CK18和CK13在含有纤毛柱状细胞的根端囊肿衬里上皮中的表达

目的:探讨细胞角蛋白CK13和CK18在含有纤毛柱状细胞的根端囊肿衬里上皮中的表达意义。方法:利用单克隆CK18和CK13抗体,采用免疫组化SP法检测32例含有纤毛柱状细胞的根端囊肿衬里上皮中CK13和CK18的表达情况。结果:CK18在此32例含纤毛柱状上皮细胞的根端囊肿衬里中呈现3种表达形式:①伴随纤毛柱状上皮细胞呈连续带状表达在根端囊肿衬里上皮的表层;②呈片段状表达在根端囊肿衬里上皮中;③呈零星散在状表达于衬里上皮细胞中。CK13表达在根端囊肿衬里上皮的鳞状细胞中。一部分基底细胞层细胞和上皮片断既不表达CK18,也不表达CK13。结论:根端囊肿衬里上皮中除含有鳞状细胞外还含有以多种形式分布存在的纤毛柱状细胞及可能处于化生过渡阶段的细胞或上皮发生细胞。根端囊肿衬里上皮中存在着多种细胞类型为根端囊肿的发生机制及组织来源的进一步研究探讨提供了一定的依据。     

细胞角蛋白(CK)13在鼻腭囊肿衬里上皮中的表达及其意义

目的 研究细胞角蛋白(CK)13在鼻腭囊肿衬里上皮中变化的规律和其表达的意义.方法 对经临床和病理诊断的36例鼻腭囊肿的衬里上皮进行常规HE染色和免疫组化染色,检测CK13在鼻腭囊肿上皮中的表达.结果 CK13在鼻腭囊肿的纤毛上皮表达呈阴性;在纤毛柱状与鳞状上皮细胞的混合型上皮的中间层表达呈弱阳性;CK13在部分鳞状上皮的基底层、中间层或表层表达为阳性;鳞状上皮的全层呈强阳性.结论 在纤毛柱状上皮细胞中出现化生鳞状上皮细胞时,同时表达CK13;CK13随着纤毛柱状上皮向鳞状上皮化生程度的逐渐增强而逐渐增多;CK13可作为判断鼻腭囊肿上皮鳞状化生的标志和检测化生程度.根据纤毛柱状上皮向鳞状上皮化生过程中细胞形态的变化和CK13的表达,推测鳞状上皮细胞是由纤毛柱状上皮化生而来.     

细胞角蛋白(CK)13在含牙囊肿衬里上皮的表达

目的 研究纤毛柱状上皮向鳞状上皮化生时细胞角蛋白13的表达，探讨其表达在上皮化生方面的意义。方法 取54例含牙囊肿和6例鼻腭管囊肿，分别采用细胞角蛋白13单克隆抗体进行免疫组织化学染色、HE染色和RT-PCR方法研究细胞角蛋白13基因的表达。结果 54例含牙囊肿中，46例为复屡鳞状上皮细胞衬里，CK13均呈阳性表达；8例混合性上皮衬里，其中的鳞状上皮细胞部分CK13也呈阳性表达；纤毛柱状上皮部分和6例鼻腭管囊肿的纤毛柱状上皮细胞均呈阴性表达。RT-PCR结果显示细胞角蛋白13基因在复屡鳞状上皮中表达最强，在混合性上皮中表达减弱，在纤毛柱状上皮中表达极弱。结论 细胞角蛋白13基因（CK13-mRNA）在纤毛柱状上皮、混和性上皮和复屡鳞状上皮三种不同上皮衬里中均有表达，但表达强度不同。呈由弱到强的变化。在鳞状上皮细胞中细胞角蛋白13呈强阳性表达；在纤毛柱状上皮细胞中细胞角蛋白13呈阴性；可是，当纤毛柱状上皮中出现鳞状上皮细胞时，可见细胞角蛋白13呈阳性表达，因而，细胞角蛋白13是纤毛柱状上皮细胞向鳞状上皮细胞化生过程中的标志性产物，但是有待进一步验证。     

细胞角蛋白18及其基因在牙源性角化囊肿衬里上皮中表达的意义

目的检测细胞角蛋白18（CK18）及其基因在牙源性角化囊肿（OKC）衬里上皮中的表达。方法选取32例OKC的衬里上皮组织,分别进行CK18、CK8和CK19单克隆抗体的免疫组织化学染色。对其中12例使用RT- PCR法检测CK18 mRNA,观察其在衬里上皮中的表达;同时使用CK18基因探针进行原位杂交,检测CK18 mRNA在衬里上皮细胞层的定位表达。结果在免疫组织化学染色中, 17例CK18蛋白在OKC衬里上皮的表层细胞层表达为弱阳性;27例CK18蛋白在棘细胞层上层染色为阳性;14例CK18蛋白在棘细胞层染色为阳性;所有标本基底细胞层染色呈阴性。RT- PCR法检测见4例CK18 mRNA表达为强阳性,8例表达为弱阳性。原位杂交法检测见8例CK18 mRNA在棘细胞层和棘细胞层上层呈阳性,4例在上皮基底细胞层和角化层呈阳性。CK8蛋白在所有32例OKC衬里上皮基底细胞层均有表达。CK19蛋白在23例OKC衬里上皮表层均有表达。结论CK18在OKC衬里上皮的表达由基底细胞层向棘细胞层迁移,CK18蛋白免疫组织化学染色阳性表达与CK18 mRNA原位杂交法阳性表达不同,提示CK18可能与衬里上皮的增殖活性有关,OKC衬里上皮中可能存在CK18蛋白和CK18 mRNA表达的调控因子。     

细胞角蛋白18和13在术后性上颌囊肿化生上皮中的表达

目的研究口腔术后性颌骨囊肿（POMC）化生上皮起源以及细胞角蛋白（CK）在鳞状化生细胞中的表达.方法采用免疫组化SP法、原位杂交法和RT-PCR法,分别检测46例POMC中细胞角蛋白的表达情况.其中13例囊肿衬里上皮只含有假复层纤毛柱状上皮细胞;30例囊肿衬里既含有纤毛柱状上皮细胞,又含化生的鳞状上皮细胞;3例囊肿衬里只含有化生的鳞状上皮细胞.结果43例纤毛柱状上皮细胞中,39例表达CK8,9例表达CK13,43例均有CK18表达.发生鳞状上皮化生的细胞中CK13表达较多,CK8和CK18表达较少.在33例发生化生的囊肿衬里中,24例表达CK8,23例表达CK13,26例表达CK18.CK13和CK18蛋白的表达与CK13、CK18-mRNA表达水平相关.原位杂交方法检测CK1g-mRNA表达时发现,26例CK18蛋白阳性的化生囊肿衬里上皮以及7例发生化生但不表达CK18蛋白的囊肿衬里上皮都有CK18-mRNA的表达.RT-PCR结果进一步证明所有发生鳞状化生的囊肿衬里上皮都有CK18-mRNA的表达,但是其表达水平较未发生化生的柱状细胞囊肿衬里上皮弱,而且CK13-mRNA的表达与CK18-mRNA表达相反.结论在发生上皮化生的全过程中,CK18-mRNA保持不变,但是其蛋白的表达水平下降,而且发生鳞状化生时CK18表达减少,CK13表达增加.     

细胞角蛋白(CK)13及其基因在术后性上颌囊肿衬里上皮中的表达

目的研究CK13蛋白和CK13-mRNA在术后性上颌囊肿(post operative maxillary cysts,POMCs)衬里上皮细胞中的表达,进而探讨CK13在纤毛柱状上皮向鳞状上皮化生过程中的意义.方法本研究采用免疫组织化学染色法和RT-PCR法分别检测CK13蛋白和CK13-mRNA在32例POMCs衬里上皮中的表达.结果免疫组织化学染色显示CK13蛋白在鳞状上皮中呈阳性表达,在纤毛柱状上皮中呈阴性;RT-PCR法检测出CK13-mRNA在所有囊肿衬里上皮均呈阳性表达,其中13例纤毛柱状上皮呈弱阳性,14例既有纤毛柱状上皮又有鳞状上皮的混合上皮呈中等强度表达,5例鳞状上皮呈强阳性.结论 POMCs衬里的纤毛柱状上皮中存在CK13-mRNA,提示化生的鳞状上皮细胞可能来源于纤毛柱状上皮.在纤毛柱状上皮向鳞状上皮化生时CK13蛋白可能起着重要作用.     

牙源性发育囊肿和牙囊中上皮的细胞角蛋白表达模式

人体的细胞角蛋白共有19种,已经证实不同类型的上皮组织其细胞角蛋白的表达模式亦不同。含牙囊肿和牙源性角化囊肿均来源于牙胚中的牙源性上皮,但对它们的角蛋白表达模式和上皮分化过程中的细胞角蛋白变化了解甚少。本研究目的是检查牙源     

细胞角蛋白10、13、14和19在OLP中的表达研究

目的:研究口腔扁平苔藓(OLP)中细胞角蛋白(CK)的表达变化.方法:采用免疫组化SP法检测7例正常颊黏膜和17例OLP颊部病变中细胞角蛋白的表达部位和强度.结果:在对照组和OLP颊部病变中CK19表达阴性,CK10、CK13表达于复层上皮的基底上层.在对照组中CK14主要表达于基底层,在OLP颊部病变中CK14可扩展至基底上层;与对照组比较OLP中CK10、CK14出现过度表达,而CK13表达下调,有时甚至无表达.结论:OLP中CK表达的转变与OLP病变上皮的异常角化、增殖和分化有关.     

口腔粘膜上皮角蛋白的细胞生物学

角蛋白是上皮细胞主要的结构蛋白和分化的标志产物。本文综述了角蛋白多肽的结构、功能和调节机制,重点介绍了口腔粘膜生长发育过程中及成人阶段角蛋白表达的特征及影响因素,为认识正常口腔粘膜生理及研究上皮发病机制提供了依据。     

腺牙源性囊肿两例报告及其细胞角蛋白18的表达

目的探讨腺牙源性囊肿衬里上皮的组织学特征及其细胞角蛋白18、19、CK18-mRNA的表达和该囊肿的组织来源。方法对2例腺牙源性囊肿采用常规HE切片、免疫组织化学染色和原位杂交的方法分别进行组织学观察，检测CK18、CK19和CK18-mRNA的表达。结果囊壁内有微小子囊存在，子囊周围有黏液细胞为主的混合性腺体结构。CK18在囊肿衬里上皮呈阳性表达；在子囊的衬里上皮CK18呈阴性，而CK19呈阳性表达；腺体结构中CK18和CK19均呈阳性表达。在原位杂交中CK18-mRNA在所有上皮中均呈不同程度的阳性表达。结论CK18-mRNA及其蛋白的表达差异可能与囊肿上皮细胞的分化有关，腺牙源性囊肿的角蛋白表达谱存在牙源性上皮和腺源性上皮的交叉，可能同时存在牙源性和腺源性分化。     

Immunohistochemical expression of vascular endothelial growth factor and matrix metalloproteinase-9 in radicular and residual radicular cysts

Objective

This study assessed and compared the immunoexpression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in radicular cysts (RCs) and residual radicular cysts (RRCs), relating them to the angiogenic index and the intensity of the inflammatory infiltrate.

Material and Methods

Twenty RCs and 10 RRCs were evaluated by immunohistochemistry using anti-VEGF and anti-MMP-9 antibodies. The angiogenic index was determined by microvessel count (MVC) using anti-von Willebrand factor antibody.

Results

The expression of both VEGF and MMP-9 was higher in RCs than in RRCs. RCs and RRCs presented strong epithelial expression of VEGF, irrespective of the intensity of the inflammatory infiltrate. Lesions with strong expression of MMP-9 showed significantly higher number of immunopositive cells for VEGF (p<0.05) and higher MVC (p<0.05). Lesions with dense inflammatory infiltrate exhibited significantly higher MVC (p<0.05) and higher number of immunopositive cells for VEGF (p<0.05). There was a positive correlation between both MVC (p<0.05) and the quantity of immunopositive cells for VEGF (p<0.05), with intensity of the inflammatory infiltrate. In addition, it was observed a positive correlation between the number of immunopositive cells for VEGF and MVC (p<0.05).

Conclusions

VEGF and MMP-9 might play important roles in the angiogenesis in RCs and RRCs. In these lesions, the expression of these molecules and the MVC is closely related to the intensity of the inflammatory infiltrate. The expression of VEGF in the epithelial lining of RCs and RRCs might be important for the enlargement of these lesions.     

Comparative immunohistochemical expression of RANK, RANKL and OPG in radicular and dentigerous cysts

Objective

Receptor activator of nuclear factor-κB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG) are members of the superfamily of ligands and receptors of tumour necrosis factor family involved in bone metabolism. The formation, differentiation and activity of osteoclasts are regulated by these proteins. To clarify the roles of osteoclast regulatory factors in cystic expansion of odontogenic cysts, expression of these proteins were analysed in radicular and dentigerous cysts.

Design

The immunohistochemistry expression of these biomarkers were evaluated and measured in lining epithelium and fibrous capsule of the radicular (n = 20) and dentigerous cysts (n = 20).

Results

A similar expression in lining epithelium was observed in the lesions. The fibrous capsule of dentigerous cyst showed a higher content of RANK-positive and RANKL-positive cells than fibrous capsule of radicular cyst. In the lining epithelium the RANKL/OPG ratio showed higher numbers of OPG-positive than RANKL-positive cells, whereas fibrous capsule of the cysts had a tendency to present a similar expression (OPG = RANKL).

Conclusion

Ours findings indicate the presence of RANK, RANKL and OPG in cysts. Moreover, increased expression of OPG compared to RANKL in the lining epithelium could contribute to the differential bone resorption activity in theses lesions.     

Apoptosis in epithelial cells of apical radicular cysts

AIM: To investigate the occurrence of apoptotic cell death in the epithelium of radicular cysts and to compare its frequency in lesions presenting a distinct functional state. METHODOLOGY: Twenty radicular cysts were selected and arranged into two groups with 10 lesions in each group: atrophic (quiescent) and hyperplastic (active) epithelium. Morphologic investigations of apoptosis were conducted by means of optic microscopy in haematoxylin and eosin slides. Immunohistochemical techniques to detect the bcl-2 protein were carried out by streptavidin-biotin-peroxidase assay. In both instances, 30 sequential high-power microscopic fields were observed to determine apoptotic (AI) and bcl-2 immunostaining (bcl-2I) indexes. The presence of AI and bcl-2I within the two groups was compared using the t-test. Correlation between the AI and the bcl-2I was investigated using the Spearman test. RESULTS: Apoptosis was detected in the epithelium of all cysts. Higher AI levels were found in lesions with an atrophic (0.17 +/- 0.19) rather than a hyperplastic (0.10 +/- 0.10) epithelium. The same was found for the bcl-2I levels (0.06 +/- 0.03 vs. 0.04 +/- 0.01, respectively). However, these differences were not statistically significant. A positive and significant correlation was found between AI and bcl-2I. CONCLUSIONS: Apoptosis was always present in the epithelium of the lesions and was more frequent in lesions with atrophic (quiescent) epithelium.     

根尖囊肿上皮细胞增殖与郎格罕细胞分布的关系

目的 分析根尖囊肿衬里上皮细胞增殖及其与郎格罕细胞分布的关系。方法 收集23例经组织病理学确诊的根尖囊肿石蜡标本，分别进行增殖细胞标记Ki-67和郎格罕细胞标记S-100的免疫组化染色，观察根尖囊肿衬里上皮的细胞增殖及其与郎格罕细胞分布的关系。结果 衬里上皮内的Ki-67阳性的细胞数目与结缔组织囊壁内的S-100阳性的细胞数目呈正相关（P〈0．05），但与衬里上皮内的S-100阳性的细胞数目无相关关系（P〉0．05）。结论 结缔组织囊壁中的郎格罕细胞数目与衬里上皮的增殖活性呈正相关，提示囊壁内由郎格罕细胞参与介导的免疫反应可能有促进上皮增殖的作用。     

T lymphocyte activation and cytokine expression in periapical granulomas and radicular cysts

Introduction

Radicular cysts (RCs) are periapical lesions resulting in jaw bone destruction. The inflammatory dental periapical granuloma (PG) is considered to be the origin of RC formation; however the mechanism of RC development remains unclear.

Methods

Cell suspension from the surgically extirpated tissue of 27 RCs and 25 PGs was obtained. Bacteriological analysis of the PG tissue samples was performed in order to define two major groups of PG according to the prevailing causative bacterial infection: the streptococcal PG (PG-S, n = 10) and the anaerobe PG (PG-A, n = 9) group. The inflammatory response of tissue infiltrating lymphocytes was assessed by following T lymphocyte activation (HLA-DR expression) as well as interferon γ (IFN-γ) and interleukin 4 (IL-4) production which were evaluated by the flow cytometry.

Results

In comparison to RC both types of PG contained a higher proportion of activated T cells (HLA-DR) and lower proportion of IL-4 producing cells. PG-A tissue contained increased percentage of CD3 cells and increased percentage of T helper 1 (Th1) cells in comparison with PG-S. In RC the IFN-γ production is higher than in streptococcal PG-S but similar as in PG-A.

Discussion

Tissue infiltration by Th2 cells and IL-4 production is likely to play an etiopathogenic role in RC formation.     

Oral lichen planus: an immunohistochemical study of heat shock proteins (HSPs) and cytokeratins (CKs) and a unifying hypothesis of pathogenesis

The expression of heat shock proteins HSP60 and HSP70 and cytokeratins CK1/10 and CK7/18 were compared in epithelium of oral lichen planus (OLP) lesions and oral fibromas using an avidin-biotin-peroxidase complex (ABC) immunohistochemical method. An immunostaining intensity distribution (IID) index was developed to assess staining intensity and the proportion of positively stained cells in different layers of the epithelium. The expression of HSP60 in the basal layer was significantly higher in OLP than in fibromas. No difference in HSP70 expression was evident between OLP and fibromas. The expression of CK1/10 in the epithelial basal and suprabasal layers was significantly higher in OLP than in fibromas. There was no demonstrable staining for CK7/18 in either OLP or fibromas. A significant correlation was evident between the expression of HSP60 and CK1/10 in the basal epithelial cells in OLP. The findings support a role for HSP60 in the pathogenesis of OLP. A unifying hypothesis of the pathogenesis of OLP, involving two sequential immune reactions, is proposed.     

Vacuolated cells and mucous metaplasia in the epithelial linings of radicular and residual cysts

The purpose of this study was to determine whether a consistent association exists between mucous cells and clear or vacuolated cells in the epithelial lining of radicular and residual cysts and to consider whether the vacuolated cells may represent a stage in the histogenesis of mucous metaplasia in these linings. Single sections from each of 154 mandibular radicular and residual cysts were stained with periodic acid-Schiff (PAS) after diastase digestion. Fifteen cases which showed mucous metaplasia were included in the study and were examined for the presence of vacuolated cells associated with the mucous cells. Mucous cells were present singly or in groups within all layers of the stratified squamous epithelial lining except the basal cell layer. In nearly all instances small to large ovoid vacuolated cells were found closely associated with the mucous cells. Occasional vacuolated cells contained sparse mucin granules or a delicate network of PAS-positive, diastase-resistant material. It is suggested that the clear cells may represent a stage in the histogenesis of mucous metaplasia.     

Effects of the patient's age and the size of the primary lesion on the speed of shrinkage after marsupialisation of keratocystic odontogenic tumours,dentigerous cysts,and radicular cysts

Marsupialisation, by which a surgical window is created in the cavity of a cystic lesion, has been recommended to avoid the formation of a bony defect in the jaw and a reduction in a patient's quality of life. However, information about the factors that affect the reduction in the size of a cyst after marsupialisation is limited. We have studied the effects of the patient's age and the size of the primary lesion on the speed of shrinkage after marsupialisation of keratocystic odontogenic tumours (KCOTs), dentigerous cysts, and radicular cysts. The speed of shrinkage (mm²/month) was evaluated by measuring the radiolucent area on panoramic radiographs taken before and after marsupialisation for KCOT (n = 28), dentigerous cysts (n = 26), and radicular cysts (n = 18) in the mandibular molar regions. The mean duration of marsupialisation for each type of cyst was 11 (5), 8 (5), and 5 (2) months, respectively. The radiolucent area decreased linearly in the 3 types depending on the time after marsupialisation: r = −0.86 (p < 0.01), r = −0.73 (p < 0.01), and r = −0.72 (p < 0.01), respectively. The relative speed of shrinkage did not correlate with the age of the patients, though it did correlate with the size of the radiolucent area before marsupialisation in KCOT (r = 0.69, p < 0.01) and radicular cysts (r = 0.73, p < 0.01). These results suggest that the size of a cyst before marsupialisation may affect the speed of shrinkage in KCOT and radicular cysts, while the age of the patient does not.     

The use of digital subtraction radiography to evaluate bone healing after surgical removal of radicular cysts

Objective The purpose of this study was to evaluate the bone healing process after surgical removal of radicular cysts by using preoperative and postoperative panoramic radiographs, which were digitized and subtracted using a projective standardization software program (Emago). Methods Seventeen patients with large radicular cysts treated by surgical enucleation were included in the study. All surgical procedures were performed by one of the authors (D.K.). Each patient had a panoramic preoperative radiograph (plain film) and a panoramic postoperative radiograph (plain film), which was taken 6 to 12 months after surgery. All radiographs were taken with the same panoramic unit. The part of the radiograph that included the lesion in the preoperative radiograph was digitized using a CCD digital camera at a standard distance. The postoperative radiograph was also digitized using the same standardized parameters. The preoperative and postoperative images were then manipulated by means of the projective standardization software program Emago to reveal the regenerated area. This area was calculated in pixels, and the percentage of bone healing was determined for each patient. The data were analyzed using Student's t test and the Wilcoxon test for pair differences. Results The percentage of bone healing ranged from 55.14% to 95.68% with a mean of 72.27%. In all cases, the differences were significantly different at P = 0.01. Conclusions Digitizing the part of the panoramic radiograph that included the lesion area and subsequently performing projective standardization is a suitable method for analyzing the healing process by means of subtraction radiography. The projective standardization software program performs the geometric reconstruction and subtraction process. An evaluation of the healing process can be obtained by calculating the regenerated bone area in the subtracted images.