口腔黏膜异种移植模型巨噬细胞浸润及他克莫司干预的实验研究

目的:观察大鼠口腔黏膜异种移植模型急性排斥反应过程中巨噬细胞浸润记数变化及他克莫司的干预作用。方法:Wistar大鼠接受BALB/c小鼠舌体移植建立口腔黏膜异种移植模型,免疫组化法观察颊黏膜急性排斥反应过程中3个时点巨噬细胞记数变化情况。结果:大鼠颊黏膜术后即出现急性排斥反应并随时间推移排斥反应减弱,移植区巨噬细胞浸润程度随病程呈递减趋势。术后第7天移植组巨噬细胞记数为580.00个/HPF,移植用药组325.80个/HPF,术后第14天移植用药组246.45个/HPF,移植组220.00个/HPF,术后第28天移植组173.00个/HPF,移植用药组73.00个/HPF。不同处理组术后3个时点记数差别均有统计学意义(P<0.05),移植组术后3个时点记数差别有统计学意义(P<0.05),移植组与移植用药组3个时点两两比较差别无统计学意义(P>0.05)。结论:成功建立大鼠口腔黏膜异种移植模型,巨噬细胞浸润程度与急性排斥反应病程有关,他克莫司对于巨噬细胞浸润无明显抑制作用。     

缺氧诱导因子-1α和环氧合酶-2在大鼠放射性口腔黏膜炎中的表达及意义

目的检测缺氧诱导因子-1α（HIF-1α）和环氧合酶-2（COX-2）mRNA及蛋白在大鼠放射性口腔黏膜炎组织中的表达水平．并探讨二者之间的关系。方法建立放射性口腔黏膜炎SD大鼠模型，切取左、右侧颊黏膜，用半定量RT—PCR检测各标本HIF—1α和COX-2的mRNA相对表达值．采用免疫组化SABC法检测HIF—1α和COX-2蛋白在黏膜炎组织中的表达部位及表达强度。结果成功地建立了SD大鼠放射性口腔黏膜炎动物模型：RT—PCR结果显示ROM黏膜组织能够表达HIF—1α mRNA和COX-2 mRNA．而右侧自身对照组基本不表达HIF—1α和COX-2 mRNA；免疫组化结果显示：HIF—1α蛋白和COX-2蛋白在ROM黏膜组织中均能广泛表达，HIF—1α和COX-2蛋白的表达基本处于平行的关系．并且表达的部位和细胞类型也基本一致。结论放射性口腔黏膜炎组织有HIF—1α和COX-2 mRNA及蛋白的表达．两者的表达呈高度相关．HIF—1α和COX-2的表达与放射性口腔黏膜炎的严重程度有关。     

甲状旁腺激素对去势大鼠骨髓间充质干细胞增殖及凋亡的影响

目的：研究甲状旁腺激素（ PTH）对去势大鼠骨髓间充质干细胞增殖以及凋亡特性的影响。方法：建立去势大鼠骨质疏松模型,皮下注射甲状旁腺激素,通过流式细胞仪技术检测去势大鼠骨髓间充质干细胞细胞周期以及凋亡率的改变。结果：加PTH刺激大鼠骨髓间充质干细胞组的S期细胞比例为（8．87±0．11）％,显著高于去势大鼠骨髓间充质干细胞组的S期细胞比例（5．23±0．20）％,差异具有统计学意义（P＜0．05）;加PTH刺激大鼠骨髓间充质干细胞组凋亡率为（4．17±0．15）％,显著低于去势大鼠骨髓间充质干细胞组凋亡率（8．13±0．61）％,差异具有统计学意义（P＜0．05）。结论：甲状旁腺激素可明显提高去势大鼠骨髓间充质干细胞的增殖能力,并抑制其凋亡。     

脂肪干细胞移植治疗化疗导致的口腔黏膜炎的初步观察

目的 探讨脂肪干细胞(ADSCs)移植治疗化疗导致的口腔黏膜炎的效果。方法 将10只金黄地鼠随机分为2组,制备化疗性金黄地鼠口腔黏膜炎模型,实验组(M组)于金黄地鼠口腔颊囊溃疡周边局部多点注射被标记的ADSCs+PBS细胞混悬液;对照组(N组)于相同部位注射等体积PBS。每天观察记录金黄地鼠口腔黏膜情况,比较实验组和对照组颊囊粘膜愈合情况。分别于实验第9天、第14天处死实验动物,制备组织标本,行大体观察、HE染色、冰冻切片、荧光显微镜下观察。利用SAS软件对数据进行分析。结果 大体观察及组织学染色显示M组黏膜愈合程度及质量明显优于N组;荧光显微镜观察显示移植一周后CM-Dil染色的ADSCs能在该部位定居且存活良好。结论 脂肪干细胞移植对口腔黏膜溃疡具有一定的促愈作用,可以快速治疗化疗导致的口腔黏膜炎。     

造血干细胞移植患者口腔黏膜炎预防及局部干预的研究进展

口腔黏膜炎(oral mucositis, OM)是造血干细胞移植过程中常见的并发症。OM发生时随着口腔黏膜屏障功能的破坏, 加速口腔细菌环境的形成, 进而对患者生活质量和临床结局产生负面影响。保持口腔清洁、正确使用漱口液、限制细胞毒性药物向口腔组织输送能预防OM的发生和发展。本文对造血干细胞移植患者OM预防和局部干预的研究进行综述, 以期为临床干预及OM管理策略的选择提供思路, 为探索有效的OM管理方法提供借鉴。     

造血干细胞移植后发生口腔黏膜炎71例临床分析

目的 探讨造血干细胞移植(HSCT)后口腔黏膜炎(0M)的发生情况、影响因素及处理措施.方法 对北京大学人民医院2009年行HSCT的166例患者中发生OM的71例患者的资料进行临床分析.结果 OM的愈合率为95.77%,平均愈合时间为移植后(17.61±8.36)d.患者性别、年龄、HLA配型及预处理方案对OM愈合时...     

中药含服液防治放射性口腔炎的疗效观察

目的评价中医药防治放射性口腔炎的疗效。方法将134例鼻咽癌放疗后出现口腔黏膜炎的患者随机分成两组，以金银花、玄参、水牛角、麦冬等中药制剂含服治疗患者为实验组，复方硼砂溶液含漱治疗的患者作为对照组，按WHO放射性口腔黏膜反应标准记录患者的口腔黏膜炎程度。结果中药含服液治疗67例，出现Ⅰ、Ⅱ、Ⅲ、Ⅳ级口腔黏膜炎分别为37、21、6、3例：对照组67例，出现Ⅰ、Ⅱ、Ⅲ、Ⅳ级口腔黏膜炎分别为4、26、28、9例；两组比较差异具有显著性（P〈0．01）。结论中药含服液防治放射性口腔炎具有独特的疗效。     

骨髓间充质干细胞移植对舍格伦综合征大鼠模型的治疗作用

目的:探讨骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)移植对舍格伦综合征(Sjǒgren’s syndrome)大鼠模型的治疗作用及其在下颌下腺(submandibular gland,SMG)内存活、迁徙及分化情况。方法:建立大鼠舍格伦综合征动物模型;分离、培养大鼠BMSCs并对其进行鉴定和标记;下颌下腺局部注射移植经绿色荧光蛋白标记的BMSCs;测定大鼠静态唾液总流率,记录正常组、模型组、模型治疗组及模型治疗对照组大鼠每日饮水量;荧光显微镜下观察移植干细胞在模型治疗组和模型治疗对照组下颌下腺内的存活、迁徙及分化情况。所得实验数据采用SPSS12.0软件包进行t检验。结果:模型治疗组静态唾液总流率和每天饮水量与模型治疗对照组相比,差异有统计学意义(P<0.05)。经标记的骨髓间充质干细胞移植后第1周,BMSCs主要分布于针道附近,第2周主要分布于腺泡之间的间质中,第4周开始在下颌下腺其他区域出现。术后第1周,免疫组化染色移植干细胞未见淀粉酶表达,第8周可见在少量移植细胞的胞质内有淀粉酶表达,具备了类似腺泡细胞的分泌功能。模型治疗对照组未发现以上现象。结论:BMSCs移植对舍格伦综合征大鼠有一定治疗作用。     

狗牙周缺损处移植自体骨髓干细胞的骨化实验观察

目的 对应用自体骨髓干细胞移植的动物骨化实验的观察进行评价。方法 将6只成年狗的36颗牙，分为实验组和对照组（各18颗牙）。有人工制造的牙周缺损中进行体外培养的自体骨髓干细胞移植，结合引导组织再生（guided tissue regeneration，GTR）方法（实验组）和单纯GTR方法（对照组）。6周后对切片行牙周组织学观察。结果 实验组新牙槽骨骨化效果明显好于对照组。实验组建立了正常钙化骨结构，Masson染色为红色；对照组虽然建立了正常骨结构，但钙化程度极低，基本为胶原性骨结构，Masson染色为蓝绿色。结论 应用自体骨髓干细胞移植结合聚四氟乙烯膜引导牙周组织再生可促进牙周组织的再生、加快正常骨组织结构的建立，并缩短修复再生时间。     

造血干细胞移植引起的慢性移植物抗宿主病口腔黏膜及牙周状况的调查分析

目的: 观察造血干细胞移植（hematopoietic stem cell transplantation,HSCT）后有慢性移植物抗宿主病患者的早、晚期口腔黏膜及牙周改变的临床表现,探讨慢性移植物抗宿主病对口腔状况的影响。方法: 收集108例造血干细胞移植后患者,按移植后治疗时间不同分为移植时间小于1年(n=32)和超过1年(n=43) 2组。采用问卷调查、临床检查,收集临床资料,确定罹患口腔慢性移植物抗宿主病患者的基本信息,记录口腔黏膜及牙周病损及严重程度,采用SPSS21.0软件包进行统计学分析。结果: HSCT后出现口腔表征发生率为78.95%,HSCT小于1年组与HSCT超过1年组的轻度症状发生率分别为59.38%和18.60%,差异显著（ χ²=11.525,P=0.001）;HSCT超过1年的苔藓样反应与HSCT小于1年的发生率分别为60.47%和31.25%,差异有统计学意义（χ²=6.274,P=0.012）;浅、中度牙周袋患者检出率分别为49.38%和40.74%;可视牙菌斑、牙结石、牙龈出血检出率分别为62.96%、64.20%和50.62%。结论: HSCT后黏膜及牙周改变的发生率较高,早期口腔黏膜慢性移植物抗宿主病轻度症状明显,晚期口腔表现则以苔藓样反应为主。     

Oral mucositis after bone marrow transplantation. A marker of treatment toxicity and predictor of hepatic veno-occlusive disease

Oral mucositis and hepatic veno-occlusive disease (HVOD) are common complications after bone marrow transplantation (BMT). Forty-seven patients were prospectively examined for development of ulcerative oral mucositis (UOM) and HVOD after allogeneic BMT. In 17 patients (36%) UOM developed between 2 days before and 18 days after transplant (median 4 days after BMT). In seven patients (15%) HVOD developed with onset between 3 and 21 days after transplant (median 18 days after BMT). In a time-dependent analysis, in patients given busulfan plus cyclophosphamide or busulfan, etoposide, and cyclophosphamide UOM was 19 times more likely to develop than in patients treated by cyclophosphamide plus total body irradiation or by cyclophosphamide alone (p less than 0.001). Patients in whom UOM developed were 6.5 times more likely to develop HVOD than those in whom UOM did not develop (p less than 0.03). The sensitivity (86%), specificity (73%), and negative predictive value (97%) of UOM to predict HVOD were high, but the positive predictive value (35%) was low. The association of UOM and HVOD support the concept that both are toxic effects of treatment. Patients with hepatic abnormalities but without UOM are unlikely to have HVOD, and other causes of the hepatic dysfunction should be investigated.     

GM-CSF mouthrinses in the treatment of severe oral mucositis: a pilot study.

OBJECTIVE: The aim of this open trial was to test the efficacy of granulocyte-macrophage colony-stimulating factor (GM-CSF) mouthrinses as a potential treatment in reducing the duration of severe oral mucositis in patients undergoing bone marrow transplantation for hematologic malignancies. STUDY DESIGN: The study group was composed of 10 consecutive patients suffering from severe oral mucositis during bone marrow transplantation procedures. The control group was similar to the study group in age and gender and comprised 29 historical patients with similar clinical characteristics. Freshly prepared GM-CSF mouthwash (0.5 microg/mL) was administered to the study population for 1 minute 3 times per day after oral hygiene procedures, starting from the first day of mucositis until clinical improvement of oral lesions. The study and control populations were compared with respect to duration of severe oral mucositis (1-9 days, 10-19 days, > or =20 days). RESULTS: There was no statistically (chi2 exact test) significant difference in mean mucositis score between the study group (11.9+/-6.1) and the control group (16.6+/-8.9). However, the duration of severe mucositis appeared to be reduced; 60% of the GM-CSF mouthrinse patients had severe mucositis for less than 9 days, whereas only 28% of the controls had severe mucositis for less than 9 days. In addition, 10% of the GM-CSF mouthrinse patients experienced severe mucositis lasting 20 or more days, whereas 34% of the controls experienced severe mucositis for 20 or more days. CONCLUSIONS: These findings suggest that GM-CSF may reduce the duration of severe mucositis, but controlled, double-blind clinical trials are now required.     

The use of tretinoin as oral mucositis prophylaxis in bone marrow transplantation patients: a preliminary study

OBJECTIVE: The examination of prophylactic efficacy of tretinoin on oral mucositis, post bone marrow transplantation (BMT).
PATIENTS AND METHODS: The study population consisted of 11 patients undergoing BMT.Six tretinoin topically-treated patients (0.25 mg daily of 0.1 % tretinoin cream) were matched with five non-treated control patients comparing mucositis severity, duration and analgetic (morphine) requirementS. Concomitant follow-up included conditioning parameters associated with mucositis and engraftment.
RESULTS: The mean of oral mucositis peak scores was significantly lower in the tretinoin-treated patients vs the non-treated patients (score 1.5 vs 3.6; P < 0.02). In the majority of cases the duration of the most severe phase of oral mucositis was shorter in the tretinoin-treated group as compared with the control. Only one patient in the experimental group required morphine analgesics compared with four patients in the control group.
CONCLUSIONS: This preliminary study indicates that the severity of oral mucositis, both objective and subjective, in BMT patients may be reduced by 0.1% topical tretinoin cream, 0.25 mg, administered daily from the beginning of the BMT conditioning regimen until marrow engraftment.     

Oral recurrent human herpes virus infection and bone marrow transplantation survival

OBJECTIVE: This study was conducted to compare the survival rates of bone marrow transplantation (BMT) patients who were affected with the survival rates of those who were not affected by oral recrudescent human herpes virus-1 infection (HHV-1) after transplantation. STUDY DESIGN: Fifty-two consecutive patients who underwent BMT were included in the study. The time of death after BMT was displayed, by means of the Kaplan-Meier method, for the following parameters: age and gender of the patient, donor gender, primary disease, stem cells, conditioning regimen, platelet number after day 100, acute and chronic graft-versus-host disease, oral recurrent HHV-1 infection post-BMT, oral lichenoid lesions of graft-versus-host disease, graft-versus-host disease at the salivary glands, parenteral nutrition, and oral mucositis. The data were initially analyzed by means of the log-rank test and then included in the Cox proportional hazards model. RESULTS: The multivariate analysis demonstrated a significance of 5% for only the platelet numbers and oral recurrent HHV-1 infection. CONCLUSION: The present study provides evidence that platelet numbers below 100,000 cells/mm(3) after day 100 and oral recurrent HHV-1 infection are independent negative prognostic variables in BMT patients' 24-month survival rates.     

Systemic absorption of lidocaine after topical application for the treatment of oral mucositis in bone marrow transplantation patients

This paper evaluates lidocaine absorption via oral mucosa following its topical application for symptomatic treatment of bone marrow transplantation (BMT)-induced oral mucositis. Five patients with high-grade oral mucositis after allogeneic BMT were entered consecutively into the study. Five healthy individuals served as controls. All 10 participants rinsed their mouth with 5 ml of a 2% lidocaine solution for 1 min, after which they expectorated the liquid. Blood samples were drawn at 1, 5, 10, 20, 30 and 60 min after rinsing and centrifuged. Plasma lidocaine levels were measured by fluorescence polarization immunoassay. In the BMT patients, plasma lidocaine levels were lower than the therapeutic range of this drug (0.2 microg/ml vs 1.5-5.5 microg/ml), while in the controls no detectable lidocaine levels were noted. The data from this preliminary study indicate that lidocaine prescribed as an anesthetic mouthwash in BMT patients with oral mucositis results in minor systemic absorption of the drug.     

Relationship of oral complications to peripheral blood leukocyte and platelet counts in patients receiving cancer chemotherapy.

Patients undergoing cancer chemotherapy often suffer from oral complications as a result of their disease and its treatment. The effects of the chemotherapy on the bone marrow and oral mucosa, coupled with the patient's immunosuppressed state and altered oral microbial flora, predispose these patients to oral mucositis, infection, and hemorrhage. The oral mucosa appears to mirror the effects of the chemotherapy on the bone marrow, as there appears to be a direct relationship between the changing peripheral blood counts and the status of the oral mucosa.     

Vascular endothelial growth factor (VEGF) and chronic graft-versus-host disease (cGVHD) in salivary glands of bone marrow transplant (BMT) recipients

BACKGROUND: The purpose of this study was to compare the immunolocalization of vascular endothelial growth factor (VEGF) in salivary glands of bone marrow transplant (BMT) recipients with normal controls and between the different stages of chronic graft-versus-host disease (cGVHD). In addition, the impact of the immunolocalization of VEGF on the survival rate of BMT patients was investigated as well. METHODS: Labial salivary glands obtained at the day 100+ from 36 consecutive patients, who underwent BMT, were included in the study. The streptavidin-biotin-peroxidase complex stain was used to detect VEGF in the salivary glands. Time of death after BMT was displayed by means of the Kaplan-Meier method for the following parameters: age and gender of the patients, donor gender, acute GVHD, cGVHD staging at the labial salivary glands, primary disease, platelet and neutrophils counts on day of biopsy, stem cell, oral mucositis, parenteral nutrition, oral lichenoid lesions of GVHD, conditioning regimen and immunolocalization degree of VEGF in labial salivary glands. The data were initially analyzed by means of the log-rank test and then included in the Cox's proportional hazard model. RESULTS: No differences on the immunolocalization of VEGF in the labial salivary glands of BMT recipients and control group or between the different stages of glandular cGVHD were noted. Both univariate and multivariate analysis of the survival rate showed significance of 5% only for platelet count over 100 x 109/l on the day of biopsy and male donor gender. CONCLUSIONS: Platelet count over 100 x 109/l and male donor gender are positive predictive factors on the survival rate after BMT. In addition, the immunolocalization of VEGF in salivary glands is not altered in BMT recipients at day 100+ and is not influenced by the stage of cGVHD.     

Acyclovir prophylaxis for oral herpes simplex virus infection in patients with bone marrow transplants

Reactivation of herpes simplex virus (HSV) appears to play a significant role in oral mucositis resulting from bone marrow transplantation. The acyclic guanosine derivative acyclovir has been shown to be effective in treating and protecting against HSV infection in this group. The purpose of this study was to determine the role of HSV reactivation in oral mucositis in patients undergoing bone marrow transplantation who were seronegative for HSV or who received acyclovir prophylaxis. The results suggest that HSV plays an insignificant role in oral mucositis in these patients.     

Oral condition of patients with leukemia and severe aplastic anemia. Follow-up 1 year after bone marrow transplantation

Twenty patients with leukemia and seven patients with severe aplastic anemia who had been treated with bone marrow transplantation were investigated 1 year after transplantation to assess their oral condition. Ten patients had mild clinical chronic graft-versus-host disease (GVHD), and three patients had moderate or severe GVHD. Histopathologic changes in the oral mucosa or the minor salivary glands were observed in nineteen patients. Clinical changes in the oral mucosa were observed in sixteen of those patients. Colonization with Candida albicans was more frequent in patients with advanced histopathologic changes in oral mucosa. The numbers of Streptococcus mutans and lactobacilli and the whole salivary flow rate were not correlated to high or low caries incidence. Whole salivary flow rate were not correlated to GVHD but rather, to conditioning with total body irradiation (p less than 0.05).     

Effect of local application of the antimicrobial peptide IB-367 on the incidence and severity of oral mucositis in hamsters.

OBJECTIVE: The purpose of this animal study was to determine whether IB-367, an antimicrobial peptide, is able to ameliorate oral mucositis by reducing microflora densities on the mucosal surfaces of the mouth. STUDY DESIGN: Oral mucositis was induced in hamsters by intraperitoneal injection of 5-fluorouracil followed by superficial abrasion of the buccal mucosa. A test formulation was applied topically to the buccal mucosa 5 or 6 times per day starting 6 to 8 hours before abrasion. RESULTS: Mucositis scores were significantly lower (P < .05) in hamsters given formulations containing 0.5 or 2.0 mg/mL of IB-367 than in placebo-treated controls. Treatment with IB-367 produced a more than 100-fold reduction in oral microflora densities. In a second experiment, treatment of hamsters with a formulation containing IB-367 at 0.12, 0.5 or 2.0 mg/mL resulted in a dose-dependent reduction in mucositis severity. CONCLUSION: The results indicate that reduction of local microflora densities through use of IB-367 may improve clinical outcomes in patients at risk for the development of oral mucositis.