell infiltration. An observational, single-centre, and prospective study was conducted from January 2018 to July 2022. Clinical characteristics and anthropometric measures were registered. Consecutive PAD patients with surgical indications for a common femoral artery approach and patients with varicose veins with an indication for surgical ligation of the saphenofemoral junction were included. In both groups, samples of sartorius skeletal muscle, subcutaneous adipose tissue (SAT), and perivascular adipose tissue (PVAT) were collected from the femoral region. We analysed the characteristics of adipocytes and the presence of haemorrhage and inflammatory cells in the samples of PVAT and SAT via haematoxylin-eosin staining. We found that patients with PAD had significantly more inflammatory cells in PVAT [16 (43.24%) vs. 0 (0%) p = 0.008]. Analysing SAT histology, we observed that patients with PAD had significantly more CD45+ leucocytes upon immunohistochemical staining [32 (72.73%) vs. 3 (27.27%) p = 0.005]. Upon analysing skeletal muscle histology with haematoxylin-eosin staining, we evaluated skeletal fibre preservation, as well as the presence of trauma, haemorrhage, and inflammatory cells. We registered a significantly higher number of inflammatory cells in patients with PAD [well-preserved skeletal fibres: PAD = 26 (63.41%) vs. varicose veins = 3 (37.50%) p = 0.173; trauma: PAD = 4 (9.76%) vs. varicose veins = 2 (25.00%) p = 0.229; haemorrhage: PAD = 6 (14.63%) vs. varicose veins = 0 (0%) p = 0.248; inflammatory cells: PAD = 18 (43.90%) vs. varicose veins = 0 (0%) p = 0.018]. Patients with PAD had a higher number of inflammatory cells in skeletal muscle and adipose tissue (PVAT and SAT) when compared with those with varicose veins, emphasizing the role of inflammation in this group of patients.

The effect of bradykinin 1 receptor antagonist BI 1026706 on pulmonary inflammation after segmental lipopolysaccharide challenge in healthy smokers

BackgroundBradykinin 1 receptor (B1R) signalling pathways may be involved in the inflammatory pathophysiology of chronic obstructive pulmonary disease (COPD). B1R signalling is induced by inflammatory stimuli or tissue injury and leads to activation and increased migration of pro-inflammatory cells. Lipopolysaccharide (LPS) lung challenge in man is an experimental method of exploring inflammation in the lung whereby interference in these pathways can help to assess pharmacologic interventions in COPD. BI 1026706, a potent B1R antagonist, was hypothesized to reduce the inflammatory activity after segmental lipopolysaccharide (LPS) challenge in humans due to decreased pulmonary cell influx. MethodsIn a monocentric, randomized, double-blind, placebo-controlled, parallel-group, phase I trial, 57 healthy, smoking subjects were treated for 28 days with either oral BI 1026706 100 mg bid or placebo. At day 21, turbo-inversion recovery magnitude magnetic resonance imaging (TIRM MRI) was performed. On the last day of treatment, pre-challenge bronchoalveolar lavage fluid (BAL) and biopsies were sampled, followed by segmental LPS challenge (40 endotoxin units/kg body weight) and saline control instillation in different lung lobes. Twenty-four hours later, TIRM MRI was performed, then BAL and biopsies were collected from the challenged segments. In BAL samples, cells were differentiated for neutrophil numbers as the primary endpoint. Other endpoints included assessment of safety, biomarkers in BAL (e.g. interleukin-8 [IL-8], albumin and total protein), B1R expression in lung biopsies and TIRM score by MRI as a measure for the extent of pulmonary oedema. ResultsAfter LPS, but not after saline, high numbers of inflammatory cells, predominantly neutrophils were observed in the airways. IL-8, albumin and total protein were also increased in BAL samples after LPS challenge as compared with saline control. There were no significant differences in cells or other biomarkers from BAL in volunteers treated with BI 1026706 compared with those treated with placebo. Unexpectedly, neutrophil numbers in BAL were 30% higher and MRI-derived extent of oedema was significantly higher with BI 1026706 treatment compared with placebo, 24 h after LPS challenge. Adverse events were mainly mild to moderate and not different between treatment groups. ConclusionsTreatment with BI 1026706 for four weeks was safe and well-tolerated in healthy smoking subjects. BI 1026706 100 mg bid did not provide evidence for anti-inflammatory effects in the human bronchial LPS challenge model. Trial registrationThe study was registered on January 14, 2016 at ClinicalTrials.gov (NCT02657408).

Hydrogel-Containing Biogenic Silver Nanoparticles: Antibacterial Action, Evaluation of Wound Healing, and Bioaccumulation in Wistar Rats.

Wound infections are feared complications due to their potential to increase healthcare costs and cause mortality since multidrug-resistant bacteria reduce treatment options. This study reports the development of a carbomer hydrogel containing biogenic silver nanoparticles (bioAgNPs) and its effectiveness in wound treatment. This hydrogel showed in vitro bactericidal activity after 2 h, according to the time-kill assay. It also reduced bacterial contamination in rat wounds without impairing their healing since the hydrogel hydrophilic groups provided hydration for the injured skin. The high number of inflammatory cells in the first days of the skin lesion and the greater degree of neovascularization one week after wound onset showed that the healing process occurred normally. Furthermore, the hydrogel-containing bioAgNPs did not cause toxic silver accumulation in the organs and blood of the rats. This study developed a bioAgNP hydrogel for the treatment of wounds; it has a potent antimicrobial action without interfering with cicatrization or causing silver bioaccumulation. This formulation is effective against bacteria that commonly cause wound infections, such as Pseudomonas aeruginosa and Staphylococcus aureus, and for which new antimicrobials are urgently needed, according to the World Health Organization's warning.

Angiogenesis and Tissue Repair Depend on Platelet Dosing and Bioformulation Strategies Following Orthobiological Platelet-Rich Plasma Procedures: A Narrative Review.

Angiogenesis is the formation of new blood vessel from existing vessels and is a critical first step in tissue repair following chronic disturbances in healing and degenerative tissues. Chronic pathoanatomic tissues are characterized by a high number of inflammatory cells; an overexpression of inflammatory mediators; such as tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1); the presence of mast cells, T cells, reactive oxygen species, and matrix metalloproteinases; and a decreased angiogenic capacity. Multiple studies have demonstrated that autologous orthobiological cellular preparations (e.g., platelet-rich plasma (PRP)) improve tissue repair and regenerate tissues. There are many PRP devices on the market. Unfortunately, they differ greatly in platelet numbers, cellular composition, and bioformulation. PRP is a platelet concentrate consisting of a high concentration of platelets, with or without certain leukocytes, platelet-derived growth factors (PGFs), cytokines, molecules, and signaling cells. Several PRP products have immunomodulatory capacities that can influence resident cells in a diseased microenvironment, inducing tissue repair or regeneration. Generally, PRP is a blood-derived product, regardless of its platelet number and bioformulation, and the literature indicates both positive and negative patient treatment outcomes. Strangely, the literature does not designate specific PRP preparation qualifications that can potentially contribute to tissue repair. Moreover, the literature scarcely addresses the impact of platelets and leukocytes in PRP on (neo)angiogenesis, other than a general one-size-fits-all statement that "PRP has angiogenic capabilities". Here, we review the cellular composition of all PRP constituents, including leukocytes, and describe the importance of platelet dosing and bioformulation strategies in orthobiological applications to initiate angiogenic pathways that re-establish microvasculature networks, facilitating the supply of oxygen and nutrients to impaired tissues.

Revealing of stomach histology of beta-agonist free, hygienic, and natural swine

Currently, the occurrence of diseases in the digestive system in swine has increased. Environmental stress and gastric bacteria were associated with body imbalance and stress ulcer, which are related to pathological changes in the stomach. This study aimed to investigate stomach histological characteristics of beta-agonist free, hygienic, and natural swine in different farming methods. Three types of swine’s stomachs were bought from three different farms. The samples in each part of the stomach, including the fundus, body, antrum, and pylorus, were processed through histopathological techniques and dyed with haematoxylin and eosin. The results showed that in the fundus of the stomach, beta agonist-free swine had the thinnest mucosa and submucosa layers with the highest number of inflammatory cells in both layers (p < 0.001, all). The hygienic swine showed the highest amount of red blood cells in the body, antrum, and pylorus of the stomachs (p < 0.001). In conclusion, we showed the histological characteristics of stomachs in three kinds of swine, which presents swine stomach health status. This could be information for consumers.

The Role of Local Inflammation and Hypoxia in the Formation of Hypertrophic Scars—A New Model in the Duroc Pig

Hypertrophic scars continue to be a major burden, especially after burns. Persistent inflammation during wound healing appears to be the precipitating aspect in pathologic scarring. The lack of a standardized model hinders research from fully elucidating pathophysiology and therapy, as most therapeutic approaches have sparse evidence. The goal of this project was to investigate the mechanisms of scar formation after prolonged wound inflammation and to introduce a method for generating standardized hypertrophic scars by inducing prolonged inflammation. Four wound types were created in Duroc pigs: full-thickness wounds, burn wounds, and both of them with induced hyperinflammation by resiquimod. Clinical assessment (Vancouver Scar Scale), tissue oxygenation by hyperspectral imaging, histologic assessment, and gene expression analysis were performed at various time points during the following five months. Native burn wounds as well as resiquimod-induced full-thickness and burn wounds resulted in more hypertrophic scars than full-thickness wounds. The scar scale showed significantly higher scores in burn- and resiquimod-induced wounds compared with full-thickness wounds as of day 77. These three wound types also showed relative hypoxia compared with uninduced full-thickness wounds in hyperspectral imaging and increased expression of HIF1a levels. The highest number of inflammatory cells was detected in resiquimod-induced full-thickness wounds with histologic features of hypertrophic scars in burn and resiquimod-induced wounds. Gene expression analysis revealed increased inflammation with only moderately altered fibrosis markers. We successfully created hypertrophic scars in the Duroc pig by using different wound etiologies. Inflammation caused by burns or resiquimod induction led to scars similar to human hypertrophic scars. This model may allow for the further investigation of the exact mechanisms of pathological scars, the role of hypoxia and inflammation, and the testing of therapeutic approaches.

Increased macrophage M2/M1 ratio is associated with intracranial aneurysm rupture.

Intracranial aneurysm (IA) rupture results in one of the most severe forms of stroke, with severe neurological sequelae. Inflammation appears to drive aneurysm formation and progression with macrophages playing a key role in this process. However, less is known about their involvement in aneurysm rupture. This study is aimed at demonstrating how relationship between the M1 (pro-inflammatory) and M2 (reparative) macrophage subtypes affect an aneurysm's structure resulting in its rupture. Forty-one saccular aneurysm wall samples were collected during surgery including 13 ruptured and 28 unruptured aneurysm sacs. Structural changes were evaluated using histological staining. Macrophages in the aneurysm wall were quantified and defined as M1 and M2 using HLA-DR and CD163 antibodies. Aneurysm samples were divided into four groups according to the structural changes and the M2/1 ratio. Data were analyzed using the Mann-Whitney U test. This study has demonstrated an association between the severity of structural changes of an aneurysm with inflammatory cell infiltration within its wall and subsequent aneurysm rupture. More severe morphological changes and a significantly higher number of inflammatory cells were observed in ruptured IAs (p < 0.001). There was a prevalence of M2 macrophage subtypes within the wall of ruptured aneurysms (p < 0.001). A subgroup of unruptured IAs with morphological and inflammatory changes similar to ruptured IAs was observed. The common feature of this subgroup was the presence of an intraluminal thrombus. The degree of inflammatory cell infiltration associated with a shift in macrophage phenotype towards M2 macrophages could play an important role in structural changes of the aneurysm wall leading to its rupture.

Pentoxifylline Reduces Inflammation And Myocardial Perfusion Derangements In Experimental Chronic Chagas Cardiomyopathy In Hamsters.

Introduction Microvascular perfusion disturbance (MPD) is common in chronic Chagas cardiomyopathy (CCC) and may cause myocardial damage. Hypothesis Pentoxifylline (PT) is a phosphodiesterase inhibitor with anti-inflammatory properties that may lead to reduction of inflammation and MPD and attenuate the progression of the myocardial dysfunction in CCC. Methods We investigated 3 groups of hamsters: controls and treated with saline (CO, n=9), infected with T. cruzi and treated with saline (CH+SL, n=12), infected and treated with PT (CH+PT, n=12). After 6 months of infection and 60 days of PT (20 mg/kg/day, ip) or saline, the animals were submitted to echocardiography and to rest Sestamibi-Tc99m SPECT myocardial perfusion scintigraphy (MPS). The area of myocardial perfusion defects (PD) was assessed through the calculation of polar maps. After euthanasia, we performed a histopathological study of cardiac inflammation and gene expression of TNF-α. Results At baseline all groups showed similar areas of PD (CO: 0.0±0.0%, CH+SL:0.8±1.6%, CH+PT:1.9±3.0%, p>0.05) and LVEF (CO:53.4±5.6%, CH+SL:47.0±10.1%, CH+PT:53.9±6.5%, p>0.05). After treatment, there was a significant increase of the PD only in the CH+SL group (0.8±1.6 to 9.4±9.7%, p>0.05), and CH+PT animals exhibited PD area similar to CO animals 2.7±2.7% and 0.0±0.0%, respectively (p>0.05). Similar decrease of LVEF was seen in both infected groups: (CH+SL:40.0±13.7% and CH+PT:42.7±9.0%). No change of the LVEF was observed in CO (53.4±5.6%). Quantitative histological analysis revealed a larger number of nuclei of mononuclear inflammatory cells in CH+SL, 615.1±133.0 cel/mm² and CH+PT groups, 395.7±88.3 cel/mm² when compared to CO, 193.0 ± 25.7 cel/mm², p<0.001. A higher number of inflammatory cells was found in the CH+SL group when compared to the CH+PT group, p<0.001. TNF-α expression was higher in the CH+SL group (2.5±1.7) when compared to the CO (0.9±0.4), but similar to the CH+PT (2.4±1.7), p>0.9. There was a moderate correlation between PD and inflammatory cells (R=0.6) and LVEF (R=-0.7), p<0.0001 for both; and between LVEF and inflammatory cells (R=-0.4, p=0.01). Conclusions The prolonged use of PT in CCC animals was effective in reducing inflammation and progression of MPD evaluated in vivo. However, PT did not prevent the progression of left ventricular systolic dysfunction in this model. These results suggest that PD may represent a surrogate marker for inflammation and its potential use to monitor disease progression. Microvascular perfusion disturbance (MPD) is common in chronic Chagas cardiomyopathy (CCC) and may cause myocardial damage. Pentoxifylline (PT) is a phosphodiesterase inhibitor with anti-inflammatory properties that may lead to reduction of inflammation and MPD and attenuate the progression of the myocardial dysfunction in CCC. We investigated 3 groups of hamsters: controls and treated with saline (CO, n=9), infected with T. cruzi and treated with saline (CH+SL, n=12), infected and treated with PT (CH+PT, n=12). After 6 months of infection and 60 days of PT (20 mg/kg/day, ip) or saline, the animals were submitted to echocardiography and to rest Sestamibi-Tc99m SPECT myocardial perfusion scintigraphy (MPS). The area of myocardial perfusion defects (PD) was assessed through the calculation of polar maps. After euthanasia, we performed a histopathological study of cardiac inflammation and gene expression of TNF-α. At baseline all groups showed similar areas of PD (CO: 0.0±0.0%, CH+SL:0.8±1.6%, CH+PT:1.9±3.0%, p>0.05) and LVEF (CO:53.4±5.6%, CH+SL:47.0±10.1%, CH+PT:53.9±6.5%, p>0.05). After treatment, there was a significant increase of the PD only in the CH+SL group (0.8±1.6 to 9.4±9.7%, p>0.05), and CH+PT animals exhibited PD area similar to CO animals 2.7±2.7% and 0.0±0.0%, respectively (p>0.05). Similar decrease of LVEF was seen in both infected groups: (CH+SL:40.0±13.7% and CH+PT:42.7±9.0%). No change of the LVEF was observed in CO (53.4±5.6%). Quantitative histological analysis revealed a larger number of nuclei of mononuclear inflammatory cells in CH+SL, 615.1±133.0 cel/mm² and CH+PT groups, 395.7±88.3 cel/mm² when compared to CO, 193.0 ± 25.7 cel/mm², p<0.001. A higher number of inflammatory cells was found in the CH+SL group when compared to the CH+PT group, p<0.001. TNF-α expression was higher in the CH+SL group (2.5±1.7) when compared to the CO (0.9±0.4), but similar to the CH+PT (2.4±1.7), p>0.9. There was a moderate correlation between PD and inflammatory cells (R=0.6) and LVEF (R=-0.7), p<0.0001 for both; and between LVEF and inflammatory cells (R=-0.4, p=0.01). The prolonged use of PT in CCC animals was effective in reducing inflammation and progression of MPD evaluated in vivo. However, PT did not prevent the progression of left ventricular systolic dysfunction in this model. These results suggest that PD may represent a surrogate marker for inflammation and its potential use to monitor disease progression.

Periodontal ligament repair after active splinting of replanted dogs' teeth.

The high rate of root resorption resulting from tooth replantation represents a serious clinical problem. In order to prevent ankylosis and replacement resorption, the contemporary literature highlights the importance of using a flexible stabilization for traumatized teeth. For this purpose, orthodontic devices may be promising for obtaining a better prognosis and periodontal repair. The aim of this study was to evaluate the effect of an active splinting protocol with controlled force in dog's teeth following replantation. Sixty premolar roots from three dogs were used. They were submitted to endodontic treatment, hemisected, atraumatically extracted and subsequently replanted. They were divided into four groups: Passive Stabilization (n=20)-after 20min in a dry medium; Active Stabilization (n=20)-after 20min in a dry medium; Negative control (n=10)-immediate replantation and passive Stabilization; and Positive control (n=10)-90min of extra-alveolar time and passive Stabilization. The samples were collected and submitted to histologic processing. They were then evaluated for the count of inflammatory cells, expression of neurotrophin 4, osteoclasts, apoptotic cells and collagen fibres. The results were submitted to ANOVA or Kruskal-Wallis statistical tests followed by Tukey or Dunn post-tests (α=5%). Passive Stabilization with orthodontic brackets without traction used after replantation had the highest number of inflammatory cells (p=.0122), osteoclasts (p=.0013) and percentage of collagen fibres in the periodontal ligament (p<.0001) when compared to Active Stabilization with orthodontic brackets applying amild tensile force. Neurotrophin 4had no statistically significant difference (p=.05), regardless of the treatment. The apoptotic cells count revealed statistical differences (p<.0001) between Active Stabilization (189.70±47.99) and Positive Control (198.90±88.92) when compared to Passive Stabilization (21.19±32.94). The active splinting protocol using orthodontic appliances generating a light and controlled force favoured periodontal ligament repair of replanted teeth.

Biocompatibility and biomineralization of the experimental nanoparticulate mineral trioxide aggregate (MTA)

To investigate the tissue response and the biomineralization ability of the experimental nanoparticulate mineral trioxide aggregate compared to grey MTA and Fillapex MTA. Polyethylene tubes containing materials or empty tubes for control were inserted into the subcutaneous tissues of 30 rats. After 7, 15, 30, 60, and 90 days, the rats were killed and the tubes were removed for analysis using hematoxylin-eosin staining, von Kossa staining, and under polarized light. Inflammation was graded through a score system; the biomineralization ability was recorded as present or absent. The results were statistically analyzed using the Kruskal-Wallis test (p&lt;0.05). On days 7 and 15 there was a significant difference between the Nano MTA (median score of 3) and MTA Fillapex groups (median score of 4), being MTA Fillapex the material with the highest number of inflammatory cells. At 30, 60, and 90 days there was no difference between the Nano MTA, Grey MTA, and MTA Fillapex groups. All materials induced the formation of mineralized tissue in all experimental periods. Nano MTA showed biocompatibility and biomineralization similar to grey MTA Angelus.

Equine herpesvirus 1 elicits a strong pro-inflammatory response in the brain of mice.

Equine herpesvirus type 1 (EHV-1) is an emerging pathogen that causes encephalomyelitis in horses and non-equid species. Several aspects of the immune response in the central nervous system (CNS), mainly regarding the role of inflammatory mediators during EHV-1 encephalitis, remain unknown. Moreover, understanding the mechanisms underlying extensive neuropathology induced by viruses would be helpful to establish therapeutic strategies. Therefore, we aimed to evaluate some aspects of the innate immune response during highly neurovirulent EHV-1 infection. C57BL/6 mice infected intranasally with A4/72 and A9/92 EHV-1 strains developed a fulminant neurological disease at 3 days post-inoculation with high viral titres in the brain. These mice developed severe encephalitis with infiltration of monocytes and CD8+ T cells to the brain. The inflammatory infiltrate followed the detection of the chemokines CCL2, CCL3, CCL4, CCL5, CXCL2, CXCL9 and CXCL-10 in the brain. Notably, the levels of CCL3, CCL4, CCL5 and CXCL9 were higher in A4/72-infected mice, which presented higher numbers of inflammatory cells within the CNS. Pro-inflammatory cytokines, such as interleukins (ILs) IL-1α, IL-1β, IL-6, IL-12β, and tumour necrosis factor (TNF), were also detected in the CNS, and Toll-like receptor (TLR) TLR2, TLR3 and TLR9 genes were also upregulated within the brain of EHV-1-infected mice. However, no expression of interferon-γ (IFN-γ) and IL-12α, which are important for controlling the replication of other herpesviruses, was detected in EHV-1-infected mice. The results show that the activated innate immune mechanisms could not prevent EHV-1 replication within the CNS, but most likely contributed to the extensive neuropathology. The mouse model of viral encephalitis proposed here will also be useful to study the mechanisms underlying extensive neuropathology.

Cytotoxicity, inflammation, biomineralization, and immunoexpression of IL-1β and TNF-α promoted by a new bioceramic cement.

Aim To evaluate the cytotoxicity, biocompatibility and mineralization capacity of BIO-C PULPO, and MTA.Methodology L929 fibroblasts were cultured and MTT assay was used to determine the material cytotoxicity on 6, 24, and 48 h. A total of 30 male rats (Wistar) aged between 4 and 6 months, weighing between 250 and 300 g were used. Polyethylene tubes containing BIO-C PULPO, MTA, and empty tubes were implanted into dorsal connective tissue. After the experimental periods (7, 15, 30, 60, and 90 days) the tubes were histologically analyzed using hematoxylin-eosin (H&E), immunolabeling of IL-1β and TNF-α, and von Kossa staining, or without staining for polarized light analysis. The average number of inflammatory cells was quantified; the mineralization assessment was determined by the area marked in μm2 and semiquantitative immunolabeling analyses of IL-1β and TNF-α were performed. Then, data underwent statistical analysis with a 5% significance level.Results It was observed that BIO-C PULPO and MTA presented cytocompatibility at 6, 24, and 48 similar or higher than control for all evaluated period. On periods 7 and 15 days, BIO-C PULPO was the material with the highest number of inflammatory cells (p<0.05). On periods 30, 60, and 90 days, BIO-C PULPO and MTA presented similar inflammatory reactions (p>0.05). No statistical differences were found between Control, BIO-C PULPO, and MTA for immunolabeling of IL-1β and TNF-α in the different periods of analysis (p<0.05). Positive von Kossa staining and birefringent structures under polarized light were observed in all analyzed periods in contact with both materials, but larger mineralization area was found with BIO-C PULPO on day 90 (p<0.05).ConclusionBIO-C PULPO was biocompatible and induced mineralization similar to MTA.

Obesity worsens the outcome of influenza virus infection associated with impaired type I interferon induction in mice

Increasing evidence indicates that obesity is a risk factor for increased severity of influenza virus infection. However, its precise immunological mechanism is not fully understood. To investigate this, diet-induced obese (DIO) mice were established by feeding C57BL/6 male mice a high-fat diet for 16 weeks. DIO and lean control mice were infected intranasally with 3000 pfu of influenza A virus (IAV) (PR8/H1N1). Interestingly, we found adipose tissue located along the bronchus in naïve DIO mice. In addition, the Nos2 level was significantly higher and Arg1 level was significantly lower in lung macrophages of naïve DIO mice, consistent with an M1-skewed phenotype. The survival rate and body weight of DIO mice infected with IAV were significantly lower than those of lean control mice and associated with higher viral load in the lungs of DIO mice. Histopathological analysis demonstrated higher numbers of inflammatory cells in the lungs of DIO mice after IAV infection. Levels of cytokines, including TNF-α, IL-6, IL-10, and type I IFN (IFN-α and IFN-β), in bronchoalveolar lavage fluid (BALF) were altered after IAV infection; in particular, IFN-α and IFN-β levels were significantly suppressed in the BALF of DIO mice. In vitro, bone marrow-derived macrophages were stimulated with ligands of toll-like receptor (TLR) 7/8, a pattern recognition receptor for single-stranded RNA, and levels of TNF-α, IL-6, and IL-10 were similarly altered. In addition, levels of IFN-α and IFN-β were significantly lower in culture supernatants of alveolar macrophages sorted from naïve DIO mice and infected with IAV, compared to those in macrophages sorted from lean control mice. Collectively, these results suggest that macrophages may be the main contributors to poor outcomes of influenza virus infection in obesity.

Untersuchung des Tränenfilms und histologische Untersuchung der Augenlider von Möpsen

To determine if qualitative tear film and histological changes of the eyelid margins in pugs compared to other brachycephalic dogs could be potential contributing factors to the high prevalence of corneal diseases in this breed. Ophthalmic examin ation (including tear film break-up time [TFBUT] and meibometry) was undertaken on three groups (pugs with and without ophthalmologic abnormalities as well as on other brachycephalic breeds with history of ophthalmologic abnormalities). Histology of eyelid tissue obtained during medial canthoplasty was performed, using hematoxylin-eosin and oil-red-O-staining. Seventy-eight pugs and 11 brachycephalic dogs were included. Mean ages were 3.54 and 5.5 years respectively. STT 1 values below 15 mm/min were found in 12 of 150 eyes of pugs and in three of 18 eyes of other brachycephalic dogs. Tear film break-up time values below 20 seconds were determined in 118 of 126 eyes of pugs, and in eight of 18 eyes of other brachycephalic dogs. Meibometry values over 100 MU were only identified in 20 of 147 eyes of pugs and 12 of 20 eyes in other brachycephalic dogs. Eyelid tissue of 21 pugs and 11 brachycephalic dogs was obtained. All pugs had a higher number of inflammatory cells in eyelid tissue than other brachycephalic breeds. Young pugs are often presented with STT 1 values within the reference range and low TFBUT and meibometry values. As other brachycephalic breeds are often presented with STT 1 values within reference ranges as well as low TFBUT values, the only marked difference between pugs and other brachycephalic breeds are low meibometry values and the higher number of inflammatory cells in the medial canthal lid margins.

The Subcutaneous Tissue Reaction on Poly (L-lactide-co-caprolactone) based Threads

The thread-lifting procedure is gaining popularity all over the globe. It is a mini-invasive, rejuvenate procedure with a short downtime. Aesthetic practitioners use various medical devises and thread materials during thread lifting procedures, but thread degradation and tissue reaction on thread implants are currently poorly understood. Objective: This article will describe tissue reaction and thread changes from implantation to full degradation. Methods: Tissue reaction on L-lactide-co-caprolactone threads was observed on 0, 4, 13, 26, 34, 52, 64, and 72 weeks after implantation in 21 rabbit models. In all groups, except the 26-week group, eight implants were placed in each model. Specifically, four Test Articles-Aptos Excellence Visage poly (L-lactide-co-caprolactone) threads were implanted on the right side in the cranial-caudal direction, and four control articles were implanted on the left side in the cranial-caudal direction. Results: Analysis of the local tissue reaction showed that the Test Article and the control article caused the same tissue reaction. However, compared with the control article, Test Article was associated with higher numbers of inflammatory cells on 13 and 34–72 weeks. At 72 weeks, the average area of Test Article had decreased by 41.1%. Conclusion: After subcutaneous placement of the APTOS thread, the thread was progressively surrounded by fibrous tissue and exhibited slow degradation (41.1% over 72 weeks). The prolonged tissue reaction guarantees stable and durable tread-lifting procedure results. The ability of Photic tissue repositions, nucleogenesis and neovascularization has a strong influence on skin texture, structure, color and body couture which is provided by APTOS threads composition and structure.

Randomized controlled clinical study of veneered zirconia abutments for single implant crowns: Clinical, histological, and microbiological outcomes.

To analyze the effect of veneering of the submucosal part of zirconia abutments and the type of retention (cemented vs screw-retained) on clinical, microbiological, and histological outcomes of single-tooth implant crowns. A total of 44 patients with a single missing tooth to be replaced by an implant in the anterior region participated in the study. Implants were randomly assigned to receive zirconia-based CAD/CAM reconstructions using either one of four treatment modalities: cement-retained with submucosal veneering (CR-P), cement-retained without submucosal veneering (CR-W), screw-retained with submucosal veneering (SR-P), and screw-retained without submucosal veneering (SR-W). Clinical parameters were assessed at baseline (after crown insertion), at 6 and 12 months. Histological and microbiological analyses were performed at 6 months. Descriptive statistics and the Kruskal-Wallis test were applied. The clinical evaluation revealed, in general, stable peri-implant soft tissues with minimal differences for all measured parameters between the four groups, except for bleeding on probing with the two cemented groups exhibiting higher values at 12 months (35.0% ± 26.5% for CR-W and 25.0% ± 38.8% for CR-P versus 13.1 ± 14.8 for SR-W and 13.0 ± 18.2 for SR-P). The descriptive and semi-quantitative histology showed a trend for a higher inflammatory reaction in the two cemented (a medium to high number of inflammatory cells) compared to the screw-retained groups (low number of inflammatory cells) at 6 months. The microbiological test demonstrated low bacterial counts and a similar distribution in between the groups except for two species (Tannerella forsythia and Peptostreptococcus micros) that were found in higher counts in the cemented groups at 6 months. Submucosal veneering of zirconia abutments did not negatively affect the health of the peri-implant tissues. The cemented groups, though, did show a clinical and histological trend to higher levels of inflammation.

Morphological Classification of Corrupted Colonic Crypts in Ulcerative Colitis

In ulcerative colitis (UC) the colonic mucosa shows, in addition to a high number of inflammatory cells, crypts with architectural distortions, called corrupted colonic crypts (CCC). Here we classify the histologic repertoire and assess the frequency of CCC in UC. Five-hundred and sixteen histologic sections from 29 colectomy specimens with UC (24 having adenocarcinoma and five, high-grade dysplasia, HGD) were reviewed. The vast majority of the colonic mucosa portrayed countless crypts with normal shapes (CNS) lined with normal epithelium, except for 45 CNS: 28 showed inconclusive-suspected cellular changes (ISCC), and 17, high-grade dysplasia (HGD). CCC were subdivided into four groups: i) Crypts with fission distortions, ii) Crypts with length distortions, iii) Crypts with outline distortions and iv) Crypts with axial polarity distortions. The most frequent CCC group had axial polarity distortions (33.4%), and the less frequent CCC group, outline distortions (21.1%) (p<0.05). No apparent differences in frequency between groups were found in colectomies with HGD/carcinoma, or in colectomies preformed for medically-refractory UC without HGD/carcinoma. Out of the 902 CCC present in the specimens, 343 (38.0%) displayed ISCC, 186 (20.6%) HGD, and the remaining 373 (41.4%) normal epithelium. Hence, of the 203 crypts exhibiting HGD, 186 (91.6%) were CCC and the remaining 17 (8.4%) CNS (p<0.05). Based on these findings it is suggested that the microscopic search for HGD in UC colectomy-specimens should preferentially be focused on mucosal areas exhibiting CCC. This view is validated by recent findings showing that p53 overexpression (a biomarker of epithelial carcinogenesis) significantly correlated with architectural distortions of the crypts in UC.

Reduced urothelial regeneration in rat bladders augmented with permeable porcine small intestinal submucosa assessed by magnetic resonance imaging.

Augmentation enterocystoplasty remains the gold standard surgical bladder reconstruction procedure to increase the capacity and compliance of dysfunctional bladders. Since the use of the patient's intestine has severe risks of complications, alternative biodegradable matrices have been explored. Porcine small intestinal submucosa (SIS) has gained immense interests in bladder reconstruction due to its favorable properties. However, trials have shown inconsistent regeneration with SIS, attributed to the heterogeneity in microstructures and mechanical properties. We hypothesize that uneven SIS permeability to urine is a factor responsible for the inconsistency. We measured permeability to urine in situ using a contrast enhanced-magnetic resonance imaging (MRI), and evaluated urothelium regeneration using immunohistochemical staining of urothelial cell markers in SIS-augmented rat bladders. Results showed significant differences in permeability among SIS-augmented rat bladders. Commercial SIS scaffolds were then categorized into nonleaky and leaky groups based on MRI results. Hematoxylin and eosin staining showed higher numbers of inflammatory cells in leaky SIS on day 14 relative to nonleaky SIS. In addition, trichrome staining showed major changes in the distribution of collagen on day 28 between SIS-augmented bladder groups. Furthermore, expressions of urothelium-associated markers (cytokeratins AE1/AE3, claudin 4, and uroplakin III) were completed in bladders augmented with nonleaky SIS, whereas limited urothelial differentiation was noticed in leaky SIS-augmented bladders at post-augmentative day 14. These results show that scaffold permeability to urine may be responsible for variations in regenerative capacity of porcine SIS. Applications of MRI technique will be helpful to understand a relationship between biomaterial property and regenerative capacity. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1778-1787, 2018.

Melatonin suppresses eosinophils and Th17 cells in hamsters treated with a combination of human liver fluke infection and a chemical carcinogen.

The combination of Opisthorchis viverrini (OV) infection and chemical carcinogen induces cholangiocarcinoma (CCA) in hamsters via inflammation-mediated mechanisms. Thus, suppression of inflammatory cells at the initial stages of CCA development would be of benefit. We aimed to investigate whether IL-17-producing CD4+ T cells (Th17) and CD4+ Foxp3+ T cells (Treg) are involved in the early stages of CCA genesis and can be targeted for suppression by melatonin. Inflammation, an initial stage of CCA development, was induced in hamsters by a combination of O. viverrini infection and N-nitrosodimethylamine (NDMA) administration. Melatonin (50mg/kg) was additionally administered to one group for the 30days of the experiment. Liver tissue-resident T cells were investigated using immunostaining, western blotting, and real-time PCR. OV+NDMA-induced CCA tissues showed significantly higher numbers of inflammatory cells, especially eosinophils, bile duct proliferation and IL-17+ cell infiltration compared to normal livers. Expression of Foxp3 was localized in the bile duct epithelial cells, and especially in the bile duct hyperplasia. Accumulation of CD4+ and IL-17+ cells and intense staining of the Foxp3+ marker were consistent with their protein levels. Infiltration of IL-17+ inflammatory cells and Foxp3+ cells, as well as increases in their transcription expression levels, were significantly lower in the melatonin-treated group. In contrast, increased CD4+ cell infiltration and TNF-α expression were also observed through melatonin treatment. Melatonin exerts an immunomodulatory effect, suppressing eosinophils and Th17 cells and expression of Foxp3, but enhancing CD4+ cells and TNF-α. This suggests that melatonin may be used for CCA chemoprevention.

Alterations in biomechanical properties and microstructure of colon wall in early-stage experimental colitis.

The aim of the current study was to investigate the effects of early-stage dextran sodium sulfate (DSS)-induced mouse colitis on the biomechanical properties and microstructure of colon walls. In the present study, colitis was induced in 8-week-old mice by the oral administration of DSS, and then 10 control and 10 experimental colitis samples were harvested. Uniaxial tensile tests were performed to measure the ultimate tensile strength and ultimate stretches of colon tissues. In addition, histological investigations were performed to characterize changes in the microstructure of the colon wall following treatment. The results revealed that the ultimate tensile stresses were 232±33 and 183±25 kPa for the control and DSS groups, respectively (P=0.001). Ultimate stretches at rupture for the control and DSS groups were 1.43±0.04 and 1.51±0.06, respectively (P=0.006). However, there was no statistically significant difference in tissue stiffness between the two groups. Histological analysis demonstrated high numbers of inflammatory cells infiltrated into the stroma in the DSS group, leading to significant submucosa edema. Hyperplasia was also identified in the DSS-treated submucosa, causing a disorganized microstructure within the colon wall. Furthermore, a large number of collagen fibers in the DSS-treated muscular layer were disrupted, and fiber bundles were thinner when compared with the control group. In conclusion, early-stage experimental colitis alters the mechanical properties and microstructural characteristics of the colon walls, further contributing to tissue remodeling in the pathological process.