微弧氧化和碱处理技术在多孔钽修复兔颅骨缺损中的应用

目的观察微弧氧化和碱处理对多孔钽表面性状、生物相容性和成骨能力的影响。方法微弧氧化和碱处理多孔钽片后,扫描电镜观察表面微孔数量、表面钙磷沉积和接触角。植入钽片修复兔颅骨缺损模型,在4周和12周观察骨愈合情况。结果扫描电镜显示处理组表面有更多的微孔和钙磷沉积以及更小的接触角(P<0.05)。植入多孔钽片后,所有动物均生长良好,伤口愈合佳。CT观察多孔钽片和周围骨组织耦合良好;钙黄绿素标记检测显示12周时有新生骨长入多空钽材料内部;扫描电镜观察发现4周时多空钽材料内部有新生血管,12周时有骨小梁长入材料内部。结论微弧氧化和碱处理能改变多孔钽材料表面形状,处理后多孔钽片具有良好的生物相容性和成骨能力。     

γ-PGA/HEMA/PEG聚合物晶胶支架修复兔眼眶骨折缺损的实验研究

目的 近年来,眶骨骨折发生率逐年增加,其治疗关键旨在修复缺损的眶骨,聚（γ-谷氨酸）/2-羟乙基甲基丙烯酸酯/聚（乙二醇）（γ-PGA/HEMA/PEG）聚合物晶胶是一种具有互连多孔结构的新型支架材料,研究旨在检验其在眼眶骨折缺损修复中的骨修复效果。方法 采用低温凝胶技术制备了γ-PGA/HEMA/PEG聚合物晶胶。制备了兔的眼眶骨缺损模型,根据植入支架材料的不同分为3组,空白对照组、聚合物晶胶组（Gel组）、矿化聚合物晶胶组（M-gel组）。植入后8周和16周标本取材进行大体观察,通过影像学和组织学检查观察其血管生成和成骨效果。结果 影像学检查结果表明,支架材料能有效促进眶骨缺损的修复,缺损区被骨组织完全替代。组织学结果表明,支架材料可以增加Runt相关转录因子2（Runx-2）、碱性磷酸酶（ALP）、骨桥蛋白（OPN）和血小板内皮细胞黏附分子1（CD31）的表达,这表明支架材料植入后血管生成和成骨能力增强。结论 矿化聚合物晶胶是一种良好的眼眶骨折缺损修复的支架材料。     

两种壳聚糖仿生复合材料对家兔桡骨骨缺损修复的实验研究

目的:评价壳聚糖/碳酸钙三维复合材料(CS/CaCO3)和壳聚糖/羟基磷灰石复合材料(CS/HA)用于骨缺损修复的可行性.方法:家兔24只,随机分为对照、CS/CaCO3、CS/HA三组.左前肢去毛后,2%巴比妥钠(30mg/kg,iv)麻醉,距桡骨远端3cm处截骨1cm,形成骨缺损,分别植入相应材料.术后4w、8w、12w分别处死动物,X线摄片后,取骨缺损标本,进行大体与组织学观察.结果:术后4周植入块颜色变红,周围有较多量的新生骨样组织包裹,骨痂增多,向植入块内移行;术后8周,植入块周围有明显新骨生成,将材料分隔包围,新骨中央区可见材料呈蜂窝状残留.术后12周缺损区大部分编织骨被成熟的板层骨组织替代,并形成髓腔.结论:CS/CaCO3和CS/HA两种仿生复合材料能明显促进兔桡骨骨缺损修复,诱导骨痂生成.     

多孔钛合金不同孔径大小对新骨长入的影响

目的:评价不同孔径多孔钛合金植入物在骨缺损区对新骨长入的影响。方法:采用电子束熔融(EBM)技术制备三种不同孔径(孔径分别为1.0 mm,2.0 mm,3.0 mm)的多孔钛合金材料,其孔隙率依次为73%,79%,86%。将18只家犬随机分为1.0 mm孔径材料组,2.0 mm孔径材料组,3.0 mm孔径材料组,每组6只。制备家犬双侧股骨外侧髁缺损模型,然后植入各孔径组材料,于术后4周,8周,12周分别行大体标本观察,X线片观察,组织形态学观察三组不同孔径材料与周围骨的整合情况及孔隙中的新骨长入情况。结果:通过大体标本观察和X线片观察显示,12周后三组材料均与周围紧密骨连接。其中1.0 mm孔径组材料中心明显成骨,2.0 mm孔径组和3.0 mm孔径组中心仍为较多白色组织填充。组织学观察显示,12周时2.0 mm孔径组和3.0 mm孔径组材料周围有骨质包绕,但中心空洞,基本无骨质形成。1.0 mm孔径组材料周围骨质包绕紧密,孔中新生骨形成较多,且有大量纤维母细胞和软骨细胞形成。各时间点1.0 mm孔径组新生骨面积百分比明显高于2.0 mm孔径组和3.0 mm孔径组,P<0.01,差异具有统计学意义。2.0 mm孔径组和3.0 mm孔径组相比,P>0.05,无显著差异。结论:孔径大小影响多孔钛合金材料的骨长入,适当孔径的设计将更有利于材料的传导成骨。     

低强度脉冲超声波辐照与泡沫TiC/Ti对犬节段性骨缺修复的促进作用

目的探讨低强度脉冲超声波辐照对节段性骨缺损修复效果的影响。方法将直径12 mm长20mm泡沫TiC/Ti植入6只Beagle犬的左侧胫骨节段性骨缺损区。随机分为超声组和对照组,超声组采用低强度脉冲超声波辐照（频率1.5 MHz、强度30 mW/cm2、脉冲宽度200μs、脉冲周期1 kHz、20 min/次、1次/d）,对照组为不开功率源的假辐照,术后4、8周后分别行X线检查及骨密度测定,观察及分析材料周围骨愈合情况。结果 6只beagle犬均进入结果分析。术后4周超声组骨早期成熟度优于对照组,表现在材料周围骨痂影密度增高,骨痂影由两端向中央生长;对照组仅见骨痂区密度低,还可见部分骨痂缺如。术后8周超声组新生骨痂面积优于对照组,骨干结构相对稳定;对照组骨缺损区未闭合,在骨干两侧看到少量骨痂,愈合较差。骨密度测定结果显示,4周时超声组高于对照组,两组间存在统计学差异;8周时超声组略高于对照组,但两组间没有统计学差异。结论通过联合应用低强度脉冲超声波辐照与人工骨材料修复可提高新骨形成速度及骨组织密度,缩短节段性骨缺损的骨愈合时间。     

石墨化炭/ 炭复合材料+ 羟基磷灰石涂层(C/C+HA) 复合骨植入材料研究

目的:通过建立兔股骨缺损的动物实验模型,对采用等温化学气相沉积法和等离子喷涂技术所制备的石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料进行骨植入实验的的生物相容性进行评价,探索该复合材料作为植入机体骨组织的可行性依据.方法:采用骨科钻在实验动物股骨髁上钻孔的方法建立骨缺损的动物实验模型,将待研究比较的实验材料分别植入实验动物的股骨髁内,持续观察8周,在术后第2、4、8周时应用X线照片、组织学染色和扫描电镜技术,分别观察所研究材料在机体内对骨缺损愈合及其对机体的影响,进行组间比较和相关性分析.结果:石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料的骨植入实验生物相容性良好,材料与骨组织结合牢固,界面中成骨细胞生长明显,且炭颗粒脱落现象少,未见炎症细胞浸润.植入动物体内的材料在植入期未引起机体局部的炎症浸润反应且表面脱落的碳颗粒在机体组织中也未引起局部严重的炎症反应.在实验动物植入材料后的连续8周观察期中,组织学观察显示:表面涂有HA的炭/炭复合材料对骨组织形态改建上表现良好,其与骨组织接界处所形成的纤维结缔组织膜层厚度明显比未涂HA的材料要小,与骨组织结合更为紧密和牢固;碳颗粒出现脱落游离的现象明显减少.结论:在炭/炭复合材料表面涂以HA生物涂层对骨的形态改建和促进骨小梁生长等方面具有良好的作用,是一种具有发展潜力的骨修复材料.     

石墨化炭/炭复合材料＋羟基磷灰石涂层（C/C＋HA）复合骨植入材料研究

目的：通过建立兔股骨缺损的动物实验模型,对采用等温化学气相沉积法和等离子喷涂技术所制备的石墨化炭/炭复合材料＋羟基磷灰石涂层（C/C＋HA）复合骨植入材料进行骨植入实验的的生物相容性进行评价,探索该复合材料作为植入机体骨组织的可行性依据。方法：采用骨科钻在实验动物股骨髁上钻孔的方法建立骨缺损的动物实验模型,将待研究比较的实验材料分别植入实验动物的股骨髁内,持续观察8周,在术后第2、4、8周时应用X线照片、组织学染色和扫描电镜技术,分别观察所研究材料在机体内对骨缺损愈合及其对机体的影响,进行组间比较和相关性分析。结果：石墨化炭/炭复合材料＋羟基磷灰石涂层（C/C＋HA）复合骨植入材料的骨植入实验生物相容性良好,材料与骨组织结合牢固,界面中成骨细胞生长明显,且炭颗粒脱落现象少,未见炎症细胞浸润。植入动物体内的材料在植入期未引起机体局部的炎症浸润反应且表面脱落的碳颗粒在机体组织中也未引起局部严重的炎症反应。在实验动物植入材料后的连续8周观察期中,组织学观察显示：表面涂有HA的炭/炭复合材料对骨组织形态改建上表现良好,其与骨组织接界处所形成的纤维结缔组织膜层厚度明显比未涂HA的材料要小,与骨组织结合更为紧密和牢固;碳颗粒出现脱落游离的现象明显减少。结论：在炭/炭复合材料表面涂以HA生物涂层对骨的形态改建和促进骨小梁生长等方面具有良好的作用,是一种具有发展潜力的骨修复材料。     

植入了骨修复材料的小型猪腰椎椎体不脱钙病理组织切片制备方法

摘要 目的：建立植入了骨修复材料小型猪腰椎椎体骨组织标本的不脱钙病理组织切片制备方法。方法：将含骨修复材料的腰椎椎体骨组织标本进行分割暴露组织切面,梯度浓度乙醇脱水后经Technovit 7200 VLC光聚树脂浸润,经黄蓝光共同辐照进行光聚合包埋,借助硬组织病理切磨系统制备含骨修复材料不脱钙病理组织切片。结果：结果显示通过上述方法制备的病理组织切片,经苏木精-伊红（HE）染色及甲苯胺蓝染色后光学显微镜下观察能较好地显示骨的各种组织细胞结构,可清晰的观察到骨小梁的走向及连接情况。结论：研究建立了含骨修复材料骨组织标本病理组织切片制备方法,实现了含骨修复材料不脱钙骨组织病理切片的制备,经病理染色后实现了带植入物的组织学观察,为生物材料及医疗器械动物试验研究提供了新的病理检测手段及组织学评价途径。     

改良Masquelet技术治疗胫骨感染性缺损的临床疗效分析

摘要 目的：探讨和分析使用改良Masquelet技术在胫骨感染性缺损中的临床疗效。方法：回顾性分析2018.01-2022.01范围内,使用改良Masquelet治疗胫骨感染性缺损的共计21患者。通过骨愈合(Samantha X 线评分),软组织愈合,感染控制,下肢功能,并发症等方面对临床疗效进行评价。结果：21例患者均获得完整随访,随访时间为12-36个月,平均为18.6个月。所有患者均达到骨愈合标准（3.2±8.3个月）。Samantha X 线评分：1例为4分,8例为5分,12例6分。PALEY功能评分为：优9人,良11人,可1人,差0人,优良率为95.2%。患者中均未出现严重并发症(P>0.05)。结论：在胫骨感染性缺损的治疗中,改良Masquelet技术是一种简单,安全,有效的治疗方法,在临床中可以进一步推广。     

多孔n-HA/PA66支架复合rhBMP-2修复兔桡骨缺损的实验研究

目的 ：研究多孔纳米羟基磷灰石/聚酰胺66（nHA/PA66）骨修复材料作为骨组织工程支架复合基因重组人骨形态发生蛋白2（rhBMP2）后的成骨能力的变化,探讨加速nHA/PA66人工骨与受体骨愈合的方法。方法：选用新西兰大白兔双侧桡骨制作骨缺损模型,将nHA/PA66/rhBMP2复合材料植入左侧骨缺损处,右侧骨缺损以nHA/PA66植入作为实验对照,另做不植入任何材料的骨缺损空白对照。在1、2、4、8、12周各时相点分别进行大体观察、X线照片、组织学切片、免疫组化原位杂交进行检测图象分析。结果：nHA/PA66/BMP2与nHA/PA66组骨缺损均完全修复,而空白对照组骨缺损未见修复;2周时nHA/PA66与nHA/PA66/rhBMP2两组间原位杂交阳性细胞表达有统计学意义（ P<0.05）, 4周时nHA/PA66与nHA/PA66/rhBMP2两组间原位杂交阳性细胞表达无统计学意义（ P>0.05）,2周及4周实验和实验对照两组分别与空白对照组比较均无统计学意义（ P>0.05）,nHA/PA66/rhBMP2组较nHA/PA66组可加速人工骨/植入体/受体界面骨愈合。 结论：多孔nHA/PA66作为骨组织工程支架复合具有诱导成骨活性的rhBMP2后,增强了早期成骨能力,加速了其与受体骨的愈合。     

Engineering of bone tissue with porcine bone marrow stem cells in three-dimensional trabecular metal: in vitro and in vivo studies

The aim of this study was to investigate capability of cell attachment and ectopic bone formation in pigs after either ex vivo transplantation and expansion of bone marrow stem cells (BMSc) into three-dimensional porous tantalum, or porous tantalum supplemented with BMSc. After 24 hours incubation, cells adhering to the porous tantalum discs were quantified by means of scintillation counting of 3H-thymidine-labeled cells. After 7 days of incubation, the cell-loaded porous tantalum discs were harvested for histological analysis or implanted in the infrasternal muscle; an empty disc and disc implanted immediately after cell loading served as controls. All implants were taken out after 8 weeks of implantation and histological examination was performed. The results of in vitro cell attachment to the porous tantalum discs were not improved significantly with gelatin, collagen or fibronectin coatings. Histological analysis of cell loaded discs in vitro demonstrated viable BMSc within the 3-D tantalum structure. In vivo bone induction was demonstrated when the porous tantalum discs were cultured with BMSc. Our findings indicated that porous tantalum was suitable for cell attachment, and ectopic bone formation in pigs was achieved by means of BMSc cultured with porous tantalum. The present study suggests that cell-mediated hard bone tissue repair technology makes it possible to prefabricate autologous BMSc into three-dimensional trabecular metal in order to engineer bone tissue.     

人工骨表面覆盖口腔修复膜对颌骨囊性缺损人工骨植入术患者植入腔感染的预防作用

目的:探讨人工骨表面覆盖口腔修复膜对颌骨囊性缺损人工骨植入术患者植入腔感染的预防作用。方法:将我院2012年4月～2016年2月收治的78例颌骨囊性缺损患者按治疗时间分为对照组38例与观察组40例,均行开窗人工骨植入术,其中对照组采取常规抗感染,观察组采取人工骨表面覆盖口腔修复膜。比较两组的囊腔体积、面积缩减率、人工骨植入量、骨厚度及植入腔感染率。结果:两组的囊腔体积、面积缩减率及人工骨植入量比较差异无统计学意义(P0.05)。但观察组的骨厚度明显大于对照组(P0.05),植入腔感染率明显低于对照组(P0.05)。结论:在开窗人工骨植入术的基础上使用口腔修复膜不仅能引导骨组织再生,而且能够显著降低植入腔的感染率。     

冲击波诱导人骨髓基质细胞体内成骨研究

为了研究冲击波(SW)诱导人骨髓基质细胞(hMSCs)在动物体内成骨作用,根据前期工作结果,应用适宜能量冲击波(10kV,500次)处理体外培养的hMSCs,将SW组和对照组hMSCs与羟基磷灰石(HA)载体复合后体外培养2周,应用扫描电镜(SEM)检测细胞在载体表面的生长情况.将hMSCs-HA载体复合体植入裸鼠皮下,分别于术后4周、8周取材进行组织学、四环素荧光标记、SEM观察、碱性磷酸酶测定、RT-PCR检测骨钙素mRNA表达.结果表明,SW组及对照组细胞与HA载体体外复合后生长良好,且SW组细胞分泌较多的细胞基质;细胞载体复合体植入动物体内后,SW组载体表面有类骨组织形成,而对照组HA载体表面无骨组织形成;SW组与对照组的hMSCs-HA载体复合体碱性磷酸酶表达有显著性差异(P<0.01);SW组hMSCs-HA载体复合体术后4周与8周表达骨钙素mRNA,而对照组则无表达.提示hMSCs经适宜能量冲击波作用后与HA载体复合植入裸鼠体内具有成骨作用,适宜能量的冲击波作为一种新的促进hMSCs成骨分化的方法,可应用于组织工程领域.     

Bone regeneration potential of allogeneic or autogeneic mesenchymal stem cells loaded onto cancellous bone granules in a rabbit radial defect model

For developing a clinically effective bone regeneration strategy, we compare the bone regeneration potential of cultured allogeneic bone marrow-derived mesenchymal stem cells (BM-MSCs) and of autologous BM-MSCs loaded onto allogeneic cancellous bone granule scaffolds. A critical-sized segmental bone defect was made at the mid-shaft of both radiuses in 19 New Zealand White rabbits (NWRs). In the experimental group, allogeneic BM-MSCs loaded onto small-sized allogeneic cancellous bone granules (300~700 um in diameter) were implanted in one side of a bone defect. In the control group, autologous BM-MSCs loaded onto allogeneic cancellous granules were grafted in the other side. Bone regeneration was assessed by radiographic evaluation at 4, 8, 12 and 16 weeks post-implantation and by micro-computed tomography (micro-CT) and histological evaluation at 8 and 16 weeks. The experimental groups showed lower bone quantity indices (BQIs) than the control groups at 12 and 16 weeks (p?<?0.05), although no significant difference was observed at 4 and 8 weeks (p?>?0.05). Micro-CT analysis revealed that both groups had similar mean total bone volume and other parameters including trabecular thickness, number and separation at either 8 or 16 weeks. Only bone surface area revealed less area in the experimental group at 16 weeks. Histological evaluation of 8-week and 16-week specimens showed similar biologic processes of new bone formation and maturation. There was no inflammatory reaction indicating an adverse immune response in both allogeneic and autologous MSC groups. In conclusion, allogeneic BM-MSCs loaded onto allogeneic cancellous bone granules had comparable bone regeneration potential to autologous BM-MSCs in a rabbit radial defect model.     

A preliminary study in osteoinduction by a nano-crystalline hydroxyapatite in the mini pig

To test the probable osteoinductive properties of NanoBone, a new highly non-sintered porous nano-crystalline hydroxylapatite bone substitute embedded into a silica gel matrix, granules were implanted subcutaneously and intramuscularly into the back region of 18 mini pigs. After periods of 5 and 10 weeks as well as 4 and 8 months, implantation sites were investigated using histological and histomorphometric procedures. Signs of early osteogenesis could already be detected after 5 weeks. The later periods were characterized by increasing membranous osteogenesis in and around the granules leading to the formation of bone-like structures showing periosteal and tendon-like structures with bone marrow and focal chondrogenesis. Bone formation was better in the subcutaneous than in the intramuscular implantation sites. This ectopic osteogenesis is discussed with regard to the nanoporosity and microporosity of the material, physico-chemical interactions at its surface, the differentiation of osteoblasts, the role of angiogenesis and the probable involvement of growth factors. The results of this preliminary study indicate that this biomaterial has osteoinductive potential and induces the formation of bone structures, mainly in subcutaneous adipose tissue in the pig.     

Combination of bone tissue engineering and BMP-2 gene transfection promotes bone healing in osteoporotic rats

OBJECTIVE: The aim of this study was to develop a feasible approach to promote bone healing in osteoporotic rats using autogenous bone tissue-engineering and gene transfection of human bone morphogenetic protein 2 (hBMP-2). METHODS: Bone marrow stromal cells (BMSCs) from the left tibia of osteoporotic rats were transfected with the hBMP-2 gene in vitro which was confirmed by immunohistochemistry, in situ hybridization and Western blotting. Autogenous transfected or untransfected BMSCs were seeded on macroporous coral hydroxyapatite (CHA) scaffolds. Each cell-scaffold construct was implanted into a defect site which was created in the ramus of the mandible of osteoporotic rats. Four or eight weeks after implantation in situ hybridization was performed in BMSCs transfected with hBMP-2, X-ray examinations, histological and histomorphological analyses were used to evaluate the effect of tissue-engineered bone on osseous defect repair. RESULTS: Newly formed bone was observed at the margin of the defect 4 weeks after implantation with BMSCs transfected with BMP-2. Mature bone was observed 8 weeks after treatment. In the control group there was considerably less new bone and some adipose tissue was observed at the defect margins 8 weeks after implantation. CONCLUSIONS: Autogenous cells transfected with hBMP-2 promote bone formation in osteoporotic rats. BMSC-mediated BMP-2 gene therapy used in conjunction with bone tissue engineering may be used to successfully treat bone defects in osteoporotic rats. This method provides a powerful tool for bone regeneration and other tissue engineering.     

Porous Silk Scaffolds for Delivery of Growth Factors and Stem Cells to Enhance Bone Regeneration

Stem cell-based tissue engineering shows promise for bone regeneration and requires artificial microenvironments to enhance the survival, proliferation and differentiation of the seeded cells. Silk fibroin, as a natural protein polymer, has unique properties for tissue regeneration. The present study aimed to evaluate the influence of porous silk scaffolds on rat bone marrow stem cells (BMSCs) by lenti-GFP tracking both in vitro and in vivo in cranial bone defects. The number of cells seeded within silk scaffolds in rat cranial bone defects increased from 2 days to 2 weeks after implantation, followed by a decrease at eight weeks. Importantly, the implanted cells survived for 8 weeks in vivo and some of the cells might differentiate into endothelial cells and osteoblasts induced by the presence of VEGF and BMP-2 in the scaffolds to promote angiogenesis and osteogenesis. The results demonstrate that porous silk scaffolds provide a suitable niche to maintain long survival and function of the implanted cells for bone regeneration.     

Surface porous fibre-reinforced composite bulk bone substitute

The aim of this study was to describe and evaluate the significance of a porous surface with bioactive glass granules (S53P4) covering an artificial bulk material based on polymethylmetacrylate (PMMA) and fibre-reinforced composite (FRC) technology. Effort was focused particularly on characters of the porous surface and biomechanical properties of the material in vitro, and test in vivo the implant in reconstruction in an experimental long bone segment defect model. The defect, 10 mm in length, created in the shaft of rabbit tibia, was reconstructed by the implant and fixed by intramedullary K-wires. The implant was incorporated within 4 weeks by new bone growth from the host bone covering particularly its posterior surface and cortex/implant junctions with bridging trabecular bone. Later, at 8 weeks, new bone was found also at the cortex/implant interface and in the medullary canal of the implant. Histometric measurements revealed direct bone/implant surface contact in 34% at the interface. Bioactive glass granules in the porous surface evoked the most direct contact with bone. The implants manufactured from PMMA only served as a control group, and showed significantly lower osteoconductive properties. Biomechanical measurements in vitro of fibre-reinforced PMMA specimens revealed values for bending strength and the flexural modulus to match them to human bone. This artificial bulk bone material based on PMMA/FRC technology seems to have proposing properties to be used as a bone substitute on load-bearing conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of tantalum implants used for avascular necrosis of the femoral head: a review of five retrieved specimens

Aim: The effective results shown in the porous systems of tantalum employed for the use of osseointegrates has been demonstrated by means of animal experimentation. However, there is a total lack of any research studies on the osseointegration of tantalum implants from retrieval of the same after a period of time whereby the material had been implanted within the human body. Materials and Methods: For this study, five rod implants used for the treatment of avascular necrosis of the femoral head were retrieved following collapse of the femoral head and conversion to total hip arthroplasty. The time of implantation ranged between six weeks and twenty months. Results: Observation during this study has confirmed the effectiveness of osseointegration within this period of time. New bone was observed around and within the porous system of the on rod devices at retrieval date. The bone ingrowth, however, proved to be slower and less intense than that resulting within animal species during the first few months after implantation. Conclusions: Nevertheless, the results obtained in the quantitative assessment of this process proved to be similar to those results achieved by other authors in previous experimental work studies.