Evidence Of Inflammatory Cell Infiltrate Research Articles

Atmospheric pollutant black carbon induces ocular surface damage in mice

The threats of air pollution to human health have been gradually discovered, including its effects on eyes. The purpose of the study is to investigate the potential correlation between ocular surface exposure to black carbon and ocular surface structural damage as well as tear film dysfunction. To achieve this goal, 60 6-8-week-aged male BALB/C mice were randomly divided into 4 groups (n = 15). 0.5 mg/ml (group A), 1 mg/ml (group B), 5 mg/ml (group C) black carbon suspension droplets and PBS solution (group D) were used in the right eyes, 4 μl per time of three times per day. Tear break-up time, corneal fluorescein staining scores, and tear volume were assessed before treatment (day 0) and on days 4, 7, 10, and 14 after treatment. On day 14, the mice were sacrificed, and corneal and conjunctival tissues were collected for histological analysis. As the exposure time increased, there were no significant changes in the measured parameters from PBS-treated group of mice (P > 0.05). However, in the black carbon-treated group, there were significant decreases in tear film break-up time, significant increases in corneal fluorescein staining scores, and significant reductions in tear secretion (all P < 0.05). After 14 days, H&E staining of the corneal epithelium showed that in the PBS-treated group of mice, the corneal epithelial cells were neatly arranged, with no inflammatory cell infiltration, while in the black carbon-treated group, the corneal epithelium was significantly thickened, the basal cell arrangement was disrupted, the number of cell layers increased, and there was evidence of inflammatory cell infiltration. In the ultrastructure of the corneal epithelium, it could be observed that the black carbon-treated group had an increased amount of corneal epithelial cell detachment compared to the PBS-treated group, at the same time, the intercellular connections were looser, and there was a decrease in the number of microvilli and desmosomes in the black carbon-treated group. The results indicate that the ocular surface exposure to black carbon can result in a decrease in tear film stability and tear secretion in mice. Moreover, it can induce alterations in the corneal structure.

Ketamine Analog Methoxetamine Induced Inflammation and Dysfunction of Bladder in Rats.

The novel synthetic psychoactive ketamine analog methoxetamine is reportedly being used for recreational purposes. As ketamine use can result in urinary dysfunction, we conducted the present study to investigate how methoxetamine affects the bladder. A cystometry investigation showed that female Sprague-Dawley rats experienced increased micturition frequency bladder dysfunction after receiving a daily intraperitoneal injection of 30 mg/kg methoxetamine or ketamine for periods of 4 or 12 weeks. Histologic examinations of rat bladder tissue revealed damaged urothelium barriers, as well as evidence of inflammatory cell infiltration and matrix deposition. The drug-treated rats showed significantly upregulated levels of pro-inflammatory cytokines such as IL-1β, IL-6, CCL-2, CXCL-1, CXCL-10, NGF, and COX-2. In addition, interstitial fibrosis was confirmed by increased levels of collagen I, collagen III, fibronectin and TGF-β. Besides direct toxic effect on human urothelial cells, methoxetaminealso induced the upregulation related cytokines. Our results indicate that long term methoxetamine treatment can induce bladder dysfunction and inflammation in rats. Methoxetamine was confirmed to produce direct toxic and pro-inflammatory effects on human urothelial cells. Methoxetamine-associated bladder impairment may be similar to ketamine-induced cystitis.

486 Prenatal Allergen Exposure Facilitated Airway Remodeling by Airborne Allergen Stimuli in Postnatal Life

BackgroundMurine asthma models are mainly created through adulthood sensitization, but lack airway remodeling hallmarks.AimsThis study aimed to examine airway remodeling in the prenatally-sensitized murine asthma model.MethodsFVB/N fetuses were exposed to...

The relationship between components of tumour inflammatory cell infiltrate and clinicopathological factors and survival in patients with primary operable invasive ductal breast cancer

Background:The importance of the components of host local inflammatory response in determining outcome in primary operable ductal invasive breast cancer is not clear. The aim of this study was to examine the relationship between components of the tumour inflammatory cell infiltrate and standard clinicopathological factors including hormone status (oestrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor (HER)-2), Ki-67 and survival in patients with primary operable invasive ductal breast cancer.Methods:Tumour inflammatory cell infiltrate, hormone status (ER, PR and HER-2), Ki-67 and standard clinicopathological factors were determined using routine pathological and immuno-histochemical techniques in 468 patients.Results:The large majority (94%) of ductal tumours had evidence of inflammatory cell infiltrate. The general inflammatory cell infiltrate was positively associated with high grade (P<0.001), the absence of ER (P<0.001), the absence of PR (P<0.01), the presence of vascular invasion (P<0.05) and high lymphocytic infiltrate, plasma cell infiltrate, other inflammatory cell infiltrate and macrophage infiltrate (all P<0.001). The median follow-up of the survivors was 165 months. During this period, 93 patients died of their cancer. On univariate analysis, stratified for ER status, tumour size (P<0.01), lymph node involvement (P<0.001), tumour plasma cell infiltrate (P<0.001), other inflammatory cell infiltrate (P<0.05) and treatment (P<0.05) were associated with poorer cancer-specific survival whereas lymphocyte infiltrate (P<0.001) was associated with improved cancer-specific survival. On multivariate analysis, stratified for ER status, lymph node involvement (P<0.05) was independently associated with poorer cancer-specific survival whereas increased tumour lymphocyte infiltrate (P<0.001) was independently associated with improved cancer-specific survival.Conclusion:The results of this study show that, using routine histology, the general inflammatory cell infiltrate was a common feature and was positively associated with high grade, the absence of ER, the absence of PR, the presence of vascular invasion and high-grade infiltration of lymphocytes, plasma cells, other inflammatory cells and macrophages. Also, that within a mature cohort of patients, a high lymphocytic infiltrate was associated with improved survival, independent of clinicopathological characteristics including ER status, in primary operable ductal invasive breast cancer. These results rationalise previous work and provide a sound basis for future studies in this important area of breast cancer research.

T helper17 Cells Are Sufficient But Not Necessary to Induce Acute Graft-Versus-Host Disease

T helper (Th)1 cells were considered responsible for the induction of graft-versus-host disease (GVHD), but recently the concept has been challenged. Th17 cells play a critical role in mediating autoimmune diseases, but their role in the pathogenesis of GVHD remains unclear. Herein we compare the ability of in vitro generated Th1 and Th17 cells from C57BL/6 mice to induce GVHD in lethally irradiated BALB/c recipients. Allogeneic Th17 cells had superior expansion and infiltration capabilities in GVHD target organs, which correlated with their increased pathogenicity when compared with naïve or Th1 controls. Th17 cells caused no pathology in the syngeneic recipients, indicating that antigen-activation was required for their pathogenicity. Polarized Th17 cells could not maintain their phenotype in vivo as they produced a significant amount of interferon (IFN)-gamma after being transplanted into allogeneic recipients; however, IFN-gamma was not required for Th17 cell-induced GVHD. Further, we evaluated the pathogenesis of Th17 cells in GVHD by using polyclonal nonprimed CD4T cells in a clinically relevant allogeneic bone marrow transplantation (BMT) setting. We found that disruption of Th17-differentiation alone by targeting RORgammat (Th17-specific transcription factor) had no significant effect on GVHD development. We conclude that Th17 cells are sufficient but not necessary to induce GVHD.

Sinus augmentation using human mesenchymal stem cells loaded into a β-tricalcium phosphate/hydroxyapatite scaffold

Implant placement in the posterior maxilla may often be contraindicated because of insufficient bone volume and presence of the maxillary sinus. In these situations, sinus floor augmentation frequently has been proposed as the best treatment. This clinical study was based on the hypothesis that the clinical effectiveness of adult mesenchymal stem cells (MSCs) loaded to the biphasic scaffold. In this report, the clinical and radiographic results are presented on 6 consecutively treated patients using MSCs in combination with biphasic hydroxyl apatite/ beta-tricalcium phosphate (HA/TCP) for sinus elevation. All the patients in the study had less than 3 mm initial bone height in the posterior maxillary area (IBH). MSCs were cultured and expanded from bone marrow aspirate for each patient. Three months after sinus elevation, radiographic evaluation was performed for the patients and the secondary bone height was measured (SBH(1)). In the second stage surgery, 30 implants were placed. Trephine bur was used as a pilot drill and a core biopsy was obtained from each implant site. Prosthetic rehabilitation of the patients was performed after 4 months. Secondary bone height was measured 9 months after implant placement (SBH(2)). Of 30 implants, 28 (93%) were considered clinically successful. Two implants were removed due to mobility at the time of surgical exposure. Histologic evaluation of the biopsy specimens revealed numerous areas of osteoid and bone formation HA/TCP, with no evidence of inflammatory cell infiltrate. Mean bone regenerate was 41.34%. Clinically, no complications were observed, and all implants were considered clinically osseointegrated after 4 months. Mean bone height was measured 3 and 12 months after sinus grafting (mean of SBH(1)= 12.08 mm and mean of SBH(2)= 10.08 mm). These clinical and histological findings suggest that sinus grafting with HA/TCP in combination with MSCs provide a viable therapeutic alternative for implant placement. The findings suggest that the addition of MSCs to bone derivative/substitute materials may enhance bone formation in the maxillary sinus area. Of course more studies with the control groups are needed for the evaluation of this method as a clinical solution for the patients.

Evaluation of bone healing enhancement by lyophilized bone grafts supplemented with platelet gel: A standardized methodology in patients with tibial osteotomy for genu varus

Orthopedic practice may be adversely affected by an inadequate bone repair that might compromise the success of surgery. In recent years, new approaches have been sought to improve bone healing by accelerating the rate of new bone formation and the maturation of the matrix. There is currently great interest in procedures involving the use of platelet gel (PG) to improve tissue healing, with satisfactory results both in vitro and in maxillofacial surgery. Otherwise, to our knowledge, only a preliminary clinical study was undertaken in the orthopedic field [Kitoh et al., Bone 2004;35:892-898] and the efficacy of PG is still controversial. Our paper focuses on the effect on bone regeneration by adding PG to lyophilized bone chips used for orthopedic applications. The clinical model and the laboratory methodology were standardized. As a clinical model, we employed the first series of patients of a randomized case-control study undergoing high tibial osteotomy (HTO) for genu varus. Ten subjects were enrolled: in 5 patients lyophilized bone chips supplemented with PG were inserted during tibial osteotomy (group A); 5 patients were used as a control (group B) and lyophilized bone chips without gel were applied. Forty-five days after surgery, computed tomography scan guided biopsies of grafted areas were obtained and the bone maturation was evaluated by a standardized methodology: the osteogenic and angiogenic processes were semi-quantitatively characterized by using histomorphometry, and the mineral component of the lyophilized and host bone was analyzed by using X-ray diffraction technique with sample microfocusing and microradiography. Lyophilized bone with PG seems to accelerate the healing process, as shown by new vessel formation and deposition of newly formed bone, with no evidence of inflammatory cell infiltrate, when compared with lyophilized bone without gel. On the contrary, lyophilized bone undergo a resorption process, and a fibrous tissue often fills the spaces between chips. A histiocytic/giant-cell reaction is sometimes present. Otherwise, no differences have been found concerning microstructure. Our findings show the reliability of the methodology used to monitor early bone repair. The completion of the study and the evaluation of the ultimate clinical outcome are necessary in order to verify PG in vivo effects in orthopedic surgery.

Chronic stress induces mast cell–dependent bacterial adherence and initiates mucosal inflammation in rat intestine

Background & Aims: Chronic psychological stress is an important factor in relapses of intestinal disorders, but it remains unclear if stress can induce primary gut inflammation in a previously healthy host. Methods: Mast cell-deficient (Ws/Ws) rats and wild-type control (+/+) rats were submitted to water avoidance stress or sham stress (1 h/day) for 10 consecutive days, as a model of ongoing life stress. Results: Both rat groups had similar systemic responses to stress, as assessed by changes in weight, corticosterone levels, and defecation. In +/+ rats, chronic stress induced barrier dysfunction in the ileum and colon (increased macromolecular permeability and depletion of mucus) and ultrastructural changes in epithelial cells (enlarged mitochondria and presence of autophagosomes) associated with bacterial adhesion and penetration into enterocytes. Moreover, hyperplasia and activation of mast cells, infiltration of neutrophils and mononuclear cells, and increased myeloperoxidase (MPO) activity were documented in the mucosa. In intestine of Ws/Ws rats, epithelial function and morphology were unchanged by chronic stress, bacterial-epithelial cell interaction was not demonstrated, and there was no evidence of inflammatory cell infiltration. Conclusions: These findings suggest that chronic psychological stress can be an initiating factor in intestinal inflammation by impairing mucosal defenses against luminal bacteria and highlight the importance of mast cells in this process.GASTROENTEROLOGY 2002;123:1099-1108

Alveolar ridge and sinus augmentation utilizing platelet-rich plasma in combination with freeze-dried bone allograft: case series.

Alveolar bone regeneration is frequently necessary prior to placement of implants. Efforts to improve wound healing have focused on factors that may enhance bone formation following guided bone regeneration (GBR) techniques alone or in combination with bone replacement graft materials. Recent reports suggest that platelet-rich plasma (PRP), presumably high in levels of peptide growth factors, may enhance the formation of new bone when used in combination with autogenous graft material. In this report, the clinical and radiographic results are presented on 15 consecutively treated patients using autologous PRP in combination with freeze-dried bone allograft (FDBA) for sinus elevation and/or ridge augmentation. FDBA and PRP (0.5 g/2cc PRP) were mixed and placed as a composite graft material. A gel formed by mixing autologous thrombin-rich plasma with PRP (1:4 ratio) was used to cover the graft material. Core biopsies of grafted areas were obtained in several patients as part of implant site preparation and were evaluated histologically to determine site maturation. Of 36 implant fixtures, 32 (89%) were considered clinically successful demonstrating complete bone coverage of the implant, no mobility, and a normal radiographic appearance at the time of re-entry and 12 months post-implant exposure. Four implants were removed due to mobility at the time of surgical exposure. Histologic evaluation of biopsy specimens revealed numerous areas of osteoid and bone formation around FDBA particles, with no evidence of inflammatory cell infiltrate. These clinical and histological findings suggest that ridge augmentation and sinus grafting with FDBA in combination with PRP provide a viable therapeutic alternative for implant placements. Future studies are necessary to determine whether PRP enhances new bone formation or maturation with bone replacement allografts.

Evaluation of the inhalation toxicity of diethylethanolamine (DEEA) in rats

Diethylethanolamine (DEEA), an industrial anticorrosive additive, was evaluated in rats for its potential toxicity. A 2-week inhalation exposure to 10 ppm revealed no signs of toxicity. At 56 ppm, rats exhibited signs of nasal irritation and corneal opacities. Significant mortality (males, 90%; females, 50%) occurred at 301 ppm. Histopathology revealed inflammatory cell infiltrations of the nasal turbinate mucosa and squamous metaplasia after exposure to 56 ppm. In the 14-week subchronic study, rats were exposed to 0, 11, 25, or 76 ppm. During exposure to DEEA, transient signs of mild to moderate respiratory irritation (noises or rales) were observed with a frequency and severity that was dose dependent. These signs usually cleared within 1 hr postexposure. However, at 76 ppm some rats continued to exhibit these respiratory signs overnight. There were no significant DEEA-induced changes in either blood chemistry or neurobehavioral parameters. Nasal cavities of rats exposed to 25 or 76 ppm revealed evidence of inflammatory cell infiltration, focal hyperplasia, and squamous metaplasia in the respiratory epithelium of the anterior nasal turbinates. Hypertrophic goblet cells, focal necrosis, and exudate in the nasal lumen were also seen at 76 ppm. DEEA vapors failed to produce persistent ocular or respiratory tract toxicity below 26 ppm. The no-observed-effect-level in this study was 10 ppm.

Immunohistochemical Analysis of Iris Biopsy Specimens From Patients With Fuchs' Heterochromic Cyclitis

Using immunohistochemical techniques, we analyzed iris biopsy specimens from eight patients with Fuchs' heterochromic cyclitis, seven patients with various other types of uveitis, and eight glaucoma patients without uveitis. No specific abnormalities related to Fuchs' heterochromic cyclitis could be detected. Four of the patients with Fuchs' heterochromic cyclitis and four of the patients with uveitis showed evidence of an inflammatory cell infiltrate, which was a mixture of interleukin-2 receptor-negative T helper and suppressor cells, B lymphocytes, and plasma cells. Only an occasional T lymphocyte could be seen in two of the patients without uveitis. The class II antigen HLA-DR was expressed on iris stromal cells in every patient in the Fuchs' heterochromic cyclitis group and uveitis group and in six of the patients in the nonuveitis group. In six of the Fuchs' heterochromic cyclitis patients, including two without immunohistochemical evidence of inflammatory cell infiltrate, histologic abnormalities were present on hematoxylin and eosin sections.

Sulfuric acid-induced changes in the physiology and structure of the tracheobronchial airways.

Sulfuric acid aerosols occur in the ambient particulate mode due to atmospheric conversion from sulfur dioxide (SO2). This paper describes the response of the rabbit tracheobronchial tree to daily exposures to sulfuric acid (H2SO4) aerosol, relating physiological and morphological parameters. Rabbits were exposed to filtered air (sham control) or to submicrometer-sized H2SO4 at 250 micrograms/m3 H2SO4, for 1 hr/day, 5 days/week, with sacrifices after 4, 8, and 12 months of acid (or sham) exposure; some rabbits were allowed a 3-month recovery after all exposures ended. H2SO4 produced a slowing of tracheobronchial mucociliary clearance during the first weeks of exposure; this change became significantly greater with continued exposures and did not improve after exposures ended. Airway hyperresponsiveness was evident by 4 months of acid exposure; the condition worsened by 8 months of exposure and appeared to stabilize after this time. Standard pulmonary mechanics parameters showed no significant trends with repeated acid exposure, except for a decline in dynamic lung compliance in animals exposed to acid for 12 months. Lung tissue samples obtained from exposed animals showed a shift toward a greater frequency of smaller airways compared to control, an increase in epithelial secretory cell density in smaller airways, and a shift from neutral to acidic glycoproteins in the secretory cells. The effect on airway diameter resolved after the exposures ceased, but the secretory cell response did not return to normal within the recovery period. No evidence of inflammatory cell infiltration was found due to H2SO4 exposure. Thus, significant alterations in the physiology of the tracheobronchial tree have been demonstrated due to repeated 1-hr exposures to a concentration of H2SO4 that is one-fourth the current 8-hr threshold limit value for exposure in the work environment. The cumulative dose inhaled by the rabbits is similar to current peak daily doses from ambient exposure in North America. The results obtained in the rabbit model provide insight into early changes in the tracheobronchial tree due to repeated irritant exposure and may be involved in the pathogenesis of chronic airway disease.

Sulfuric Acid-Induced Changes in the Physiology and Structure of the Tracheobronchial Airways

Sulfuric acid aerosols occur in the ambient particulate mode due to atmospheric conversion from sulfur dioxide (SO2). This paper describes the response of the rabbit tracheobronchial tree to daily exposures to sulfuric acid (H2SO4) aerosol, relating physiological and morphological parameters. Rabbits were exposed to filtered air (sham control) or to submicrometer-sized H2SO4 at 250 micrograms/m3 H2SO4, for 1 hr/day, 5 days/week, with sacrifices after 4, 8, and 12 months of acid (or sham) exposure; some rabbits were allowed a 3-month recovery after all exposures ended. H2SO4 produced a slowing of tracheobronchial mucociliary clearance during the first weeks of exposure; this change became significantly greater with continued exposures and did not improve after exposures ended. Airway hyperresponsiveness was evident by 4 months of acid exposure; the condition worsened by 8 months of exposure and appeared to stabilize after this time. Standard pulmonary mechanics parameters showed no significant trends with repeated acid exposure, except for a decline in dynamic lung compliance in animals exposed to acid for 12 months. Lung tissue samples obtained from exposed animals showed a shift toward a greater frequency of smaller airways compared to control, an increase in epithelial secretory cell density in smaller airways, and a shift from neutral to acidic glycoproteins in the secretory cells. The effect on airway diameter resolved after the exposures ceased, but the secretory cell response did not return to normal within the recovery period. No evidence of inflammatory cell infiltration was found due to H2SO4 exposure. Thus, significant alterations in the physiology of the tracheobronchial tree have been demonstrated due to repeated 1-hr exposures to a concentration of H2SO4 that is one-fourth the current 8-hr threshold limit value for exposure in the work environment. The cumulative dose inhaled by the rabbits is similar to current peak daily doses from ambient exposure in North America. The results obtained in the rabbit model provide insight into early changes in the tracheobronchial tree due to repeated irritant exposure and may be involved in the pathogenesis of chronic airway disease.

The response of the dental pulp to mechanical exposure in gnotobiotic rats mono-infected with a strain of Streptococcus mutans.

Summary. The maxillary molar pulps of gnotobiotic rats mono‐infected with a strain of Streptococcus mutans and maintained on a high sugar diet, were mechanically exposed and left open to the oral environment. No evidence of inflammatory cell infiltration of pulp tissue apart from superficial cells was observed. Seven days after pulpal exposure, necrosis was detected in the superficial layers of the pulp. After 28 days, the necrosis was more extensive, and repair, manifested by dentine bridge formation, was observed in only a limited number of teeth. The lack of inflammation and slow progress of necrosis in pulps left open to the oral cavity appeared to indicate that the test strain of Streptococcus mutans had limited pathogenicity to the pulp. These observations provide support for the practice of indirect pulp capping in deep carious lesions.

Histologic changes in gastroesophageal reflux.

Three aspects of the esophageal mucosal injury that occur in gastroesophageal reflux disease are discussed in this article. First, the histologic changes in the squamous epithelium-lined esophagus are variable. Some patients with reflux symptoms do not have histologic evidence of inflammatory cell infiltration within the mucosa, but rather may have more subtle effects of mucosal injury, namely, thickening of the basal regenerative layer, elongation of the papillae, or intraepithelial eosinophils. However, these more subtle findings often are merely confirmatory of other clinical indicators of gastro-esophageal reflux; they may have their most practical application to the research investigation of reflux disease. Second, the presence of esophageal mucosal inflammation may adversely affect lower esophageal sphincter function. This phenomenon can be demonstrated in the cat model. Whether it is operative in humans is unclear. Finally, the development of columnar metaplasia (Barrett's epithelium) is more than a histologic curiosity because this condition confers an increased risk of developing adenocarcinoma of the esophagus. Among several unanswered questions about Barrett's epithelium is what is an appropriate surveillance program for patients with Barrett's epithelium to detect the presence of, or predict the development of, cancer?