Periostin and osteopontin are overexpressed in chronically inflamed sinuses.

OBJECTIVE: In chronic rhinosinusitis (CRS), the inflammation leads to a proliferative response in the extracellular matrix (ECM). Periostin and osteopontin are 2 ECM proteins which have received attention for their roles in tissue remodeling in inflammatory diseases of the upper and lower airways. Transforming growth factor beta-1 (TGFß1) is an inflammatory cytokine that has been implicated in fibrotic conditions affecting virtually every organ. In this study we seek to evaluate the differential expression of periostin, osteopontin, and TGFß1 in the ethmoid sinus and nasal floor of patients with CRS. Furthermore, we seek to determine if a correlation exists between their differential expression in the nose and sinuses of patients with CRS. METHODS: Biopsies from ethmoid sinus and nasal floor mucosa were taken from 15 patients undergoing functional endoscopic sinus surgery (FESS) for CRS. Complementary DNA (cDNA) microarray analysis demonstrated upregulation of periostin and osteopontin in the ethmoid sinus samples. Quantitative real-time polymerase chain reaction (RT-PCR) was performed for periostin, osteopontin, and TGFß1. Statistical analysis was undertaken to examine correlations between the 3 genes of interest. RESULTS: RT-PCR confirmed that periostin and osteopontin were overexpressed in the ethmoid samples compared to the nasal floor. The differential expression of both periostin and osteopontin showed significant correlation with TGFß1. CONCLUSION: Periostin and osteopontin appear to be involved in ECM remodeling seen in CRS. TGFß1 may be an upstream inducer of CRS-related changes in ethmoid sinus mucosa.

Establishment of osteoblast culture from human ethmoidal sinus.

OBJECTIVE: Chronic sinusitis is characterized by persistent chronic inflammation of the sinus system and local expression and release of various cytokines, such as IL-1 beta, IL-4, IL-6, IL-8, TGF-beta and TNF-alpha are deeply involved in the pathogenesis of the disease. We hypothesized that not only the sinus mucosa-containing cells, such as epithelial cells and fibroblasts, but also osteoblasts, of which sinus bone structure consists, may contribute to this inflammatory network. This study evaluates the development and establishment of an osteoblasts culture system derived from human ethmoidal sinus as an initial step toward verifying this hypothesis. METHODS: Ethmoidal sinus bone was obtained from patients at the time of sinus surgery for chronic sinusitis and outgrowth cell sheets were obtained according to the explant-outgrowth cell culture technique. In order to examine the specific characteristics of osteoblasts in the obtained cells, four major features of osteoblasts (collagen type I, osteocalcin synthesis, alkaline phosphatase activity and extracellular matrix mineralization ability) were investigated at the third passage of the culture and productions of TGF-beta 1, which modulate osteoblast proliferation and maturation in an autocrine fashion, in the cultured medium was also investigated in time of culture up to 20 days. RESULTS: The cells obtained in our study clearly show collagen type I synthesis, osteocalcin synthesis, alkaline phosphatase activity and production of visible extracellular matrix mineralization. Production of TGF-beta 1 in the medium did not significantly different in time of culture up to 20 days. CONCLUSION: Our results, the first of their kind, indicate that osteoblast-like cells can be cultured from adult human ethmoidal sinus bones. It suggested that proliferation and maturation of the osteoblast were continuously modulated in an autocrine fashion by producing TGF-beta 1.

Eosinophilic apoptosis in sinus mucosa: relationship to tissue eosinophilia and its resolution in allergic sinusitis.

BACKGROUND: Apoptosis, which is regulated by both cell survival and death signals, is important for the swift clearance of unwanted cells. OBJECTIVE: We sought to elucidate whether eosinophilic apoptosis is associated with tissue eosinophilia and to determine its resolution in allergic sinusitis (AS). METHODS: Numbers of eosinophils, numbers of IL-5(+) cells, and the apoptosis index of eosinophils were calculated in the submucosa (both superficial and deep layers) of patients with AS by using histochemical methods before and after prednisolone treatment. Patients without AS were used for control groups. Anti-EG2 antibody was used to identify eosinophils. IL-5, Fas, or Bax expression of eosinophils was evaluated to elucidate the role of the factors affecting eosinophilic apoptosis. RESULTS: EG2 and IL-5(+) cells were abundant in the submucosa of patients with AS, especially in the superficial layer. About 50% to 60% of the IL-5-producing cells were eosinophils. Apoptotic eosinophils were less numerous in the superficial layer than the deep layer in these diseases. After prednisolone treatment, an induction of eosinophilic apoptosis was accompanied by a significant decrease in the number of EG2(+) and IL-5(+) cells. No remarkable difference was observed in the Fas or Bax expression of eosinophils after prednisolone treatment. CONCLUSION: Autocrine secretion of IL-5 from eosinophils may be one reason why eosinophilic disease is difficult to manage. Induction of eosinophilic apoptosis is critical for reversing tissue eosinophilia in patients with AS.

Expression of mucin genes in chronic ethmoiditis.

We have investigated the profiles of MUC genes expressed in chronic ethmoiditis mucosa and normal ethmoid mucosa using RT-PCR, and the morphology of chronic ethmoiditis by a combination of light and electron microscope was observed. In the light- and electron-microscopic studies, chronic ethmoiditis mucosa revealed increased numbers of goblet cells with higher production of mucus in comparison to normal ethmoid mucosa. RT-PCR of cDNAs from three normal ethmoid mucosa revealed the same pattern of mucin gene expression, such as MUC5AC and MUC8. However, RT-PCR of cDNAs from eight chronic ethmoiditis mucosa showed the expression of two MUC1, six MUC4, eight MUC5AC, five MUC5B, seven MUC7, and eight MUC8. MUC2 and MUC6 were not detected. These results suggest that MUC4, MUC5AC, MUC5B, MUC7, and MUC8 are major mucins in the ethmoid mucosa and are up-regulated by chronic inflammation.

Monocyte chemotactic protein expression in allergy and non-allergy-associated chronic sinusitis.

OBJECTIVE: Chronic sinusitis (CS) is characterized by inflammatory mucosal thickening and polyp formation with a predominantly eosinophilic infiltrate. Chemokines are a novel group of inflammatory mediators capable of attracting specific inflammatory cell populations. Monocyte chemotactic proteins (MCP) are a subfamily of chemokines (MCP-1, MCP-2, MCP-3, and MCP-4) that share a number of functional properties including chemotactic activity for eosinophils. The purpose of this study was to investigate the expression of the MCP family of chemokines in allergy and non-allergy-associated chronic sinusitis using the technique of immunocytochemistry. METHOD: We examined the expression of MCP-1, MCP-3, and MCP-4 in biopsies from the ethmoid sinuses of patients with CS and normal controls. RESULTS: MCPs were localized to the epithelial cells and a subset of inflammatory cells within the mucosa. The expression of both MCP-3 and MCP-4 immunoreactivity were significantly increased in patients with both allergy and non-allergy-associated CS compared to normal controls (p < .001). There was no significant difference in the expression of MCP-1 in nasal biopsies from individuals with CS and normals. The level of expression of MCP-3 and MCP-4 correlated with eosinophil (p < .001) and CD4-positive T-cell infiltrate (p < .001) but not with CD8-positive T-cell infiltration. CONCLUSIONS: Our data suggest biologic redundancy in the expression of eosinophil chemoattractants in CS and a potential role for MCP-3 and MCP-4, but not MCP-1, in the pathophysiology of this disorder. Further, chemokines may be a common link between the eosinophilia of allergy-associated and non-allergy-associated CS, a finding that may have therapeutic implications.

Comparison of inflammatory cell profile and Th2 cytokine expression in the ethmoid sinuses, maxillary sinuses, and turbinates of atopic subjects with chronic sinusitis.

Chronic sinusitis is a common disease characterized by persistent inflammation of the sinus mucosa. This study was undertaken to investigate immunopathologic findings in biopsy specimens from the ethmoid sinuses, maxillary sinuses, and inferior nasal turbinates of 14 allergic subjects with chronic sinusitis. The composition of the inflammatory infiltrate in the three tissue sites was examined by immunocytochemistry with anti-CD3 (total T cells), anti-CD4 (helper T cells), anti-CD8 (suppressor T cells), anti-MBP (eosinophils), antitryptase (mast cells), and antichymase (mast cells) antibodies. These revealed a significant increase in the T-cell helper/suppressor ratio and eosinophils in the ethmoid sinus mucosa compared with those in the maxillary sinus mucosa and the inferior turbinate. Eosinophil numbers were also higher in the maxillary sinus than in the inferior turbinate. Mast cells were present in significantly higher numbers in the ethmoid sinus and inferior turbinate biopsy sections than in the maxillary sinus. With antisense, radiolabeled riboprobes, we used in situ hybridization to examine the expression of interleukin-4 and interleukin-5 transcripts. The density of cells expressing interleukin-4 transcripts was significantly higher in the inferior turbinate biopsy sections than in those from the ethmoid and maxillary sinuses. In addition, the number of interleukin-4 mRNA-positive cells was higher in the ethmoid than in the maxillary sinus mucosa. The density of interleukin-5 mRNA-positive cells was significantly higher in the ethmoid and maxillary sinuses than in the inferior turbinate. The results of this study indicate (1) a more intense inflammatory response in the ethmoid sinus than in the maxillary sinus and inferior turbinate in allergic chronic sinusitis and (2) different inflammatory responses in the upper airways that are dependent on the anatomic site. These findings have potential implications in the design of new therapeutic interventions for allergic chronic sinusitis.

Histology and histomorphometry of ethmoid bone in chronic rhinosinusitis.

Mucosal changes have been well described in chronic sinusitis, yet little is known about the underlying bone, despite clinical and experimental evidence suggesting that bone may be involved in chronic sinusitis. Techniques of undecalcified bone analysis were used for detailed histologic examination of ethmoid bone in chronic sinusitis compared with controls. Bone synthesis, resorption, and inflammatory cell presence were specifically assessed. Additionally, histomorphometry techniques were used to determine ethmoid bone physiology in individuals undergoing surgery for chronic sinusitis. Overall, individuals undergoing surgery for chronic sinusitis were found to have evidence of marked acceleration in bone physiology with histologic changes including new bone formation, fibrosis, and presence of inflammatory cells. These findings are compared with osteomyelitis in long bone and the jaw. The suggestion that underlying bone may serve as a catalyst for chronic sinusitis is supported and implications for therapy are discussed.

Chronic hyperplastic sinusitis: association of tissue eosinophilia with mRNA expression of granulocyte-macrophage colony-stimulating factor and interleukin-3.

BACKGROUND: We investigated the association among tissue eosinophilia, cellular infiltration, and cytokine mRNA expression in chronic hyperplastic sinusitis (CHS). METHODS: Percutaneous biopsies of the maxillary sinuses and nasal polyps were performed in 12 adult patients (six men and six women) of whom seven were nonallergic and 11 were asthmatic. Tissues were compared with biopsy specimens from the inferior and middle turbinates of normal control subjects. RESULTS: Histologically, an eosinophil-predominant inflammatory infiltrate was seen in 10 of 12 patients, whereas a mild to moderate neutrophilic infiltrate was seen in 4 of 12 patients. As determined by immunocytochemistry, diseased tissues and normal control tissues differed significantly in terms of the number of activated (EG2+) eosinophils (p = 0.005) but not in terms of CD3+ or CD4+ T lymphocytes, elastase-positive neutrophils or CD68+ macrophages. The number of eosinophils did not correlate with that of any other cell type. By in situ hybridization, CHS tissues showed significantly higher numbers of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-3 mRNA-positive cells than normal control tissues (p = 0.002 and 0.0005, respectively) per high-powered field. There was a significant correlation between the number of infiltrating EG2+ eosinophils and cells that expressed mRNA for GM-CSF (r = 0.60, p = 0.041) or IL-3 (r = 0.69, p = 0.013). Furthermore, epithelial cells did not show detectable mRNA expression for GM-CSF or IL-3. No significant correlation was found between IL-5 mRNA expression and infiltrating EG2+ eosinophils in diseased tissues. However, the IL-5 density was significantly higher in the five patients with CHS who had positive allergy skin test results than in the seven patients with negative skin test results (p = 0.017) or in normal control subjects. CONCLUSIONS: Our data support a role for GM-CSF and IL-3 in the eosinophilia characteristic of CHS and show that IL-5 mRNA expression is not a prominent feature of nonallergic inflammation. The cellular sources of GM-CSF and IL-3 in CHS remain to be definitely determined.