人工关节无菌性松动动物模型的建立和二膦酸盐复合HA涂层假体的抗假体周围骨溶解作用

发布时间：2018-02-26 22:06

本文关键词： 人工关节无菌性松动骨溶解动物模型二膦酸盐涂层　出处：《第四军医大学》2009年硕士论文　论文类型：学位论文

【摘要】： 关节置换手术随着人群的老龄化逐渐增加。随之而来就出现了一个大问题,即越来越多的患者需要进行至少一次的返修手术。翻修的首要原因除了感染就是假体无菌性松动。以UHMWPE颗粒为主,引起的假体周围骨溶解是无菌性松动的首要原因。许多研究已经指出非骨水泥假体的寿命很大程度上取决于生物学固定效果。假体的早期移动会增加假体无菌性松动的危险。因此假体的初始稳定性至关重要,迅速而持续的骨长入可以为假体提供良好的生物学固定,预防假体移动。同时,假体周围新生骨组织的密度越高,骨-假体界面抵抗磨屑颗粒引起的骨溶解作用越强。 HA涂层可以增强假体的稳定性、骨-假体界面的结合强度和抗疲劳性能、骨矿化水平和骨长入速率。同时二膦酸盐类药物可以改善假体的生物学固定效果。最近很多实验关注将二膦酸盐与假体结合,结果表明二膦酸盐复合涂层可以刺激假体周围新骨的形成和力学性能的改善。目前需要研究的问题是: 1.缺少一种既符合人工关节假体无菌性松动病理特点又简便有效的动物模型。 2.阿仑膦酸钠复合HA涂层的非多孔钛假体是否可以抑制UHMWPE颗粒诱导的假体周围骨溶解。 3.阿仑膦酸钠在预防磨屑诱导的假体周围骨溶解过程中是否能够影响成骨细胞的活性。 4.阿仑膦酸钠的局部释放是否具有于系统性治疗效果。 5.阿仑膦酸钠复合HA涂层假体是否能够作为一种缓释载体为假体固定提供支持。因此,本实验通过UHMWPE颗粒诱导建立了新西兰大白兔的人工关节无菌性松动动物模型。同时在此模型基础上,通过病理学、影像学、骨组织形态学、生物力学等方面的观测,进一步研究以上问题。具体内容如下: 1.X线测量动物处死后即刻拍摄双侧胫骨正侧位X线,观察左侧胫骨假体周围骨溶解的影像学改变,并测量双侧胫骨髓腔峡部骨皮质厚度。 2.病理学检测膝关节滑膜HE染色,分别用普通光镜和偏振光镜观察滑膜结构,包括是否增生肥厚、是否有炎细胞浸润以及UHMWPE颗粒是否被滑膜组织包绕吞噬等。 3.生物力学测量通过假体拔出实验(pull-out test)描绘负荷-位移曲线,采集好负荷-位移曲线后计算极限抗剪强度、表观抗剪强度和总能量吸收值。 4.骨组织形态学测量进行四环素荧光双标记,分别通过荧光切片和甲苯胺蓝染色切片来计算骨组织形态学动态和静态指标。依照ASBMR命名法来计算以下参数:骨密度(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)、类骨质表面(OS/BS)、类骨质厚度(O.Th)以及骨形成速率(BFR/BS)、松质骨骨矿沉积率(MAR)和皮质骨骨矿沉积率(E-MAR)。通过改良丽春红三色切片来计算骨-假体接触率和骨体积分数两项指标。通过VON KOSSA染色切片来计算骨皮质处钙盐沉积岛平均面积、钙盐沉积岛数量和钙盐沉积岛面积分数(钙盐沉积岛总面积/骨组织面积)。 5.扫描电镜测量分别在普通模式和背散射模式下观测。左侧胫骨标本分别测量距离假体表面200μm、200-500μm和500-800μm处的骨体积分数,以及距离骨内膜表面800μm处的骨体积分数,右侧测量距离骨内膜表面800μm处的骨体积分数并于普通模式下观测标本表面显微形态。 6.Ash Weight 测定右侧尺骨在灼烧前后的重量,标本称湿重(Ww),然后于密闭坩埚内480℃烧24h,称干重(Wa),计算干湿比(Wa/Ww),以此反映标本内钙盐含量。 7.DEXA 测量第2腰椎骨密度值。 8.阿仑膦酸钠体外释放周期绘制阿仑膦酸钠体外释放曲线,用来评估此新型复合涂层假体的药物缓释能力。以上检测结果均显示阿仑膦酸钠的局部应用可以增加术侧和对侧的骨皮质厚度,骨小梁参数,成骨细胞活性,骨矿化水平和骨密度,并能显著增加术侧骨皮质厚度,假体生物力学固定强度,骨-假体接触率,假体周围骨体积分数,骨矿化水平和假体周围骨密度。结论: 1.我们建立的新西兰大白兔人工关节无菌性松动动物模型无论从病理机制还是造模效果上都能为研究相关课题提供基础,并且造模简便可靠。 2.在体内,阿仑膦酸钠复合HA涂层的非多孔钛假体可以明显抑制UHMWPE颗粒诱导的假体周围骨溶解,甚至比在没有磨屑颗粒的情况下单独使用HA涂层假体取得的效果都要好。 3.在体内,阿仑膦酸钠在预防磨屑诱导的假体周围骨溶解过程中能够促进成骨细胞的活性。 4.阿仑膦酸钠的局部释放具有于系统性治疗效果。 5.阿仑膦酸钠复合HA涂层作为一种缓释载体,一方面能够为假体的初始稳定性提供早期短时间的高药物浓度释放,另一方面又能够提供长时间的低浓度药物缓释。
[Abstract]:Joint replacement surgery increased gradually with the aging of the population. It will be a big problem that more and more patients need surgical repair. At least one of the leading causes of revision surgery in addition to infection is aseptic loosening. The UHMWPE particles mainly caused by periprosthetic osteolysis is the primary cause of aseptic loosening. Many studies have pointed out that the life of non bone cement prosthesis depends largely on the biological fixation. Early mobile prosthesis would increase the risk of aseptic loosening of the prosthesis. The prosthesis initial stability is crucial, rapid and sustained bone ingrowth canprovide good biological fixation, prevention of prosthesis movement. At the same time, around the prosthesis of newborn the higher the density of bone tissue, bone prosthesis interface resistance effect of wear debris induced osteolysis is stronger.
HA coating can enhance the stability of the prosthesis, bone prosthesis interface bonding strength and anti fatigue performance, bone mineralization and bone ingrowth rate. The effect of biological fixation and bisphosphonates can improve the prosthesis. Recently a lot of experiments concerning bisphosphonates and prosthesis combined with, results show that the composite coating can stimulate the formation of bisphosphonates and mechanical properties around the prosthesis the new bone improvement. The current need to study the problem is:
1. there is a lack of an animal model which is not only in accordance with the pathological characteristics of aseptic loosening of artificial joint prosthesis, but also simple and effective.
Whether the non porous titanium prosthesis 2. alendronate HA composite coating can inhibit UHMWPE particles induced osteolysis.
3. of alendronate sodium in the prevention of grinding affect the activity of osteoblasts to periprosthetic osteolysis induced by process chip.
Whether the local release of 4. alendronate has a systemic therapeutic effect.
5. alendronate compound HA coated prosthesis can provide support for prosthesis fixation as a kind of sustained-release carrier.
Therefore, this experiment established by the artificial joint aseptic loosening animal model in New Zealand rabbits by UHMWPE particles. Based on this model, the pathology, imaging, histological observation, biomechanical aspects of the further research on the above problems. The specific contents are as follows:
1. X ray measurement
The animal were sacrificed immediately after the shooting of bilateral tibial lateral X ray imaging observation, left tibial periprosthetic osteolysis pathology, and measurement of bilateral tibial bone marrow cavity isthmus thickness of cortical bone.
2. pathological examination
The synovial membrane of knee joint was stained with HE. The synovial membrane structure was observed by ordinary light microscope and polarized light microscope, including whether hypertrophy, inflammatory cell infiltration and UHMWPE particles were surrounded by synovial tissue.
3. biomechanical measurement
The load displacement curve is depicted through the prosthesis pull out test (pull-out test), and the ultimate shear strength, apparent shear strength and total energy absorption value are calculated after the load displacement curve is collected.
4. morphologic measurement of bone histomorphology
For tetracycline double labeling respectively to calculate the bone morphology index and the static and dynamic slicing through fluorescence sectioning and toluidine blue staining. According to ASBMR nomenclature to calculate the following parameters: bone mineral density (BV/TV), Liang Houdu (Tb.Th), trabecular bone trabecular bone volume (Tb.N), osteoid surface (OS/BS), osteoid thickness (O.Th) and bone formation rate (BFR/BS), cancellous bone mineral deposition rate (MAR) and cortical bone mineral deposition rate (E-MAR).
Two indexes of bone prosthesis contact rate and bone volume fraction were calculated by improving the tricolor slice of Li Chun Hong.
VON KOSSA staining slices were used to calculate the average area of calcium salt deposition Island, the number of calcium deposition islands and the area of calcium deposition island area (the total area of calcium deposition Island / bone tissue area).
5. scanning electron microscopy
Were observed in the normal mode and the back scattering mode. The left tibia specimens were measured from the surface of the artificial bone volume fraction 200 m, 200-500 m and 500-800 m, and the distance between the endosteal surface 800 m bone volume fraction, bone volume fraction on the right side of the measurement distance the endosteal surface of 800 m. Observation of surface morphology and specimens in normal mode.
6.Ash Weight
The weight of the right ulna is measured before and after burning. The specimen is called wet weight (Ww), then 24h is burned at 480 degree C in the closed crucible, and the dry weight (Wa) is calculated. The dry wet ratio (Wa/Ww) is calculated to reflect the calcium content in the specimen.
7.DEXA
The BMD of the second lumbar vertebrae was measured.
In vitro release cycle of 8. alendronate
The release curve of alendronate in vitro was designed to evaluate the drug release ability of the new composite coating prosthesis.
The results above showed that local application of alendronate can increase the operative side and contralateral cortical bone thickness, trabecular bone parameters, the activity of osteoblasts, bone mineralization and bone density, and can significantly increase the operative side of cortical bone thickness, the fixation strength of prosthesis biomechanics, bone implant contact, prosthesis around the bone volume fraction, bone density around the prosthesis and bone mineralization.
Conclusion:
1., the animal models of aseptic loosening of artificial joints in New Zealand white rabbits established by us can provide foundation for research related topics, no matter from pathological mechanism or modeling effect, and the models are simple and reliable.
2. in vivo, non porous titanium implant alendronate HA composite coating can inhibit UHMWPE particles induced osteolysis, even more than in the absence of UHMWPE particles used alone HA coated prosthesis works better.
3. in vivo, alendronate sodium in the prevention of wear debris induced osteolysis process can promote the activity of osteoblasts.
The local release of 4. alendronate has a systemic therapeutic effect.
5. alendronate sodium composite HA coating as a slow release carrier, on the one hand, can provide early, short time high drug concentration release for the initial stability of the prosthesis, and on the other hand, it can provide long time low concentration drug release.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332;R687

【相似文献】