医用钛板表面涂层2-MPC对细菌生物膜形成的影响

目的探讨医用钛板表面涂层2-甲基丙烯酰氧基乙基磷酰胆碱(2-methacryloyloxyethyl phosphorylcholine,2-MPC)对金黄色葡萄球菌生物膜形成的影响。方法利用化学接枝法制备钛板表面2-MPC涂层并用扫描电镜观察其表面形貌,以金黄色葡萄球菌(ATCC25923)标准菌株作为实验用菌,将已消毒的2-MPC涂层钛板(实验组)及普通钛板(对照组)置入5 mL细菌浓度为1.5×10~6cfu/mL的菌液中共同培养。分别于体外培养后24 h、48 h行膜内活菌计数和扫描电镜观察。结果扫描电镜观察显示通过化学接枝法制备的2-MPC涂层钛板表面形貌光滑,钛板原先的纹理消失。体外培养24 h、48 h实验组钛板表面活菌计数数量均显著少于对照组(P0.05);扫描电镜观察发现24 h、48 h实验组钛板表面细菌黏附量均明显少于对照组(P0.05),且在48 h时实验组钛板表面菌膜形成量明显少于对照组。结论化学接枝法能有效将2-MPC涂层于钛板表面,且2-MPC涂层可以有效抑制钛板表面细菌黏附,预防钛板表面细菌生物膜的形成。     

载银硅灰石涂层抗菌性能实验研究

目的 通过动物实验验证载银硅灰石涂层体内抗菌性能.方法 将22只新西兰大白兔随机分为两组,随机取一侧股骨髁,试验组植入载银硅灰石钛板,对照组植入普通硅灰石钛板.术后第12天于术侧膝关节腔内注射金黄色葡萄球菌(ATCC25923)菌液108CFU.两组实验钛板的抗菌性能经实验兔肛温、体重变化,X线摄片结果,钛板、螺钉、骨...     

载银与载庆大霉素硅灰石涂层体外抗菌性能的对比研究

目的 制备具有长效药物缓释效果的骨科抗菌植入材料表面涂层.方法 采用等离子体喷涂技术在钛合金表面沉积硅灰石涂层.一组试样在浓度为重量百分率5%的硝酸银溶液中浸泡24 h获得载银硅灰石涂层,另一组试样通过与装载庆大霉素的胶原溶液发生接枝反应制得载庆大霉素硅灰石涂层.两种涂层的性能通过抗菌剂体外溶液释放实验、抗会黄色葡萄球菌的抑菌环实验和成骨细胞接触培养毒性实验进行表征.结果 成功制备载银与载庆大霉素硅灰石涂层.载银硅灰石涂层均匀释放银离子达50 d;涂层抗金黄色葡萄球菌能力可维持40 d以上.载庆大霉素硅灰石涂层呈爆发性释放庆大霉素,涂层抗会黄色葡葡球菌能力可以维持18 d.两种涂层均不影响成骨细胞在其表面的黏附、增殖以及碱性磷酸酶活性的表达.结论 载银硅灰石涂层与载庆大霉素硅厌石涂层相比,抗菌剂释放较均匀、抗菌时效较长,更具临床应用前景.     

反应时间对聚多巴胺/纳米银修饰多孔钛生物学性能的影响

目的 探讨制备新型钛基材料过程中,纳米银颗粒载入反应时间对其体外抗菌性能和细胞相容性等生物学性能的影响。方法 通过聚合作用以多巴胺在纳米孔钛表面构建聚多巴胺(PDA)膜层,通过调节纳米银颗粒载入反应时间(15 min,30 min,60 min),得到不同纳米银颗粒浓度的聚多巴胺修饰钛。利用扫描电镜(SEM)、细胞黏附、细胞毒性及杀菌率等检测方法,对载不同纳米银颗粒浓度的聚多巴胺修饰钛进行体外生物相容性及抗菌性能评价。结果 载入纳米银反应时间为30 min的钛基材料较其他反应时间的材料表现出良好的细胞黏附能力、较低的细胞毒性和满意的抗菌性能。结论 通过调整纳米银沉积反应时间,可调节材料表面银离子释放量,可得到既有良好细胞相容性,又有较强抗菌能力的聚多巴胺/纳米银修饰多孔钛。     

钛合金表面聚合氨基阳离子纳米薄膜的制备及抗菌性能的初步研究

目的制备钛合金表面聚合氨基阳离子纳米薄膜并初步评价其抗菌性能。方法通过低温等离子体聚合技术(Plasma)使烯丙胺单体于钛合金材料表面引发聚合,制备带表面涂层的钛合金样品。采用扫描电子显微镜(SEM)观察样品表面形貌,水接触角测量仪分析表面亲水性,X线光电子能谱仪(XPS)分析表面元素含量,体外抗菌实验测试样品对金黄色葡萄球菌及大肠杆菌的抑菌率。在新西兰大白兔胫骨的骨髓腔内注入金黄色葡萄球菌,然后置入带涂层的钛合金螺钉,观察体内抗菌实验效果。结果采用Plasma方法制备的带涂层的钛合金样品表面散在颗粒样聚合物,水接触角由不带涂层的对照组的100.3°±3.4°下降至13.6°±3.0°,差异具有统计学意义(P0.05);X线光电子能谱仪显示氮(N)元素含量由对照组的2.75%提高至11.80%。体外抗菌实验中,与对照组相比,涂层组对金黄色葡萄球菌的抑菌率为(86.1±3.8)%,对大肠杆菌的抑菌率为(75.5±2.6)%,差异具有统计学意义(P0.05)。动物实验中,与对照组相比,涂层组螺钉周围中性粒细胞明显减少。结论低温等离子体聚合技术可以在钛合金表面成功制备聚合氨基阳离子纳米薄膜,制品具有良好的抗菌性,为聚合阳离子进一步的抗菌研究提供基础。     

二氧化钛纳米管负载蒲公英多糖对牙龈卟啉单胞菌活性的影响

目的:通过阳极氧化二氧化钛纳米管负载不同浓度的蒲公英多糖,研究该复合涂层对牙龈卟啉单胞菌活性的影响,评价其抗菌性能。方法:设置阳极二氧化钛片为对照组(A组),阳极二氧化钛片负载不同浓度的蒲公英多糖为实验组:多糖浓度50mg/ml(B组),100mg/ml(C组),200mg/ml(D组)。场发射扫描电镜(FESEM)观察钛片表面形貌结构,X射线光电子能谱仪(XPS)检测钛片表面元素组成,台盼蓝染色荧光显微镜下计数活菌/死菌比例数。结果:FESEM下观察到各组钛片表面管径均一的纳米管阵列,管径80～100nm,实验组纳米管表面可见白色粟粒状颗粒沉积,D组C组B组;XPS显示C含量显著增加,特征性C峰值增高,D组C组B组;台盼蓝染色镜下观察牙龈卟啉单胞菌活菌呈现透明,死菌被染成蓝色,单视野下死菌数量D组C组B组A组,细菌活性(活菌/死菌×100%)D组C组B组A组,差异有统计学意义(P0.05)。结论:二氧化钛纳米管负载蒲公英多糖可抑制牙龈卟啉单胞菌的活性,该复合涂层有一定的抗菌效应。     

富血小板凝胶对金黄色葡萄球菌的体外抗菌性研究

目的探究人体富血小板凝胶(PRG)对金黄色葡萄球菌的体外抗菌作用;对比观察不同浓度富血小板凝胶对金黄色葡萄球菌的体外抗菌作用。方法分别取20例健康志愿者外周静脉血30 mnl,制备不同浓度富血小板凝胶,血小板浓度依次为1.8×10~(12)/L(PRG_1组)、1.2×10~(12)/L(PRG_2组)、0.6×10~(12)/L(PRG_3组)。通过体外抗菌实验观察不同浓度PRG在甲氧西林敏感金黄色葡萄球菌(MSSA)、耐甲氧西林金黄色葡萄球菌(MRSA)溶液中孵育1、2、4、8、12、24 h后的抗菌效果。结果在孵育开始后,各浓度PRG菌落计数明显少于PBS对照组,PRG对MRSA和MSSA均有明显的抗菌作用(P0.05),各浓度PRG抗菌作用均在孵育2 h后最明显,随后逐渐下降;同一时间PRG_1、PRG_2和PRG_3的抑菌率依次下降,与其含血小板浓度呈正相关关系,各组间菌落计数差异均有统计学意义(P0.05)。在孵育12 h后,细菌生长进入稳定期,各组抑菌率明显下降,各组菌落计数差异无统计学意义(P0.05)。结论在一定时间内,PRG对MSSA和MRSA均有明显的抑制作用,PRG的抗菌作用与血小板浓度呈正相关关系。     

载银纳米抗菌复合骨填充材料体外抗菌性能及生物相容性研究

[目的]探讨载银纳米抗菌复合骨填充材料其体外抗菌性能及生物相容性。[方法]采用抑菌环试验、菌落数试验检测材料对金黄色葡萄球菌及大肠杆菌的体外抗菌效果,采用扫描电镜观察材料对金黄色葡萄球菌及大肠杆菌的抗粘附效果。并采用MTT法检测细胞毒性及急性溶血实验评价载银纳米抗菌复合骨填充材料的生物相容性。[结果]抑菌环试验显示第1 d时载银纳米抗菌复合骨填充材料对金黄色葡萄球菌和大肠杆菌的抑菌环直径均最大,分别为(23.6±1.14)mm和(18.8±0.84)mm,随后抑菌环直径随时间延长而缩小,抑菌环在金黄色葡萄球菌和大肠杆菌中的持续时间分别为33、24 d;菌落数试验显示细菌与载银纳米抗菌复合骨填充材料接触24 h后,对金黄色葡萄球菌和大肠杆菌的抗菌率分别为94.18%和85.96%;扫描电镜发现实验组材料表面黏附的细菌明显少于对照组。MTT法测定示载银纳米抗菌复合骨填充材料毒性分级为1级,急性溶血实验示载银纳米抗菌复合骨填充材料溶血率为0.28%。[结论]载银纳米抗菌复合骨填充材料有良好的生物相容性,对金黄色葡萄球菌及大肠杆菌有明显抗菌作用,无明显细胞毒性和红细胞破坏性。     

聚维酮碘溶液对钛板表面金黄色葡萄球菌细菌生物膜的体外抗菌活性

目的观察两种不同浓度的聚维酮碘溶液对金黄色葡萄球菌(SA)细菌生物膜(BBF)的体外抗菌活性。方法在45枚钛板表面建立SA的BBF后,将其随机分成3组,每组15枚钛板。其中两组随机选取10枚钛板分别置于0.25%、0.5%浓度的聚维酮碘溶液中作用5 min,另外一组作为空白对照组;各组中的其余5枚钛板经超声裂解后做细菌培养,观察是否有细菌生长。3组活菌率比较以SPSS 17.0软件采用单因素方差分析,组内不同层的活菌率比较采用t检验。结果对照组培养24 h后成熟BBF中层活菌率为(67.4±1.1)%,底层活菌率为(39.4±2.2)%。中层和底层的BBF活菌率具有明显的统计学意义(t=35.81,P0.01)。0.5%、0.25%聚维酮碘溶液作用5 min后的BBF的中层、底层活菌率分别为(66.7±1.4)%,(38.2±3.2)%、(66.5±1.0)%,(39.1±2.4)%;0.5%聚维酮碘溶液作用5 min后的BBF的中层、底层活菌率与0.25%聚维酮碘溶液作用5 min后的BBF的中层、底层活菌率和空白对照组相比差异都无统计学意义(P0.05)。对照组、0.25%和0.5%聚维酮碘组钛板经超声裂解后的血平板细菌培养均可见SA菌落形成。结论 0.25%、0.5%浓度的聚维酮碘溶液对钛板表面SA的BBF作用5 min无明显抗菌活性。     

能谱CT分析进展期胃癌碘含量与脉管癌栓的相关性

目的应用能谱CT增强扫描分析进展期胃癌碘含量与脉管癌栓的关系。方法对40例经病理证实的进展期胃癌患者于术前2周内行能谱CT三期增强扫描,应用物质分离技术观察碘基图上增强扫描各期肿瘤黏膜侧高强化区及整体碘含量(ICs)、相同层面腹主动脉碘含量(ICao),计算标准化碘含量(NICs)、静脉期碘含量升高率(ICERvp),分析其与脉管癌栓的相关性。结果脉管癌栓阳性者胃癌动脉期黏膜侧高强化区、整体碘含量明显高于阴性者[(3.27±0.91)mg/ml vs(2.32±0.79)mg/ml,P=0.004;(1.98±0.65)mg/ml vs(1.41±0.75)mg/ml,P=0.023];标准化后脉管癌栓阳性者碘含量仍明显高于阴性者[(0.30±0.08)mg/ml vs(0.23±0.07)mg/ml,P=0.013;(0.18±0.06)mg/ml vs(0.14±0.07)mg/ml,P=0.037];阴性者黏膜侧高强化区及整体ICERvp明显高于脉管癌栓阳性者(0.39±0.26vs 0.12±0.26,P=0.005;0.55±0.46vs 0.22±0.28,P=0.047)。结论不同增强时相中胃癌碘含量与脉管癌栓存在一定相关性。     

Effect on infection resistance of a local antiseptic and antibiotic coating on osteosynthesis implants: an in vitro and in vivo study.

The purpose of this study was to acquire information about the effect of an antibacterial and biodegradable poly-L-lactide (PLLA) coated titanium plate osteosynthesis on local infection resistance. For our in vitro and in vivo experiments, we used six-hole AO DC minifragment titanium plates. The implants were coated with biodegradable, semiamorphous PLLA (coating about 30 microm thick). This acted as a carrier substance to which either antibiotics or antiseptics were added. The antibiotic we applied was a combination of Rifampicin and fusidic acid; the antiseptic was a combination of Octenidin and Irgasan. This produced the following groups: Group I: six-hole AO DC minifragment titanium plate without PLLA; Group II: six-hole AO DC minifragment titanium plate with PLLA without antibiotics/antiseptics; Group III: six-hole AO DC minifragment titanium plate with PLLA + 3% Rifampicin and 7% fusidic acid; Group IV: six-hole AO DC minifragment titanium plate with PLLA + 2% Octenidin and 8% Irgasan. In vitro, we investigated the degradation and the release of the PLLA coating over a period of 6 weeks, the bactericidal efficacy of antibiotics/antiseptics after their release from the coating and the bacterial adhesion of Staphylococcus aureus to the implants. In vivo, we compared the infection rates in white New Zealand rabbits after titanium plate osteosynthesis of the tibia with or without antibacterial coating after local percutaneous bacterial inoculations at different concentrations (2 x 10(5)-2 x 10(8)): The plate, the contaminated soft tissues and the underlying bone were removed under sterile conditions after 28 days and quantitatively evaluated for bacterial growth. A stepwise experimental design with an "up-and-down" dosage technique was used to adjust the bacterial challenge in the area of the ID50 (50% infection dose). Statistical evaluation of the differences between the infection rates of both groups was performed using the two-sided Fisher exact test (p < 0.05). Over a period of 6 weeks, a continuous degradation of the PLLA coating of 13%, on average, was seen in vitro in 0.9% NaCl solution. The elution tests on titanium implants with antibiotic or antiseptic coatings produced average release values of 60% of the incorporated antibiotic or 62% of the incorporated antiseptic within the first 60 min. This was followed by a much slower, but nevertheless continuous, release of the incorporated antibiotic and antiseptic over days and weeks. At the end of the test period of 42 days, 20% of the incorporated antibiotic and 15% of the incorporated antiseptic had not yet been released from the coating. The antibacterial effect of the antibiotic/antiseptic is not lost by integrating it into the PLLA coating. The overall infection rate in the in vivo investigation was 50%. For Groups I and II the infection rate was both 83% (10 of 12 animals). In Groups III and IV with antibacterial coating, the infection rate was both 17% (2 of 12 animals). The ID50 in the antibacterial coated Groups III and IV was recorded as 1 x 10(8) CFU, whereas the ID50 values in the Groups I and II without antibacterial coating were a hundred times lower at 1 x 10(6) CFU, respectively. The difference between the groups with and without antibacterial coating was statistically significant (p = 0.033). Using an antibacterial biodegradable PLLA coating on titanium plates, a significant reduction of infection rate in an in vitro and in vivo investigation could be demonstrated. For the first time, to our knowledge, we were able to show, under standardized and reproducible conditions, that an antiseptic coating leads to the same reduction in infection rate as an antibiotic coating. Taking the problem of antibiotic-induced bacterial resistance into consideration, we thus regard the antiseptic coating, which shows the same level of effectiveness, as advantageous.     

Prevention of pin tract infection with iodine-supported titanium pins

Background

Pin tract infection is one of the most common complications of external fixation. We developed techniques to coat titanium implant surfaces with iodine. This study clinically evaluated the infection-preventive effects and biological safety of iodine-coated external fixation pins.

Patients and methods

Iodine-supported pins were placed in 39 limbs of 38 patients. The mean age of the patients was 33.6 years. Twenty-six patients were men and 12 were women. In all patients, the iodine-coated pins were used to prevent infection. There were 476 pin insertion sites. Pin sites were classified according to the Checketts–Otterburn classification (grade 1–6). White blood cells (WBC) and C-reactive protein (CRP) were measured pre- and postoperatively in all patients. To confirm whether iodine from the implant affected physiological functions, thyroid hormone levels in the blood were monitored. The change in the amount of iodine deposited in the body over time was calculated by examining the removed pins.

Results

External fixation was used for a mean duration of 6 months. Grade 1 infection was found in 2.5 % of patients, and grade 2 infection in 1.1 %. There was no patient with an infection of grade 3 or higher. Median WBC levels were in the normal range, and median CRP levels returned to <0.3 mg/dl within 3 weeks after surgery. Abnormalities of thyroid gland function were not detected. The amount of iodine was maintained for a long time, with approximately 40 % remaining after 1 year.

Conclusions

Iodine-supported titanium pins were able to decrease the pin tract infection rate and had no impact on thyroid function. These results suggest that iodine-coated titanium pins are biologically safe and effective at preventing pin tract infections.     

Antibacterial Activity in Iodine-coated Implants Under Conditions of Iodine Loss: Study in a Rat Model Plus In Vitro Analysis

BackgroundWe developed iodine-coated titanium implants to suppress microbial activity and prevent periprosthetic joint infection (PJI); their efficacy was demonstrated in animal and in vitro models. The iodine content in iodine-coated implants naturally decreases in vivo. However, to our knowledge, the effect of reduced iodine content on the implant’s antimicrobial activity has not been evaluated to date.Questions/purposes(1) How much does the iodine content on the implant surface decrease after 4 and 8 weeks in vivo in a rat model? (2) What effect does the reduced iodine content have on the antimicrobial effect of the implant against multiple bacteria in an in vitro model?MethodsThis experiment was performed in two parts: an in vivo experiment to determine attenuation of iodine levels over time in rats, and an in vitro experiment in which we sought to assess whether the reduced iodine content observed in the in vivo experiment was still sufficient to deliver antimicrobial activity against common pathogens seen in PJI. For the in vivo experiment, three types of titanium alloy washers were implanted in rats: untreated (Ti), surface-anodized to produce an oxide film (Ti-O), and with an iodine layer on the oxidation film (Ti-I). The attenuation of iodine levels in rats was measured over time using inductively coupled plasma-mass spectrometry. Herein, only the Ti-I washer was used, with five implanted in each rat that were removed after 4 or 8 weeks. For the 4- and 8-week models, two rats and 15 washers were used. For the in vitro study, to determine the antibacterial effect, three types of washers (Ti, Ti-O, and Ti-I) (nine washers in total) were implanted in each rat. Then, the washers were removed and the antibacterial effect of each washer was examined on multiple bacterial species using the spread plate method and fluorescence microscopy. For the spread plate method, six rats were used, and five rats were used for the observation using fluorescence microscopy; further, 4- and 8-week models were made for each method. Thus, a total of 22 rats and 198 washers were used. Live and dead bacteria in the biofilm were stained, and the biofilm coverage percentage for quantitative analysis was determined using fluorescence microscopy in a nonblinded manner. Ti-I was used as the experimental group, and Ti and Ti-O were used as control groups. The total number of rats and washers used throughout this study was 24 and 213, respectively.ResultsIodine content in rats implanted with Ti-I samples decreased to 72% and 65% after the in vivo period of 4 and 8 weeks, respectively (p = 0.001 and p < 0.001, respectively). In the in vitro experiment, the Ti-I implants demonstrated a stronger antimicrobial activity than Ti and Ti-O implants in the 4- and 8-week models. Both the median number of bacterial colonies and the median biofilm coverage percentage with live bacteria on Ti-I were lower than those on Ti or Ti-O implants for each bacterial species in the 4- and 8-week models. There was no difference in the median biofilm coverage percentage of dead bacteria. In the 8-week model, the antibacterial activity using the spread plate method had median (interquartile range) numbers of bacteria on the Ti, Ti-O, and Ti-I implants of 112 (104 to 165) × 10⁵, 147 (111 to 162) × 10⁵, and 55 (37 to 67) × 10⁵ of methicillin-sensitive Staphylococcus aureus (Ti-I versus Ti, p = 0.026; Ti-I versus Ti-O, p = 0.009); 71 (39 to 111) × 10⁵, 50 (44 to 62) × 10⁵, and 26 (9 to 31)× 10⁵ CFU of methicillin-resistant S. aureus (Ti-I versus Ti, p = 0.026; Ti-I versus Ti-O, p = 0.034); and 77 (74 to 83) × 10⁶, 111 (95 to 117) × 10⁶, and 30 (21 to 45) × 10⁶ CFU of Pseudomonas aeruginosa (Ti-I versus Ti, p = 0.004; Ti-I versus Ti-O, p = 0.009). Despite the decrease in the iodine content of Ti-I after 8 weeks, it demonstrated better antibacterial activity against all tested bacteria than the Ti and Ti-O implants.ConclusionIodine-coated implants retained their iodine content and antibacterial activity against methicillin-sensitive S. aureus, methicillin-resistant S. aureus, and P. aeruginosa for 8 weeks in vivo in rats. To evaluate the longer-lasting antibacterial efficacy, further research using larger infected animal PJI models with implants in the joints of both males and females is desirable.Clinical RelevanceIodine-coated titanium implants displayed an antibacterial activity for 8 weeks in rats in vivo. Although the findings in a rat model do not guarantee efficacy in humans, they represent an important step toward clinical application.     

Iodine-supported titanium implants have good antimicrobial attachment effects

BackgroundWe have developed iodine-supported titanium implants, which were shown to have good anti-bacterial effects for Methicillin-sensitive Staphylococcus aureus (MSSA) in our past basic research. However, PJI can be caused by various bacteria including MRSA, Pseudomonas aeruginosa, MSSE, and fungus. The purpose of this study was to investigate whether these implants also have good antibacterial attachment effects for MRSA, P. aeruginosa, MSSE, and fungus.MethodsTi–6Al–4V titanium plates were either left untreated (Ti), treated with oxide film on the Ti surface by anodization (Ti–O), or treated with an iodine coating on oxidation film (Ti–I). The antibacterial activity of the TiI was measured by experimental methods according to Japanese Industrial Standard (JIS) protocols. Implants in this study were exposed to MRSA (ATCC43300), P. aeruginosa (ATCC27853), MSSE (ATCC35984), and Candida Albicans (ATCC10231). Colonies were counted immediately after the bacteria attached to the metal surface and again after 24 h incubation. The difference in the number of bacteria on each metal plate was statistically investigated and an antibacterial activity value was calculated. An effective antibacterial active value of more than 2.0 was judged to be effective according to JIS protocol.ResultsNo countable viable bacteria were observed on the Ti–I surface. For all bacteria there was a significant difference in the mean number of viable bacteria between Ti–I and Ti or Ti–O. Antibacterial activity value in Ti–I and Ti–O was more than 5.9 and 3.6 respectively for MRSA, more than 2.8 and zero for P. aeruginosa, more than 4.3 and zero for MSSE, and more than 4.7 and zero for C. Albicans.ConclusionsThis study showed that iodine-supported titanium implants have good antimicrobial attachment effects for MRSA, P. aeruginosa, MSSE, and C. Albicans. Iodine-supported titanium implants could have great potential as innovative antibacterial implants that can prevent early onset periprosthetic joint infection.     

壳聚糖表面改性钛合金材料的研究进展

钛合金材料生物性能良好,是骨科常用的内植入材料,但其骨整合性及抗菌性能较差,需进行表面改性以弥补其不足之处。壳聚糖具有良好的生物相容性及成膜性,且可作为载体将目标药物引入钛合金表面,可有效改善钛合金材料的生物学性能,增加其使用范围。本文对近几年壳聚糖表面改性钛合金材料的相关研究进行归纳总结,从壳聚糖涂层改性的方式、钛合金材料成骨性及抗菌性的改善3个方面展开论述,以期为钛合金材料涂层改性在临床中的应用提供指导依据。     

纯钛种植体表面不同化学组成微弧氧化膜的结构与成分分析

目的：探讨采用微弧氧化技术在不同电解液中对纯钛种植体表面进行生物改性。方法：试件预处理后分别在不同化学组成的电解液中进行微弧氧化处理。扫描电镜（SEM）观察微弧氧化膜层的表面形貌，X射线能谱（EDX）和X射线衍射（XRD）仪分析膜层的元素构成和晶相结构。结果：电镜观察表明在2种电解液中试件表面都形成了内层致密，外层多孔的陶瓷膜；能谱分析证实这2种陶瓷膜分别由Ti、Ca、P、0 4种元素和Ti、Mg、03种元素组成；X射线衍射表明陶瓷膜由锐钛矿型与金红石型TiO2及钙钛矿和钙镁矿组成。结论：通过不同电解液中的微弧氧化处理在纯钛试件表面形成了内层致密，外层多孔的TiO2陶瓷膜，并分别含有Ca，P和Mg活性成分，在提高材料耐磨性，耐腐蚀性的同时，提高了生物活性。     

仿生电活性涂层钛植入材料促进骨结合的实验研究

目的评价仿生电活性钛酸钡(BaTiO3,BTO)/聚偏氟乙烯-三氟乙烯P(VDF-TrFE)涂层钛植入材料促进骨结合性能的效果。方法首先将直径为2 mm、长度为5 mm的医用纯钛圆柱进行表面喷砂-酸蚀处理,然后将BaTiO3/P(VDF-TrFE)溶液均匀涂覆在钛柱表面。待烘干后对涂层表面进行电晕极化处理。采用扫描电镜、能谱分析、原子力显微镜、水接触角测量仪分别对材料表面形貌、元素组成、表面粗糙度和亲疏水性进行表征检测,PLLA涂层钛柱作为对照材料。选取实验兔4只,在双侧胫骨位置各制备3个间隔为1 cm的圆形缺损,在左侧胫骨植入PLLA涂层钛柱,右侧胫骨植入仿生电活性涂层钛柱,术后4周和12周分别取材进行硬组织的骨形态检测分析。采用SPSS15.0软件对数据进行统计分析。结果理化性能表征检测显示,BaTiO3/P(VDF-TrFE)涂层和PLLA涂层均匀附着在钛柱表面,涂层厚度约50μm,且表面结构致密。电活性涂层可见钛酸钡纳米颗粒均匀分布在P(VDF-TrFE)基体内。两种涂层表面的粗糙度和水接触角无明显差异。电活性涂层具有稳定的压电性能,且压电常数接近生理量级。动物实验显示,术后4周,仿生电活性涂层材料表面和新骨结合紧密,涂层材料稳定无降解;而PLLA涂层材料表面由于材料有部分降解导致新骨结合较差,电活性涂层的骨结合率明显高于PLLA涂层。术后12周,两组的新骨成熟程度均增加,骨陷窝明显,仿生电活性涂层仍然保持稳定状态;而PLLA涂层进一步发生降解,和新骨结合程度弱于电活性涂层组。结论仿生电活性BaTiO3/P(VDF-TrFE)涂层钛可能作为一种具有促进骨整合功能的种植体涂层材料。     

钽涂层对骨髓间充质干细胞的黏附增殖及成骨分化的影响

目的探讨钽涂层金属在体外促进大鼠骨髓间充质干细胞(BMSCs)黏附、增殖及成骨分化方面的作用。方法在体外取6只6周龄SD大鼠BMSCs进行原代培养至第3代,使用化学气相沉积系统自行制备钽涂层金属。然后进行流式细胞术鉴定、BMSCs三向诱导、荧光染色、细胞增殖检测及实时荧光定量聚合酶链反应技术(Q-PCR)试验。观察比较BMSCs在钛合金圆片(Ti6Al4V组)和钽金属圆片(Ta组)表面黏附的BMSCs数量、增殖速率、成骨细胞特异性转录因子(OSX)、Runt相关转录因子2(RUNX2)、骨粘连蛋白(OSN)和骨桥蛋白(OPN)的表达情况。结果流式细胞术结果显示CD44(94.55%)、CD90(95.01%)、CD34(0.06%)。诱导成骨分化14 d后碱性磷酸酶(ALP)染色阳性;诱导成骨分化21 d后出现茜素红钙化结节;成脂诱导21 d后油红O染色呈阳性;成软骨诱导21 d后阿利新蓝染色评估有软骨形成能力。激光共聚焦显微镜结果显示BMSCs在Ti6Al4V组和Ta组金属圆片表面贴附生长,细胞间互相接触、聚集成片,Ta组金属圆片上黏附的BMSCs数量明显多于Ti6Al4V组,并且具有更好的延展性能。细胞增殖检测结果发现,分别共培养1、3、5、7 d后Ta组BMSCs的增殖速率明显快于Ti6Al4V组,差异均有统计学意义(P<0.05)。Q-PCR结果发现,与Ti6Al4V组相比,体外培养7 d后Ta组金属圆片更能促进OSN和OPN的表达,差异有统计学意义(P<0.05);体外共培养21 d后,Ta组金属圆片更能促进OSX、RUNX2、OSN和OPN的表达,差异均有统计学意义(P<0.05)。结论相比于传统的骨科植入物钛合金而言,钽涂层金属能够更好地促进BMSCs的黏附,增殖和成骨分化。     

颈椎前路微型记忆加压合金板的体外生物力学研究

目的:采用体外生物力学研究方法评估颈椎前路椎间盘切除融合内固定手术的微型记忆加压合金板(GYZ记忆合金板)置入后的即刻稳定性。方法:6具新鲜颈椎尸体标本,男性,年龄55.2±7.1岁,身高175.4±3.8cm,体重72.3±6.0kg。通过薄层CT及骨密度检查,排除畸形、骨折、骨质疏松等疾病。所有标本剔除脊柱周围血管、肌肉等软组织,保留韧带、关节囊后,先后制作为生理组、损伤组、新型合金板组、传统钛板组4组:生理组为空白对照组;损伤组标本仅将C3/4椎间盘破坏;新型合金板组将C3/4椎间盘切除后置入椎间融合器,前路辅助GYZ记忆合金板融合固定;传统钛板组前路辅助传统钛板固定。每组标本均在前屈、背伸、左右侧弯、左右扭转方向上给予1.5N·m的载荷。记录第3次试验的结果并对四组标本C3/4椎间隙在前屈、背伸、左右侧弯及左右扭转方向上的活动度进行对比分析。结果:术后影像学检查显示置入物位置良好。生理组C3/4在前屈、背伸、左侧弯、右侧弯、左旋转、右旋转方向上的活动度分别为4.8°±0.5°、2.5°±0.1°、2.4°±0.3°、2.6°±0.3°、2.0°±0.2°与3.9°±0.1°;损伤组分别为6.0°±0.7°、3.4°±0.3°、3.4°±0.5°、3.8°±0.6°、2.4°±0.9°与5.7°±1.3°,较生理组明显增大(P<0.05)。新型合金板组C3/4椎间隙在上述六个方向上的活动度分别为0.6°±0.1°、0.4°±0.1°、0.5°±0.1°、0.7°±0.1°、0.6°±0.1°及1.0°±0.1°,传统钛板组分别为0.6°±0.1°、0.5°±0.1°、0.5°±0.1°、0.7°±0.1°、0.5°±0.1°与1.0°±0.1°,两组在各方向上的活动度无明显差异(P>0.05)。结论:微型记忆合金板可以获得与传统钛板相似的固定强度和重建手术部位的即刻稳定性。     

零切迹桥形锁定融合器与前路cage联合钛板固定治疗脊髓型颈椎病的疗效比较

目的探讨零切迹桥形锁定融合器(ROI-C)与前路cage联合钛板固定治疗双、三节段脊髓型颈椎病的疗效差异。方法回顾性分析2017年6月至2019年10月接受手术治疗的85例双、三节段脊髓型颈椎病患者的病历资料,男43例,女42例;年龄(52.3±8.0)岁(范围28~66岁);双节段63例、三节段22例。采用ROI-C治疗45例(ROI-C组),采用前路cage联合钛板固定40例(钛板组)。主要观察指标包括手术时间、术中出血量、颈椎Cobb角、融合节段Cobb角、平均椎间高度、疼痛视觉模拟评分(visual analogue scale,VAS)、日本骨科协会(Japanese Orthopaedic Association,JOA)评分和颈椎功能障碍指数(neck disability index,NDI)。结果85例患者随访时间为(16.9±2.0)个月(范围12~22个月)。双节段ROI-C组手术时间为(110.37±8.25)min,较钛板组(139.5±10.54)min短;术中出血量为(15.74±8.10)ml,较钛板组(23.71±9.70)ml少。三节段ROI-C组手术时间为(130.00±5.70)min,较钛板组(162.83±5.59)min短;而术中出血量的差异无统计学意义。术后1年ROI-C组双、三节段颈椎Cobb角分别为15.31°±1.55°、15.20°±0.42°,优于术前11.23°±2.03°、9.20°±1.14°;钛板组为15.89°±1.13°、16.08°±1.88°,优于术前11.25°±2.01°、9.00°±1.60°;术前及术后1年两组间的差异均无统计学意义。术后1年ROI-C组双、三节段VAS评分分别为(1.83±0.66)分、(2.60±0.52)分,低于术前(7.49±0.51)分、(7.60±0.52)分;钛板组为(1.79±0.50)分、(2.41±0.51)分,低于术前(7.61±0.63)分、(7.42±0.52)分;术前及术后1年两组间的差异均无统计学意义。术后1年ROI-C组双、三节段JOA评分分别为(15.00±0.84)分、(14.70±0.95)分,优于术前(7.20±0.87)分、(6.60±1.27)分;钛板组为(15.29±0.85)分、(14.83±0.58)分,优于术前(6.89±1.03)分、(6.92±0.67)分;术前及术后1年两组间的差异均无统计学意义;两组术后JOA改善率均为优。ROI-C组术后发生吞咽困难1例(2.22%,1/45),钛板组发生吞咽困难8例(20.00%,8/40),发生率的差异有统计学意义(χ2=5.32,P=0.02)。结论ROI-C与前路cage联合钛板固定治疗双、三节段脊髓型颈椎病均可取得良好的近期临床疗效,但采用ROI-C手术时间较短、术后吞咽困难发生率低。