硬组织锯割机制作带种植体不脱钙骨磨片的方法研究

目的探讨LeicaSP1600硬组织锯割机制作带种植体不脱钙骨磨片的方法要点及注意事项。方法使用LeicaSP1600硬组织锯割机将常规塑料包埋后的带种植体骨标本沿种植体长轴锯割出厚150～200μm骨片，将其磨至60-80μm厚，行荧光染色观察，再行亚甲兰～酸性品红染色观察。结果制得的带种植体不脱钙骨磨片，光学显微镜和荧光显微镜下能观察到种植体周骨结合情况。结论正确使用LeicaSP1600硬组织锯割机，能较快制作出高质量的带种植体不脱钙骨磨片，可相对延长锯片使用寿命，节省骨标本。     

塑料包埋技术在口腔硬组织切片中的应用研究

目的 利用国内现有病理切片设备，通过对塑料包埋技术制作口腔砭组织切片及磨片的方法、步骤以及染色法的研究，确定其操作过程中的关键步骤以及需注意的问题。方法 选取带牙颌骨以及带种植体的动物下颌骨为材料，进行塑料包埋，用德国Leica公司生产的切片机制作薄切片或磨片，进行染色观察。结果 与用石蜡包理相比，不脱钙骨组织经塑料包埋后，可以制作出1μm的薄切片，以及30μm左右的磨片。切片染色后，可以观察到矿化骨矿化的过程，以及种植体周围骨组织的生长情况和二者的结合程度。结论 利用塑料包埋并配合重型切片机制作切片，可以进行骨组织矿化程度的研究与分析、牙齿龋病研究，以及种植体与骨组织结合情况的研究。这些都是石蜡切片所不能完成的。     

种植体骨界面组织形态学研究方法探讨

目的:探讨利用Leitz-1600型锯割切片机及骨磨片技术进行种植体骨结合状况的组织学研究方法.方法:选取植入种植体的颌骨或其他部位的骨组织,经过X线定位、塑料包埋、锯割切片机分切、手工磨片及特殊染色等组织学手段,考察种植体骨结合情况,以及植入骨再生材料的成骨情况.结果:植入种植体的骨组织经过完善的组织学处理后,采用手工磨片可以磨制30μm～50μm的骨种植体磨片,并能清楚地观察到种植体骨结合的状况,以及骨再生材料成骨的变化过程.结论:塑料包埋,手工磨片,以及特殊染色等组织学方法,可以成为研究种植体骨结合及其他与种植体相关研究的重要手段.     

手工制作带种植体骨磨片的常见问题与处理

目的：分析手工制作带种植体骨磨片易出现的问题,探讨并提出最佳的处理与解决方法。方法：针对制作带种植体骨磨片时出现的诸如定位不准确、脱水浸润不彻底、包埋温度过高、染色方法使用不当等制作过程中的诸多问题,通过采用不同脱水浸润时间、二次包埋聚合法、顺序研磨以及选择合适的染色方法等规范化操作,以提高手工制作带种植体骨磨片的成功率。结果：通过改进方法,规范化操作,经完善脱水处理并采用分段式包埋聚合,包埋块组织内无气泡,研磨后的切片内无金属碎屑,经甲苯胺蓝及亚甲基蓝一碱性品红两种方法染色,新生骨与老骨区分明显。结论：尽管在骨磨片厚度、染色方法的适用范围、劳动强度等方面,手工制作带种植体骨磨片与磨片机的制作水平还有一定差距。但通过标准化的操作流程,我们所制作的骨磨片已完全能满足目前国内研究的需求。     

带种植体骨磨片的标准化制作

目的：探讨带种植体骨磨片的标准化制作流程，为进行人工种植体相关研究提供病理实验依据。方法：回顾自1998年以来制作的850张带种植体骨磨片，按照塑料包埋前、包埋、粘结、切片、研磨、染色等几大步骤，对磨片进行评价。以（1）种植体完整，种植体长轴与组织面平行且在同一平面上；（2）组织经脱水处理包埋聚合后组织内无气泡；（3）组织与载物片粘贴紧密无气泡；（4）镜下观察磨片组织内无金属碎屑，种植体无脱落；（5）染色均匀，新生骨组织区分明显等5个方面作为标准化制作流程的评价指标，且5个指标需同时达标才能认为磨片制作成功。结果：对照5项指标对850张磨片进行回访式检查，早期因制作经验以及方法不当等原因，合格率约为50％，后经改进工艺与提高技术水平，2000年后所制作磨片的成功率提高到85％。结论：通过改进方法，规范化操作，选择适宜的染色方法，可以提高种植体骨磨片制作的成功率，扩大带种植体骨组织的研究范围。     

带种植体骨磨片技术

带种植体骨联合磨片已成为研究种植体与骨结合情况的常用与必备的方法，本文从标本取材处理、包埋、切片及染色等方面对带种植体骨磨片制作方法，特别是浸润与包埋过程中易出现的问题进行了详尽的论述，并对手工与机器两种磨片制作方法进行了比较。对国内开展带种植体骨磨片的制作具有一定的参考意义。     

手工制作带金属种植体骨组织磨片的方法

金属种植体坚硬,普通硬组织切片机不能满足要求,自动磨片机价格昂贵,且国内没有同类机器。我们经过试验初步掌握了手工制作带金属种植体骨组织磨片的方法,现介绍如下。1材料:Ｌｅｉｔｚ1600锯割切片机(德国产),360目、500目、1000/1200目金钢砂耐水砂纸,石蜡油,氰基丙烯酸酯,甲基丙烯酸β羟乙酯,过氧化苯甲酰,二甲基亚枫。2组织处理:将带有钛合金种植体的骨组织,锯成15ｃｍ×10ｃｍ×08ｃｍ的骨块,于10%甲醛中固定24ｈ,入70%酒精中脱水并脱至95%酒精为止。3配制浸泡液:取甲基丙烯酸β羟乙酯溶液100ｍｌ,过氧化苯甲酰1ｇ混合,在磁力搅拌器上…     

荧光标记法比较4种表面处理方式种植体骨矿化沉积率的研究

目的荧光标记法观察比较4种表面处理方式种植体周围骨矿化沉积率的差异。方法微创拔除4只Beagle犬双侧下颌4颗前磨牙,待拔牙创愈合3个月后,植入奥齿泰种植体40颗,其中机械形态表面(ma-chined morphology,MM)组4颗,喷砂加酸蚀(sand blasted with alumina and acid etched,SA)组、可吸收性研磨介质(resorbable blasting media,RBM)组和生物化学喷砂酸蚀组(biochemistry sand blasted with alumina and acid etched,Bio-SA)组各12颗,愈合3个月,处死前13、14 d和3、4 d分别注射盐酸四环素和钙黄绿素,制取标本并置于70%酒精固定,采用塑料包埋技术制作骨磨片,荧光显微镜下观察,测量骨矿化沉积率。结果螺纹间及远离螺纹区域新骨矿化沉积率在4组间的差异均没有统计学意义(P>0.05);单独比较每组种植体螺纹间及远离区域骨矿化沉积率的差异性,RBM组及SA组组内差异没有统计学意义(P>0.05),而MM组和Bio-SA组组内差异具有统计学意义(P<0.05)。结论采用塑料包埋技术,通过荧光标记法能成功观察到新骨矿化沉积的情况;种植体植入3个月后,4种不同粗糙程度的种植体螺纹间及远离螺纹区域骨矿化沉积率没有差异性,而经机械形态表面处理的种植体和生物化学喷砂酸蚀处理的种植体,螺纹间骨矿化沉积比远离螺纹区域快。     

改进超硬组织切片技术用于钛种植体骨界面研究

目的 研究改进的超硬组织切片技术应用于钛种植体骨界面研究中的关键步骤及需注意的问题。方法 本研究于2012 年8—11月在山西医科大学动物实验中心完成。选用6只成年雄性Beagle犬,在上颌牙槽骨分别植入6枚种植体后,锯下带有种植体的牙槽骨,采用改进超硬组织切片技术制作切片并进行染色观察。结果 60～80 μm厚的切片上通过甲苯胺蓝染色可以观察到骨组织的微细结构、种植体与骨组织的结合情况以及胶原纤维和类骨质。结论 改进的超硬组织切片技术适合种植体与骨组织结合情况研究。     

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目的研究改进的超硬组织切片技术应用于钛种植体骨界面研究中的关键步骤及需注意的问题。方法本研究于2012年8—11月在山西医科大学动物实验中心完成。选用6只成年雄性Beagle犬,在上颌牙槽骨分别植入6枚种植体后,锯下带有种植体的牙槽骨,采用改进超硬组织切片技术制作切片并进行染色观察。结果60～80μm厚的切片上通过甲苯胺蓝染色可以观察到骨组织的微细结构、种植体与骨组织的结合情况以及胶原纤维和类骨质。结论改进的超硬组织切片技术适合种植体与骨组织结合情况研究。     

Bone-tissue formation and integration of titanium implants: an evaluation with newly developed enzyme and immunohistochemical techniques

Background: Examination of the tissue surrounding retrieved implants involve routine investigations on cut and ground sections. Undecalcified sections with implants in situ are histologically stained followed by qualitative and quantitative observations of the tissue response to the implants by light microscopy. Purpose: A novel technique that allows for the accurate definition and quantification of enzymes involved in bone formation (alkaline phosphatase) and resorption (acid phosphatase) in the tissue is presented. Materials and Methods: Commercially pure titanium and titanium alloy (Ti6A14V) implants were retrieved after 6 and 12 weeks of healing in rabbit bone. In addition, 4-week specimens from commercially pure titanium bone harvest chambers placed in rabbit bone were used. Undecalcified cut and ground sections were produced and evaluated with enzyme and immunohistochemical staining techniques. Results: The titanium implants retrieved after 6 weeks of insertion in rabbit bone revealed a higher activity of both alkaline phosphatase and acid phosphatase activity compared to the implants followed for 12 months. The former samples revealed ongoing bone-tissue remodeling in the interface, whereas the latter ones showed steady-state bone conditions. Applying the new technique allowed for investigation of various bone proteins present in the tissue that had formed inside titanium canals of harvest chambers at various times of follow-up. Conclusions: The combination of routine histologic stainings with enzyme and immunohistochemical technique of cut and ground specimens is a valuable tool in the investigations of retrieved implants from humans and animals. This novel technique now may be used to describe the state of bone regeneration in the interface zone associated with implant research.     

Histomorphometric evaluation of the dimensional changes in rat-incisor tissues following histological processing and embedding in paraffin wax and glycol methacrylate

A three-dimensional reconstruction of undemineralized and paraffin wax- or or glycol methacrylate-embedded demineralized rat incisors was made using a computerized histomorphometry. Compared to the undemineralized ground sections, the two demineralizing and embedding procedures grossly distorted the tooth sections. In general, embedding in wax caused shrinkage and in methacrylate caused swelling. Direction and magnitude of the deformation varied in the different tissues of the tooth and its surrounding structures. The distortion was influenced by the ratio between the soft and hard tissues at different cross-sectional levels of the tooth.     

Immunohistochemistry of soft tissues surrounding late failures of Brånemark implants

The objective of the present investigation was to characterize the cellular composition of the soft tissues surrounding consecutively retrieved late failures of Brånemark implants. Criteria for implant failure were signs of loss of osseointegration (radiographic peri-fixtural radiolucency and mobility). The clinical history of the implants did not include adverse symptoms. At the time of retrieval, percussion-induced pain was experienced at 4/S implants, but no macroscopical signs of inflammation or infection was observed. Immunohistochemistry was applied on 6 marginal peri-implant specimens and on specimens of deeper tissues associated with the previously load-bearing implant surface from 8 failed implants, whereas 6 clinically healthy mucosal specimens and 4 hyperplastic biopsies from stable implants served as controls. The immunohistochemical evaluation showed that the soft tissues surrounding failed implants contained a large number of macrophages (CD68), HLA-DR positive cells, lymphocytes and plasma cells preferentially accumulated towards the removed implant surface. PMNs were a rare finding. Downgrowth of epithelium, in some cases encapsulating the whole fixture, was observed in sections where an intact implant/soft tissue interface was preserved. Healthy control mucosal specimens always contained labelled cells, albeit in a low amount, whereas hyperplastic control samples displayed an intense inflammatory and immunological response with numerous positive cells and PMNs scattered throughout the biopsy. In conclusion, failed implants were characterized by a chronic inflammatory response of the surrounding tissues with macrophages as the predominant labelled cell type, while hyperplastic tissues around stable implants were distinguished by an acute inflammatory process. These findings suggest that an on-going infection is unlikely to be the etiological factor for the late failures of dental implants examined in this study.     

正畸微种植体表面阳极氧化处理对周围骨组织的影响

目的通过组织形态学及组织形态测量学评价正畸微螺旋种植体表面阳极氧化处理对周围骨组织愈合情况的影响。方法24枚钛合金正畸用微螺旋种植体种植于两只新西兰大白兔的后腿,在闭合环境下生长6周。12枚微种植体经过表面阳极氧化处理作为研究组;另外12枚表面没有经过任何处理作为对照组。兔处死后,部分微种植体及周围骨组织进行组织学苏木精-伊红染色（hematoxylin-eosin staining,HE）处理,每组有4枚微种植体及周围骨组织块采用维拉努埃瓦（Villanueva）染色。光学显微镜评价种植体的组织学变化并计算种植体的骨结合率。结果两组种植体均与周围骨组织有一定的结合,其间无结缔组织可见。螺旋区骨纤维走向紊乱,表面阳极氧化组微种植体周围有较多的骨组织可见。阳极氧化组微种植体的骨结合率略高于对照组。结论阳极氧化处理可能促进钛合金微种植体与周围骨组织的结合。     

A method for the study of undecalcified bones and teeth with attached soft tissues*

A new sawing-grinding method is described for the histological evaluation of jaw bones with teeth or bones containing implants (ceramic or metallic). The undecalcified bone is embedded in acrylic resin and sawed at 100 to 150 μm. The slices are ground automatically by a special machine to a thickness of 5–10 μm. The usually employed staining procedures for hard plastic embedded tissues may be used. Plaque, fillings, crowns, bridges, implants and soft tissues are preserved in situ. Macroscopic and microscopic detail of good quality is preserved for histological and morphometrical evaluation.     

Implant site development: clinical realities of today and the prospects of tissue engineering

The success of dental implants depends on their placement in bone of adequate density and volume in order to achieve primary stability. Optimal esthetics of implants requires their placement in a position approximating that of the natural teeth they replace. However, there is generally at least some degree of atrophy in the sites where implants are to be placed. This atrophy may occur either before or after tooth extraction. Following extraction of teeth, there is commonly alveolar ridge resorption in horizontal and vertical dimensions. Alternatively, some of the oral hard and soft tissues may be destroyed by pathologic conditions such as periodontitis, endodontic infections, or trauma. All of these conditions may potentially compromise the final esthetics and function of implant-supported restorations. During the initial years of the development of the osseointegration protocol, implants were placed with little or no modification of implant sites. Though osseointegration was successfully achieved, esthetic outcome was not a primary objective of therapy at that time. A gradual paradigm shift has occurred in implant dentistry from merely achieving successful osseointegration to achieving final restorative outcomes that mimic natural dentition and their surrounding oral tissues. These objectives have been materialized by advancements in surgical techniques, as well as availability of biomaterials to enable predictable regeneration of oral hard and soft tissues. The objective of the present review is to briefly discuss some of the techniques that are currently available for implant site development.     

中空种植体-骨间隙愈合界面的组织学观察

目的:从组织学上观察中空种植体与旧骨之间间隙的愈合情况。方法:经双酸蚀处理后的T形中空纯钛种植体,植入7只SD大鼠的股骨远端。术后8d和16d分别取3只大鼠的股骨远端做硬组织切片,观察种植体-骨愈合情况。结果:术后8d,骨组织或骨样组织在一些区域与种植体内外表面直接接触,中空部分的骨小梁较稀疏;16d,骨组织或骨样组织呈网状充填种植体中空部分,较8d组,骨小梁显著增粗,种植体表面局部与骨组织直接接触。结论:骨组织可生长并充满于中空种植体的中空间隙。