Cytometric analysis of oral scrapings of patients with oral lichen planus

A. Acha‐Sagredo, Y. Jiménez, J. V. Bagán, M. A. Echebarria‐Goicouria and J. M. Aguirre‐Utric analysis of oral scrapings of patients with oral lichen planus Objective: Aneuploidy has been associated with malignant and premalignant oral lesions. In the past few years, its application in oral precancerous lesions and its prognostic meaning have been controversial issues. The aim of our study was to characterize alterations in DNA content by automated DNA image cytometry in oral scrapings of patients with oral lichen planus. Methods: Cytological samples from 40 patients clinicopathologically diagnosed with oral lichen planus were analysed by DNA image cytometry. Results: All the cases were classified as diploid, showing a predominant population of cells with normal DNA content (DNA index, 0.85–1.15). Atrophic/erosive lesions showed a higher percentage of tetraploid cells when compared with reticular/papular lesions but this was not statistically significant (P = 0.09). Conclusions: Aneuploidy does not seem a common event in oral lichen planus lesions. However, we consider that the use of DNA image cytometry of oral scrapings may be an easy and helpful methodology in the follow‐up of patients with these lesions.     

肥厚性扁平苔藓皮损中浸润淋巴细胞的研究

目的探讨淋巴细胞浸润在肥厚性扁平苔藓发病机制中的意义。方法采用免疫组织化学SP法对10例肥厚性扁平苔藓患者皮损、20例非肥厚性扁平苔藓患者皮损及10例正常人皮肤中CD3、CD4、CD8、CD20进行检测。结果所有患者皮损均以CD3^＋’T、CD4^＋T、CD8^＋T细胞浸润为主,散在CD20^＋B细胞浸润。肥厚性皮损中浸润T淋巴细胞数少于非肥厚性皮损（P〈0．01）,CD20^＋B细胞占浸润淋巴细胞百分率高于非肥厚性皮损（P〈0．05）。正常人皮肤仅在真皮偶见CD3^＋T细胞。结论肥厚性扁平苔藓皮损T淋巴细胞的减少,B淋巴细胞的增多表达可能与疾病的持续存在及局部过度增殖性改变密切相关。     

Tissue expression of proliferative antigens (PCNA and Ki-67) in oral lichen ruber related to clinical status

The aim of this study was to determine the expression intensity of PCNA and Ki-67 tissue antigens related to pathologically modified oral mucosa in OLR lesions, and to determine the reaction intensity of these antigens in individual clinical forms, i.e. lichen ruber planus (LRP) and lichen ruber erosivus (LRE) comparing the reaction intensity with the inflammation grade and the degree of hyperkeratosis in lesions of 30 patients. Control group included patients (n = 15) with oral leukoplakia simplex. Tissue antigens were observed by immunohistochemical analysis using APAAP and LSAB methods. The reaction on tested tissue antigens was focal positive and of mosaic type. The reaction of the PCNA antigen was intensely high in OLR lesions regardless on the clinical form of the lesion. The reaction intensity positively correlated with the inflammation grade and the degree of hyperkeratosis in lesions. The reaction on Ki-67 tissue antigen ranged from low to moderately high intensity. Intensely high reaction was observed in lesions of lichen ruber erosivus. The reaction positively correlated with the inflammation grade and the degree of hyperkeratosis in lesions. Observed modified reaction of analyzed tissue antigens related with individual clinical forms of OLR might be the indicator of transformed nature of these lesions.     

Evaluation of angiogenesis in normal and lichen planus skin by CD34 protein immunohistochemistry: preliminary findings

Angiogenesis is a critical component of both neoplastic and chronic inflammatory disorders, but whether angiogenesis also occurs in inflammatory skin diseases (such as lichen planus) has yet to be established. This study tests the hypothesis that the development of cutaneous lichen planus is associated with alterations of dermal vascularization (microvessel density, MVD). Thirty cases of cutaneous lichen planus and 40 cases of normal skin were studied. Dermal microvessels were immunostained for CD34 and counted in 10 areas with the highest numbers of microvessels; the mean value represented the final MVD. Compared with normal skin, dermal microvessel density was increased in cutaneous lichen planus [mean, 2.50 (SEM, 0.09) versus 1.39 (SEM, 0.12)]. The microvessel number was higher in the dermal inflammatory infiltrate (intralichenoid infiltrate) and at dermoepidermal junction (below Max-Josef space) compared with adventitial dermis [mean, 2.50 (SEM, 0.09) versus 1.6 (SEM, 0.10)]. The higher MVD values in cutaneous lichen planus suggest a possible link between angiogenesis, and development of these cutaneous lesions. These results provide a morphological evidence of potent angiogenic activity in cutaneous lichen planus, indicating that the local microvasculature may undergo an intense process of inflammation-dependent angiogenesis.     

Trace metals in oral mucosa in relation to the lichen ruberplanus pathology. A preliminary study carried out by neutron activation analysis.

Lichen ruberplanus, contact allergy and hypersensitivity can be linked to oral exposure to metals released from metal alloys commonly used in dentistry. In this context neutron activation analysis was developed for the microdetermination of 36 elements in oral mucosa biopsies of two patients affected by lichen ruber planus and of five subjects as control group. In order to minimise metal contamination during sample collection, biopsies were taken by laser bistoury technique and then submitted to radiochemical neutron activation analysis (RNAA). Among the metals analysed, chromium showed obvious high concentration in gingival biopsies of the two pathological subjects compared to the corresponding tissues of control group. Cobalt and nickel were also determined in higher concentrations, but only in one of the oral mucosa of the two patients. The present findings way support the hypothesis concerning a potential link of lichen ruber planus condition with the exposure to Cr, Co and Ni as released into oral cavity from prosthodontic alloys.     

低功率氩离子激光治疗口腔癌前病变的临床研究

目的对氩离子激光治疗口腔癌前病变的治疗机理、照射功率、方法及疗效差异进行探讨。方法 对５３例口腔白斑和扁平苔藓患者应用Ａｒ＾＋激光治疗,检查比较患者治疗前后的临床变化。结果 口腔白斑３１例全部治愈,治愈率１００％;口腔扁平苔藓２２例治愈１３例,显效７例,无效２例,治疗有效率９０．９％,总有效率达９６．２％。结论 氩离子激光治疗口腔白斑和扁平苔藓等清表的口腔慢性病损,不仅疗效显著,操作简便,并发症少     

Quantitative cytomorphologic analyses of Papanicolaou-stained smears from oral lichen planus

OBJECTIVE: To apply quantitative techniques to cytologic smears in order to improve the diagnostic sensitivity of cytology in the detection of malignant change in the oral cavity. STUDY DESIGN: Eighty-two patients with lesions of oral lichen planus were investigated. A total of 247 Papanicolaou-stained buccal smears underwent nuclear and cytoplasmic area measurements using a TV image analyzer. RESULTS: The cytomorphologic results of this study suggest that lesions of oral lichen planus contain smaller cells and nuclei than those observed in clinically normal oral mucosal smears. There was a statistically significant reduction in nuclear area (P < .001) and cytoplasmic area (P < .05) for the oral lichen planus smears. CONCLUSION: The TV image analysis system could provide a valuable method of quantifying cell changes in oral smears collected from oral mucosa as a method of monitoring lesions of oral lichen planus.     

Image cytometric evaluation of nuclear texture features and DNA content of the reticular form of oral lichen planus

OBJECTIVE: To analyze image cytometric chromatin changes reflected in nuclear texture features and DNA ploidy of oral lichen planus in relation to the normal buccal mucosa and buccal mucosa expressing malignancy-associated changes in cancer patients. STUDY DESIGN: Twenty-eight patients with the reticular form of oral lichen planus, with a follow-up period of 25 years, 50 healthy controls and 50 lung cancer patients were included in the study. Scrapings of buccal mucosa were suspended in transport medium. Monolayer filter preparations were Feulgen-thionin stained. Image cytometric analysis was performed by Cyto-Savant. RESULTS: All oral lichen planus specimens in our study were diploid. In univariate analysis, differences between the normal buccal mucosa and oral lichen planus were found in several nuclear texture features, which gave an 80% correct classification rate in multivariate analysis. In the second part of the study, the classifier that recognizes malignancy-associated changes on the buccal mucosa of patients with lung cancer correctly recognized > 80% of oral lichen planus samples as normal buccal mucosa. CONCLUSION: Our results indicate that chromatin changes in oral lichen planus exist compared to normal cells; however, the chromatin structure of the reticular form of oral lichen planus does not express malignancy-associated changes and is more similar to normal squamous cells.     

Candida biotypes in patients with oral leukoplakia and lichen planus

Prevalence of yeasts in 35 leukoplakia and 34 oral lichen planus patients was compared with that observed in persons without oral diseases. Serotype and morphotype were determined on Candida albicans isolates. Yeasts were isolated from the oral cavity specimens of 43.7% of the patients. C. albicans (serotype A) was the predominant species (76% in leukoplakia, 88.2% in lichen planus and 60.8% in healthy persons). Sixteen morphotypes were encountered on malt extract agar, being 732, 733, 734, 753 and 754 the most frequently found. Morphotypes SP1N and SP1Y were the most common on Sabouraud-trypheniltetrazolium agar (68.4% of the isolates from leukoplakia and 73.3% from lichen planus, but only 46.6% of the isolates from healthy oral mucosa showed SP1N morphotype). Presence of oral lesions was associated with a marked reduction in the yeast species and C. albicans biotypes, suggesting that C. albicans and particularly some of its biotypes, show a high potential of adaptation to the changes associated with the development of oral leukoplakia and lichen planus.     

Lichen planus and liver diseases: a multicentre case-control study. Gruppo Italiano Studi Epidemiologici in Dermatologia (GISED).

OBJECTIVE--To assess the association of lichen planus with liver complaints and with known aetiological factors of liver diseases. DESIGN--Multicentre case-control study. Interviews were conducted by trained medical investigators on the basis of a structured questionnaire. At the interview patients and controls were asked for consent to blood samples being taken to determine transaminase activities and the presence of hepatitis B virus surface antigen. SETTING--Outpatient departments of 27 Italian general and teaching hospitals that were collaborating in the Gruppo Italiano Studi Epidemiologici in Dermatologia (GISED). SUBJECTS--Incident cases and controls were eligible. A total of 577 patients with lichen planus and 1031 controls with dermatological diseases other than lichen planus were interviewed. Less than 1% of the people contacted refused to participate. Patients and controls were matched for sex and age in five year intervals. RESULTS--The risk of lichen planus was higher in patients with a history of liver diseases requiring hospital admission or specialist consultation (relative risk = 1.6; 95% confidence interval = 1.2 to 2.2), those who had had liver biopsy (5.5; 1.9 to 15.6), and those with a history of viral hepatitis (1.9; 1.1 to 3.1). High activities of liver enzymes and positive results of tests for hepatitis B virus surface antigen were also associated with lichen planus. The association with alcohol consumption was not clearly confirmed by a dose-risk relation. CONCLUSION--This study adds quantitative epidemiological evidence to the clinical observation that liver disease is a risk factor for lichen planus although not a specific marker of it.     

DNA content as a prognostic marker of oral lichen planus with a risk of cancer development

OBJECTIVE: To assess the presence of aneuploidy in oral lichen planus (OLP) and its usefulness as a prognostic marker. STUDY DESIGN: Eighty-one formalin-fixed, paraffin-embedded biopsy samples taken from atrophic-erosive OLP from 70 patients were studied. Approximately 150 random nuclei in basal and/or parabasal epithelia were analyzed with static cytometry. RESULTS: Aneuploidy was detected in 41% of samples. OLPs with ulcerations or location in the tongue had significantly higher values, respectively, for the 2.5c exceeding rate (ER) (p<0.001 and 0.001) and proliferation index (PI) (p = 0.012 and 0.013) than did lesions without ulcerations or at other locations. 2.5c ER was significantly higher in dysplastic OLP lesions (p < 0.001), and the significant value (p = 0.001)for 2.5c ER discriminating DNA aneuploidy was 15.3%. In multivariate analysis only the G2/M ER (G2/MER) was a significant independent predictor of developing cancer in OLP (OR 2.349, 95% CI 1.39-3.97, p = 0.001). CONCLUSION: Ulcerated atrophic-erosive OLPs of the tongue and with dysplasia are at increased risk of cancer development. 2.5c ER, PI and G2/MER might be useful in prognosticating the increased risk of malignancy in OLP.     

蕈样肉芽肿与扁平苔藓、慢性皮炎湿疹浸润细胞的免疫组化比较研究

目的探讨免疫表型对蕈样肉芽肿与扁平苔藓、慢性皮炎湿疹鉴别诊断的意义。方法本研究应用ABC免疫组化技术检测15例蕈样肉芽肿,17例慢性皮炎湿疹,17例扁平苔藓,6例正常皮肤的CD1a、CD8、ICAM-1和CD7的表达情况。结果1.蕈样肉芽肿表皮CD1a,CD8,ICAM-1的阳性细胞密度明显高于扁平苔藓、皮炎湿疹和正常皮肤。2.蕈样肉芽肿真皮中CD1a阳性细胞的线性密度高于扁平苔藓、慢性皮炎湿疹和正常皮肤,蕈样肉芽肿真皮内CD8阳性细胞百分比较慢性皮炎湿疹增多,蕈样肉芽肿真皮表达ICAM-1抗原的细胞百分比高于扁平苔癣和慢性皮炎湿疹。3.慢性皮炎湿疹、扁平苔藓真皮内CD7阳性细胞百分比高于蕈样肉芽肿和正常皮肤。结论CD1a、CD8、ICAM-1、CD7免疫表型的研究可能为蕈样肉芽肿和扁平苔藓、慢性皮炎湿疹等一些炎症性皮肤病的鉴别诊断提供一定的线索。     

Affections dermatologiques de la muqueuse genitale masculine et de la verge

A wide variety of infectious, inflammatory or dysplasic disorders may affect both male genital skin and mucous membranes. We review here the clinical patterns or the most common disorders. The diagnosis is based upon the existence of erythematous balanitis, erosions or ulcerations. Among infections, candidiosis, genital herpes, HPV papillomas are the most common. Most of inflammatory skin disorders may affect the male organ, especially psoriasis, lichen planus or erosive lichen planus, allergic dermatitis, auto-immune bullous dermatoses, drug-reactions. Chronic balanitis, or recurrent inflammatory balanitis, lichen sclerosus may lead to the emergence of intraepithelial neoplasias (PIN) or invasive carcinomas. The management of genital lesions needs in fact: the observation or oral mucous membrances and of the skin it-self with a special attention to very peculiar body sites for the presence of typical skin disease lesions; a surgical biopsy for microscopic observation; in some cases immunopathology with direct and indirect immunofluorescence for the diagnosis of auto-immune disorders.     

Biochemical Changes Observed After PUVA Versus PUVA Plus Methotrexate Therapy in Mycosis Fungoides Using Synchrotron Infrared Microspectroscopy

Mycosis fungoides (MF) is the most common type of cutaneous T cell lymphoma in which the distinction between early stage MF and other inflammatory dermatosis remains difficult. Twenty patients of early stage MF and nine patients with psoriasis and lichen planus were included in this study. Ten MF patients were treated with psoralen plus UVA (PUVA) and the other 10 MF patients were treated with PUVA plus methotrexate (MTX) until complete clinical remission. Synchrotron infrared microspectroscopy (SIRM) found that MF lesions were biochemically different compared to inflammatory diseases. After treating MF with either therapeutic modality, the lymphocytic count decreased significantly in both the epidermis and dermis (P < 0.001) but no biochemical changes were observed in the remaining lymphocytes after treatment, indicating the disease process was slowed by treatment but not eradicated. In conclusion SIRM is a promising method for distinguishing MF from other inflammatory diseases such as psoriasis and lichen planus. A significant reduction in lymphocyte count indicated that PUVA therapy is an effective treatment for early stage MF, and MTX could be reserved for more advanced cases that are not PUVA responsive. However, SIRM evidence of persistent disease suggests that maintenance therapy is recommended after clinical remission.     

Exfoliative-cytologic diagnosis of basal-cell carcinoma, with the use of DNA image cytometry as a diagnostic aid

Eighty-four skin lesions clinically suspected of being basal-cell carcinomas were investigated by exfoliative cytology. Specific criteria were found for the cytologic diagnosis of basal-cell carcinoma. All 37 cases of basal-cell carcinoma were correctly classified cytologically and could be differentiated from the 8 cases of squamous-cell carcinoma. There were no false-positive or false-negative diagnoses. Insufficient cellular material was obtained in 17% of the cases. The technique for collecting exfoliated epidermal cells with a new swab is described. DNA image cytometry was used as a diagnostic aid in doubtful cases. DNA image cytometry showed that 83% of the basal-cell carcinomas had an aneuploid nuclear DNA content.     

Comparison of diversity of torque teno virus 1 in different mucosal tissues and disorders

Diversity of TTV1 was assessed in the head and neck region in patients with potentially malignant (oral lichen planus, oral leukoplakia) and malignant lesions (oral and laryngeal squamous cell cancers) and was compared to that found in the uterine cervix (cervical atypia and cervical cancer) by directly sequencing the NG061-063 segment of ORF1. These sequences were classified by the formerly used genogroup-genotype system as well as by the newly accepted species classification by aligning with the corresponding region of the type sequences of the 29 TTV species. All sequences obtained during the study clustered together with the TTV1 type sequence; to express diversity within TTV1, genotypes and subtypes of the former classification were used.The commonest subtypes were 2c followed by 2b, 1a and 1b. Subtypes 2b and 2c were evenly distributed among cervical samples; subtype 1a was more frequent in patients with cervical atypia or cancer. Subtypes 2c was more frequent than 2b in head and neck lesions. In conclusion, genotype and even subtype distribution may be important in association with diseases, therefore using this classification for characterization of intraspecies diversity of TTV1 is proposed.     

清热解毒祛风中药治疗口腔扁平苔癣的临床疗效及对患者外周血T淋巴细胞亚群的影响观察

目的：探讨养血解毒祛风中药治疗40例口腔扁平苔藓的临床疗效以及其对OLP患者外周血T淋巴细胞亚群的影响。方法：将80例扁平苔癣患者随机分为中药组40例和西药对照组40例。治疗组采用中药养血解毒祛风法辨证施治,对照组患者以西药治疗,用左旋咪唑50mg／次,3次／d,口服3天停4天一个月。结果：治疗组与对照组统计学上有显著性差异,中药治疗对患者T细胞亚群有很好的调节作用。结论：中药清热解毒祛风治疗口腔扁平苔癣有显著疗效。     

白色念珠菌分泌型天冬氨酸蛋白酶在口腔扁平苔藓患者唾液中的表达

研究白色念珠菌分泌型天冬氨酸蛋白酶(SAP)家族在扁平苔藓患者唾液中的表达,进而探讨白色念珠菌病与扁平苔藓的关系。选取白色念珠菌携带者、白色念珠菌病患者、携带白色念珠菌的扁平苔藓患者、伴有白色念珠菌病的扁平苔藓患者各20例,念珠菌培养阴性的扁平苔藓患者10例。从唾液标本中直接提取RNA,进行RT-PCR。白色念珠菌携带者唾液中有SAP2、SAP4-6、SAP7的不同表达,携带白色念珠菌的口腔扁平苔藓患者唾液中有SAP2、SAP4-6、SAP7、SAP8、SAP9的不同表达,所有携带者中均未表现SAP1、SAP3和SAP10的阳性扩增。白色念珠菌患病者及伴有白色念珠菌病的口腔扁平苔藓患者唾液中有SAP1-10的不同表达。念珠菌培养阴性的扁平苔藓患者均无SAP1-10的阳性扩增。SAP9与扁平苔藓密切相关。     

Oxidative and nitrative DNA damage in animals and patients with inflammatory diseases in relation to inflammation-related carcinogenesis

Infection and chronic inflammation are proposed to contribute to carcinogenesis through inflammation-related mechanisms. Infection with hepatitis C virus, Helicobacter pylori and the liver fluke, Opisthorchis viverrini (OV), are important risk factors for hepatocellular carcinoma (HCC), gastric cancer and cholangiocarcinoma, respectively. Inflammatory bowel diseases (IBDs) and oral diseases, such as oral lichen planus (OLP) and leukoplakia, are associated with colon carcinogenesis and oral squamous cell carcinoma (OSCC), respectively. We performed a double immunofluorescence labeling study and found that nitrative and oxidative DNA lesion products, 8-nitroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), were formed and inducible nitric oxide synthase (iNOS) was expressed in epithelial cells and inflammatory cells at the site of carcinogenesis in humans and animal models. Antibacterial, antiviral and antiparasitic drugs dramatically diminished the formation of these DNA lesion markers and iNOS expression. These results suggest that oxidative and nitrative DNA damage occurs at the sites of carcinogenesis, regardless of etiology. Therefore, it is considered that excessive amounts of reactive nitrogen species produced via iNOS during chronic inflammation may play a key role in carcinogenesis by causing DNA damage. On the basis of our results, we propose that 8-nitroguanine is a promising biomarker to evaluate the potential risk of inflammation-mediated carcinogenesis.     

Methodologic aspects of DNA assessment by means of image cytometry in tumors of the salivary glands. A comparison between the results obtained using sections and cytospin preparations from the same paraffin-embedded specimens

The image cytometric nuclear DNA assessments on paraffin-embedded tissue sections and on Cytospin preparations of disaggregated specimens from the same cases were compared in 98 salivary gland tumors, including 21 acinic cell carcinomas, 29 mucoepidermoid carcinomas, 21 adenocarcinomas and 27 adenoid cystic carcinomas. The histogram type (diploid, tetraploid or aneuploid) and the number of cells with DNA values greater than 2.5c (expressed in relative units) were considered as variables in the correlation. A high correlation between the results in different specimens was found in acinic cell carcinomas, mucoepidermoid carcinomas and adenocarcinomas; the histogram type and the number of cells with DNA values greater than 2.5c were essentially the same between specimen types in these three tumor entities. The cases of adenoid cystic carcinomas showed a considerably lower degree of correlation: in 8 of the 27 cases, the Cytospin preparations yielded diploid histograms, while the tissue sections yielded aneuploid histograms. The number of cells with DNA values greater than 2.5c was notably lower in the Cytospin preparations from adenoid cystic carcinoma; the reasons for this exceptional behavior of the cells of adenoid cystic carcinoma are discussed. These findings demonstrate that paraffin-embedded specimens of different tumor entities, even from the same organ, can be affected differently by disaggregation procedures. While retrospective studies on disaggregated paraffin-embedded specimens can yield reliable results, comparative assessments using both DNA analysis techniques, as in this study, should be performed before a large number of cases is evaluated.