Pokorný D, Slouf M, Horák Z, Jahoda D, Entlicher G, Eklová S, Sosna A
Ortopedická Klinika 1, LF UK a FN, Motol, Praha.
Acta Chir Orthop Traumatol Cech. 2006 Aug;73(4):243-50.
Aseptic loosening of implants is the main complication affecting the longevity of joint prostheses. The highest proportion of loosening occurs due to osteolysis produced by the presence of ultra-high molecular weight polyethylene (UHMWPE) wear particles smaller than 1 microm. These can be identified by microscopic, spectroscopic or light-scattering methods. Here we describe our method for counting wear particles, based on the principle of light scattering.
Between 2002 and 2004, we collected samples of polyethylene granuloma in 19 patients who underwent revision total hip arthroplasty (THA) for aseptic loosening. The samples were obtained from strictly defined areas corresponding to the radiographic presentation of periprosthetic zones describes by Gruen and DeLee in THA. The frozen samples were lyophilized and subjected to delipidation and hydrolyzation procedures in 65 % HNO3. The top part of solution containing wear particles was blended with isopropanol, and the mixture was filtered through a 10-microm polycarbon membrane. Subsequently, the filtrate was filtered through a 0.1-microm membrane. Membranes with trapped particles, 0.1 to 1.0 microm in size, were sent for particle characterization and quantification. The number of wear particles was measured by the method based on light scattering with calibration (LSC), using a Beckman Coulter LS230 analyzer that can express particle size distribution in a given volume in percent. The method was based on the fact that each particle reflects rays that can be measured. The medium measured contained an unknown number of UHMWPE particles and a known number of calibration glass beads varying in size. The number of UHMWPE particles was calculated from the known number of calibration beads.
Because the collected samples were also used to develop the method, comprehensive data was obtained in six patients only. Particle distribution recorded in the periprosthetic zones in THA varied greatly; up to a five-fold difference in particle concentration was observed between the zones. In five of the six patients, the highest particle concentration was found in zone III.
Seeking a method that would be exact, quick and cheap and would eliminate particle aggregation remains the subject of study for researchers cooperating with clinical practice. At present methods based on weighing isolated particles are used most frequently. We developed the LSC method that, for quantification, utilizes the ability of particles to disperse light, and allows us to calculate the real numbers of UHMWPE wear particles in a medium containing a known number of calibration particles. Although this is an indirect method, it gives more accurate results than the direct weighing of particles. The advantages of the LSC method involve less demand on sample purity, greater speed and low limits of detection. The method is useful for statistical evaluation of a larger number of samples. The variation in particle distribution in THA found in this study is in agreement with the relevant literature data; it is also in agreement with our assumptions and clinical findings.
The authors developed an original method for assessment of UHMWPE wear particles in tissue samples, which is quicker than the methods so far used. In the periprosthetic tissues studied, particles about 1 microm in size were detected; their numbers (about 1010 particles per gram dry tissue) are in agreement with the literature data. The distribution of particles in periprosthetic zones in THA was uneven. The highest number of particles was found in the neighboring zone III and zone 7, as described by Gruen and DeLee. Key words: wear, polyethylene, total hip arthroplasty, light scattering, aseptic loosening, wear particles.
植入物的无菌性松动是影响关节假体使用寿命的主要并发症。松动的最大比例是由小于1微米的超高分子量聚乙烯(UHMWPE)磨损颗粒导致的骨溶解引起的。这些颗粒可以通过显微镜、光谱或光散射方法识别。在此,我们描述基于光散射原理的磨损颗粒计数方法。
在2002年至2004年期间,我们收集了19例因无菌性松动接受翻修全髋关节置换术(THA)患者的聚乙烯肉芽肿样本。样本取自与Gruen和DeLee在THA中描述的假体周围区域的影像学表现相对应的严格定义区域。将冷冻样本冻干,并在65%硝酸中进行脱脂和水解处理。含有磨损颗粒的溶液上层与异丙醇混合,混合物通过10微米的聚碳酸酯膜过滤。随后,滤液通过0.1微米的膜过滤。截留了尺寸在0.1至1.0微米之间颗粒的膜被送去进行颗粒表征和定量分析。使用能够以百分比表示给定体积内颗粒尺寸分布的贝克曼库尔特LS230分析仪,通过基于光散射校准(LSC)的方法测量磨损颗粒数量。该方法基于每个颗粒都会反射可测量光线这一事实。所测量的介质中含有数量未知的UHMWPE颗粒和数量已知、尺寸各异的校准玻璃珠。根据已知的校准珠数量计算UHMWPE颗粒的数量。
由于收集的样本也用于方法开发,仅在6例患者中获得了全面数据。THA中假体周围区域记录的颗粒分布差异很大;不同区域之间观察到颗粒浓度相差高达五倍。在6例患者中的5例中,III区颗粒浓度最高。
寻找一种准确、快速且廉价并能消除颗粒聚集的方法仍然是与临床实践合作的研究人员的研究课题。目前最常使用基于称量分离颗粒的方法。我们开发了LSC方法,该方法在定量时利用颗粒散射光的能力,并使我们能够计算含有已知数量校准颗粒的介质中UHMWPE磨损颗粒的实际数量。尽管这是一种间接方法,但它比直接称量颗粒能给出更准确的结果。LSC方法的优点包括对样品纯度要求较低、速度更快以及检测限低。该方法对于大量样本的统计评估很有用。本研究中发现的THA中颗粒分布的变化与相关文献数据一致;也与我们的假设和临床发现一致。
作者开发了一种评估组织样本中UHMWPE磨损颗粒的原创方法,该方法比迄今使用的方法更快。在所研究的假体周围组织中,检测到了约1微米大小的颗粒;其数量(约每克干组织10¹⁰个颗粒)与文献数据一致。THA中假体周围区域的颗粒分布不均匀。如Gruen和DeLee所描述,在相邻的III区和7区发现颗粒数量最多。关键词:磨损、聚乙烯、全髋关节置换术、光散射、无菌性松动、磨损颗粒