激光酸蚀与喷砂酸蚀的纯钛种植体表面生物效能的对比性研究
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摘要
研究背景和目的
早在1981年,Albrektsson等已认为种植体表面形貌结构是影响种植体骨结合的六大因素之一,他们认为粗糙的表面比光滑表面更有利于促进骨结合。随后便有大量的学者对种植体的表面形貌进行了相关的体内外研究及改性处理,大大的缩短了种植体临床的骨愈合时间；至今已有30年余,但对种植体表面形貌及理化性质的研究仍然是当今口腔种植学领域十分热门的研究之一!仍然有大量的学者在研究种植体的表面处理方法,因为种植体表面形貌和理化特征不仅可以决定骨组织细胞的附着、增殖和分化,而且在很大程度上决定种植体与骨组织间的愈合速率,也是评估一个种植体系统优劣的重要标准之一。尽管目前的种植表面处理方法已经达到了很好的临床骨结合效果,但每当我们误以为种植体表面处理技术的研究已经到了没有什么值得再去研究时,一些学者及机构对种植体表面处理技术的进一步发掘及改进,这却又让整个口腔种植界发生重要的变化,譬如SLActive表面、全粗糙表面的种植体及粗糙度逐级过渡的表面等等。故再进一步缩短种植体的临床骨愈合时间,最大限度的减少患者牙体缺损的痛苦仍然是基础和临床研究人员的一个追求与期望。
在种植体的表面处理方法中,主要是有涂层和非涂层两种处理方法,但多年的研究表明非涂层的种植体表面处理更优于涂层的表面处理方法,成为了种植体表面处理的主流。目前非涂层表面处理经常采用物理学、化学以及电化学等方法直接粗化钛种植体表面,如喷砂、阳极氧化、酸蚀等获得了进一步改善种植体骨结合的数量与质量的效果。近些年来,喷砂酸蚀(Sand Blast and Acid Etching, SLA)表面已成为国际上使用最为广泛的种植体表面之一,其表面良好的机械生物相容性和骨组织引导性已经得到了口腔种植界的公认。种植体表面能在大颗粒喷砂下获得10～30μm孔坑,这些大孔坑具有促成骨的作用,再经过酸蚀后,种植体表面又形成大量的微孔(大小主要为1-3μm)；同时酸蚀能在一定程度上清洁纯钛种植体的表面杂质,增加其表面的亲水性,这样便形成了二级粗糙度。可以说,一级粗糙度类似于骨陷窝有利于成骨细胞的附着,二级粗糙度可以刺激成骨细胞增殖分化,有利于骨结合的形成；此外,体内试验已经证实：喷砂加酸蚀明显增加了种植体的扭矩值。因此,目前世界上一些著名品牌的种植体也采用了喷砂酸蚀的表面处理方法,如瑞士的straumman种植体系统(SLA/SLActive)、德国的Ankylos系统等,这些系统在临床上获得了良好的骨结合,进一步缩短了口腔种植的临床治疗时间。尽管喷砂加酸蚀的表面处理方法具有许多优点,但其也具有一定的缺点：如喷砂操作的不可控性,其形成的孔坑大小不同可直接影响种植体表面的成骨效能,而影响孔坑大小的因素包括颗粒大小、气压及喷砂距离等；此外,SLA的表面处理技术未能达到真正的防污染和表面净化的作用,其表面会嵌留下一些不利于骨结合的颗粒如Al2O3等,以上这些不足促使了口腔种植学的学者们对种植体表面处理的方法继续深入研究!
近年来,随着激光加工和口腔种植学的不断进步,激光表面处理新技术逐渐被引进种植体表面处理的领域。将光应用于材料加工和医学已有悠久的历史,但正式将激光应用于粗化种植体表面也是近10年多来才开始的。与上述传统的种植体表面处理技术不同的是,激光表面处理是一种非接触性的干洁技术,其具有准确可控、高度重复性、高效率性等特点,它能在纯钛的表面产生独特的表面形貌和表面物质构成,还能增加钛表面的耐腐蚀性等。利用不同的激光表面处理技术对钛合金进行表面改性,可达到不同的改善合金表面性能的效果,或赋予合金生物活性的不同目的,如激光表面合金化(Laser Surface Alloying, LSA)、激光表面熔覆陶瓷涂层及脉冲激光沉积(Pulsed Laser Deposition,PID)等技术已经广泛应用[19]；Shan-hui Hsu[20]等利用激光照射种植体表面后来达到杀菌、清洁的作用,再加酸蚀处理的细胞和动物实验,发现明其显增加了种植体表面骨细胞的吸附和骨结合；最近有学者用Nd:YAG激光轰击种植体表面来增加其表面积和粗糙度,研究其骨结合效应,得出了更利于骨细胞生长,明显增加了扭力及种植体骨接触面积的结论；Itala等的实验结果提示：表面孔坑的直径为40或501μm比100μm以上的孔坑表面更有利于骨结合；Davide Berardi等在种植表面对不同直径及洞深的表面进行了研究,他们得出的结论是直径为30μm、深度为20μm的激光处理表面更有利于种植体骨结合。目前的喷砂处理技术一般在种植体表面喷出的直径为10～30μm。而最近Sang-Hwan Kang等[25]进行了激光表面处理与微弧氧化表面处理的研究,他们利于激光在种植表面处理出孔径为40gm、深度为23μm大量孔坑后植入动物体内的实验,发现该表面比微弧表面处理更能增加种植体的骨扭出力。根据以上文献,可以推测直径为20～50μm的孔坑比大于100μm直径更有利于骨结合。此外,粗糙度的大小也是影响喷砂酸蚀表面骨结合效果的一个重要的指标,由于喷砂本身的不可控性,不同的直径的喷颗粒和喷砂时间等都会有不同的喷砂效果,其所形成的表面特征大也不相同,如Straumann牙种植体公司推出的SLA表面ITI牙种植体表面的Ra值为2.93μm,孔坑的直径约2μm；美国3i牙种植体Osseotite种植体喷砂出表面Ra值为0.86μm,孔坑的直径约2μm；Frialit-Xive系酸蚀出表面Ra值为2.75μm,孔坑直径主要范围在3～5μm,坑深2～3μm。结合口腔种植的临床效果和以上的表面粗糙度的差异,可以推断由喷砂酸蚀处理所产生的不同表面,其生物活性也将各有差异,目前尚没有一个统一的数值或观点,只有一个范围,较多的研究结果多倾向于表面小孔坑的直径为1-4gm、粗糙度为1～3.62μm的种植体表面较利于骨细胞附着生长,而粗糙度大于3.62μm就不利于骨组织附着结合。两者具体为何值更利于骨结合仍需进一步的研究。
根据文献报道可知,目前用于种植体表面轰击处理的激光器主要有YAG激光、CO2激光两种,而YAG激光器比CO2激光器在改变种植体表面形貌方面方面显得更有优势,主要体现在打孔用的平均输出功率高、重复频率高,并且具有高亮度、高聚焦性、容易实现精细加工等特点。国外已有研究表明：经Nd:YAG激光照射后的钛表面没有细胞毒性,比光滑钛表面更能促进成骨细胞的附着和骨结合；此外,激光轰击与喷砂处理相比,具有清洁和可控性强等优点,可以在种植体上准确的形成所需孔径大小、均匀一致的三维窝洞,甚至有学者认为,在可行的表面处理技术中,只有激光能达到不污染种植体表面的一项新技术。经处理的种植体表面的形貌及化学性质的改变与激光照射的能量、脉冲、角度及波长等有密切的关系。而最近光纤激光器作为第三代激光技术的代表,具有其他激光器无可比拟的技术优越性：如产品优势光束质量高,分辨率为1微米,是传统产品的10倍以上,其打标速度是传统产品的3-5倍以上,并且耗电量低,是传统产品的1/25-1/10；值得可喜的是,光纤激光器的价格比以前大大降低,这为本实验对它的引进和研究提供了可能性。最近Cho SA[21]及Sang-Hwan等使用光纤激光来处理种植体的实验,均获得良好的种植体表面形貌及较好的骨结合效果。值得一提的是,结合多种处理方法对种植体表面进行处理是当今口腔种植体表面处理发展的一大趋势,例如最近国内外报道将喷砂酸蚀与微弧氧化、微弧载银和非载银等综合处理方法相互比较种植体的表面活性和骨结合效果；又有学者将种植体表面先经过喷砂加酸蚀,再进行激光处理等方法研究其表面生物效应,以寻求出一种更好的表面处理方法；而在今年最新的有关激光处理种植体表面的研究报道中,Prodanov L[36]在激光处理与喷砂酸蚀的表面对比性研究中,得出激光处理与喷砂酸蚀均获得了良好骨结合的结论。这为我们进一步的激光表面处理研究提供及时的实验基础及理论依据。
综上所述,尽管现在已经有少数的有关激光和喷砂酸蚀的研究报道,但仍未见有对激光加酸蚀和喷砂加酸蚀的对比性研究的报道,究竟激光酸蚀能不能获得与喷砂酸蚀这一经典的表面处理一样的骨结合效果呢?又或者比该经典方法更好?关于这些,仍需要相关的对比性研究；此外,本科研组前期已经对激光加酸蚀的表面和光滑的表面进行了对比性研究,发现激光加酸蚀表面能明显的促进成骨的效果。故本次的研究拟根据目前激光表面处理的一些新进展,改变激光处理的一些条件,缩小蚀刻孔坑的直径等,再经过酸蚀处理的方法,与喷砂加酸蚀这双重处理的表面在理化性质和动物实验方面进行对比,对这两种表面处理方法作一个初步的综合性研究,为激光加酸蚀的深入研究提供实验基础及理论依据,以最终寻求出一种更好的种植体表面处理技术为目标。
方法
1.实验纯钛种植体的制备：通过查阅大量国内外文献及结合本实验的研究目的,使用精密的数控机床生产出合乎实验要求的实验种植体,根据预先确定的表面处理方法及相关实验参数,制备出较好的两种种植体表面：激光加酸蚀表面和喷砂加酸蚀表面。
2.形貌及理化性质的检测：通过扫描电子显微镜、X线光电子能谱仪和BMT Experi3D表面形貌仪分别对喷砂加酸蚀和激光加酸蚀这两种表面的形貌特征、粗糙度及元素成分等进行分析,评估这两处理组表面的理化性能特性的异同。
3.实验种植体的植入：先使用新西兰大白兔的胫骨上段研究不同植入位点是否会对种植体早期骨结合效果产生影响,然后再根据其结果指导下一步的动物实验；然后选择beagle犬作为SLA和LA表面对比性研究的实验动物,按随机分配的原则,分别在其胫骨上段植入这两种表面的植体,每侧胫骨共植入8枚种植体,编号从上往下编号为1-8号,每种处理组各4枚。按一定的时间间隔在beagle犬的皮下注射钙黄绿素和四环素双色标记,分别在2、4周两个时间点处死实验动物,5～8号种植体行扭出力的测量,1-4号种植体行骨组织切片的制作。
4.组织学观察：将解剖组的大白兔胫骨标本脱钙后制作石蜡切片,行HE染色观察测量；将带种植体的骨组织标本制作成不脱钙的骨组织切片(大白兔和Beagle犬),按实验目的行荧光双标记及染色观察分析,评估骨结合的生长速率和计算BIC%。
5.测量记录好骨扭出力值和BIC%(Bone-to-implant Contact, BIC)等数据,进行统计分析,得出实验结论。
统计学处理
实验数据以均数±标准差(x±s)来表示,样本量以n表示,应用SPSS v13.0软件(SPSS Inc.,Chicago, USA)进行统计分析。当满足正态分布且方差齐性时,采用两独立样本t检验或单因素方差来进行整体的均数比较；当不满足正态分布或方差不齐齐性时,采用两独立样本t’检验或秩和检验,再以Bonffronni检验比较组间差异；而对于需要分析交互效应的数据,则以析因设计的方差分析来检测处理因素的主效应和交互作用。假设检验为双侧检验,检验水准为0.05。当P>0.05时,差异被认为无统计学意义；当P<0.05时,差异被认为具有统计学意义。
结果
1.通过查阅大量国内外文献及结合本实验的研究目的,使用精密的数控机床生产出合格的实验种植体,然后根据预先设定的表面处理方法及相关实验参数,制备出较好的两种种植体表面：激光加酸蚀表面和喷砂加酸蚀表面。
2.两种表面理化性质检测结果：喷射加酸蚀组和激光加酸蚀组均具有良好的二级粗糙结构,LA表面的粗糙度大于SLA表面的粗糙度(LA:Ra=2.11μm; SLA:Ra=1.53μm),两者具有统计学差异(P<0.01)；激光加酸蚀的表面仍可见孔洞周围存留突起的熔融物,但酸蚀能酸洗去激光表面的碳物质；喷砂加酸蚀表面可见尖锐的边缘,其中仍有散在的一些三氧化二铝颗粒。
3.经过对大白兔胫骨上段的不同位点植入种植体的研究,发现不同的植入位点会对种植体早期的骨结合效果会产生明显的影响(P<0.01)。
4.LA组和SLA组在2周和4周的骨扭出力测量结果未见统计学差异(P>0.05),这两种在2周和4周时均获得良好的骨结合效果,亦未见统计学差异(P>0.05)。
结论
通过测量分析得出,新西兰大白兔的胫骨上段主要存在3种不同骨密度结构的区域,这3处的种植体骨结合率具有统计学差异；激光加酸蚀与喷砂加酸蚀的表面均具有二级粗糙的表面,前者较后者干洁而规则,这两种表面均具有良好的生物相容性和良好的骨组织引导性,激光加酸蚀组在2、4周均获得了与喷砂加酸蚀相似的BIC%和骨扭出力值,均能增强种植体早期骨结合的效能；激光加酸蚀处理均是一种可行的、效果较好的表面处理方法,但仍需要进一步的深入研究。
Background and Objection:
In the1980s, surface structure of dental implant was identified as one of the six factors which were very important for implant incorporation into bone, So far,30years had been past, the studies of the implant surface topography and physicochemical properties are still very popular. Today, they are the hot topic in the field of Oral Implantology yet! The implant surface treatment methods are still studied by a large number of scholars, because the physical and chemical characteristics, the morphology of implant surface can not only determine the bone tissue cell attachment, proliferation and differentiation,even the speed of healing rate between bone and implant were determined by it in a large extent. It was one of the important criterions for assessment of an implant system merits. Although the planting surface treatment method had reached an good effect in osseointegration, but each significant change in oral Implantology areas would usual be happed followed the discovery of surface treatment. It was endless in academic study of implant surface treatments. Shorten the implant healing time and reduced pains in dentition defect patients in futher are still a pursuit and expectations of basic and clinical researchers. In the methods of implant surface treatment, they mainly divided into coated and uncoated processings, but many researches show that the non-coated implant surface treatment was better than the coated one, it has become the the main stream of implant surface treatment. Of course, adding the active substances in the non-coated surface has also appeared in the treatment study in recent years, but these are still in preliminary study stage. Non-coating surface treatment was frequently used with physics, chemistry and electrochemical methods such as Sand blasting, Anodizing and Etching to roughened titanium implant surface directly, so as to further improve the quantity and quality of the implant osseointegration.
In recent years, the Sandblasted and Acid-etched (SLA) surface has been widely used in implant surface treatment, it's mechanical biocompatibility has been recognized. The holes with diameter10-30μm in the implant surface can be obtained by the large particles blasting, these holes were good for the bone formation. Meanwhile, a large number of pores (size at about1-3μm) were formed by the acid-etched method. To some extent, titanium implant surface was also cleaned, that would increase their surface hydrophilicity,so it had primary and secondly roughness. It could be said that, the primary roughness was helpful to the osteoblasts adhesion, and proliferation and differentiation of osteoblasts could be stimulate by secondly roughness. In addition, the fact that sandblasting plus etching could increase the the implant torque values significantly had been confirmed. Therefore, from some famous implant systems in the world, we found that they dealed the surface with a sandblasted etching method, such as Switzerland straumman implant system (SLA/SLActive), Ankylos system in Germany. All these systems had the good osseointegration in clinical and the clinical treatment time was further shorten. Although the sandblasted and acid-etched surface had so many advantages, but it also had some certain disadvantages:such as sandblasting was out of control, the size of the holes directly affected osteogenic performance of implant surface. The size of the holes was mainly affected by the particle size, pressure and sandblasting distance. SLA surface treatment technology failed to meet the real pollution prevention and surface purification. The sandblasting particle such as Al2O3embedded in the surface was harmful for osseointegration, these inadequacies prompted scholars to do continue in-depth study!
In recent years, with the continuous advancement of laser science and oral implantology, laser surface treatment as a new technology was gradually being introduced to the field of implant surface treatment Laser used in material processing and medicine had a long history, but officially applied to etched and roughened implant surface just began nearly10years. Different from the above-described conventional surface treatment technology, The laser surface treatment was a non-contact technique, having accurately controllable, highly reproducible and efficient and other characteristics. It could produce unique surface morphology and surface material and increase corrosion resistance of the titanium surface. Using different laser surface treatment technology for treating titanium alloys could achieve the purpose of improving the titanium alloy surface properties, or could add the bioactive, such as the laser alloying surface (LSA), Pulsed laser deposition (PID) and laser cladding ceramic coatings and other technology had been widely used. Shan-hui was successed in using the laser irradiation for sterilizating and cleaning the implant surface. Through the vitro and vivo experiments, they found that it can increase the adsorption of osteoblasts and bone formation on implant surface significantly. Recently, the Nd:YAG laser was used to increase the surface areas and roughness, the study suggested that the surface was more suit for osteoblasts to grow and increased torque and implant-bone contact significantly. Itala's experimental results suggest that the diameters of the hole in40or50μm were more more conducive to bone than the diameters of100μm; The different diameter and deep holes in the implant surface were studies by Davide Berardi, it concluded that a hole with a diameter of30μm, depth of20μm in the surface by the laser processing was more conducive to implant osseointegration; The holes on the implant surface by Sandblasting techniques were generally in the diameter of10to30μm. Recently, the pore size with the diameter of40μm, a depth of23μm treated by laser, Sang-Hwan Kang found that laser surface was more than the micro-arc surface can help increase the implant bone torque; According to the above literature, it can be speculated that the diameter of the holes of10-50μm is more conducive to bone ratio of greater than100pm diameter binding. In addition, the roughness values was an important indicator to affect the implant osseointegration of sandblasting and etching surface. Wihtout controllable, different diameter of spray particles and sandblasting time and other factors would have a different effect of sandblasting, Such as the SLA surface of ITI Straumann dental implants, the Ra value was2.93μm, the pit diameter was about2μm; the3i dental implant(USA) with the surface Ra value of0.86μm, the pit diameter of2μm; the Frialit-Xive with the surface Ra value of2.75μm, the main pit diameter was3to5μm, the pit depth was2to3μm. Thereby the biological activity of the generated surface of these methods could be inferred by the above will also vary, a uniform good value of Ra was not existed at the moment, just only one range. More and more studies implied the diameter of the pit from1to4μm,the roughness values from1to3.62μm implant were more conducive to bone cell adhesion, while the roughness values greater than3.62μm was not conducive to bone tissue attachment. What the value was more conducive to osseointegration needed further study.
Lasers for implant surface treatment mainly included YAG laser and CO2laser, but the YAG laser was more suitable than the CO2laser in it[291, mainly reflected in the punch with high average output power, high repetition frequency, and having high brightness, high focusability, easy to achieve the fine processing characteristics. studies abroad have shown that:no cell toxicity on the titanium surface after the Nd:YAG laser irradiation, it was better than a smooth titanium surface to promote osteoblast attachment and osseointegration. In addition, compared with the blasting treatment, laser-treated was clean and controllable, the accurate formation of the desired hole size and uniform three-dimensional cavity could be created by it. Many scholars believe that:in viable surface treatment technology, only laser treatment had no contaminate to implant surface at the moment. The energy, pulse angle and wavelength of the laser irradiation had a close relationship with implant surface morphology and chemical changes. Fiber laser as the representative of the third generation of laser technology, with unparalleled technical superiority of other lasers: the advantages beam quality,1micron resolution, which was about10times than that of traditional products, the marking speed was3-5times higher than traditional products, and low power consumption. In addition, the price of fiber lasers was greatly reduced, which was possible for us to use. In recent time, Cho SA and Sang-Hwan used the fiber laser to obtain good implant surface morphology and better osseointegration. It was worth to mention that the implant surface treatment development trended to a combination of different methods, such as the recent reported in sandblasting etching and micro-arc oxidation, micro-arc containing silver and non containing silver and other integrated approach between the surfactant and bone effects1. For example, the implant surface treated with the Sandblasted/acid-etched firstly, and laser processing secondly to change surface morphology to seek better methods. This year, Prodanov's study was the latest research in laser implant surface treatment reports, there was the comparative study of laser-treated with the Sandblasted/acid-etched. both of them obtained good osseointegration. It provided timely experimental basis and theoretical basis for further study in laser surface treatment.
In summary, although there had been numerous studies about the laser-treated and the Sandblasted/acid-etched, but a comparative studies about the laser plus acid etching with sandblasting plus etching were not seen in reports. Whether the osseointegration of Laser-treated/acid-etched surface had the some effect as or better than the Sandblasted/acid-etched surface was to be determined yet. Some relative comparative study on the urgent need; In addition, our research group had been done comparative study about Laser-treated/acid-etched surface and the acid-etched surface, we found that Laser-treated/acid-etched surface had good biocompatibility and osteoconduction. Based on some new developments of laser surface treatment, The study was intended to changed some conditions of the laser processing, narrowing the diameter of etching holes and combined with the acid-etched, and then made a comparative study with the sandblasted/acid-etched surface by animal experiments, so as to provid experimental basis and theoretical basis for in-depth study of the Laser-treated/acid-etched, to seek a better implant surface treatment technology eventually was also the experimental target.
Methods and materials
1. Preparation of laboratory samples:based on a large number of domestic and foreign literature and research purposes, using precision CNC machine tools to produce an experiment implant; By surface treatment method, according to pre-determined and related experimental parameters, prepared two good implant surfaces:the Laser-treated/acid-etched surface and the sandblasted/acid-etched surface.
2. The test of physical and chemical properties:measure the morphology, roughness, elemental composition with scanning electron microscopy, X-ray photoelectron spectroscopy and BMT Experi3D surface topography instrument, respectively, to assess the physical and chemical properties of SLA and LA surfaces.
3. Animal model and implantation:New Zealand rabbits were used in the study of different implantation sites of the proximal tibia to osseointegration, implant were installed in3different sites-,4beagle dogs as experimental animals. According to the principle of random allocation, two different surface implants were installed into each side of the tibia, numbered them from the top to down with No.1to8, each treatment group had4implants in one tibia. At a certain time interval, made some subcutaneous injection with a hypodermic calcein and tetracycline color marker, experimental animals were sacrificed at2,4weeks,1-4implants were used to production of bone tissue slices,5-8implant for torque measurement.
4. Making implant bone tissue sections and measurement of experimental:After decalcification, the anatomy of the rabbit tibia were made into paraffin section for HE staining observing; The bone tissues with implant were used to made sections of undecalcified, osseointegration growth rate and the BIC%was assessment and calculated by the fluorescence labeling and staining.
5. The measurement recorded torque measurement value and BIC%data, statistical analysis obtained experimental results.
Statistical analyses:
The experimental data are expressed as mean±standard deviation (SD). Statistical analysis was carried out by SPSS±v13.0software (SPSS Inc., Chicago, USA). For the single factor data, when meet the normal distribution and homogeneity of variance, using two independent sample t test and single factor variance to the overall mean comparison; When the data not meet the normal distribution or variance not neat homogeneous, using two independent sample t'test or rank sum test; For two factors of single variable data with normal distribution and homogeneity of variance, to analyse the main effect and interaction by analysis of variance of factorial design. Hypothesis test for two-sided test, the test level was0.05, probabilities (P)>0.05was considered to be no statistically significant; Probabilities (P)<0.05was considered to be statistically significant.
Results:
1. Through access to a large number of domestic and foreign literatures and combined the purpose of the present study, using precision CNC machine tools produced some experimental implant. According to pre-set surface treatment methods and experimental parameters, two different implant surfaces were made: Laser-treated/acid-etched surface and Sandblasted/acid-etched surface.
2. Results of the physical and chemical nature detecting:both of SLA and LA obtainted two rough structure, the surface roughnesses of LA was bigger than the SLA (LA:Ra=2.1μm; SLA:Ra=1.53μm)(P<0.01). Some melt was still obserted in LA surface, but the carbon material could been cleaned by the acid-etched; the sharp edge of the etching was visible on the surface of SLA, few oxide aluminum particles was still existed.
3. The different implantation sites in the selected20mm area of rabbit's tibias demonstrated different early osseointegration (P<0.05).
4. No significant difference was found in torque measurement value and the BIC%in two and four weeks between two groups (P>0.05), both groups obtained good osseointegration.
Conclusion:
In the present study, the different implantation sites in the selected20mm area of rabbit's tibias demonstrated different early osseointegration; the sites located7±1.5mm below the epiphyseal line were best suited for observing the effectiveness of early osseointegration among the3sites; The surface treatment methods of the Laser-treated/acid-etched and the Sandblasted/acid-etched could produce the rough surface, the former was more cleaner and more uniform than the latter, but both of them had good biocompatibility and promoted a good implant osseointegration, these two methods were feasible for implant surface treatment. Further studies were still needed.
早在1981年,Albrektsson等已认为种植体表面形貌结构是影响种植体骨结合的六大因素之一,他们认为粗糙的表面比光滑表面更有利于促进骨结合。随后便有大量的学者对种植体的表面形貌进行了相关的体内外研究及改性处理,大大的缩短了种植体临床的骨愈合时间；至今已有30年余,但对种植体表面形貌及理化性质的研究仍然是当今口腔种植学领域十分热门的研究之一!仍然有大量的学者在研究种植体的表面处理方法,因为种植体表面形貌和理化特征不仅可以决定骨组织细胞的附着、增殖和分化,而且在很大程度上决定种植体与骨组织间的愈合速率,也是评估一个种植体系统优劣的重要标准之一。尽管目前的种植表面处理方法已经达到了很好的临床骨结合效果,但每当我们误以为种植体表面处理技术的研究已经到了没有什么值得再去研究时,一些学者及机构对种植体表面处理技术的进一步发掘及改进,这却又让整个口腔种植界发生重要的变化,譬如SLActive表面、全粗糙表面的种植体及粗糙度逐级过渡的表面等等。故再进一步缩短种植体的临床骨愈合时间,最大限度的减少患者牙体缺损的痛苦仍然是基础和临床研究人员的一个追求与期望。
在种植体的表面处理方法中,主要是有涂层和非涂层两种处理方法,但多年的研究表明非涂层的种植体表面处理更优于涂层的表面处理方法,成为了种植体表面处理的主流。目前非涂层表面处理经常采用物理学、化学以及电化学等方法直接粗化钛种植体表面,如喷砂、阳极氧化、酸蚀等获得了进一步改善种植体骨结合的数量与质量的效果。近些年来,喷砂酸蚀(Sand Blast and Acid Etching, SLA)表面已成为国际上使用最为广泛的种植体表面之一,其表面良好的机械生物相容性和骨组织引导性已经得到了口腔种植界的公认。种植体表面能在大颗粒喷砂下获得10～30μm孔坑,这些大孔坑具有促成骨的作用,再经过酸蚀后,种植体表面又形成大量的微孔(大小主要为1-3μm)；同时酸蚀能在一定程度上清洁纯钛种植体的表面杂质,增加其表面的亲水性,这样便形成了二级粗糙度。可以说,一级粗糙度类似于骨陷窝有利于成骨细胞的附着,二级粗糙度可以刺激成骨细胞增殖分化,有利于骨结合的形成；此外,体内试验已经证实：喷砂加酸蚀明显增加了种植体的扭矩值。因此,目前世界上一些著名品牌的种植体也采用了喷砂酸蚀的表面处理方法,如瑞士的straumman种植体系统(SLA/SLActive)、德国的Ankylos系统等,这些系统在临床上获得了良好的骨结合,进一步缩短了口腔种植的临床治疗时间。尽管喷砂加酸蚀的表面处理方法具有许多优点,但其也具有一定的缺点：如喷砂操作的不可控性,其形成的孔坑大小不同可直接影响种植体表面的成骨效能,而影响孔坑大小的因素包括颗粒大小、气压及喷砂距离等；此外,SLA的表面处理技术未能达到真正的防污染和表面净化的作用,其表面会嵌留下一些不利于骨结合的颗粒如Al2O3等,以上这些不足促使了口腔种植学的学者们对种植体表面处理的方法继续深入研究!
近年来,随着激光加工和口腔种植学的不断进步,激光表面处理新技术逐渐被引进种植体表面处理的领域。将光应用于材料加工和医学已有悠久的历史,但正式将激光应用于粗化种植体表面也是近10年多来才开始的。与上述传统的种植体表面处理技术不同的是,激光表面处理是一种非接触性的干洁技术,其具有准确可控、高度重复性、高效率性等特点,它能在纯钛的表面产生独特的表面形貌和表面物质构成,还能增加钛表面的耐腐蚀性等。利用不同的激光表面处理技术对钛合金进行表面改性,可达到不同的改善合金表面性能的效果,或赋予合金生物活性的不同目的,如激光表面合金化(Laser Surface Alloying, LSA)、激光表面熔覆陶瓷涂层及脉冲激光沉积(Pulsed Laser Deposition,PID)等技术已经广泛应用[19]；Shan-hui Hsu[20]等利用激光照射种植体表面后来达到杀菌、清洁的作用,再加酸蚀处理的细胞和动物实验,发现明其显增加了种植体表面骨细胞的吸附和骨结合；最近有学者用Nd:YAG激光轰击种植体表面来增加其表面积和粗糙度,研究其骨结合效应,得出了更利于骨细胞生长,明显增加了扭力及种植体骨接触面积的结论；Itala等的实验结果提示：表面孔坑的直径为40或501μm比100μm以上的孔坑表面更有利于骨结合；Davide Berardi等在种植表面对不同直径及洞深的表面进行了研究,他们得出的结论是直径为30μm、深度为20μm的激光处理表面更有利于种植体骨结合。目前的喷砂处理技术一般在种植体表面喷出的直径为10～30μm。而最近Sang-Hwan Kang等[25]进行了激光表面处理与微弧氧化表面处理的研究,他们利于激光在种植表面处理出孔径为40gm、深度为23μm大量孔坑后植入动物体内的实验,发现该表面比微弧表面处理更能增加种植体的骨扭出力。根据以上文献,可以推测直径为20～50μm的孔坑比大于100μm直径更有利于骨结合。此外,粗糙度的大小也是影响喷砂酸蚀表面骨结合效果的一个重要的指标,由于喷砂本身的不可控性,不同的直径的喷颗粒和喷砂时间等都会有不同的喷砂效果,其所形成的表面特征大也不相同,如Straumann牙种植体公司推出的SLA表面ITI牙种植体表面的Ra值为2.93μm,孔坑的直径约2μm；美国3i牙种植体Osseotite种植体喷砂出表面Ra值为0.86μm,孔坑的直径约2μm；Frialit-Xive系酸蚀出表面Ra值为2.75μm,孔坑直径主要范围在3～5μm,坑深2～3μm。结合口腔种植的临床效果和以上的表面粗糙度的差异,可以推断由喷砂酸蚀处理所产生的不同表面,其生物活性也将各有差异,目前尚没有一个统一的数值或观点,只有一个范围,较多的研究结果多倾向于表面小孔坑的直径为1-4gm、粗糙度为1～3.62μm的种植体表面较利于骨细胞附着生长,而粗糙度大于3.62μm就不利于骨组织附着结合。两者具体为何值更利于骨结合仍需进一步的研究。
根据文献报道可知,目前用于种植体表面轰击处理的激光器主要有YAG激光、CO2激光两种,而YAG激光器比CO2激光器在改变种植体表面形貌方面方面显得更有优势,主要体现在打孔用的平均输出功率高、重复频率高,并且具有高亮度、高聚焦性、容易实现精细加工等特点。国外已有研究表明：经Nd:YAG激光照射后的钛表面没有细胞毒性,比光滑钛表面更能促进成骨细胞的附着和骨结合；此外,激光轰击与喷砂处理相比,具有清洁和可控性强等优点,可以在种植体上准确的形成所需孔径大小、均匀一致的三维窝洞,甚至有学者认为,在可行的表面处理技术中,只有激光能达到不污染种植体表面的一项新技术。经处理的种植体表面的形貌及化学性质的改变与激光照射的能量、脉冲、角度及波长等有密切的关系。而最近光纤激光器作为第三代激光技术的代表,具有其他激光器无可比拟的技术优越性：如产品优势光束质量高,分辨率为1微米,是传统产品的10倍以上,其打标速度是传统产品的3-5倍以上,并且耗电量低,是传统产品的1/25-1/10；值得可喜的是,光纤激光器的价格比以前大大降低,这为本实验对它的引进和研究提供了可能性。最近Cho SA[21]及Sang-Hwan等使用光纤激光来处理种植体的实验,均获得良好的种植体表面形貌及较好的骨结合效果。值得一提的是,结合多种处理方法对种植体表面进行处理是当今口腔种植体表面处理发展的一大趋势,例如最近国内外报道将喷砂酸蚀与微弧氧化、微弧载银和非载银等综合处理方法相互比较种植体的表面活性和骨结合效果；又有学者将种植体表面先经过喷砂加酸蚀,再进行激光处理等方法研究其表面生物效应,以寻求出一种更好的表面处理方法；而在今年最新的有关激光处理种植体表面的研究报道中,Prodanov L[36]在激光处理与喷砂酸蚀的表面对比性研究中,得出激光处理与喷砂酸蚀均获得了良好骨结合的结论。这为我们进一步的激光表面处理研究提供及时的实验基础及理论依据。
综上所述,尽管现在已经有少数的有关激光和喷砂酸蚀的研究报道,但仍未见有对激光加酸蚀和喷砂加酸蚀的对比性研究的报道,究竟激光酸蚀能不能获得与喷砂酸蚀这一经典的表面处理一样的骨结合效果呢?又或者比该经典方法更好?关于这些,仍需要相关的对比性研究；此外,本科研组前期已经对激光加酸蚀的表面和光滑的表面进行了对比性研究,发现激光加酸蚀表面能明显的促进成骨的效果。故本次的研究拟根据目前激光表面处理的一些新进展,改变激光处理的一些条件,缩小蚀刻孔坑的直径等,再经过酸蚀处理的方法,与喷砂加酸蚀这双重处理的表面在理化性质和动物实验方面进行对比,对这两种表面处理方法作一个初步的综合性研究,为激光加酸蚀的深入研究提供实验基础及理论依据,以最终寻求出一种更好的种植体表面处理技术为目标。
方法
1.实验纯钛种植体的制备：通过查阅大量国内外文献及结合本实验的研究目的,使用精密的数控机床生产出合乎实验要求的实验种植体,根据预先确定的表面处理方法及相关实验参数,制备出较好的两种种植体表面：激光加酸蚀表面和喷砂加酸蚀表面。
2.形貌及理化性质的检测：通过扫描电子显微镜、X线光电子能谱仪和BMT Experi3D表面形貌仪分别对喷砂加酸蚀和激光加酸蚀这两种表面的形貌特征、粗糙度及元素成分等进行分析,评估这两处理组表面的理化性能特性的异同。
3.实验种植体的植入：先使用新西兰大白兔的胫骨上段研究不同植入位点是否会对种植体早期骨结合效果产生影响,然后再根据其结果指导下一步的动物实验；然后选择beagle犬作为SLA和LA表面对比性研究的实验动物,按随机分配的原则,分别在其胫骨上段植入这两种表面的植体,每侧胫骨共植入8枚种植体,编号从上往下编号为1-8号,每种处理组各4枚。按一定的时间间隔在beagle犬的皮下注射钙黄绿素和四环素双色标记,分别在2、4周两个时间点处死实验动物,5～8号种植体行扭出力的测量,1-4号种植体行骨组织切片的制作。
4.组织学观察：将解剖组的大白兔胫骨标本脱钙后制作石蜡切片,行HE染色观察测量；将带种植体的骨组织标本制作成不脱钙的骨组织切片(大白兔和Beagle犬),按实验目的行荧光双标记及染色观察分析,评估骨结合的生长速率和计算BIC%。
5.测量记录好骨扭出力值和BIC%(Bone-to-implant Contact, BIC)等数据,进行统计分析,得出实验结论。
统计学处理
实验数据以均数±标准差(x±s)来表示,样本量以n表示,应用SPSS v13.0软件(SPSS Inc.,Chicago, USA)进行统计分析。当满足正态分布且方差齐性时,采用两独立样本t检验或单因素方差来进行整体的均数比较；当不满足正态分布或方差不齐齐性时,采用两独立样本t’检验或秩和检验,再以Bonffronni检验比较组间差异；而对于需要分析交互效应的数据,则以析因设计的方差分析来检测处理因素的主效应和交互作用。假设检验为双侧检验,检验水准为0.05。当P>0.05时,差异被认为无统计学意义；当P<0.05时,差异被认为具有统计学意义。
结果
1.通过查阅大量国内外文献及结合本实验的研究目的,使用精密的数控机床生产出合格的实验种植体,然后根据预先设定的表面处理方法及相关实验参数,制备出较好的两种种植体表面：激光加酸蚀表面和喷砂加酸蚀表面。
2.两种表面理化性质检测结果：喷射加酸蚀组和激光加酸蚀组均具有良好的二级粗糙结构,LA表面的粗糙度大于SLA表面的粗糙度(LA:Ra=2.11μm; SLA:Ra=1.53μm),两者具有统计学差异(P<0.01)；激光加酸蚀的表面仍可见孔洞周围存留突起的熔融物,但酸蚀能酸洗去激光表面的碳物质；喷砂加酸蚀表面可见尖锐的边缘,其中仍有散在的一些三氧化二铝颗粒。
3.经过对大白兔胫骨上段的不同位点植入种植体的研究,发现不同的植入位点会对种植体早期的骨结合效果会产生明显的影响(P<0.01)。
4.LA组和SLA组在2周和4周的骨扭出力测量结果未见统计学差异(P>0.05),这两种在2周和4周时均获得良好的骨结合效果,亦未见统计学差异(P>0.05)。
结论
通过测量分析得出,新西兰大白兔的胫骨上段主要存在3种不同骨密度结构的区域,这3处的种植体骨结合率具有统计学差异；激光加酸蚀与喷砂加酸蚀的表面均具有二级粗糙的表面,前者较后者干洁而规则,这两种表面均具有良好的生物相容性和良好的骨组织引导性,激光加酸蚀组在2、4周均获得了与喷砂加酸蚀相似的BIC%和骨扭出力值,均能增强种植体早期骨结合的效能；激光加酸蚀处理均是一种可行的、效果较好的表面处理方法,但仍需要进一步的深入研究。
Background and Objection:
In the1980s, surface structure of dental implant was identified as one of the six factors which were very important for implant incorporation into bone, So far,30years had been past, the studies of the implant surface topography and physicochemical properties are still very popular. Today, they are the hot topic in the field of Oral Implantology yet! The implant surface treatment methods are still studied by a large number of scholars, because the physical and chemical characteristics, the morphology of implant surface can not only determine the bone tissue cell attachment, proliferation and differentiation,even the speed of healing rate between bone and implant were determined by it in a large extent. It was one of the important criterions for assessment of an implant system merits. Although the planting surface treatment method had reached an good effect in osseointegration, but each significant change in oral Implantology areas would usual be happed followed the discovery of surface treatment. It was endless in academic study of implant surface treatments. Shorten the implant healing time and reduced pains in dentition defect patients in futher are still a pursuit and expectations of basic and clinical researchers. In the methods of implant surface treatment, they mainly divided into coated and uncoated processings, but many researches show that the non-coated implant surface treatment was better than the coated one, it has become the the main stream of implant surface treatment. Of course, adding the active substances in the non-coated surface has also appeared in the treatment study in recent years, but these are still in preliminary study stage. Non-coating surface treatment was frequently used with physics, chemistry and electrochemical methods such as Sand blasting, Anodizing and Etching to roughened titanium implant surface directly, so as to further improve the quantity and quality of the implant osseointegration.
In recent years, the Sandblasted and Acid-etched (SLA) surface has been widely used in implant surface treatment, it's mechanical biocompatibility has been recognized. The holes with diameter10-30μm in the implant surface can be obtained by the large particles blasting, these holes were good for the bone formation. Meanwhile, a large number of pores (size at about1-3μm) were formed by the acid-etched method. To some extent, titanium implant surface was also cleaned, that would increase their surface hydrophilicity,so it had primary and secondly roughness. It could be said that, the primary roughness was helpful to the osteoblasts adhesion, and proliferation and differentiation of osteoblasts could be stimulate by secondly roughness. In addition, the fact that sandblasting plus etching could increase the the implant torque values significantly had been confirmed. Therefore, from some famous implant systems in the world, we found that they dealed the surface with a sandblasted etching method, such as Switzerland straumman implant system (SLA/SLActive), Ankylos system in Germany. All these systems had the good osseointegration in clinical and the clinical treatment time was further shorten. Although the sandblasted and acid-etched surface had so many advantages, but it also had some certain disadvantages:such as sandblasting was out of control, the size of the holes directly affected osteogenic performance of implant surface. The size of the holes was mainly affected by the particle size, pressure and sandblasting distance. SLA surface treatment technology failed to meet the real pollution prevention and surface purification. The sandblasting particle such as Al2O3embedded in the surface was harmful for osseointegration, these inadequacies prompted scholars to do continue in-depth study!
In recent years, with the continuous advancement of laser science and oral implantology, laser surface treatment as a new technology was gradually being introduced to the field of implant surface treatment Laser used in material processing and medicine had a long history, but officially applied to etched and roughened implant surface just began nearly10years. Different from the above-described conventional surface treatment technology, The laser surface treatment was a non-contact technique, having accurately controllable, highly reproducible and efficient and other characteristics. It could produce unique surface morphology and surface material and increase corrosion resistance of the titanium surface. Using different laser surface treatment technology for treating titanium alloys could achieve the purpose of improving the titanium alloy surface properties, or could add the bioactive, such as the laser alloying surface (LSA), Pulsed laser deposition (PID) and laser cladding ceramic coatings and other technology had been widely used. Shan-hui was successed in using the laser irradiation for sterilizating and cleaning the implant surface. Through the vitro and vivo experiments, they found that it can increase the adsorption of osteoblasts and bone formation on implant surface significantly. Recently, the Nd:YAG laser was used to increase the surface areas and roughness, the study suggested that the surface was more suit for osteoblasts to grow and increased torque and implant-bone contact significantly. Itala's experimental results suggest that the diameters of the hole in40or50μm were more more conducive to bone than the diameters of100μm; The different diameter and deep holes in the implant surface were studies by Davide Berardi, it concluded that a hole with a diameter of30μm, depth of20μm in the surface by the laser processing was more conducive to implant osseointegration; The holes on the implant surface by Sandblasting techniques were generally in the diameter of10to30μm. Recently, the pore size with the diameter of40μm, a depth of23μm treated by laser, Sang-Hwan Kang found that laser surface was more than the micro-arc surface can help increase the implant bone torque; According to the above literature, it can be speculated that the diameter of the holes of10-50μm is more conducive to bone ratio of greater than100pm diameter binding. In addition, the roughness values was an important indicator to affect the implant osseointegration of sandblasting and etching surface. Wihtout controllable, different diameter of spray particles and sandblasting time and other factors would have a different effect of sandblasting, Such as the SLA surface of ITI Straumann dental implants, the Ra value was2.93μm, the pit diameter was about2μm; the3i dental implant(USA) with the surface Ra value of0.86μm, the pit diameter of2μm; the Frialit-Xive with the surface Ra value of2.75μm, the main pit diameter was3to5μm, the pit depth was2to3μm. Thereby the biological activity of the generated surface of these methods could be inferred by the above will also vary, a uniform good value of Ra was not existed at the moment, just only one range. More and more studies implied the diameter of the pit from1to4μm,the roughness values from1to3.62μm implant were more conducive to bone cell adhesion, while the roughness values greater than3.62μm was not conducive to bone tissue attachment. What the value was more conducive to osseointegration needed further study.
Lasers for implant surface treatment mainly included YAG laser and CO2laser, but the YAG laser was more suitable than the CO2laser in it[291, mainly reflected in the punch with high average output power, high repetition frequency, and having high brightness, high focusability, easy to achieve the fine processing characteristics. studies abroad have shown that:no cell toxicity on the titanium surface after the Nd:YAG laser irradiation, it was better than a smooth titanium surface to promote osteoblast attachment and osseointegration. In addition, compared with the blasting treatment, laser-treated was clean and controllable, the accurate formation of the desired hole size and uniform three-dimensional cavity could be created by it. Many scholars believe that:in viable surface treatment technology, only laser treatment had no contaminate to implant surface at the moment. The energy, pulse angle and wavelength of the laser irradiation had a close relationship with implant surface morphology and chemical changes. Fiber laser as the representative of the third generation of laser technology, with unparalleled technical superiority of other lasers: the advantages beam quality,1micron resolution, which was about10times than that of traditional products, the marking speed was3-5times higher than traditional products, and low power consumption. In addition, the price of fiber lasers was greatly reduced, which was possible for us to use. In recent time, Cho SA and Sang-Hwan used the fiber laser to obtain good implant surface morphology and better osseointegration. It was worth to mention that the implant surface treatment development trended to a combination of different methods, such as the recent reported in sandblasting etching and micro-arc oxidation, micro-arc containing silver and non containing silver and other integrated approach between the surfactant and bone effects1. For example, the implant surface treated with the Sandblasted/acid-etched firstly, and laser processing secondly to change surface morphology to seek better methods. This year, Prodanov's study was the latest research in laser implant surface treatment reports, there was the comparative study of laser-treated with the Sandblasted/acid-etched. both of them obtained good osseointegration. It provided timely experimental basis and theoretical basis for further study in laser surface treatment.
In summary, although there had been numerous studies about the laser-treated and the Sandblasted/acid-etched, but a comparative studies about the laser plus acid etching with sandblasting plus etching were not seen in reports. Whether the osseointegration of Laser-treated/acid-etched surface had the some effect as or better than the Sandblasted/acid-etched surface was to be determined yet. Some relative comparative study on the urgent need; In addition, our research group had been done comparative study about Laser-treated/acid-etched surface and the acid-etched surface, we found that Laser-treated/acid-etched surface had good biocompatibility and osteoconduction. Based on some new developments of laser surface treatment, The study was intended to changed some conditions of the laser processing, narrowing the diameter of etching holes and combined with the acid-etched, and then made a comparative study with the sandblasted/acid-etched surface by animal experiments, so as to provid experimental basis and theoretical basis for in-depth study of the Laser-treated/acid-etched, to seek a better implant surface treatment technology eventually was also the experimental target.
Methods and materials
1. Preparation of laboratory samples:based on a large number of domestic and foreign literature and research purposes, using precision CNC machine tools to produce an experiment implant; By surface treatment method, according to pre-determined and related experimental parameters, prepared two good implant surfaces:the Laser-treated/acid-etched surface and the sandblasted/acid-etched surface.
2. The test of physical and chemical properties:measure the morphology, roughness, elemental composition with scanning electron microscopy, X-ray photoelectron spectroscopy and BMT Experi3D surface topography instrument, respectively, to assess the physical and chemical properties of SLA and LA surfaces.
3. Animal model and implantation:New Zealand rabbits were used in the study of different implantation sites of the proximal tibia to osseointegration, implant were installed in3different sites-,4beagle dogs as experimental animals. According to the principle of random allocation, two different surface implants were installed into each side of the tibia, numbered them from the top to down with No.1to8, each treatment group had4implants in one tibia. At a certain time interval, made some subcutaneous injection with a hypodermic calcein and tetracycline color marker, experimental animals were sacrificed at2,4weeks,1-4implants were used to production of bone tissue slices,5-8implant for torque measurement.
4. Making implant bone tissue sections and measurement of experimental:After decalcification, the anatomy of the rabbit tibia were made into paraffin section for HE staining observing; The bone tissues with implant were used to made sections of undecalcified, osseointegration growth rate and the BIC%was assessment and calculated by the fluorescence labeling and staining.
5. The measurement recorded torque measurement value and BIC%data, statistical analysis obtained experimental results.
Statistical analyses:
The experimental data are expressed as mean±standard deviation (SD). Statistical analysis was carried out by SPSS±v13.0software (SPSS Inc., Chicago, USA). For the single factor data, when meet the normal distribution and homogeneity of variance, using two independent sample t test and single factor variance to the overall mean comparison; When the data not meet the normal distribution or variance not neat homogeneous, using two independent sample t'test or rank sum test; For two factors of single variable data with normal distribution and homogeneity of variance, to analyse the main effect and interaction by analysis of variance of factorial design. Hypothesis test for two-sided test, the test level was0.05, probabilities (P)>0.05was considered to be no statistically significant; Probabilities (P)<0.05was considered to be statistically significant.
Results:
1. Through access to a large number of domestic and foreign literatures and combined the purpose of the present study, using precision CNC machine tools produced some experimental implant. According to pre-set surface treatment methods and experimental parameters, two different implant surfaces were made: Laser-treated/acid-etched surface and Sandblasted/acid-etched surface.
2. Results of the physical and chemical nature detecting:both of SLA and LA obtainted two rough structure, the surface roughnesses of LA was bigger than the SLA (LA:Ra=2.1μm; SLA:Ra=1.53μm)(P<0.01). Some melt was still obserted in LA surface, but the carbon material could been cleaned by the acid-etched; the sharp edge of the etching was visible on the surface of SLA, few oxide aluminum particles was still existed.
3. The different implantation sites in the selected20mm area of rabbit's tibias demonstrated different early osseointegration (P<0.05).
4. No significant difference was found in torque measurement value and the BIC%in two and four weeks between two groups (P>0.05), both groups obtained good osseointegration.
Conclusion:
In the present study, the different implantation sites in the selected20mm area of rabbit's tibias demonstrated different early osseointegration; the sites located7±1.5mm below the epiphyseal line were best suited for observing the effectiveness of early osseointegration among the3sites; The surface treatment methods of the Laser-treated/acid-etched and the Sandblasted/acid-etched could produce the rough surface, the former was more cleaner and more uniform than the latter, but both of them had good biocompatibility and promoted a good implant osseointegration, these two methods were feasible for implant surface treatment. Further studies were still needed.
引文
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