Presence Of Inflammatory Cell Infiltrate Research Articles

Comparative assessment of tissue response to a mesh implant made of polypropylene modified with carbon nanotubes and silver nanoparticles

Treatment of abdominal hernias remains one of the most urgent problems of modern surgery. A large number of complications after hernia operations require the development of new types of implants for tissue plastic surgery. The purpose of the study is to carry out an experimental comparative assessment of tissue reaction to the implantation of the developed polypropylene mesh implant modified with carbon nanotubes and silver nanoparticles. Research was conducted on 105 sexually mature laboratory rats in three series of experiments (35 rats each). In the first series, polypropylene mesh implants were implanted in the tissues of the anterior abdominal wall, in the second – polypropylene implants coated with an antiseptic, and in the third – polypropylene implants modified with carbon nanotubes and silver nanoparticles. Animals were removed from the experiment after preliminary anesthesia 3, 5, 7, 14, 21, 30 and 90 days after the operation. The tissues of the abdominal wall were taken together with the implants, histological preparations were made, which were stained with hematoxylin and eosin and according to Van Gieson. We studied the composition and ratio of elements of cell infiltration in tissues with further statistical processing of the obtained data. It was established that during the implantation of mesh edoprostheses, regardless of their type, necrotic changes with reactive inflammation, the presence of inflammatory cell infiltrate, tissue swelling and microcirculation disorders were detected in the tissues 3-5 days after the operation. Under the condition of implantation of the developed mesh, a less pronounced exudative phase of inflammation and an earlier onset of the reparation phase were detected. Depending on the type of implant used, the subsequent reaction differed between the groups of experimental animals, which was evidenced by the regression of inflammatory phenomena in the tissues and the processes of formation of the fibrous capsule around the implants. During the implantation of nanomodified mesh implants by the 7th day of the experiment, the exudative phase of inflammation ended and the formation of a thin connective tissue capsule began, the formation of which was completed by the 21st day of observation, while when the mesh was implanted with polypropylene and polypropylene with an antimicrobial coating, the formation of the capsule lasted up to 30 days. Thus, it was established that the exudative phase of inflammation continues in the tissues around the implantation of classic polypropylene and polypropylene meshes with an antimicrobial coating up to the 14th day of the experiment, and the connective tissue capsule is formed up to the 30th day. At the same time, in the tissues around the mesh implants made of polypropylene modified with carbon nanotubes and silver nanoparticles, the exudative phase of inflammation ended by the 7th day of the experiment. This ensured the intensity of reparative regeneration processes and the separation of the implant from the surrounding tissues by a thin connective tissue capsule for up to 21 days of observation.

Influence of antiresorptive/antiangiogenic therapy on changes in periodontal and oral tissue structures: a histomorphometrical analysis in rats

ObjectiveThe objective of this study was to investigate the influence of various antiresorptive and antiangiogenic medications on morphological changes in periodontal and oral tissue structures.Materials and methodsFifty-five Wistar rats randomly received dual application (i.e., at baseline and after 12-weeks) one of the following medications: (1) amino-bisphosphonate [zoledronate (Zo)], (2) RANKL inhibitor [denosumab (De)], (3) antiangiogenic [bevacizumab (Be)], (4) Zo + Be, (5) De + Be or (6) no medication [Control (Co)]. Periodontal and oral tissue biopsies were obtained at 17 (n = 21 animals, Phase 1, (De = 3, De + Be = 3, Zo = 5, Be = 3, Zo + Be = 2, Co = 5) and 29 (n = 34 animals, (De = 8, De + Be = 6, Zo = 2, Be = 7, Zo + Be = 4, Co = 7, Phase 2) weeks after the second drug application. The following outcomes were histomorphometrically assessed: periodontal space width in the coronal (PLS-C, mm) and apical sections (PLS- A), number of empty alveolar bone lacunae in the coronal, apical sections and at the apex at respective tooth sites (EL – C, EL- A, EL- Ap), mucosal thickness at edentulous alveolar ridge areas (MT, mm), and, when present, associated areas of inflammatory cell infiltrates (ICI, mm2).ResultsComparable mean PLS-C, PLS-A, ET-A, ET-C, ET-Ap, and MT values were observed in all experimental groups after Phases 1 and 2. The presence of ICI was identified in 3 animals in the Co group (Phase 1: 1, Phase 2: 2), and 17 animals in the test groups (Phase 1: 4; Phase 2: 14). The estimated ICI surface area was significantly higher in the Zo + Be group, followed by the Zo and Be groups compared to that measured in the Co group. The time (i.e., Phases 1 and 2) was not found to be a predictor for the extent of the ICI area. In all groups, the EL-C, EL-A, and EL-Ap values were significantly higher after Phase 2 compared to those assessed after Phase 1. The MT values were significantly reduced in all groups after Phase 2 compared to those measured after Phase 1.ConclusionsThe present evaluation was not able to find any morphological effects of different antiresorptive and antiangiogenic medications on periodontal and oral tissue structures. The presence of inflammatory cell infiltrates was more frequently observed in the animals administered with antiresorptive and antiangiogenic medications as well as combinations thereof.Clinical relevanceAdministration of antiresorptive and antiangiogenic medications may be capable of inducing inflammatory reactions in periodontal tissues.

EXPERIMENTAL AND MORPHOLOGICAL ASSESSMENT OF THE INFLUENCE OF HYDROXYAPATITE-CONTAINING OSTEOTROPIC MATERIAL AND ELECTRICAL STIMULATION ON REPARATIVE OSTEOGENESIS OF THE LOWER JAW.

Aim: The aim of the study was to reveal the peculiarities of reparative osteogenesis in experimental lower jaw defect under the conditions of hydroxyapatite-containing osteotropic material application and electrical stimulation. Materials and Methods: An experiment was conducted on 48 mature male rats of the WAG population. All animals were divided into 4 groups (12 animals in each group). Group 1 included rats that were not subjected to any manipulations. Group 2 included rats that were modeled with a perforated defect of the lower jaw body. Group 3 included rats that were modeled with a perforated defect similar to group 2, the cavity of which was filled with synthetic bone graft "Biomin GT" (RAPID, Ukraine). Group 4 included animals that were modeled with a perforated defect similar to groups 2-3, the cavity of which was filled with synthetic bone graft "Biomin GT". In animals of group 4, a microdevice for electrical action was implanted subcutaneously in the neck area on the side of the simulated bone defect. Morphological and statistical methods were used. Results: The research carried out by the authors proved that the use of the above-mentioned bone replacement material helps to increase the regenerative potential of the bone tissue of the lower jaw, but does not lead to the formation of a full-fledged bone regenerate, as evidenced by the results of the morphometry of the regenerate (the specific volume of lamellar bone tissue accounted for 54.9%); disordered localization of bone beams, which were characterized by reduced signs of mineralization; the presence in connective, osteogenic fibroreticular and lamellar bone tissues the encapsulated bone graft granules with the presence of inflammatory cell infiltration. In cases the combined use of synthetic bone graft "Biomin GT" and electrical stimulation, the authors noted more intensive reparative osteogenesis processes in the bone defect of the lower jaw compared to cases when only one bone graft was used, but they also did not lead to the formation of a full-fledged bone regenerate. Conclusions: The experimental and morphological study conducted by the authors proved that the use of hydroxyapatite-containing osteotropic material ("Biomin GT"), especially in cases of its combined use with electrical stimulation, significantly activates reparative osteogenesis in the bone defect of the lower jaw, which does not lead to the formation of a full-fledged bone regenerate.

Primary infection of BALB/c mice with a dengue virus type 4 strain leads to kidney injury

Dengue is a disease caused by dengue virus (DENV-1 through -4). Among the four serotypes, DENV-4 remains the least studied. Acute kidney injury is a potential complication of dengue generally associated with severe dengue infection.The goal of this study was to investigate the alterations caused by experimental dengue infection in the kidney of adult BALB/c mice.In this study, BALB/c mice were infected through the intravenous route with a DENV-4 strain, isolated from a human patient. The kidneys of the mice were procured and subject to histopathological and ultrastructural analysis.The presence of the viral antigen was confirmed through immunohistochemistry. Analysis of tissue sections revealed the presence of inflammatory cell infiltrate throughout the parenchyma. Glomerular enlargement was a common find. Necrosis of tubular cells and haemorrhage were also observed. Analysis of the kidney on a transmission electron microscope allowed a closer look into the necrotic tubular cells, which presented nuclei with condensed chromatin, and loss of cytoplasm.Even though the kidney is probably not a primary target of dengue infection in mice, the inoculation of the virus in the blood appears to damage the renal tissue through local inflammation.

PS-C28-6: HIGH TESTOSTERONE AND LOW ESTRADIOL SERA LEVELS ARE ASSOCIATED WITH UNSTABLE PLAQUE COMPOSITION IN MEN WITH SEVERE CAROTID ATHEROSCLEROSIS

Background and Objectives: Ischemic strokes caused by the rupture of unstable atherosclerotic plaques in the carotid arteries, are leading causes of mortality and disability worldwide. Sex differences exist in plaque composition, where men are more likely to develop unstable plaques than women. Sex hormones affect vascular reactivity and inflammation, but it remains unclear if they are involved in plaque instability and how their role differs in men and women. Herein, we investigated the association between circulating endogenous sex hormone levels and plaque composition/instability in men and women with severe carotid atherosclerosis. Design and method: Pre-operative fasting blood samples and plaque specimens were collected from post-menopausal women and men greater than 55 years old with severe carotid atherosclerosis who underwent a carotid endarterectomy at McGill-affiliated hospitals in Montreal, Canada. Plaque composition and stability were examined by experienced vascular pathologists according to gold-standard histological classifications. Liquid chromatography-tandem mass spectrometry-based methods were developed to quantify endogenous sex hormones, 17ß-estradiol (E2), testosterone, androstenedione, and dehydroepiandrosterone, in sera samples. Results: Subjects (n = 460) had a mean age of 71.0 ± 9.0, were 68.9% male, and had various comorbidities (hypercholesterolemia [82.6%], hypertension [84.6%], type 2 diabetes [34.5%]). Clinical characteristics were similar among post-menopausal women (n = 143) and men (n = 317) with stable versus unstable plaques. Men had a significantly greater proportion of unstable versus stable plaques compared to women (men unstable: 61.5%, women unstable: 41.3%; P < 0.001), characterized predominantly by a large lipid-rich core, hemorrhage, and high presence of inflammatory cell infiltration. Men with unstable plaques had significantly higher levels of testosterone than men with stable plaques (P = 0.004). In men, testosterone levels were inversely associated with fibrosis (P < 0.05), while directly associated with unstable plaque features, including greater lipid core size (P < 0.05), and a greater presence of foam cells (P < 0.05), neovascularization (P < 0.01), and plaque (P < 0.01) and cap inflammation (P < 0.05). No associations were observed between testosterone levels and plaque features in women. Endogenous E2 levels were inversely associated with neovascularization (P < 0.05) and plaque and cap inflammation (P < 0.001) in men, while inversely associated with fibrosis levels in women (P < 0.05). Conclusion: Our findings suggest that higher testosterone and lower E2 levels are associated with unstable plaque composition in men with carotid atherosclerotic disease and may partly explain the higher proportion of unstable plaques observed among men than women of similar age and comorbidity status. Instead, in women, estradiol appears to play a detrimental role in plaque instability by inversely affecting fibrous tissue levels (a key stabilizing feature of plaques).

Myocarditis-like Episodes in Patients with Arrhythmogenic Cardiomyopathy: A Systematic Review on the So-Called Hot-Phase of the Disease.

Arrhythmogenic cardiomyopathy (ACM) is a genetically determined myocardial disease, characterized by myocytes necrosis with fibrofatty substitution and ventricular arrhythmias that can even lead to sudden cardiac death. The presence of inflammatory cell infiltrates in endomyocardial biopsies or in autoptic specimens of ACM patients has been reported, suggesting a possible role of inflammation in the pathophysiology of the disease. Furthermore, chest pain episodes accompanied by electrocardiographic changes and troponin release have been observed and defined as the “hot-phase” phenomenon. The aim of this critical systematic review was to assess the clinical features of ACM patients presenting with “hot-phase” episodes. According to PRISMA guidelines, a search was run in the PubMed, Scopus and Web of Science electronic databases using the following keywords: “arrhythmogenic cardiomyopathy”; “myocarditis” or “arrhythmogenic cardiomyopathy”; “troponin” or “arrhythmogenic cardiomyopathy”; and “hot-phase”. A total of 1433 titles were retrieved, of which 65 studies were potentially relevant to the topic. Through the application of inclusion and exclusion criteria, 9 papers reporting 103 ACM patients who had experienced hot-phase episodes were selected for this review. Age at time of episodes was available in 76% of cases, with the mean age reported being 26 years ± 14 years (min 2–max 71 years). Overall, 86% of patients showed left ventricular epicardial LGE. At the time of hot-phase episodes, 49% received a diagnosis of ACM (Arrhythmogenic left ventricular cardiomyopathy in the majority of cases), 19% of dilated cardiomyopathy and 26% of acute myocarditis. At the genetic study, Desmoplakin (DSP) was the more represented disease-gene (69%), followed by Plakophillin-2 (9%) and Desmoglein-2 (6%). In conclusion, ACM patients showing hot-phase episodes are usually young, and DSP is the most common disease gene, accounting for 69% of cases. Currently, the role of “hot-phase” episodes in disease progression and arrhythmic risk stratification remains to be clarified.

Role of Th17 Cytokines in the Liver's Immune Response during Fatal Yellow Fever: Triggering Cell Damage Mechanisms.

Yellow fever (YF) is an infectious and acute viral haemorrhagic disease that triggers a cascade of host immune responses. We investigated the Th17 cytokine profile in the liver tissue of patients with fatal YF. Liver tissue samples were collected from 26 deceased patients, including 21 YF-positive and 5 flavivirus-negative patients, with preserved hepatic parenchyma architecture, who died of other causes. Histopathological and immunohistochemical analysis were performed on the liver samples to evaluate the Th17 profiles (ROR-γ, STAT3, IL-6, TGF-β, IL-17A, and IL-23). Substantial differences were found in the expression levels of these markers between the patients with fatal YF and controls. A predominant expression of Th17 cytokine markers was observed in the midzonal region of the YF cases, the most affected area in the liver acinus, compared with the controls. Histopathological changes in the hepatic parenchyma revealed cellular damage characterised mainly by the presence of inflammatory cell infiltrates, Councilman bodies (apoptotic cells), micro/macrovesicular steatosis, and lytic and coagulative necrosis. Hence, Th17 cytokines play a pivotal role in the immunopathogenesis of YF and contribute markedly to triggering cell damage in patients with fatal disease outcomes.

Th17 Activation and Th17/Treg Imbalance in Prolonged Anterior Intraocular Inflammation after Ocular Alkali Burn.

Ocular alkali burn (OAB) is a sight-threatening disease with refractory ocular inflammation causing various blinding complications. Th17 lymphocytes account for the pathogeneses of the autoimmune disease and chronic inflammation, but their role in prolonged anterior intraocular inflammation after OAB is still unknown. A rat OAB model was established for this purpose. Anterior intraocular inflammation was observed in both the acute and late phases of OAB, and histological examination confirmed the presence of inflammatory cell infiltration and fibrin exudation in the anterior segment. Luminex xMAP technology and qPCR were used to evaluate the intraocular levels of cytokines. The levels of IL-1β, IL-6, and TNF-α were significantly elevated during the acute phase. The expression of IL-17A gradually increased from day 7 onwards and remained at a relatively high level. Immunofluorescence was performed to identify Th17 cells. CD4 and IL-17A double positive cells were detected in the anterior chamber from days 7 to 28. Flow cytometry showed that the frequency of Th17 cells increased in both lymph nodes and spleen, while the frequency of Treg cells remained unchanged, resulting in an elevated Th17/Treg ratio. The present study suggests that Th17 activation and Th17/Treg imbalance account for prolonged anterior intraocular inflammation after OAB.

Syringic acid and silymarin concurrent administration inhibits sodium valproate-induced liver injury in rats.

Sodium valproate (SV) is a well-known anti-epileptic drug, also used to control convulsions, bipolar disorders and migraines. SV has been shown to induce liver toxicity in clinical subjects. Syringic acid (SA), a natural polyphenolic compound has potential antioxidant, anti-inflammatory and several beneficial effects. Therefore, in this study, we evaluated hepatoprotective effect of SA against SV-induced liver injury in rats. Wistar rats were treated with SV orally at a dose of 500 mg/kg, once daily, for 14 days. Another three groups of rats were administered with SV and concurrently treated with SA (40 and 80 mg/kg) and silymarin (SIL) (100 mg/kg) for 14 days. SV administration for 14 days caused significant (p < .001) elevation of liver transaminases and ALP in serum. Liver MDA level was significantly (p < .001) increased with a concomitant decrease (p < .001) in enzymic antioxidants activities in SV administered rats. SV administration also caused the upregulation of proinflammatory markers such as tumor necrosis factor α, c-Jun N-terminal kinase, nuclear factor kappa B, cyclooxygenase-2 and Interleukin 6 expressions in liver tissue. Histopathological studies also revealed the presence of inflammatory cell infiltration and hepatocellular necrosis upon SV administration. At both doses, concurrent administration of SA and SIL significantly (p < .001) inhibited the liver transaminase activities in serum, oxidative stress, and proinflammatory markers expression in liver tissue. Our current results suggest that SA can be a promising herbal drug that can inhibit SV-induced hepatotoxicity when administered together due its potential anti-inflammatory effects.

Effect of the Progression of Fusobacterium nucleatum–induced Apical Periodontitis on the Gut Microbiota

IntroductionFusobacterium nucleatum, which is involved in the development of periodontal disease and apical lesions, can be transmitted to the colon and metastasize to colorectal cancer, suggesting a link between oral and systemic diseases. We analyzed the effects of F. nucleatum on bacterial flora in the gut and surrounding organs in a rat model of apical periodontitis and analyzed the infection route to the gut and distant organs. MethodsWe induced apical periodontitis in rat molars by infecting the dental pulp with F. nucleatum and then took X-ray images and performed histopathologic analyses. Next, we removed the maxilla, gut, heart, liver, and kidney from the rats at 0, 2, 4, and 8 weeks postsurgery and then extracted DNA samples and performed polymerase chain reaction and microbiome analyses using the Illumina MiSeq (Illumina Co, Tokyo, Japan). ResultsThe presence of inflammatory cell infiltration confirmed apical periodontitis from 2–8 weeks. Polymerase chain reaction and microbiome analyses revealed F. nucleatum in the rat gut from 2 weeks and in the kidney from 8 weeks. The rat gut, heart, liver, and kidney exhibited altered bacterial flora, including a marked decrease in Verrucomicrobia and an increase in Proteobacteria after 2 weeks and increases in Bacteroidetes and Firmicutes after 4 weeks. ConclusionsThe onset of F. nucleatum–induced apical periodontitis changed the bacterial flora in the rat gut, heart, liver, and kidney, with a confirmed progressing infection in the large intestines.

Sustained AWT1 expression by Dupuytren's disease myofibroblasts promotes a proinflammatory milieu.

Palmar fibromatosis, also known as Dupuytren's disease (DD), is a common and heritable fibrosis of the hand. It is characterized by the formation of myofibroblastic nodules that can progress to palmar-digital contractures and permanent loss of dexterity. The presence of inflammatory cell infiltrate within these nodules has been interpreted to suggest a pathogenesis mediated by a proinflammatory microenvironment. However, the molecular mechanisms driving the formation of pro-fibrotic microenvironments in this and other fibroses remain unclear. To gain insights into this process, we have assessed the contributions of an alternatively spliced, multi-functional transcription factor, Wilms Tumor 1 (WT1), previously shown to be upregulated in primary myofibroblasts derived from DD tissues. Proinflammatory cytokine stimuli of DD myofibroblasts enhanced the expression of several distinct WT1 variants, the most sustained being a 5' truncated version of WT1, alternative WT1 (AWT1). Constitutive adenoviral expression of AWT1 in myofibroblasts derived from phenotypically non-fibrotic palmar fascia significantly induced the expression and secretion of proinflammatory cytokines, including some with potential as novel therapeutic targets. In summary, these data implicate roles for sustained AWT1 expression in DD as a transcriptional driver of a proinflammatory fascial milieu.

Biomimetic analyses of interactions between macrophages and palmar fascia myofibroblasts derived from Dupuytren's disease reveal distinct inflammatory cytokine responses.

Dupuytren's disease (DD) is a common and heritable fibrosis of the hand. It is characterized by the shortening and thickening of the palmar fascia into myofibroblastic nodules that can progress to palmar-digital contractures and permanent loss of dexterity. Molecular analyses of DD tissues and the presence of inflammatory cell infiltrates suggest a pathogenesis initiated by a proinflammatory fascial milieu that promotes myofibroblast activation and palmar fascia contractures. However, the relative contributions of vascular and/or tissue derived immune system cells and cytokine-sensitive stromal myofibroblasts to the development of this proinflammatory microenvironment are poorly understood. To gain insights into this process, we have developed and tested a collagen-based 3D tissue biomimetic co-culture system to assess paracrine interactions between THP-1-derived pro-inflammatory macrophages and primary human palmar fascia myofibroblasts (PFMs). We observed significant and reproducible impacts of collagen-adherent macrophage and PFM co-cultures on the cytokine gene expression profiles of these cells compared to their respective monocultures, and significant changes to the resulting cytokine milieu in their shared culture media, notably TNF and IL-6. Our findings are consistent with central roles for PFMs in cytokine production and immunoregulation of the pro-inflammatory milieu hypothesized to promote DD development.

Pesticide Dichlorvos Promotes Morphological Alternations in the Rat Prostate

The complex process of carcinogenesis can be modulated by the action of environmental endocrine disruptors, such as the organophosphate pesticide Dichlorvos (DDVP), capable of interfering in homeostasis of different organs and leading to the development of neoplastic lesions. Considering that prostate cancer is the second most common neoplasm in men, worldwide, it is emphasized the importance of studying morphological alterations using animal models. In rodents, tumor initiation can be induced by the carcinogen N-methyl-N-nitrosourea (MNU) and tumor promotion can be stimulated by the administration of testosterone, combinated or not with the use of pesticides. The present study aimed to evaluate the influence of DDVP on the development of morphological alterations in the ventral prostate of rats, after chemical induction by MNU. Thereunto, 32 Fischer 344 rats, aged 90 days, were randomly separated into four experimental groups: Control, DDVP, MNU, MNU+DDVP. For chemical induction, the MNU and MNU+DDVP groups were inoculated with MNU (15 mg/kg), followed by daily subcutaneous injections of testosterone cypionate (2.5 mg/kg), for 20 days. Animals of the DDVP and MNU+DDVP groups, from 120 to 240 days old, received the basal diet supplemented with 10 mg/kg of DDVP. Subsequent to the experimental period, the prostatic ventral lobe of each animal was collected and subjected to morphometric-stereological and histopathological analysis, for the classification of the lesions found. The incidence of alterations in epithelium and stroma of all experimental groups was evaluated, such as epithelial hyperplasia (increased cell population with stratification, without atypia) (Figure A and B); atrophy (retraction of the epithelium with reduction of the secretory cytoplasm) (Figure C and D); periacinary inflammatory foci (presence of inflammatory cell infiltrate) (Figure E and F); cell atypia with presence of cytoplasmic inclusions (Figure G and H) and metaplasia with stromal hyperplasia (change in the pattern of the original epithelium without proliferative change, with an increase in the adjacent stroma) (Figure I to L). The results showed a significant increase of epithelial hyperplasia areas in MNU and MNU+DDVP groups, with 100% of the animals showing this lesion (Table 1), which demonstrates a relation between chemical induction and the proliferative process. The morphometric-stereological analysis confirmed this finding, revealing a relatively greater volume of the epithelial compartment in MNU+DDVP, compared to the Control group (Table 2). Thus, it can be concluded that DDVP, even in low concentration, associated with chemical induction by MNU, was able to disturb the morphology of the ventral prostate, promoting proliferative epithelial lesions, strongly associated with the process of carcinogenesis.

Silver Sulphadiazine- xanthan gum- hyaluronic Acid Composite Hydrogel for Wound Healing: Formulation Development and &lt;I&gt;in vivo&lt;/I&gt; Evaluation

Background: Development and modifications of hybrid hydrogels have been done to improve biological properties or to decrease the disadvantages of biomaterials.Objectives: The efficacy of hyaluronic acid in combination with silver sulphadiazine in wound healing was investigated. The retaining properties of xanthan gum to aid re- epithelialization was also explored.Materials and Method: Four hybrid hydrogels comprising of different concentrations of xanthan gum, eugenol and antimicrobial agents – hyaluronic acid and silver sulphadiazine were formulated. The physicochemical properties of the gels were assessed, and the antimicrobial effectiveness of the different hydrogel were determined using the extent of wound closure as an index.Results: The hydrogel samples had approximately 90% moisture content with rate of evaporation between 26- 32% for a 5 h period at 37oC. The pH of all formulations was between 7.59 - 8.05 considering that the formulation would be applied to underlying tissues of the skin. The swelling index after a 12 h period in distilled water was 10% for HX 1, 27% for HX 2, 29% for HX 3 and 30% for HX 4. There was no new peak observed in the FTIR analysis to indicate formation of new bonds.Conclusion: Incorporation of silver sulphadiazine at 0.1% and hyaluronic acid at 1.5% in the formulation yielded the best results with regards to least presence of inflammatory cell infiltrates and excellent wound closure at 14 days compared to the control and other formulations. Further investigation may be required for clinical use as an effective wound dressing material.&#x0D; Keywords: Silver sulphadiazine, Xanthan gum, Hyaluronic acid, Hydrogels, Wound healing.

The role of vascularization on changes in ligamentum flavum mechanical properties and development of hypertrophy in patients with lumbar spinal stenosis

BACKGROUND CONTEXTLigamentum flavum (LF) induced lumbar spinal stenosis (LSS) is conditioned not only by its "gathering" but especially by hypertrophy. Previous studies have examined the pathophysiology and biochemical changes that cause the hypertrophy. Some studies have described a link between chronic LF inflammation and neovascularization but others have reported highly hypovascular LF tissue in LSS patients. Currently, there is no practical application for our knowledge of the pathophysiology of the LF hypertrophy. Considerations for future treatment include influencing this hypertrophy at the level of tissue mediators, which may slow the development of LSS. To our knowledge, there is no study of micromechanical properties of native LF to date. PURPOSE(1) To clarify the changes in vascularization, chondroid metaplasia, and the presence of inflammatory cell infiltration in LF associated with LSS. (2) To quantify changes in the micromechanical properties associated with LF degenerative processes. STUDY DESIGN/SETTINGVascular density analysis of degenerated and healthy human LF combined with measurement of micromechanical properties. METHODSThe study involved 35 patients who underwent surgery between November 1, 2015 and October 1, 2016. The LSS group consisted of 20 patients and the control group consisted of 15 patients. LF samples were obtained during the operation and were used for histopathological and nanoindentation examinations. Sample vascularization was examined as microvascular density (Lv), which was morphometrically evaluated using semiautomatic detection in conjunction with NIS-Elements AR image analysis software. Samples were also histologically examined for the presence of chondroid metaplasia and inflammation. Mechanical properties of native LF samples were analyzed using the Hysitron TI 950 TriboIndenter nanomechanical testing system. RESULTSVascular density was significantly lower in the LSS group. However, after excluding the effect of age, the difference was not significant. There was high association between Lv and age. With each increasing year of age, Lv decreased by 11.5 mm2. Vascular density decreased up to the age of 50. Over the age of 50, changes were no longer significant and Lv appeared to stabilize. No correlation was observed between Lv and the presence of inflammation or metaplasia; however, LSS patients had a significantly increased incidence of chondroid metaplasia and inflammatory signs. The mechanical properties of control group samples showed significantly higher stiffness than those samples obtained from the LSS group. ConclusionThis study showed that Lv changes were not dependent on LSS but were age-dependent. Vascular density was found to decrease up to the age of 50. A significantly higher incidence of chondroid metaplasia and inflammation was observed in LSS patients. The mechanical property values measured by nanoindentation showed high microstructural heterogeneity of the tested ligaments. Our results showed that healthy ligaments were significantly stiffer than LSS ligaments. CLINICAL SIGNIFICANCEPrevention of the loss of LF vascularization during aging may influence stiffness of LF which in turn may slow down the LF degenerative processes and delay onset of LSS.

Protection from doxorubicin-induced nephrotoxicity by clindamycin: novel antioxidant, anti-inflammatory and anti-apoptotic roles.

This study was performed to examine whether clindamycin could protect against doxorubicin (DOX)-induced acute nephrotoxicity, and if so, what molecular mechanisms responsible for this protective effect. Male albino rats were pretreated with clindamycin once per day for 5 consecutive days at a dose of 300mg/kg, i.p, then received a single dose of DOX (15mg/kg; i.p) on the 5th day. DOX-induced marked renal injury as indicated by the presence of inflammatory cell infiltration, congestion, and edema accompanied by elevation in serum levels of creatinine and urea. These effects were alleviated by clindamycin pretreatment. DOX caused glutathione depletion and reduction in level of the antioxidant enzyme, catalase. Pretreatment with clindamycin markedly prohibited DOX-induced oxidative damage in renal tissue. Moreover, DOX provoked inflammatory responses in renal tissues as confirmed by increased expressions of NF-κB and COX-2 which were significantly reduced by clindamycin pretreatment. Besides, DOX-triggered apoptotic cascades in renal tissues as evidenced by elevated expression of pro-apoptotic proteins; Bax and cytochrome c, enhancing activity of caspase-3 enzyme whereas reducing the expression of anti-apoptotic Bcl-2 protein. Clindamycin pretreatment counteracts these apoptotic effects of DOX. Summarily, our results provide an evidence for the first time that clindamycin has a potential protective action against DOX-induced acute nephrotoxicity through inhibiting oxidative stress, inflammatory cascades, and apoptotic tissue injury.

Expression of tenascin C in cardiovascular lesions of Kawasaki disease

To examine tenascin C (TN-C) expression in coronary artery lesions (CALs) and myocardial lesions (MLs) in Kawasaki disease (KD). Twenty-five KD autopsy cases (post-KD-onset range of 6 days to 17 years) were examined in this study. Time-course analysis based on the disease day was performed of the histological findings for the CALs and MLs, as well as the localization and intensity of expression of TN-C. TN-C expression was observed to coincide with the areas where inflammatory cell infiltration was present in both coronary arteries and myocardium during the acute stage of KD, and the intensity of its expression correlated with the degree of inflammation. Obvious TN-C expression persisted in the thickened intima and media of CALs even after Disease Day 27. However, in spite of the presence of inflammatory cell infiltration, TN-C expression became weaker in the adventitia and surrounding connective tissue. After 8 months or more, TN-C was not expressed in the vasculitis scars of most cases, but expression was observed around newly formed vessels in the thickened intima and around recanalized vessels after thrombotic occlusion. The findings suggest a correlation between the degree of inflammation and TN-C expression in the cardiovascular lesions of acute-stage Kawasaki disease.

Tumor-Associated Macrophages: Oblivious Confederates in Invasive Mammary Carcinoma

The infiltrating margins of carcinomas are associated with the presence of inflammatory cell infiltrate which are an integral part of the tumor microenvironment. Among the inflammatory cells, tumor-associated macrophages (TAMs) play a key role in the tumorigenesis. This study elucidates the density of TAMs in invasive mammary carcinomas and attempts to establish an association with the following pathological variables: tumor size, histological grade, nodal status, hormonal expression status and Her2Neu overexpression. Ninety diagnosed archival cases of invasive mammary carcinomas at a tertiary care center were included. Density of TAMs was assessed by using CD68 which is a pan-macrophage marker by immunohistochemistry on the archival tissue blocks. The density of TAMs (CD68 positive cells) was dichotomized into high (> 50 CD68 positive cells/HPF) and low (< 50 CD68 positive cells/HPF) and compared with the above-mentioned pathological variables using appropriate statistical tests. The density of TAMs was significantly higher around the infiltrating edge of the carcinoma in comparison with the adjoining normal terminal duct lobular units. The density of TAMs was more in the infiltrating edge of the tumor than within the tumor nodule/nests. A higher TAM density showed a significant association in tumors having large tumor size, higher histological grade, nodal metastasis, the absence of ER and PR expression and Her2Neu overexpression (p value < 0.05). TAMs play an important role in tumor progression in invasive mammary carcinomas. This is as a result of the multiple roles enacted by TAMs in the various stages of tumor development starting from tumor growth, invasion, angiogenesis and metastases. Targeted therapy against TAMs has great potential in being important components of future treatment strategies against breast carcinomas.

Tumor-associated macrophages: Oblivious confederates in invasive mammary carcinoma

Abstract Background: The infiltrating margins of carcinomas are associated with presence of inflammatory cell infiltrate which are an integral part of the tumor microenvironment. Amongst the inflammatory cells, Tumor Associated Macrophages (TAMs) play a key role in the tumorigenesis. This study elucidates the density of TAMs in invasive mammary carcinomas and attempts to establish aa association with the following pathological variables: tumor size, histological grade, nodal status, hormonal expression status and Her2Neu overexpression. Materials and Methods: 90 diagnosed archival cases of invasive mammary carcinomas at a tertiary care centre were included. Density of TAMs was assessed by using CD68 which is a pan-macrophage marker by immunohistochemistry on the archival tissue blocks. The density TAMs (CD68 positive cells) was dichotomised into high (&gt;50 CD68 positive cells/HPF) and low (&lt;5050 CD68 positive cells/HPF) and compared with the above mentioned pathological variables using appropriate statistical tests. Results: The density of TAMs was significantly higher around the infiltrating edge of the carcinoma in comparison to the adjoining normal terminal duct lobular units. The density of TAMs was more in the infiltrating edge of the tumor than within the tumor nodule/nests. A higher TAM density showed a significant association in tumors having large tumor size, higher histological grade, nodal metastasis, absence of ER and PR expression and Her2Neu overexpression (p value &lt;0.05). Conclusion: TAMs play an important role in tumor progression in invasive mammary carcinomas. This is as a result of the multiple roles enacted by TAMs in the various stages of tumor development starting from tumor growth, invasion, angiogenesis and metastases. Targeted therapy against TAMs has great potential in the being important components of future treatment strategies against breast carcinomas.

CD8+ T Cells Contribute to the Development of Coronary Arteritis in the Lactobacillus casei Cell Wall Extract-Induced Murine Model of Kawasaki Disease.

Kawasaki disease (KD) is the leading cause of acquired heart disease among children in developed countries. Coronary lesions in KD in humans are characterized by an increased presence of infiltrating CD3+ T cells; however, the specific contributions of the different T cell subpopulations in coronary arteritis development remain unknown. Therefore, we sought to investigate the function of CD4+ and CD8+ T cells, Treg cells, and natural killer (NK) T cells in the pathogenesis of KD. We addressed the function of T cell subsets in KD development by using a well-established murine model of Lactobacillus casei cell wall extract (LCWE)-induced KD vasculitis. We determined which T cell subsets were required for development of KD vasculitis by using several knockout murine strains and depleting monoclonal antibodies. LCWE-injected mice developed coronary lesions characterized by the presence of inflammatory cell infiltrates. Frequently, this chronic inflammation resulted in complete occlusion of the coronary arteries due to luminal myofibroblast proliferation (LMP) as well as the development of coronary arteritis and aortitis. We found that CD8+ T cells, but not CD4+ T cells, NK T cells, or Treg cells, were required for development of KD vasculitis. The LCWE-induced murine model of KD vasculitis mimics many histologic features of the disease in humans, such as the presence of CD8+ T cells and LMP in coronary artery lesions as well as epicardial coronary arteritis. Moreover, CD8+ T cells functionally contribute to the development of KD vasculitis in this murine model. Therapeutic strategies targeting infiltrating CD8+ T cells might be useful in the management of KD in humans.