Effect of rhBMP‐2 and VEGF in a vascularized bone allotransplant experimental model based on surgical neoangiogenesis

We have demonstrated survival of living allogeneic bone without long‐term immunosuppression using short‐term immunosuppression and simultaneous creation of an autogenous neoagiogenic circulation. In this study, bone morphogenic protein‐2 (rhBMP‐2), and/or vascular endothelial growth factor (VEGF), were used to augment this process. Femoral diaphyseal bone was transplanted heterotopically from 46 Dark Agouti to 46 Lewis rats. Microvascular repair of the allotransplant nutrient pedicle was combined with intra‐medullary implantation of an autogenous saphenous arteriovenous (AV) bundle and biodegradable microspheres containing buffer (control), rhBMP‐2 or rhBMP‐2 + VEGF. FK‐506 given daily for 14 days maintained nutrient pedicle flow during angiogenesis. After an 18 weeks survival period, we measured angiogenesis (capillary density) from the AV bundle and cortical bone blood flow. Both measures were greater in the combined (rhBMP‐2 + VEGF) group than rhBMP‐2 and control groups (p < 0.05). Osteoblast counts were also higher in the rhBMP‐2 + VEGF group (p < 0.05). A trend towards greater bone formation was seen in both rhBMP2 + VGF and rhBMP2 groups as compared to controls (p = 0.059). Local administration of VEGF and rhBMP‐2 augments angiogenesis, osteoblastic activity and bone blood flow from implanted blood vessels of donor origin in vascularized bone allografts. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 561–566, 2013     

Local cooling reduces regional bone blood flow

Local cooling is very common after bone and joint surgery. Therefore the knowledge of bone blood flow during local cooling is of substantial interest. Previous studies revealed that hypothermia leads to vasoconstriction followed by decreased blood flow levels. The aim of this study was to characterize if local cooling is capable of inducing reduced blood flow in bone tissue using a stepwise‐reduced temperature protocol in experimental rabbits. To examine bone blood flow we utilized the fluorescent microsphere (FM) method. In New Zealand white rabbits one randomly chosen hind limb was cooled stepwise from 32 to 2°C, whereas the contra lateral hind limb served as control. Injection of microspheres was performed after stabilization of bone and muscle temperature at each temperature level. Bones were removed, dissected and fluorescence intensity was determined to calculate blood flow values. We found that blood flow of all cooled regions decreased relative to the applied external temperature. At maximum cooling blood flow was almost completely disrupted, indicating local cooling as powerful regulatory mechanism for regional bone blood flow (RBBF). Postoperative cooling therefore may lead to strongly decreased bone blood flow values. As a result external cooling has capacity to both diminish bone healing and reduce bleeding complications. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:1820–1827, 2013     

The use of a micro-bubble contrast agent to allow visualization of the biliary tree

Micro-bubble contrast agents have been used as contrast agents in the vascular system to allow visualization of blood flow by ultrasound. We examined the use of this type of contrast agent for visualization of bile duct by ultrasound. A laparotomy was performed on a domestic farm pig and the common bile duct was cannulated with a 20 gauge catheter. The contrast agent, perflutren lipid microspheres (Definity), was injected in different dilutions. Sonographic images were obtained by scanning directly on the pig liver. The use of the micro-bubble contrast in the biliary system provided excellent visualization of the bile duct down to ducts as small as fifth-order branches and demonstrated bile ducts <2 mm. The use of micro-bubble contrast agents in the biliary tree may allow for real-time imaging of the bile duct and could be helpful in living donor liver transplantation.     

人表皮生长因子与碱性成纤维细胞生长因子纳米缓释剂对成纤维细胞的影响

目的:制备碱性成纤维细胞生长因子(bFGF)、人表皮生长因子(EGF)可降解缓释微球,考察其生物活性的保存情况,以及它们对成纤维细胞的作用。方法:采用改良的乳化冷凝法交联制备复合bFGF、EGF的明胶缓释微球,将它们加入成纤维细胞的培养液中,用细胞计数法、四甲基偶氮唑盐微量反应比色法(MTT法)测定细胞增殖情况。结果:复合bFGF、EGF的缓释微球平均粒径(11.32±3.64)μm;培养1天后各组细胞计数、吸光度(A)值差异均无显著性;5天后,两种生长因子缓释微球组细胞计数、吸光度(A)值明显高于对照组;7天后,两种生长因子缓释微球组值仍高于其它组,但差异无显著性。结论:复合bFGF、EGF的缓释微球制备工艺简便,成球性好;能较长时间地持续释放活性bFGF、EGF,可促进成纤维细胞的增殖。     

庆大霉素-藻酸钙三维缓释微球的药物释放研究

目的研制庆大霉素一藻酸钙三维缓释微球并调控其庆大霉素的释放。使其达到长期局部抗菌的效果。方法制作不同浓度T、S、U组庆大霉素-藻酸钙缓释凝珠,与庆大霉素-骨水泥颗粒Y组进行庆大霉索释放情况比较。通过不同时间点抽取浸泡液,送紫外分光光度法（UV）及金黄色葡萄球菌（SA）培养检测,由此计算出各组包封率、释放率,绘制庆大霉素释放曲线。结果4组样品（微生物法）的包封率及30d药物释放率分别为：U组（53．99％、36．31％）,S组（39．62％、27．55％）。T组（34．20％、30．83％）,Y组（100．00％、48．49％）。U组的包封率较高,30d释放庆大霉素的总量较大,更接近Y组。4组间差异有统计学意义（P〈0．01）。U、Y组庆大霉素的释放明显高于S、T组的释放。各组30d内庆大霉素的浓度几乎都能超过金黄色葡萄球菌的最低抑菌浓度（MIC）．即〉2μg／mI。结论U组载药缓释凝珠30d内的庆大霉索释放较为理想。可作为BMSCs的三维培养支架。     

相变高分子微球增效高强度聚焦超声消融裸鼠骨肉瘤

目的制备相变高分子微球,观察其增效高强度聚焦超声(HIFU)消融裸鼠骨肉瘤模型的效果。方法制备包裹液态氟碳的高分子微球,检测其形态、结构、粒径电位及相变条件;建立裸鼠骨肉瘤模型,随机分为空白组(不予处理)、对照组(注射生理盐水)和实验组1、2、3(注射稀释倍数分别为5、10、20倍的Pct-MP溶液),处理后立即用海级星对肿瘤进行辐照。于辐照后即刻和72h提取标本并观察坏死面积、行HE染色,并对72h提取的标本的坏死周边组织进行细胞增殖和凋亡测定。结果成功制备相变高分子微球;HIFU辐照后即刻提取的标本,实验组1、2的坏死面积与对照组差异均有统计学意义(P均0.05),但与实验组3的差异无统计学意义(P0.05);72h后提取的标本,实验组1、2的坏死面积进一步扩大、与辐照后即刻提取的标本的差异有统计学意义(P均0.05)。HIFU辐照72h后,实验组1、2、3与对照组间坏死周边组织的凋亡和增殖指数差异均有统计学意义(P均0.05)。结论在骨肉瘤裸鼠模型中,自制相变高分子微球能够增强HIFU对于骨肉瘤的消融效果,增强作用与微球浓度有关。     

Embosphere微球靶血管栓塞治疗中晚期肺癌大咯血的疗效

目的:探讨用新型栓塞剂Embosphere微球行支气管动脉栓塞术（BAE）治疗中晚期肺癌大咯血的临床疗效。 方法:2013年1月—2016年1月对14例经内科治疗无效的中晚期肺癌大咯血行BAE治疗,均采用Embosphere微球栓塞。所有病例随访3月以上。分析患者栓塞效果和并发症发生情况。 结果:完全止血8例（57.1%）,不完全止血4例（28.6%）,无效2例（14.3%）,总有效率85.7%(12/14)。随访3个月以上,3月内复发1例（死亡）,3月以上复发2例,所有患者均无严重的并发症。 结论:采用Embosphere微球治疗中晚期肺癌大咯血是一种简单、安全、有效、并发症少、复发率低的方法,临床值得推广。     

Ophthalmic Artery Embolization as Pretreatment of Orbital Exenteration for Conjunctival Squamous Cell Carcinoma

The aim of this study is to describe the effect of transarterial embolization from the ophthalmic artery as a pretreatment for orbital exenteration. A 75-year-old Chinese man with a 7-year history of gradual increase of the left eye swelling showed a massive conjunctival tumor growing outwardly from the interpalpebral fissure and had no light perception in the left eye. Magnetic resonance imaging showed orbital invasion of the tumor around the left eyeglobe. The initial surgery for the planned orbital exenteration was discontinued after skin incision around the orbital margin due to massive hemorrhage. The patient underwent transarterial embolization with gelatin sponge (Spongel) of the feeding arteries from the left ophthalmic artery and, the next day, had orbital exenteration with well-controllable bleeding and reconstruction with free vascularized anterolateral thigh cutaneous flap transfer. Pathologically, well-differentiated squamous cell carcinoma proliferated in exophytic, papillary, and nested fashions, arising from the bulbar conjunctiva. Tumor cells were also found in the conjunctival stroma around the vessels. The sclera at the equator had a perforated site with tumor cell invasion, but no intraocular invasion was found. Hematoxylin-positive gelatin sponges were found inside the orbital vessels and large choroidal vessels. In conclusion, transarterial embolization of feeding arteries arising from the ophthalmic artery is a useful pretreatment to control bleeding at orbital exenteration for malignancy.     

钇-90微球的特性、放射栓塞的操作技术及安全防护——钇-90微球放射栓塞系列回顾(一)

钇-90(90Y)微球放射栓塞是一种局部治疗方法,它将载有发射β射线的90Y树脂或玻璃微球选择性地注射到肝动脉.90Y微球随血流被阻塞在肿瘤血管床,其发出的射线对靶肿瘤具有细胞毒性作用.该方法的安全性和有效性已经在不可切除的肝脏恶性肿瘤患者中得到证实.本文作为90Y微球放射栓塞系列综述的第一部分,将讨论90Y和90Y微球的基本特性,90Y微球放射栓塞的基本操作方法,以及放射性安全与防护.