罗丹明123介导的光动力学疗法预防急性移植物抗宿主病的实验研究

目的：探讨Rh123介导的光动力学疗法(PDT)预防异基因造血干细胞移植急性移植物抗宿主病(aGVHD)的可行性及安全性。 方法： 以C57B/6小鼠为供鼠，BALB/c小鼠为受鼠，建立小鼠异基因骨髓移植的aGVHD模型；混合脾脏淋巴细胞培养(MLC)加Rh123孵育，接受氩离子激光30 mW/cm2照射3 min，再与供者骨髓混合移植给受鼠，观察受鼠移植后造血重建、aGVHD发生情况及病理改变、生存率；流式细胞仪检测MLC细胞CD3+CD69+阳性率。 结果： 光动力学治疗组的aGVHD发生减少，肝、皮肤、肠道病理程度减轻，生存率显著高于未经光动力学治疗组；光动力学处理后的混合淋巴细胞培养24 h后，CD34+CD69+表达明显下降。 结论： Rh123介导的光动力学疗法可有效预防小鼠异基因骨髓移植的aGVHD。     

Nd:YAG温热疗法与Ar+激光PDT联合对小鼠S180肉瘤DNA含量的影响研究

目的：本文研究了Nd：YAG激光温热疗法与Ar~＋激光光动力学疗法联合对小鼠S_180．肉瘤DNA含量的影响。材料与方法：实验分为激光温热疗法组、Ar~＋激光光动力学组、联合作用组及对照组等四级。激光温热疗法用Nd：YAG激光照射肿瘤,功率密度约为424mW／cm~2,控制肿瘤边缘温度为43℃照射30min;Ar~＋先动力学疗法为激光照射前24小时小鼠腹腔注射HpD（10mg小kg）,以功率为150mW／cm~2的Ar~＋激光照射15min,联合作用为Ar~＋激光先动力学疗法后立刻行激光温热疗法。结果：在所选定的剂量下,联合作用组的肿瘤DNA含量最小。结论：联合作用对肿瘤的抑制途径之一是对肿瘤细胞中DNA光敏作用。     

Nd：YAG温热疗法与Ar^＋激光PDT联合对小鼠S180肉瘤DNA含 …

目的 本文研究了Ｎｄ：ＹＡＧ激光温热疗法与Ａｒ＾＋激光光动力学疗法联合对小鼠Ｓ１８０肉瘤ＤＮＡ含量的影响。材料与方法 实验分为激光温热疗法组,Ａｒ＾＋激光光动力学组,联合作用组及对照组等四组,激光温热疗法用Ｎｄ：ＹＡＧ激光照射肿瘤,功率密度约为４２４ｍＷ／ｃｍ＾２,控制肿瘤边缘温度为４３℃照射３０ｍｉｎ;Ａｒ＾＋光动力学疗法为激光照射前２４小时小鼠腹腔注射ＨｐＤ（１０ｍｇ／ｋｇ）以功率为１５０ｍＷ     

Nd：YAG温热疗法与Ar^＋激光光动力学疗法联合对小鼠S180肉 …

本文以小鼠Ｓ１８０肉瘤为对象，研究了Ｎｄ：ＹＡＧ激光温热疗法与Ａｒ＾＋激光光动力力学疗法联合对小鼠Ｓ１８０肉瘤的作用剂量选择及作用效果。实验分为激光温热疗法组、Ａｒ＾＋激光光动力学组、联合作用组一等四组。激光温热疗法用Ｎｄ：ＹＡＧ激光照射肿瘤，功率密度上实验约为４２４ｍＷ／ｃｍ＾２，控制肿瘤边缘温度为４３℃照射３０ｍｉｎ；Ａｒ＋光动力学疗法为激光照射前２４小时小鼠腹腔注射ＨＰＤ，以实验确定的１５０     

Nd:YAG温热疗法与Ar~ 激光光动力学疗法联合对小鼠S_(180)肉瘤作用的剂量选择研究

本文以小鼠Ｓ１８０肉瘤为对象，研究了Ｎｄ：ＹＡＧ激光温热疗法与Ａｒ＋激光光动力学疗法联合对小鼠Ｓ１８０肉瘤的作用剂量选择及作用效果。实验分为激光温热疗法组、Ａｒ＋激光光动力学组、联合作用组及对照组等四组。激光温热疗法用Ｎｄ：ＹＡＧ激光照射肿瘤，功率密度由实验确定约为４２４ｍＷ／ｃｍ２，控制肿瘤边缘温度为４３℃照射３０ｍｉｎ；Ａｒ＋光动力学疗法为激光照射前２４小时小鼠腹腔注射ＨｐＤ（１０ｍｇ／ｋｇ），以实验确定的１５０ｍＷ／ｃｍ２Ａｒ＋激光照射１５ｍｉｎ。结果表明，在所选定的剂量下，联合作用组肿瘤抑制率最高。说明联合作用方法值得进一步深入研究。     

亚甲基蓝介导的光动力疗法联合小檗碱对牙龈卟啉单胞菌抑制作用的体外研究

目的探讨亚甲基蓝介导的光动力疗法(PDT)联合小檗碱对牙龈卟啉单胞菌(P.g)的体外抑制作用。方法培养P.g至对数期中后期,将不同质量浓度亚甲基蓝加入P.g菌悬液中,作用5 min,激光(波长660 nm,功率140 mW/cm^(2))照射2 min,寻找亚甲基蓝结合激光体外抑制P.g的最佳浓度;观察亚甲基蓝介导的PDT体外抑制P.g的效果以及小檗碱对P.g生长曲线的影响;探讨亚甲基蓝介导的PDT与小檗碱先后联合应用对P.g的抑制作用;扫描电子显微镜观察亚甲基蓝介导的PDT及小檗碱对P.g形态的影响;紫外分光光度仪测量各成分吸收峰。结果在660 nm激光激发下,亚甲基蓝质量浓度为24.4141μg/ml时,抑菌效果最好。与对照组比较,亚甲基蓝组和PDT组差异均有统计学意义(均P<0.001)。0.05 mg/ml小檗碱对P.g浮游细菌具有抑制作用。与对照组比较,0.05 mg/ml小檗碱组菌落数降低,其差异有统计学意义(P<0.01);0.05 mg/ml小檗碱+光照组与对照组比较差异无统计学意义(P>0.05)。当PDT与小檗碱联用时对P.g有协同抑制作用,且先PDT后小檗碱组较先小檗碱后PDT组菌落数降低,其差异均有统计学意义(均P<0.01)。P.g经亚甲基蓝介导的PDT处理后,细菌细胞壁皱缩成团,经小檗碱处理后,细菌表面变得光滑且菌体长度较对照组增长。结论亚甲基蓝介导的PDT对P.g有抑制作用,当与小檗碱联用时,对P.g有协同抑制作用,且联合应用中先PDT后小檗碱作用时抑菌效果更好。     

亚氨基二乙酸修饰四苯基卟啉光敏剂介导的光动力疗法对H22肝癌小鼠体内抗肿瘤作用的研究

目的 评价本实验室自行合成的亚氨基二乙酸修饰四苯基卟啉光敏剂的理化性质及对H22小鼠肝癌的光动力治疗作用.方法 采用紫外分光光度法测定亚氨基二乙酸修饰四苯基卟啉光敏剂的紫外吸收光谱、脂水分配系数和单线态氧产率;以肝癌H22小鼠为研究对象,随机分为空白对照组、光照组、光敏剂组、光动力治疗1次组和光动力治疗2次组,考察亚氨基二乙酸修饰四苯基卟啉光敏剂介导的光动力疗法(PDT)对实体瘤的抑制率和荷瘤动物免疫机能的影响.结果 亚氨基二乙酸修饰四苯基卟啉光敏剂在650 nm波长处存在紫外吸收,脂水分配系数和单线态氧产率分别为2.30和0.52;单纯激光照射和单纯给予亚氨基二乙酸修饰四苯基卟啉光敏剂对肝癌H22小鼠无治疗作用(P＞0.05),而亚氨基二乙酸修饰四苯基卟啉光敏剂介导的光动力疗法对小鼠肝癌H22的生长具有极其显著的抑制作用(P＜0.001),光动力治疗2次组肿瘤抑制率高达95.15％.各治疗组对荷瘤小鼠的体质量增长、肝、脾、肺、胸腺指数几乎无影响.结论 亚氨基二乙酸修饰四苯基卟啉光敏剂单线态氧产率高、两亲性好,介导的PDT抗肿瘤活性强,适合作为光动力抗肿瘤候选新药进行开发.     

压力作用下牙周组织护骨素与Wnt-1的表达

背景：目前研究发现Wnt信号通路及相关因子对骨组织具有调节作用,Wnt-1与骨形成密切相关,但与正畸骨改建的相关性少有报道。 目的：通过建立大鼠的正畸牙移动模型,观察压力侧牙周组织Wnt-1及护骨素的表达。 方法：Wistar大鼠80只制备大鼠正畸牙移动动物模型,按牙齿移动1,3,7,14 d分别取材,每只大鼠对侧牙周组织设为空白对照组。苏木精-伊红染色观察大鼠压力侧牙周组织的形态变化,然后采用RT-PCR及免疫组织化学方法检测牙周组织Wnt-1及护骨素OPG的表达。 结果与结论：免疫组织化学结果与PCR检测结果显示,大鼠牙齿移动后1,3,7,14 d时,压力侧护骨素均有表达,在牙齿移动后3和7 d时护骨素表达明显增强(P < 0.05)。Wnt-1仅在在免疫组化中检测中显示,大鼠牙齿移动后3 d与对照组表达出现明显增强(P < 0.05)。结果证实,在正畸力作用下,护骨素参与了正畸牙周组织改建的过程,护骨素在压力区随正畸牙齿移动表达升高具有时间依赖性。Wnt-1在正畸骨改建中的作用需进一步研究证实。     

小鼠淋巴结内肥大细胞生长抑素免疫组织化学观察

目的：观察小鼠淋巴结内肥大细胞的分布特点及神经肽性质。方法：采用甲苯胺蓝染色和免疫组织化学ABC方法。结果：淋巴结内肥大细胞多为圆形和椭圆形，主要分布于淋巴窦内；经与甲苯胺蓝邻片比较，肥大细胞呈生长抑素（SS）免疫反应性，可见免疫反应阳性颗粒充满于胞质内，细胞核为阴性反应。结论：小鼠淋巴结肥大细胞呈SS免疫反应性，表明肥大细胞内神经肽SS可能通过神经－内分泌－免疫网络系统调节免疫器官的功能活动。     

大鼠膝关节软骨不同染色方法的差异

背景:国内外学者对关节软骨对软骨进行了大量研究,需要用不同的染色方法对研究结果进行分析,但将多种染色方法同时应用于软骨研究及探讨染色机制的较少。目的:探讨不同染色方法对大鼠膝关节软骨染色的优缺点。方法:取正常大鼠膝关节软骨,行苏木精-伊红、番红O、阿尔辛蓝、甲苯胺蓝、番红-阿尔辛蓝、番红-固绿染色,观察软骨结构。结果与结论:苏木精-伊红、番红O、甲苯胺蓝染色均可观察到潮线,分别为蓝色、红色和蓝色,番红O和甲苯胺蓝染色强度朝潮线方向增强,番红O染色对潮线的观察优于其他染色方法。苏木精-伊红染色关节软骨4层结构清晰,软骨细胞呈柱状排列,基质呈均匀呈嗜碱性染色。番红O染色显示4层结构层次清楚,基质深层染色最红。阿尔辛蓝染色显示,pH1.0时软骨细胞周边部分被阿尔辛蓝强烈染色,pH2.5时阿尔辛蓝染色较深。甲苯胺蓝染色显示组织结构层欠清,胞核染色清晰,胞浆几乎不着色,基质呈淡蓝紫色。番红-阿尔辛蓝染色显示软骨表面及基质呈不均一的红色,深层颜色较深,软骨细胞周围蓝染;番红-固绿染色显示软骨基质呈均匀的红色,软骨下骨呈绿色,软骨组织与骨组织分对比鲜明。提示上述各种方法均可观察到软骨的四层结构,但以番红O染色显示软骨各层和潮线结构最佳;苏木精-伊红染色观察软骨细胞形态变化较其他方法清楚。     

Promotion of Transplanted Bone Marrow-derived Cell Migration into the Periodontal Tissues due to Orthodontic Mechanical Stress

Background: Bone marrow-derived cells (BMCs) have abilities of cell migration and differentiation into tissues/organs in the body and related with the differentiation of teeth or periodontal tissue including fibroblasts. Then, we examined the effect of orthodontic mechanical stress to the transplanted BMC migration into periodontal tissues using BMC transplantation model.Material and Method: BMC from green fluorescence protein (GFP) transgenic mice were transplanted into 8-week-old female C57BL/6 immunocompromised recipient mice, which had undergone 10 Gy of lethal whole-body-irradiation. Five mice as experimental group were received orthodontic mechanical stress using separator between first molar (M1) and second molar (M2) 1 time per week for 5 weeks and 5 mice as control group were not received mechanical stress. The maxilla with M1 and M2 was removed and was immunohistochemically analyzed using a Dako Envision + Kit-K4006 and a primary anti-GFP-polyclonal rabbit antibody. Immunohistochemically stained was defined as positive area and the pixel number of positive area in the periodontal tissue was compared with the previously calculated total pixel number of the periodontal tissue.Results: The immunohistochemistry revealed that GFP positive cells were detected in the periodontal tissues, both in the experimental and control specimens. The ratio of pixel number in the examination group showed 5.77 ± 3.24 % (mean ± SD); and that in the control group, 0.71±0.45 % (mean ± SD). The examination group was significantly greater than that of control group (Mann-Whitney U test: p<0.001).Conclusion: These results suggest that orthodontic mechanical stress accelerates transplanted BMC migration into periodontal tissues.     

Prenatal asfotase alfa-mediated enzyme replacement therapy restores delayed calcification in a severe infantile form of hypophosphatasia model mice

Hypophosphatasia (HPP) is a congenital disorder caused by mutations in the tissue-nonspecific alkaline phosphatase (TNALP) gene. The pathogenesis of HPP varies, ranging from severe cases in which there is total absence of fetal bone calcification, which leads to stillbirth, to relatively mild cases in which the effects are confined to the teeth, such as early loss of the primary teeth. In recent years, the establishment of enzyme supplementation as a treatment method has prolonged survival in patients; however, this approach does not provide sufficient improvement for failed calcification. Furthermore, the effects of enzyme replacement therapy on the jawbone and periodontal tissues have not yet been studied in detail. Therefore, in this study, we investigated the therapeutic effects of enzyme replacement therapy on jawbone hypocalcification in mice. Recombinant TNALP was administered to mothers before birth and newborns immediately after birth, and the effect of treatment was evaluated at 20 days of age. The treated HPP mice had improved mandible (mandibular length and bone quality) and tooth quality (root length of mandibular first molar, formation of cementum), as well as improved periodontal tissue structure (structure of periodontal ligament). Furthermore, prenatal treatment had an additional therapeutic effect on the degree of mandible and enamel calcification. These results suggest that enzyme replacement therapy is effective for the treatment of HPP, specifically in the maxillofacial region (including the teeth and mandible), and that early initiation of treatment may have additional beneficial therapeutic effects.     

Low dose irradiation as a treatment for gingivitis

Improved dental hygiene has decreased the incidence of dental caries and focused attention on periodontal diseases. Gingivitis is the inflammation and/or ulceration of gingival tissue caused by anaerobic bacteria. Ionizing radiation produces a variety of oxygen species from the water in our tissues; each is highly toxic for anaerobic. The hypothesis is: low dose irradiation should be an effective treatment for gingivitis.     

光动力疗法防龋对釉质微量元素的影响

目的 将通体发光光纤应用于光动力疗法(PDT)防龋,探讨其对大鼠磨牙牙釉质中Ca、P含量的影响.方法 接种变形链球菌(S.mutans)制备大鼠致龋模型.将80只Wistar大白鼠随机分成5组:17mW(8 mW/cm2)PDT组(A组)、34 mW(15 mW/cm2)PDT组(B组)、68 rnW(30 mW/cm2)PDT组(C组)、20 g/LNaF溶液组(阳性对照组,D组)和生理盐水组(阴性对照组,E组),每组16只.采用650 nm半导体激光器,以质量浓度为40 μg/ml的血卟啉单甲醚为光敏剂,连续进行4周实验.应用电感耦合等离子体发射光谱法检测各组大鼠磨牙牙釉菌中的Ca、P含量.结果 实验后B、C、D组Ca、P含量明显高于A、E组,差异均具有统计学意义(均P＜0.05).A组Ca、P含量实验前后差异均无统计学意义(均P＞0.05);实验后B、C组Ca、P含量分别高于实验前,差异均具有统计学意义(均P＜0.05).A组Ca增量低于D组,差异具有统计学意义(P＜0.05);B、C两组Ca、P增量明显高于D组,差异均具有统计学意义(均P＜0.05);而B、C两组间Ca、P增量差异均无统计学意义(均P＞0.05).结论 在设定的参数范围内,PDT促进牙齿再矿化效果优于20 g/L NaF溶液.采用PDT防龋可增加大鼠磨牙牙釉质中的Ca、P含量,且Ca、P含量与PDT的功率有关.当PDT功率较低时,对釉质再矿化作用不明显;随着PDT功率的增加,Ca、P含量亦增加;当功率增加至一定数值时,两元素含量增量变化不明显,表明PDT可维持牙齿再矿化微环境.     

Bone Resorption Caused by Three Periodontal Pathogens In Vivo in Mice Is Mediated in Part by Prostaglandin

Gingival inflammation, bacterial infection, alveolar bone destruction, and subsequent tooth loss are characteristic features of periodontal disease, but the precise mechanisms of bone loss are poorly understood. Most animal models of the disease require injury to gingival tissues or teeth, and the effects of microorganisms are thus complicated by host responses to tissue destruction. To determine whether three putative periodontal pathogens, Porphyromonas gingivalis, Campylobacter rectus, and Fusobacterium nucleatum, could cause localized bone resorption in vivo in the absence of tissue injury, we injected live or heat-killed preparations of these microorganisms into the subcutaneous tissues overlying the calvaria of normal mice once daily for 6 days and then examined the bones histologically. We found that all three microorganisms (both live and heat killed) stimulated bone resorption and that the strain of F. nucleatum used appeared to be the strongest inducer of osteoclast activity. Treatment of the mice concomitantly with indomethacin reduced but did not completely inhibit bone resorption by these microorganisms, suggesting that their effects were mediated, in part, by arachidonic acid metabolites (e.g., prostaglandins). Our findings indicate that these potential pathogens can stimulate bone resorption locally when placed beside a bone surface in vivo in the absence of prior tissue injury and support a role for them in the pathogenesis of bone loss around teeth in periodontitis.     

金铂合金烤瓷冠对牙周组织的影响

背景：金属烤瓷冠的修复材料种类很多,每种材料的性能及对牙周组织细胞毒性影响均不相同。 目的：观察金铂合金烤瓷冠修复后对牙周组织的影响。 方法：选择2009-06/2010-01采用金铂合金烤瓷冠进行上颌切牙修复患者21例(26颗牙),以患牙对侧的同名健康牙为对照牙。烤瓷冠戴入6~8个月后复诊。 结果与结论：修复体边缘的深度及密合度均符合临床要求,修复后患牙的菌斑指数和龈沟液内血管内皮生长因子、肿瘤坏死因子α水平与对照牙比较,差异无显著性意义(P > 0.05),但患牙探诊深度、龈沟内出血指数及龈沟液量明显高于对照牙,差异有显著性意义(P < 0.05)。说明金铂合金烤瓷冠对患牙牙周组织有一定的不良影响。     

HMME-PDT对大鼠致龋模型中变形链球菌抑制作用的研究

目的 探讨不同参数光动力疗法(PDT)在大鼠口腔龋齿预防中的作用.方法 用变形链球菌感染Wistar大鼠口腔,建立龋齿模型.以0.9%生理盐水、0.2%氟化钠为对照组,单纯激光、单纯光敏剂、不同参数PDT为实验组.采用血卟啉单甲醚(HMME)为光敏剂,532 nm半导体激光为光源,每周处理牙齿1次,5周后处死大鼠,Ke...     

糖尿病牙周炎昆明系模型小鼠牙周组织中脂联素的表达

背景：脂联素与牙周炎和2型糖尿病发病有密切关系。 目的：检测糖尿病牙周炎昆明系小鼠牙周组织中脂联素表达的变化及对牙周组织病理形态及预后的影响。 方法：将3周龄的昆明系小鼠按不同造模方式分随机为3组：对照组正常饲养,糖尿病组通过四氧嘧啶注射法建立2型糖尿病模型小鼠,糖尿病+牙周炎组采用牙周局部结扎联合涂菌建立牙周炎模型小鼠。 结果与结论：造模后20 d,糖尿病组小鼠血糖、空腹胰岛素水平及胰岛素抵抗指数高于对照组(P < 0.05)。ELISA法、RT-PCR、western lot检测显示,造模1,3,6个月后,脂联素基因及蛋白的表达对照组>糖尿病组>糖尿病伴牙周炎组小鼠牙周组织中表达水平依次减弱(P < 0.05)。造模1,3,6个月后,取3组小鼠的牙周组织通过直接观察牙龈组织炎症变化及病理组织学检测显示,糖尿病伴牙周炎组小鼠牙周组织炎细胞浸润最明显、在糖尿病组部分小鼠可见中度牙龈组织炎症、对照组小鼠牙周组织未见炎细胞浸润。结果表明,糖尿病后牙周组织脂联素表达降低是牙周炎发病及加重的重要因素之一。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

Plasmin is essential in preventing periodontitis in mice

Periodontitis involves bacterial infection, inflammation of the periodontium, degradation of gum tissue, and alveolar bone resorption, which eventually leads to loss of teeth. To study the role of the broad-spectrum protease plasmin in periodontitis, we examined the oral health of plasminogen (Plg)-deficient mice. In wild-type mice, the periodontium was unaffected at all time points studied; in Plg-deficient mice, periodontitis progressed rapidly, within 20 weeks. Morphological study results of Plg-deficient mice revealed detachment of gingival tissue, resorption of the cementum layer, formation of necrotic tissue, and severe alveolar bone degradation. IHC staining showed massive infiltration of neutrophils in the periodontal tissues. Interestingly, doubly deficient mice, lacking both tissue- and urokinase-type plasminogen activators, developed periodontal disease similar to that in Plg-deficient mice; however, mice lacking only tissue- or urokinase-type plasminogen activator remained healthy. Supplementation by injection of Plg-deficient mice with human plasminogen for 10 days led to necrotic tissue absorption, inflammation subsidence, and full regeneration of gum tissues. Notably, there was also partial regrowth of degraded alveolar bone. Taken together, our results show that plasminogen is essential for the maintenance of a healthy periodontium and plays an important role in combating the spontaneous development of chronic periodontitis. Moreover, reversal to healthy status after supplementation of Plg-deficient mice with plasminogen suggests the possibility of using plasminogen for therapy of periodontal diseases.     

人牙周韧带细胞-聚羟基乙酸支架复合体修复牙周组织缺损

BACKGROUND: In recent years, tissue engineering technology as a new model for tissue regeneration has provided new ideas and methods for the repair of periodontal tissue defects. OBJECTIVE: To investigate the effect of human periodontal ligament cells-polyglycolic acid scaffold complex for repair of periodontal tissue defects. METHODS: Passage 4 human periodontal ligament cells at a density of 1.5×109/L were seeded onto the polyglycolic acid scaffold to prepare cell-scaffold complex. Then mongrel dogs were selected to make animal models of periodontal tissue defects and then randomly assigned into experimental group subjected to cell-scaffold complex implantation or control group subjected to direct coronal reset and suture of the gingival flap. Collagen content, new blood capillaries, new cementum, new alveolar bone and new periodontal ligament were detected within 4 weeks after operation; hematoxylin-eosin staining of periodontal tissue defects was done at 8 weeks after operation. RESULTS AND CONCLUSION: In the experimental group, the collagen content, number of newborn capillaries, amount of new cementum, new alveolar bone and new periodontal ligament tissues were significantly higher than those in the control group at postoperative 1, 2, 3, 4 weeks (P < 0.05). At 8 weeks after operation, in the experimental group, there were more vessels arranging on the connective tissue surface of new alveolar bone, the alveolar bone showed a sawtooth-like interlinking with the periodontal tissues in the presence of a thin layer of cementum; in the control group, only new alveolar bone and cementum formed below the incisure. These findings indicate that human periodontal ligament cells-polyglycolic acid scaffold complex can promote periodontal tissue regeneration.