Distribution of Mast Cells and Macrophages and Expression of Interleukin-6 in Periapical Cysts

Introduction

Mast cells and macrophages are important components of the inflammatory infiltrate found in inflammatory periapical diseases. Several cytokines participate in the mechanisms of inflammation, tissue repair, and bone resorption associated with periapical cysts. The aim of the present study was to evaluate the distribution of mast cells and macrophages and the expression of interleukin-6 (IL-6) in periapical cysts.

Methods

Thirty periapical cysts were selected for the study, and clinical, demographic, and gross information from the cases was obtained from the laboratory records. Five-micrometer sections stained with hematoxylin-eosin were reviewed for analysis of the microscopic features of the cysts, and 3-μm sections on silanized slides were used for immunohistochemical reactions with anti-tryptase, anti-CD68, and anti–IL-6.

Results

There was no statistically significant difference in the mean number of mast cells and macrophages when comparing superficial and deep regions of the fibrous capsule of the cysts. Mean number of mast cells on the superficial region of the fibrous capsule was higher in cysts showing intense superficial inflammation and exocytosis. Macrophages were more commonly found in areas showing IL-6 expression, and IL-6 was less expressed in deep regions of the fibrous capsule in cysts showing greater gross volume.

Conclusions

The results reinforced the participation of mast cells and macrophages in the pathogenesis of periapical cysts and suggested that IL-6 is not the major bone resorption mediator in larger periapical cysts.     

基质金属蛋白酶诱导因子和亲环素A在人根尖囊肿和根尖肉芽肿中的表达及临床意义

目的: 检测细胞外基质金属蛋白酶诱导因子(CD147)和亲环素A(CypA)在根尖肉芽肿和根尖囊肿中的表达,探讨CD147和CyPA在人慢性根尖周炎发生发展中的作用。方法: 收集经根尖手术切除所获得的根尖肉芽肿35例和根尖囊肿30例作为实验组,同时收集埋伏阻生智齿拔除术或行牙槽骨修整术凿下的8例健康牙槽骨作为正常对照组。CBCT图像记录病例的病损大小。运用免疫组织化学法检测所有样本中CD147和CypA的蛋白表达,分析CD147和CypA的蛋白表达水平。结果: 根尖囊肿组两种蛋白的表达水平均高于肉芽肿组(P<0.05)。CD147和CypA 的蛋白表达水平在根尖囊肿和根尖肉芽肿中呈正相关(P<0.05);CD147和CypA 的表达水平均与慢性根尖周炎的病变大小呈正相关(P<0.05)。结论: CD147和CypA可能参与根尖周病损的炎性反应和骨质吸收。CD147-CypA相互作用在人慢性根尖周病的发生发展过程中可能发挥了某种协同作用。     

Immunohistochemical Analysis of Galectins-1, -3, and -7 in Periapical Granulomas,Radicular Cysts,and Residual Radicular Cysts

Introduction

Galectins play important roles in immunoinflammatory responses, but their participation in the development of periapical lesions remains unclear. This study aimed to evaluate the expressions of galectins-1, -3, and -7 in periapical lesions, correlating them with the intensity of the inflammatory infiltrate and the pattern of the cystic epithelium.

Evaluation of the Reliability and Accuracy of Using Cone-beam Computed Tomography for Diagnosing Periapical Cysts from Granulomas

Introduction

The purpose of this study was to evaluate the reliability and accuracy of cone-beam computed tomographic (CBCT) imaging against the histopathologic diagnosis for the differential diagnosis of periapical cysts (cavitated lesions) from (solid) granulomas.

Methods

Thirty-six periapical lesions were imaged using CBCT scans. Apicoectomy surgeries were conducted for histopathological examination. Evaluator 1 examined each CBCT scan for the presence of 6 radiologic characteristics of a cyst (ie, location, periphery, shape, internal structure, effects on surrounding structure, and perforation of the cortical plate). Not every cyst showed all radiologic features (eg, not all cysts perforate the cortical plate). For the purpose of finding the minimum number of diagnostic criteria present in a scan to diagnose a lesion as a cyst, we conducted 6 receiver operating characteristic curve analyses comparing CBCT diagnoses with the histopathologic diagnosis. Two other independent evaluators examined the CBCT lesions. Statistical tests were conducted to examine the accuracy, inter-rater reliability, and intrarater reliability of CBCT images.

Results

Findings showed that a score of ≥4 positive findings was the optimal scoring system. The accuracies of differential diagnoses of 3 evaluators were moderate (area under the curve = 0.76, 0.70, and 0.69 for evaluators 1, 2, and 3, respectively). The inter-rater agreement of the 3 evaluators was excellent (α = 0.87). The intrarater agreement was good to excellent (κ = 0.71, 0.76, and 0.77).

Conclusions

CBCT images can provide a moderately accurate diagnosis between cysts and granulomas.     

Differences in Inflammation and Bone Resorption between Apical Granulomas,Radicular Cysts,and Dentigerous Cysts

IntroductionDental cysts can be of inflammatory (radicular cysts) or noninflammatory (dentigerous cysts) origin. Apical periodontitis is a necrosis of the pulp and infection of the root canal causing the development of apical granulomas or radicular cysts. The immunology of granuloma and cyst formation is important because modern root filling materials are immunologically active and can contribute to the resolution of apical granulomas. In contrast, radicular cysts often require apicectomy. A better understanding of the pathophysiology of inflammation and bone resorption in apical periodontitis could be the basis for developing new root filling materials with superior immunomodulatory properties.MethodsForty-one apical granulomas, 23 radicular cysts, and 23 dentigerous cysts were analyzed in this study. A tissue microarray of the 87 consecutive specimens was created, and human leukocyte antigen–DR isotype (HLA-DR)-, CD83-, receptor activator of nuclear factor kappa B ligand–, macrophage colony-stimulating factor (MCSF)-, galectin-3 (Gal3)-, CD4-, and CD8-positive cells were detected by immunohistochemistry. Tissue microarrays were digitized, and the expression of markers was quantitatively assessed.ResultsHLA-DR, CD83, MCSF, and Gal3 expression was significantly (P < .05) higher in radicular cysts compared with apical granulomas. HLA-DR, CD83, MCSF, receptor activator of nuclear factor kappa B ligand, and Gal3 expression in dentigerous cysts was significantly (P < .05) lower than in both periapical lesions (apical granulomas and radicular cysts). CD4 and CD8 infiltration was not statistically different between apical granulomas and radicular cysts. Dentigerous cysts showed a significantly (P < .05) lower T-cell infiltration than apical periodontitis. The CD4/CD8 ratio was not significantly different between the analyzed groups.ConclusionsThe development of radicular cysts in apical periodontitis is associated with an increased expression of myeloid inflammatory markers and bone resorption parameters. Antigen-presenting cells and myeloid cells might be more relevant for the pathogenesis of apical periodontitis than T cells. Increased inflammation might promote the formation of radicular cysts and more pronounced bone resorption.     

骨髓间充质干细胞骨向诱导分化过程中破骨细胞分化因子和细胞间粘附分子-1的表达变化

目的　观察破骨细胞分化因子(ODF)和细胞间粘附分子-1(ICAM-1)在不同分化状态成骨细胞的表达变 化,探讨正畸牙移动过程中成骨细胞对破骨细胞分化成熟的诱导机制。方法　分离培养大鼠骨髓间充质干细胞, 成骨定向诱导后获得不同分化状态的成骨细胞,RT-PCR检测不同分化状态下的成骨细胞ODF和ICAM-1的表达变 化。结果　成骨细胞在分化成熟过程中,ICAM-1 mRNA表达水平逐渐升高;ODF mRNA则在诱导后6 d开始表达, 并维持在一较稳定的水平。结论　不同分化状态的成骨细胞对破骨细胞诱导分化的能力有所差异,相对成熟的成 骨细胞的诱导能力可能更强。     

平阳霉素作用静脉畸形内皮细胞后VCAM-1、ICAM-1、ICAM-3表达

目的:研究平阳霉素(PYM)作用于人静脉畸形内皮细胞(HVMECs)后Ig粘附分子(VCAM-1、ICAM-1、ICAM-3)表达。方法:体外培养HVMECs,采用细胞ELISA和RT-PCR技术检测不同浓度PYM作用人HVMECs后Ig粘附分子表达。结果:PYM作用人HVMECs后粘附分子表达具有时间浓度效应。PYM能诱导VCAM-1在人HVMECs表达,2h后明显增高,8h后达到峰值,12h后下降,48h后呈阴性表达。PYM能促进ICAM-1在人HVMECs表达,12h后显著增高,18h最高,24h逐渐下降。PYM能促进ICAM-3表达,12h后逐渐增高,24h达到峰值,72h后1CAM-3仍高表达。0.01～1mg/L PYM能诱导或促进粘附分子表达,表达水平与药物浓度成正相关,1mg/L PYM作用人HVMECs后粘附分子表达较高,10mg/L PYM作用人HVMECs后,表达下降。RT-PCR检测PYM作用人HVMECs 2、6、12h能诱导或促进VCAM-1、ICAM-1和ICAM-3mRNA表达,8、12、18h后mRNA表达最高。1mg/L PYM作用于HVMECs后,粘附分子mRNA表达最高,mR-NA表达具有时间浓度效应。结论:低浓度PYM能诱导或促进人HVMECs粘附分子表达,具有时间浓度效应。     

Higher Expression of Galectin-3 and Galectin-9 in Periapical Granulomas than in Radicular Cysts and an Increased Toll-like Receptor-2 and Toll-like Receptor-4 Expression Are Associated with Reactivation of Periapical Inflammation

Introduction

Cysts and periapical granulomas are inflammatory reactions that develop in response to periapical infection by microbial species in dental root canal. It is known that toll-like receptors (TLRs) are pathogen recognition molecules and that galectins are lectins that can be associated with the inflammatory process, stimulating or inhibiting the immune system. The objective of this study was to evaluate the in situ expression of TLRs and galectins in radicular cysts and periapical granulomas.

Methods

We analyzed 62 cases (30 radicular cysts, 27 periapical granulomas, and 5 control cases). Indirect immunohistochemistry was used to evaluate the expression of TLRs (TRL-2 and TLR-4) and galectins (Gal-3 and Gal-9).

Results

The expression of Gal-3 and Gal-9 was significantly higher in periapical granulomas and radicular cysts than in the control group. Similarly, both Gal-3 and Gal-9 were expressed significantly more in periapical granulomas than in radicular cysts. The expression of TLR-2 was significantly higher in periapical granulomas and radicular cysts than in the control group, and it was also significantly higher in radicular cysts with sinus tract than in the cases without sinus tract. Furthermore, the expression of TLR-4 was significantly higher in the cases of periapical granulomas with sinus tract than in the cases without sinus tract.

Conclusions

Gal-3/Gal-9 and TLR-2/TLR-4 expression in the periapical granulomas and radicular cysts is associated with reactive periapical inflammation. Pathobiology of periapical disease is a very complex interplay of many bioactive molecules involved in immunoinflammatory responses. Up-regulation of these bioactive molecules might be an important modulator of inflammatory periapical lesions.     

口腔侵袭性鳞癌间质中CD34和α平滑肌肌动蛋白的表达

目的观察口腔侵袭性鳞癌间质中CD34和α平滑肌肌动蛋白（α-SMA）的表达。方法采用免疫组化SP法，检测60例口腔侵袭性鳞癌和其中20例切缘无瘤黏膜间质中CD34和α-SMA的表达。结果加例切缘无瘤黏膜间质中均有CD34的表达，主要位于口腔黏膜固有层、黏膜下层和血管旁的成纤维细胞胞质中，而无α-SMA的表达。α-SMA的表达仅见于血管壁平滑肌细胞。60例口腔侵袭性鳞癌间质中小血管壁内皮细胞中有CD34阳性表达，其中53例在肌成纤维细胞胞质中有α-SMA的表达。结论口腔侵袭性鳞癌间质中存在CD34表达缺失和α-SMA的表达获得现象，联合检测CD34和α-SMA有助于判断口腔癌的侵袭性。     

Involvement of substance P, mast cells, TNF-α and ICAM-1 in the infiltration of inflammatory cells in human periapical granulomas

BACKGROUND: Substance P (SP), a potent proinflammatory peptide present in sensory neurons, is believed to be a major mediator of neurogenic inflammation. The aim of this study was to examine the localization and involvement of SP, mast cells and tumor necrosis factor-alpha (TNF-alpha)-positive cells in human periapical granulomas. METHODS: Sections from seven periapical granulomas were stained using a variety of immunohistochemical methods. RESULTS: SP-immunoreactive nerve fibers, mast cells and TNF-alpha-positive cells were found localized in the vicinity of blood vessels in all the samples of periapical granulomas. The vascular endothelial cells stained positively for E-selectin and intercellular adhesion molecule-1. SP, TNF-alpha-positive cells and E-selectin could not be detected in clinically healthy periodontal ligament, and served as a negative control. CONCLUSION: Our findings suggest that SP, mast cells, TNF-alpha-positive cells and E-selectin may modulate the pathogenesis of apical periodontitis and may be responsible for stimulating the formation of granuloma with the resorption of periapical bone.     

Various immunostaining patterns of CD31, CD34 and endoglin and their relationship with lymph node metastasis in oral squamous cell carcinomas

BACKGROUND: Intratumoral blood vessels are known to play an important role in cancer growth and metastasis. The discrepancy in previous reports using various endothelial markers individually suggested us to investigate both normal and various tumor areas with a wide panel of vascular markers. METHODS: Here, we used a panel of three antibodies (CD31, CD34, and endoglin) as blood vessel markers to investigate the distribution and properties of blood vessels in normal oral tissues and squamous cell carcinomas. RESULTS: Many microvessels with strong remodeling activity as well as undifferentiated tumoral vascular endothelial cells and immature endothelial cells were present in the cancer cell nest and marginal area of cancer infiltration. Our results showed different vascular distribution patterns using various immunostaining markers in normal and tumoral tissues. CONCLUSION: Vascular distribution and properties of endothelial cells appear to be closely associated with metastasis.     

固定正畸患者龈沟液中可溶性细胞间粘附分子-1含量变化的研究

目的 测定固定正畸患者不同矫治时期龈沟液(GCF)中可溶性细胞间粘附分子-1(sICAM-1)的含量，探索固定矫治器对牙周组织的影响。方法 随机抽取口腔正畸科门诊牙拥挤错黯患者3l例。采用酶联法检测GCF中sICAM—l的含量，分析GCF的量与sICAM—l含量在矫治过程中的变化关系。结果 与矫治前l周比较。矫治过程中GCF的量与sICAM—l的含量均有不同程度的上升趋势。结论 正畸过程中固定矫治器对牙周组织的健康有影响。此外，sICAM—l的检测可用于监测固定正畸过程中患者的牙周健康状况。     

CD25~＋ Foxp3~＋调节性T细胞和Th17细胞在口腔白斑组织中的表达

目的：通过观察CD25＋Foxp3＋和IL-17阳性细胞在口腔黏膜白斑（oral leukoplakia,OLK）和口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）组织中的表达情况,探讨Treg细胞和Th17细胞在口腔癌发生发展过程中的变化。方法：采用免疫组织化学方法检测CD25＋Foxp3＋、IL-17阳性细胞在10例单纯增生性白斑、32例异常增生性白斑、13例OSCC和10例正常口腔黏膜中的变化情况。结果：正常口腔黏膜组、单纯增生性白斑组、异常增生性白斑组和OSCC组CD25＋Foxp3＋Treg细胞数量逐渐增高（P〈0.05）,依次为（2.5±0.65）、（5.22±1.19）、（31.4±16.8）和（42.8±12.67）个/每高倍镜视野。重度异常增生白斑组织中,CD25＋Foxp3＋Treg细胞的数量高于轻、中度异常增生白斑组织（P〈0.05）。此外,正常口腔黏膜组、单纯增生性白斑组、异常增生性白斑组和OSCC组中表达IL-17的阳性细胞虽有递增趋势,但差异无统计学意义。CD25＋Foxp3＋Treg细胞和Th17细胞数量改变在口腔白斑和OSCC组织中呈正相关（r=0.318,P〈0.05）。结论：Treg细胞的数量在白斑和OSCC组织中增多,Th17细胞协同Treg细胞,在口腔癌发生发展过程中发挥了一定的作用。     

Expression of the CD54 (ICAM-1) and CD11 a (LFA-1) adhesion molecules in oral mucosal inflammation

Previous studies of chronic dermatoses have suggested that expression of the CD54 cell surface antigen (intercellular adhesion molecule-1, ICAM-1) by keratinocytes is a feature of chronic inflammation. However, whether such expression is a prerequisite for intraepithelial migration of lymphocytes is unclear. The present study evaluated the expression of CD54 and its ligand, CD1 la (lymphocyte function-associated antigen, LFA-1) in oral lesions of lichen planus, recurrent aphthous stomatitis, secondary Sjögren's syndrome and traumatic ulceration using an immunoperoxidase technique. In 33 of 56 lesions examined, substantial numbers of CD1 la + cells were present within oral mucosal epithelium despite an absence of detectable keratinocyte CD54 antigen expression. Consequently, CD54/CD1 la adhesion interactions may not be critical in the initiation of oral mucosal inflammation.     

Differential in situ distribution of interleukin-8, monocyte chemoattractant protein-1 and Rantes in human chronic periapical granuloma

In situ distribution of three prototype chemokines interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1 and Rantes was determined in chronic human periapical granulomas by immunohistochemistry using monoclonal antibodies. IL-8 was found primarily in the cytoplasm of the Malassez epithelial cells. MCP-1 immunoreactivity was confined to the endothelial cells that lined small venules. Each of the three investigated chemokines, including Rantes, exhibited a characteristic binding pattern to the extracellular matrix of the lesion. The observed chemokines may play a role in establishing the cellular composition of chronic apical periodontitis, thus augmenting the intensity of local inflammation and tissue damage.     

Coculture of peripheral blood CD34+ cell and mesenchymal stem cell sheets increase the formation of bone in calvarial critical-size defects in rabbits

The reconstruction of large bony defects remains a clinical challenge, and angiogenesis and neovascularisation are being given more attention in bone tissue engineering. In this study we cocultured peripheral blood CD34+ cells (PB-CD34+ cells), an endothelial progenitor cell/haematopoietic stem cell-enriched population, with bone marrow-derived mesenchymal stem cells (MSC) to investigate their potential for bony regeneration. Cocultured cells showed better osteogenic differentiation than MSC alone in vitro. The cocultured cells and MSC sheets were also composited with hydroxyapatite and implanted in calvarial critical-size defects in rabbits. The rabbits were killed before microcomputed tomographic (MicroCT) and histological analysis. The results showed that cocultured cell composites had promoted bony regeneration more efficiently by 8 weeks after implantation. Our results indicate that the coculture of PB-CD34+ cells and MSC increases bony regeneration in calvarial critical-size defects in rabbits, and provide a new promising therapeutic strategy to aid skeletal healing.     

Differential in situ distribution of interleukin‐8, monocyte chemoattractant protein‐1 and Rantes in human chronic periapical granuloma

In situ distribution of three prototype chemokines interleukin (IL)‐8, monocyte chemoattractant protein (MCP)‐1 and Rantes was determined in chronic human periapical granulomas by immunohistochemistry using monoclonal antibodies. IL‐8 was found primarily in the cytoplasm of the Malassez epithelial cells. MCP‐1 immunoreactivity was confined to the endothelial cells that lined small venules. Each of the three investigated chemokines, including Rantes, exhibited a characteristic binding pattern to the extracellular matrix of the lesion. The observed chemokines may play a role in establishing the cellular composition of chronic apical periodontitis, thus augmenting the intensity of local inflammation and tissue damage.