唾液细胞外囊泡作为生物标志物诊断口腔疾病的研究进展

唾液与血液相似,含有大量生物学信息,是进行非侵入性诊断的理想体液.细胞外囊泡(EVs)是存在于大多数体液(包括唾液)内的一组细胞来源的纳米级异质囊泡,主要包括外泌体和微囊泡.细胞外囊泡是真核细胞和原核细胞共有的细胞间信号传递的媒介,可以将信号传递至受体细胞,从而介导细胞间通讯和信号传导;同时它在各种生物学功能中起着重要...     

细胞外囊泡免疫调控促组织再生的研究进展

细胞外囊泡(EV)作为生物体内的一类天然产生的用于细胞间通讯的膜性小泡,可以通过调控免疫细胞而诱导适宜的免疫微环境促进组织再生.本文就不同条件下EV对免疫功能的调控,以及EV作用下的免疫微环境在组织修复中的作用和机制进行综述,重点阐述间充质干细胞来源EV的免疫调控功能.     

干细胞细胞外囊泡在骨再生治疗中的应用

干细胞来源的外泌体及微囊泡等纳米级细胞外囊泡,通过携带生物活性物质在细胞间进行信息传递,发挥促进组织再生等作用,在再生治疗中有良好的前景。大量研究显示,干细胞细胞外囊泡能促进骨再生,其机制包括抑制缺血缺氧引起的骨细胞坏死,促进干细胞增殖、迁移和成骨分化,促进血管生成等;进一步研究证实,微RNA等成分可能在其中发挥调节作用。本文就干细胞外泌体及微囊泡等细胞外囊泡的分泌、组成、功能及其在骨再生治疗中的机制及应用研究进展进行综述。     

上皮癌来源细胞外囊泡亚群蛋白质谱分析

目的 ：揭示通过不同免疫标志物分离获得的细胞外囊泡(extracellular vesicles,EVs)亚群在蛋白质内容物方面的共性和异质性,为后续研究选择合适的分选方案提供数据支持。方法：以口腔癌CAL27细胞为主、胰腺癌细胞PANC-1以及胃癌细胞AGS为辅作为研究对象,分别通过4种EVs分子标志物CD9、CD63、CD81和EpCAM,对不同类型细胞来源的EVs进行免疫富集沉淀,通过蛋白质谱技术对不同EVs亚群进行内容物检测,通过funrich3.1.3软件分析评价EVs亚群之间的蛋白质共性和异质性。采用SPSS 19.0软件包对数据进行统计学分析。结果：在相同培养条件下,AGS细胞产生的EVs数量最多,其次是CAL27细胞,最少的是PANC-1细胞。在4种不同的EVs标志物分选方案中,基于EpCAM标志物的分选方案效能最强。主成分分析结果显示,本研究中涉及的EVs亚群在细胞来源层面以及分选标志物层面均存在异质性。蛋白质谱细胞成分分析结果表明,在口腔癌细胞系CAL27来源的EVs亚群中,CD63⁺EVs富集的蛋白中有86.11%位于外泌体生成相关途径,显著...     

细胞外囊泡的生物学功能及其临床应用

细胞外囊泡是由细胞分泌的不同大小的被膜囊泡,因其内容物包含有各种生理病理信号分子且可以稳定地在循环系统中停留,在临床上常被应用为疾病诊断和预后恢复的检测指标及有益的治疗方式。目前,EVs在多种疾病的诊疗、免疫微环境调控及组织再生中均得到了不同程度地应用。因此,通过进一步认知和研究EVs的生物学功能,有望为疾病的诊断和治疗提供新策略。     

角质形成细胞低表达Hsp90蛋白与小细胞外囊泡数量的相关性及其临床意义探讨

目的： 评价角质形成细胞中热休克蛋白90（heat shock protein 90,Hsp90）表达水平与小细胞外囊泡（small extracellular vesicles,sEVs）数量的关系。方法： 采用人角质形成细胞株（HaCaT）,分为野生型组、短发夹RNA干扰组（shRNA组,Hsp90蛋白抑制剂）和格尔德霉素抑制组（17-AAG组,Hsp90低表达）,进行体外培养。以超速离心法收集各组培养体系中的sEVs,利用透射电镜观察其形态学特征;蛋白质免疫印迹法鉴定生物学特性;纳米颗粒跟踪分析检测粒子数量。采用GraphPad Prism 8.0软件包中的t检验（非参数Mann-Whitney U检验）统计分析各组之间sEVs的数量差异。结果： 超速离心法获得的HaCaT细胞来源的sEVs符合形态学和生物学鉴定标准,sEVs中Hsp90蛋白无明显表达。shRNA干扰角质形成细胞中的Hsp90AA1表达后,其sEVs数量上升。第5天时,shRNA干扰组的sEVs粒子数为[（177.4 ±4.18）×10⁸, n=3],与空载质粒组的粒子数[（82.34±4.83）×10⁸, n=3]相比显著升高（P<0.000 1）。17-AAG抑制Hsp90蛋白5天后,17-AAG组的粒子数为[（652.5±26.73）×10⁸, n=3],对照组粒子数为[（262.22±5.44）×10⁸,n=3],差异具有统计学意义（P<0.000 1）。结论： 低表达Hsp90蛋白可促进HaCaT细胞分泌sEVs,sEVs可能与炎症状态下上皮细胞和免疫细胞间的物质传递有关。     

细胞外囊泡在组织工程研究中的新应用

细胞外囊泡(EVs)是细胞分泌释放的微小囊泡,其中包含核酸、蛋白质等生物活性分子,在细胞间通讯和信号传导中发挥着重要的作用。EVs可以通过递送生物活性分子,在多种疾病中起到治疗作用,有望成为新一代药物递送的天然载体。相较于传统纳米材料,天然EVs靶向递送具有低免疫原性、良好生物相容性、天然靶向性、长循环时间等明显优势。通过进一步组织工程手段来个性化构建EVs,提高其靶向性和目标治疗分子负载能力,为靶向分子治疗提供了一种新的治疗策略。     

唾液外泌体与口腔疾病相关研究进展

唾液外泌体是指存在于唾液中的直径在30～150 nm的细胞外囊泡。随着近年来技术手段的发展,大量研究揭示唾液外泌体在多种口腔疾病的发生发展中发挥重要作用,如唾液外泌体CD9及CD81通过调控细胞粘附及运动促进肿瘤细胞转移、唾液外泌体miR-24-3p通过作用于PER1促进肿瘤细胞增殖、唾液外泌体程序性细胞死亡配体-1(programmed cell death-ligand 1,PD-L1)mRNA抑制炎症组织的破坏等,具有作为诊断口腔癌、牙周炎等口腔疾病的生物标志物的潜能。因此,唾液外泌体可作为口腔疾病潜在的预后和诊断标志物。唾液外泌体除与口腔疾病,如口腔癌、牙周炎、口腔扁平苔藓、干燥综合征等有关外,还同远处部位肿瘤如胰腺癌、肺癌等及系统性疾病如帕金森综合征、炎症性肠病等密切相关;深入研究唾液外泌体对口腔、全身系统性疾病的诊断与治疗作用,开发唾液外泌体作为疾病诊断的生物标志物的潜力具有重要意义。     

细胞外囊泡在慢性牙周炎疾病进程及诊疗中作用的研究进展

慢性牙周炎是由多种致病因素引起的牙周组织慢性炎症。目前认为,细菌是慢性牙周炎的始动因子,宿主免疫影响牙周炎的发展和结局,其调控机制尚未完全清楚。细胞外囊泡是一类细胞旁分泌产生的亚细胞成分,可携带多种遗传信息介导细胞通信。细胞外囊泡已在肿瘤、心脑血管及免疫性疾病的诊疗中取得重要研究成果,为一些难治性疾病的早期诊断、对因治...     

miRNA-21在口腔鳞癌患者唾液中的表达及意义

目的:探讨miRNA-21在口腔鳞癌患者唾液上清中的表达及其临床意义。方法:以2012-01—2015-06期间,新余市人民医院口腔科收治的64例口腔鳞癌患者及25例健康对照人群为研究对象,收集唾液标本,采用实时定量聚合酶链反应技术,检测唾液标本中miRNA-21表达情况,并分析其与口腔鳞癌临床病理学特征之间的联系。结果:口腔鳞癌患者唾液上清液中miRNA-21的表达水平(2.75±0.94)显著高于健康对照组(0.78±0.45,P<0.01)。相关性分析结果显示,唾液上清液中miRNA-21表达水平与口腔鳞癌细胞分化程度(P<0.01)及淋巴结转移状态(P<0.05)显著相关。受试者工作特征曲线分析结果显示,唾液上清液中miRNA-21表达水平,评价口腔鳞癌细胞分化程度的敏感性和特异性为89.5%和66.7%。结论:口腔鳞癌患者唾液上清液中miRNA-21的表达上调,可能与口腔鳞癌的发生发展有关,唾液中miRNA-21可作为评价口腔鳞癌细胞分化的辅助指标。     

人工唾液中直丝弓托槽与弓丝静摩擦力的实验研究

目的探讨人工唾液条件下,不同时间,几种直丝托槽与弓丝在后牙段的静摩擦力特点。方法在人工唾液条件下,4个时间段,测试4种直丝托槽与4种弓丝组合在后牙段的静摩擦力。结果0.46mm不锈钢圆丝的静摩擦力最小,0.48mm×0.64mm不锈钢方丝的静摩擦力最大。弓丝、托槽组合在人工唾液条件下作用15d静摩擦力最大、30d静摩擦力最小。结论0.46mm不锈钢圆丝的静摩擦力最小;人工唾液作用30d静摩擦力最小。     

A preliminary study on the relationship between stimulated saliva and periodontal conditions in community-dwelling elderly people

OBJECTIVES: The purpose of this study was to explore the relationship between flow rate and spinnbarkeit of stimulated whole saliva and periodontal conditions in healthy elderly people. METHODS: Three hundred and fifty-five dentate subjects aged 76 years were included. The pocket probing depth (PD), attachment level (AL), and bleeding on probing (BOP) were measured. Stimulated whole saliva was collected and the salivary flow rate (SFR) was calculated. Then, salivary spinnbarkeit (SS) was immediately measured. RESULTS: The mean SFR and SS were 1.44 ml/min and 1.91 mm, respectively. SFR was not significantly related to each periodontal parameter. On the other hand, subjects with SS2.00 mm exhibited a significantly higher mean PD (p<0.05), % of sites with PD>or=4mm (p<0.05), mean AL (p<0.01), % of site with AL>or=4mm (p<0.05) and % of sites with AL>or=6mm (p<0.01) than subjects in the other three groups (one-way ANOVA). In the logistic regression analysis, the factors significantly associated with the highest quintile of PD>or=4mm were: the low salivary flow and the high salivary spinnbarkeit (OR 3.84), current smokers (OR 5.08), cleaning interdental spaces rarely/never (OR 2.12), and frequent BOP (OR 5.20). CONCLUSION: These findings suggest that high salivary spinnbarkeit in addition to a low salivary flow rate might be a high risk for periodontal disease in elderly people.     

An in vitro study of galvanic currents between amalgam and gold alloy electrodes in saliva and in saline solutions

Abstract – The galvanic currents between amalgam and gold alloy electrodes were higher in saline solutions than in saliva. Conventional amalgam supported higher currents than high copper amalgams when short-circuited with a gold alloy. The galvanic currents were not affected or affected only to a very small degree when whole saliva from different persons were compared. Higher currents were recorded when short-circuiting conventional amalgam and gold alloy electrodes using parotid saliva than when using whole saliva collected from one person. However, removing molecules with a molecular weight higher than 10⁴ from whole and parotid saliva did not seem to affect the galvanic currents.     

A study of the relationship between trace elements in saliva and dental caries in children

About 8 ml of saliva were collected from each of 272 children, living in rural areas in North India, equally distributed into age groups 4–7 and 12–16 yr and stratified into low, moderate and high caries groups. The salivas were analysed for Zn, Cu, Fe, Mn by atomic absorption spectrophotometry and for F with a fluoride-sensitive electrode. The elements Cu and F had a consistent, inverse relationship with caries experience. The concentration of Zn, Fe and Mn in saliva did not have any consistent relationship with caries experience.     

吸烟、饮酒与老年女性颌骨骨质疏松的相关性研究

目的：探讨吸烟、饮酒与老年女性颌骨骨质吸收间是否存在相关性。方法：在367名绝经期老年女性中进行吸烟、饮酒习惯的调查,并对受试者进行全景片检查。在全景片上测量颏孔区的下颌骨骨皮质厚度。对所得资料及数据进行多元回归分析。结果：多元回归分析显示吸烟、饮酒与下颌骨骨皮质密度相关。结论：吸烟、饮酒均是老年女性骨质疏松的危险因素,少量饮酒亦不例外。     

The use of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to detect putative breast cancer markers in saliva: a feasibility study

BACKGROUND: Technologies are now available enabling saliva to be used to diagnose disease, predict disease progression, and monitor therapeutic efficacy. This pilot study describes the use of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI) to detect putative breast cancer markers in saliva. METHODS: Salivary specimens were analyzed as either pooled cancer saliva specimens, or individual specimens from healthy women and women diagnosed with carcinoma of the breast. The specimens were applied to a variety of protein chip arrays, washed extensively to remove unbound analytes and analyzed on a SELDI mass spectrometer. RESULTS: The results of this initial study suggest that the WCX protein chip array prepared and washed at pH 3.5 yielded the most promising results. Additionally, the analyses revealed a number of proteins that were higher in intensity among the cancer subjects when compared with controls. These salivary proteins were present at the 18, 113, 170, 228 and 287 km/z ranges using SELDI analyses. CONCLUSIONS: The study suggests that saliva may be useful for high-throughput biomarker discovery.     

Longitudinal study of the influence of removable partial denture and chemical control on the levels of Streptococcus mutans in saliva

Some studies have evaluated the salivary levels of mutans streptococci (MS) in removable partial denture (RPD) users. Saliva samples (2.0 mL) were obtained from 31 patients in six periods: (T0): immediately before installation of RPD; (T8): 8 days after T0; (T48): 48 days after T0; (T92): 92 days after T0; (T140): 140 days after T0 and (T189): 189 days after T0. The samples were vortexed and serially diluted from 10(-1) to 10(-6) in 0.05 m phosphate buffer (pH 7.4). From each dilution, 0.025 mL was plated on Mitis Salivarius Bacitracin (MSB). The plates were incubated in 5% CO2 at 37 degrees C for 72 h. There was an increase (t-test, P < 0.05) in the number of MS between periods T0 and T48 (mean/s.d., CFU mL(-1) of saliva): T0: 2.26/4.43 x 10(6) and T48: 0.47/1.48 x 10(8). After this, intensive treatment with CHX was accomplished in 29 patients. Saliva samples were obtained after treatment in four periods: (T24 h): 24 h after T0; (T14): 14 days after T24 h; (T28): 28 days after T24 h, and (T63): 63 days after T24 h. The number of MS in saliva did not decrease (t-test, P > 0.05). A new CHX formulation was applied in 15 patients. Saliva samples were obtained in periods: (T0): before new CHX application; (T24 h): 24 h after T0 and (T82): 82 days after T0. The new CHX reduced MS levels in saliva: (mean/s.d., CFU mL(-1) of saliva): T0: 6.64/8.47 x 10(6) and T24 h: 3.2/4.27 x 10(5) (sign rank, P < 0.05). In conclusion, there was a significant increase in the number of MS in saliva after the installation of RPD. The intensive treatment with a properly formulated CHX was effective in the reduction of MS, between 24 h and 82 days after its application.     

牙周病与早产低出生体重儿关系的临床研究

目的比较分娩早产低出生体重儿(PLBW)的孕妇与足月妊娠正常分娩新生儿(NBW)孕妇的牙周状况及牙周病的发生率,探讨牙周病与PLBW的关系。方法随机选取分娩PLBW的孕妇206例为研究组,选取同期分娩NBW的孕妇209例为对照组,检测两组孕妇的牙周状况,记录菌斑指数(PLI)、探诊深度(PD),临床附着丧失(CAL)和出血指数(BI),并计算牙周炎位点率(PD>3 mm,CAL>2 mm)及牙周病的发生率。结果分娩早产低出生体重儿的孕妇牙周炎位点率(3.3%)和牙周病发生率(81.1%)明显高于对照组(1.4%,35.9%),差异有显著性(P<0.01);牙周炎位点率与孕周和新生儿体重均呈负相关系(P<0.05)。结论牙周病可能是PLBW发生的危险因素。     

牙周病与糖尿病足临床相关性研究

目的 调查糖尿病足的风险是否与牙周病（PD）相关。方法 选择110例糖尿病足伴牙周病患者,及项目协作组医院口腔科就诊的条件相近的102名非糖尿病的牙周病患者为对照组。采用logistic回归模型,对潜在的混杂因素进行了调整,对本研究中所见的PD危险因素如：高龄、糖尿病病程、口腔保健和患者受教育水平等,采用SPSS 12.0作统计分析。结果 年龄、糖尿病病程、口腔保健和教育水平具统计学意义。在调整性别、年龄、糖尿病病程、口腔保健和教育后,PD和牙列缺损与糖尿病足有独立相关。结论 糖尿病足的风险与牙周病高度相关。