不同温度加热自体骨成骨活性的对比实验研究

目的研究加热对自体骨成骨活性的影响.方法通过手术造成家兔胫骨1cm的骨缺损,原位分别植入新鲜自体骨、加热100℃20min及50℃20min自体骨,于手术后2、4、8、12周进行X线摄片,并对平片的相对灰度进行分析,测定骨痂胶原蛋白的含量,并与组织学检查进行对比.结果新鲜自体骨成骨活性最高,其次是加热50℃20min自体骨,加热100℃20min自体骨成骨活性最差.结论加热影响植入骨的成骨活性,而较低温度加热可以保留一定的成骨活性.     

Comparison of adipose stem cells sources from various locations of rat body for their application for seeding on polymer scaffolds

Adipose tissue yields adult adipose stem cells (ASCs) in large quantities via less-invasive methods. These cells are of interest owing to their modulating properties and paracrine activities, which can be harnessed in regenerative medicine. Many studies on the use of rat fat tissue in an autologous animal model have been conducted; however, the different locations to obtain stromal vascular fraction of rat fat depots have not been fully characterized. The purpose of the current study was to identify optimal source of ASC from various locations of rat body. Animal experiments in vitro revealed that fat depots from cervical fat are an optimal ASC source. A high ASC yield facilitates subsequent studies on autologous transplantation in rats. The secondary objective was to compare the efficiency of osteoinductive media composition and evaluate of osteogenic potential of ASCs for seeding on scaffolds for bone repair. Scaffolds were assessed in vitro, using rat adipose stem cells and three-dimensional (3D) scaffolds comprising polycaprolactone (PCL) or polycaprolactone covered with tricalcium phosphate (PCL + 5%TCP). Seeded ASCs adhere to the surface and migrate to the scaffolds. Upon staining and determining alkaline phosphatase levels, PCL + 5%TCP scaffolds performed better than PCL scaffolds. Furthermore, growth factors such as BMP2 and FGF2 significantly increased ASC mineralization and induced osteogenesis (p?<?0.05). Our results may help select and develop pre-clinical animal model for confirming the use of ASC, alone or in association with appropriate biomaterials for bone repair.     

复合rhBMP2的异种骨的研制及成骨活性

目的：研制新型具有诱导成骨活性的异种骨植骨材料。方法：在重组合异种骨（RBX）基础上,以基因重组人BMP2（rhBMP2）取代从牛皮质骨中提取的牛BMP,与去抗原牛松质骨载体（BCB）复合,制成复合rhBMP2的异种骨（rhBMP2/BCB）,并采用小鼠股部肌袋模型,通过组织学、骨计量学方法检测rhBMP2/BCB的诱导成骨活性。结果：实验组（rhBMP2/BCB组）术后7d在小鼠肌袋可诱导软骨生成,14d形成纺织骨,21d改建成板状骨并形成大量骨髓;对照组（BCB组）于术后各时间点均未见有软骨成骨形成。实验组的碱性磷酸酶活性和钙含量均高于对照组,有非常显性差异（P＜0．01）。结论：rhBMP2/BCB具有较好的骨诱导能力,是一种较理想的植骨材料。     

采用骨内型牵张器建立垂直向牙槽骨牵张成骨动物模型

【目的】应用自行研制的骨内型牙槽骨牵张器,建立犬垂直向牙槽骨牵张成骨动物模型。【方法】实验动物为中山大学动物中心提供的健康成年杂种犬12只。实验材料为自行设计,纯钛制成的骨内型垂直向牙槽骨牵张器,体部直径3.75mm,长5mm。先拔除犬双侧下颌前磨牙,形成萎缩牙槽嵴的模型。3月后随机选取一侧行骨切开术,植入两枚牵张器。1周后开始牵张,每天1次,每次1mm,共5d;分别在牵张后1、2和3月将动物处死进行临床检查,X线检查和组织学检查。【结果】骨内型垂直牙槽骨牵张器愈合良好。牙槽骨高度平均增加(4.8±0.50)mm。组织学切片显示在牵张区有骨质形成,其骨小梁方向与牵引方向一致。【结论】采用该骨内型牙槽骨牵张器成功建立稳定的、重复性好的犬牙槽骨牵张成骨动物模型,为牙槽骨牵张器的国产化和临床上应用该项技术打下基础。     

骨膜与牵张成骨关系的研究进展

骨膜(包括骨外膜与骨内膜)无论在骨的发生、形成,还是骨再生、修复中都起了一定作用。本文对在牵张成骨过程中,介入牵张应力后,骨膜尤其是骨外膜的作用等的研究现状作一综述。     

下颌骨牵引成骨过程中血运重建方式初步探讨

目的通过建立兔下颌骨牵引成骨实验动物模型,观察其动态过程,初步探讨血运重建方式及机制。方法选用新西兰白兔30只,6只动物做空白对照组;24只动物行右侧下颌骨植入外置式颌骨牵引器。经7d延迟期后,按1mm/d速率延长下颌骨7d,然后固定8周。在延迟期第7天,牵引期第2、4、7天,固定期第1、3、6、8周时分别处死3只动物,用X线片、苏木精-伊红染色、免疫组化方法检测血管和新骨生成情况,以及微血管密度变化规律。结果实验组22只兔成功进行下颌骨牵引延长,牵引区血管生成方式以芽生和血管上皮岛为主。在牵引期第2天,骨断端微血管密度为28.2±7.0,明显高于其他各组(P<0.01),在固定期下降明显,固定第8周时,微血管密度为4.3±1.3,与正常组无显著性差异(P>0.05)。结论在牵引初期牵引骨断端有强烈的微血管反应,血运重建先于新骨重建;固定期微血管密度降低,血管成熟,血运重建随新骨生成改建完成而结束。     

颧骨缝牵张成骨过程中组织超微结构变化的研究

目的:观察天然骨缝牵张成骨过程中组织和细胞超微结构的变化。方法:对牵张成骨的缝区组织经过系列处理,进行超薄切片、透射电镜观察。结果:间充质细胞在牵引早期大量增生,并不断分化为成纤维细胞和成骨细胞,后2种细胞的超微结构显示出具有活跃的合成和分泌功能。1周标本成纤维细胞沿牵引力的长轴方向排列,胞核增大,在其周围包绕着发达并扩张的内质网系统;3周标本大量增生活跃的成骨细胞和成纤维细胞,成骨细胞核仁增大、粗面内质网扩张、核糖体丰富、线粒体增多、富含紧密排列的嵴;5、8周标本中骨细胞形成并发育成熟,骨基质逐渐矿化,清晰可见新形成的胶原纤维、哈氏管,以及骨基质矿化的过程。结论:三维牵张过程中,成纤维细胞、成骨细胞活跃,缝区形成新骨。     

山羊下颌骨牵张成骨的实验研究

目的:通过山羊下颌骨牵张成骨实验动物模型,了解自行设计开发的三维骨牵张器的成骨效果。方法:成年山羊6只,建立下颌骨牵张成骨实验动物模型,术后第8天开始用自行设计的牵张器,以0.6mm/次、2次/d的速度牵张,牵张期为17～18d,牵张高度20mm左右。牵张完毕后1、2及3个月各处死2只动物,进行大体标本、组织形态学和骨密度观察。结果:成功建立山羊下颌骨较大速率牵张成骨实验动物模型。大体标本观察表明,在牵张间隙形成了很好的骨痂组织,牵张间隙达到了预期的长度。组织形态学检查结果显示:牵引后1个月,牵张区充满平行排列的骨小梁;牵张后2个月,牵张区可见排列成网状粗大的骨小梁及成熟的哈佛氏系统。结论:我们自行设计的三维牵张器,制作简便,易于控制,可以稳定成骨,具有临床应用的可能性。     

成骨生长肽羧基端片段衍生物对去卵巢大鼠骨密度和骨力学的影响

目的：探讨成骨生长肽羧基端片段衍生物G381、G38K对去卵巢大鼠骨转换生化指标、骨密度和骨力学性能的影响及各指标间相互关系。方法：将3个月龄SD雌性大鼠40只随机分为5组，设假手术组（S组）和双侧卵巢切除（OVX）模型组（M组）为阴性对照，其余3组行OVX后分别予G38I、G38K和阿仑膦酸钠（ALN）治疗。术后60d处死．测血钙、磷和碱性磷酸酶，测量腰椎体和股骨骨密度（BMD），骨灰重／干重比值和骨力学指标。结果：与S组比较，M组血钙、磷升高，骨灰重／干重比值降低，经G38I、G38K治疗后上述指标恢复至S组水平，并显著升高腰椎体BMD，提高股骨弹性应力和最大应力，而股骨和腰椎体弹性模量呈升高趋势。灰重／千重比值与BMD呈正相关，BMD和力学指标间无相关性。结论：G38I或G38K的治疗可一定程度上减轻去卵巢大鼠的骨丢失，改善骨力学性能，评价抗骨质疏松药物疗效应同时关注骨量和骨质量。     

Evaluation of the Biocompatibility and Osteogenic Properties of Metal Oxide Coatings Applied by Magnetron Sputtering as Potential Biofunctional Surface Modifications for Orthopedic Implants

Biomaterials with adequate properties to direct a biological response are essential for orthopedic and dental implants. The surface properties are responsible for the biological response; thus, coatings with biologically relevant properties such as osteoinduction are exciting options to tailor the surface of different bulk materials. Metal oxide coatings such as TiO₂, ZrO₂, Nb₂O₅ and Ta₂O₅ have been suggested as promising for orthopedic and dental implants. However, a comparative study among them is still missing to select the most promising for bone-growth-related applications. In this work, using magnetron sputtering, TiO₂, ZrO₂, Ta₂O₅, and Nb₂O₅ thin films were deposited on Si (100) substrates. The coatings were characterized by Optical Profilometry, Scanning Electron Microscopy, Energy-Dispersive X-ray Spectroscopy, X-ray Photoelectron Spectroscopy, X-ray Diffraction, Water Contact Angle measurements, and Surface Free Energy calculations. The cell adhesion, viability, proliferation, and differentiation toward the osteoblastic phenotype of mesenchymal stem cells plated on the coatings were measured to define the biological response. Results confirmed that all coatings were biocompatible. However, a more significant number of cells and proliferative cells were observed on Nb₂O₅ and Ta₂O₅ compared to TiO₂ and ZrO_2. Nevertheless, Nb₂O₅ and Ta₂O₅ seemed to induce cell differentiation toward the osteoblastic phenotype in a longer cell culture time than TiO₂ and ZrO₂.