【作者】 郭锐；

【导师】 黄翠； 杨宏业； 王亚珂； 喻健；

【作者基本信息】 武汉大学 ， 口腔医学八年制， 2022， 博士

【摘要】 目的:探索并制备合适浓度的白藜芦醇改性粘接剂,依次评估其对粘接剂转化率、生物相容性、抗菌等性能的影响和对牙本质粘接耐久性的影响,并分析其作用机制与未来临床应用前景。方法:第一部分实验中,将白藜芦醇粉末溶于通用型粘接剂,制备含有0.1、1、10 mg/mL白藜芦醇的改性粘接剂。将不含白藜芦醇的对照组粘接剂和分别含有0.1 mg/mL、1 mg/mL、10 mg/mL白藜芦醇的改性粘接剂滴于96孔板盖,制作厚度约1 mm、直径约6.4 mm的粘接剂样本片。使用傅里叶变换红外光谱(FTIR)检测粘接剂的单体转化率,使用细胞计数工具盒-8(CCK-8)评估人牙髓干细胞(h DPSCs)的增殖生长情况并比较各浓度改性粘接剂的细胞毒性,使用活死细菌染色和3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)测试评价改性粘接剂的抗变形链球菌(S.mutans)生物膜形成特性。第二部分实验中,收集并筛选无龋人第三磨牙,去除冠部釉质充分暴露牙本质部分,冲洗、抛光后随机分成4组,分别作为0 mg/mL、0.1 mg/mL、1 mg/mL、10 mg/mL浓度改性粘接剂的处理组。酸蚀、冲洗样本后,分别使用不含白藜芦醇的对照组粘接剂、0.1 mg/mL、1 mg/mL、10 mg/mL白藜芦醇改性粘接剂涂布牙本质表面并光固化15 s,随后按照临床操作方法在上方堆塑复合树脂。待粘接剂完全固化,平行于牙长轴方向切割制备树脂-牙本质片样本以及树脂-牙本质棒样本。每组样本分成3个亚组:(1)即刻组:37℃水储存24 h(2)冷热循环老化组:10000次冷热循环(3)酶老化组:37℃胶原酶溶液老化1个月。分别使用微拉伸仪器测试各组树脂-牙本质棒样本的粘接强度、场发射扫描电镜(FESEM)进行断裂模式分析、FESEM观察树脂-牙本质片样本的粘接界面纳米渗漏情况、激光共聚焦显微镜(CLSM)观测树脂-牙本质片样本粘接界面的基质金属蛋白酶(MMPs)活性。结果:第一部分实验的测试结果显示,在通用型粘接剂中加入0.1 mg/mL与1mg/mL浓度的白藜芦醇不影响粘接剂的单体转化率,但10 mg/mL组的转化率受到影响而明显低于对照组。细胞毒性实验结果表明,0.1 mg/mL、1 mg/mL和10 mg/mL组白藜芦醇改性粘接剂对h DPSCs的增殖生长均无明显不良影响,具有可接受的生物相容性。抗菌实验结果显示,0.1 mg/mL白藜芦醇改性的粘接剂抗菌能力与对照组无差异,但随着浓度提高,1 mg/mL和10 mg/mL实验组中变形链球菌生物膜的生长受到明显抑制。第二部分实验结果显示,白藜芦醇改性的粘接剂与对照组粘接剂的即刻微拉伸强度没有明显差异。经冷热循环老化或酶老化后,0.1 mg/mL与1 mg/mL白藜芦醇改性的粘接剂展现出较对照组更高的粘接强度,但10 mg/mL组的微拉伸强度仍处于较低水平。纳米渗漏实验结果表明,各浓度白藜芦醇改性的粘接剂即刻纳米渗漏均减少,且1mg/mL和10 mg/mL实验组在冷热循环老化和酶老化后纳米渗漏情况无明显改变。原位酶谱实验结果表明,0.1 mg/mL白藜芦醇改性粘接剂即可显著抑制粘接界面的MMPs活性,且随浓度升高抑制作用增强。结论:适当浓度的白藜芦醇改性牙本质粘接剂不仅不影响粘接剂的固有性能和生物相容性,还可提供优异的抗菌性能。同时适当浓度白藜芦醇改性的粘接剂通过抑制界面的MMPs活性、抵抗界面老化,从而改善牙本质粘接耐久性。临床意义:白藜芦醇改性粘接剂的性能检测和应用结果展现出其增加复合树脂修复体寿命的良好前景,对未来临床工作有借鉴与指导意义。临床工作方面,全程参与了六个病例的接诊、治疗计划、资料收集、修复治疗、术后随访,病例类型包括了牙体缺损的美学固定修复,常规活动义齿修复,复制义齿技术制作全口活动牙修复、多颗牙固定义齿修复行咬合重建、使用数字化导板的种植修复等。通过六个病例的操作、记录与讨论,加强训练了口腔修复临床技能与临床思维,为将来口腔修复临床工作打下基础。更多还原

【Abstract】 Objectives: This paper aimed to prepare suitable concentration of resveratrol-doped adhesive and evaluate the antibacterial property,DC,cytotoxicity,influence on dentin bonding,analyze the mechanism and the prospect of clinical application.Methods: Experimental adhesives were prepared by incorporating resveratrol into a universal adhesive at concentrations of 0(control),0.1,1,and 10 mg/mL.In the first part,specimens were prepared using the cover of sterile 96-well plate as a mold,and all specimens were about 1 mm in thickness and 6.4 mm in diameter.FTIR was used to test the DC of adhesive.CCK-8 were used for evaluating the growth of human dental pulp stem cells(h DPSCs)and comparing the cytotoxicity of different concentrations of modified adhesives.The antibacterial property of resveratrol-doped adhesive were evaluated by live and dead bacterial staining and MTT test.In the second part,non-carious human third molars were collected with the achievement of patients’ informed consents.The flat mid-coronal dentin of these teeth was then exposed by sectioning with a water-cooled low-speed diamond saw.These samples were burnished with water-irrigated 600-grit SIC papers and randomly distributed into four groups: A(control),0 mg/mL;B,0.1 mg/mL,C,1 mg/mL;and D,10 mg/mL.All the surfaces were rinsed with deionized water,etching with 35% phosphoric-acid gel for 15 s and then blot-dried.Polymerization via an LED light-curing unit for 15 s after bonding procedure.Then resin composite build-ups were constructed.All bonded teeth were sliced longitudinally to yield slabs or beams.Every group was divided into 3 subgroups: group 1,37 ℃ water storage for 24 hours;group 2,10,000 runs of thermocycled aging;group 3,1-month of collagenase aging.Testing MTBS of all groups.Using FESEM,CLSM to evaluate fracture mode,nanoleakage of adhesive–dentin interface and in situ zymography.Results: In the first part of experiments,DC values in the 0.1 and 1 mg/mL groups did not significantly decrease compared with the control group(p > 0.05).Whereas,the DC value significantly decreased when the incorporating amount of resveratrol reached 10mg/mL(p < 0.05).There was no significant difference in the cell viability of h DPSCs among the four tested groups,suggesting acceptable biocompatibility.S.mutans activity from the 1 and 10 mg/mL groups was significantly lower than that of the control and 0.1 mg/mL groups(p < 0.05),and the 10 mg/mL group exhibited the strongest inhibitory effects on S.mutans biofilm formation(p < 0.05).In the second part,there was no significant difference in immediate bonding strength between control group and the resveratrol-doped groups.But 0.1 and 1 mg/mL groups had better MTBS after aging and were significantly higher than those in the control and 10mg/mL group(p < 0.01).The silver precipitates showed sparse,interrupted distribution as the concentration of resveratrol increased.In situ zymography suggested that 0.1 mg/mL group could significantly inhibited MMPs activity at the bonding interface,and the degrees of green fluorescence gradually decreased with increasing incorporating amounts of resveratrol.Conclusions: This study suggested that resveratrol rarely affected the basic properties of adhesive.Resveratrol-doped adhesive achieved effective improvement on secondary caries management and biocompatibility.Meanwhile,appropriate concentration of resveratrol-doped adhesive can effectively keep the immediate dentin bonding strength,resist aging and inhibit the activity of MMPs,consequently improve the durability of the bonding interface.Clinical significance: The application of the resveratrol-doped adhesive indicates promising benefits to increase the lifetime of composite restorations.In clinical work,I joined in all the 6 cases of first visit,treatment planning,data collection,repairing,and follow-up.These cases included dental aesthetic restoration of tooth defect,removable denture repairing,duplicate denture technology,the tooth occlusal reconstruction,and implant denture using digital guide plant,etc.Through the operation,recording and discussion,my clinical skills and thinking style of oral prosthodontics were strengthened,which could benefit the clinical work in the future.更多还原