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类型 基础研究 预答辩日期 2018-03-11
开始(开题)日期 2014-05-27 论文结束日期 2017-05-17
地点 逸夫科技馆北三楼会议室 论文选题来源 国家自然科学基金项目     论文字数 6.7 (万字)
题目 PHBV纳米纤维修复骨组织及神经组织的实验研究
主题词 PHBV纳米纤维,三维支架,神经导管,骨再生,神经再生
摘要 组织工程的三要素是由种子细胞、支架材料、生长因子组成,我们选取了PHBV这种具有生物降解性、生物相容性以及无免疫源性等特征的微生物产生的聚酯作为支架材料,并用静电纺丝法制备出PHBV纳米纤维。基于PHBV纳米纤维我们分别制备出适用于骨组织工程的非定向和定向PHBV/HA纳米纤维支架,非定向和定向PHBV纳米纤维以及适宜于神经组织工程的定向PHBV/Laminin纳米纤维、定向PHBV纳米纤维以及非定向PHBV/Laminin纳米纤维、非定向PHBV纳米纤维。 通过CCK-8测试干细胞在材料上的增殖情况,通过成骨标志物检查来观察材料对成骨的影响,通过对体外施旺细胞的培养来观察材料对于神经再生功能的影响。结果表明,细胞接种1, 4, 7天后,不含HA的非定向PHBV纤维支架更适宜BMSCs的粘附和增殖;接种1, 2, 4周后,检测成骨标志物:碱性磷酸酶(ALP)、骨钙素(OCN)以及钙结节,分化实验结果表明含有HA的纳米纤维支架更有利于促进MSCs向成骨细胞的分化。而体外施旺细胞在PHBV纳米纤维膜上的培养表明,无论定向与非定向还是有无层粘蛋白修饰,均适宜于施旺细胞的黏附和增殖细胞。通过SEM扫描我们发现,BMSCs以及施旺细胞在定向纳米纤维上会顺着纤维方向生长、伸展,而在非定向纳米纤维上细胞则呈现随机排列和伸展。 在体内修复骨缺损的研究中,分别将定向及非定向PHBV纳米纤维三维支架,定向及非定向PHBV/HA纳米纤维三维支架,植入新西兰兔的桡骨缺损处,在植入术后通过大体观察,放射学检查,组织学检查,扫描电镜观察,CT扫描三维重建以及力学测试来进行评估。结果表明定向及非定向PHBV纳米纤维三维支架,定向及非定向PHBV/HA纳米纤维三维支架均可促进骨缺损的修复,其中非定向PHBV/HA纳米纤维三维支架对于骨组织的再生作用最佳,而定向 PHBV纳米纤维三维支架成骨能力最弱,HA对于材料的修饰对于体内骨组织再生的影响较为明显。 在体内神经缺损修复的研究中,我们制备了基于PHBV纳米纤维的三维神经导管,分别是定向及非定向PHBV纳米纤维三维神经导管,定向及非定向PHBV/Laminin纳米纤维三维神经导管,植入SD大鼠的坐骨神经缺损处,在植入术后通过步态测试,神经电生理,大体观察,组织学检查来进行评估。结果表明定向及非定向PHBV纳米纤维三维神经导管,定向及非定向PHBV/Laminin纳米纤维神经导管均可促进神经缺损的修复,但是定向PHBV纳米纤维神经导管及定向PHBV/Laminin纳米纤维神经导管对于神经缺损的效果更好。定向纳米纤维对于神经再生的影响更为明显。
英文题目 A Study of Electrospun PHBV nanofibers for bone regeneration and nerve regeneration
英文主题词 PHBV nanofibers, 3D scaffolds, nerve guide conduits, osteogenisis, nerve regeneration
英文摘要 Tissue engineering comprised three elements: cells, growth factors and scaffolds. The PHBV(poly(3-hydroxybutyrate-co-3-hydroxyvalerate)) was a bio-absorbable polymeric material that is synthesized by microorganisms that use determined substrates as carbon sources, under conditions oflimiting nutrients. The biomaterial was recognized for its biodegradability, biocompatibility and non-immunogenicity. To mimic the natural structure of extracellular matrices, electrospinning was applied to fabricate biomaterials into ultrafine fibers. Based on the electrospun PHBV nanofibers, four kinds of nanofibers were fabricated for bone regeneration including aligned PHBV nanofibers, aligned PHBV/HA nanofibers, random-oriented PHBV nanofibers and random-oriented PHBV/HA nanofibers, another four kinds of nanofibers were fabricated for nerve regeneration including aligned PHBV nanofibers, aligned PHBV/Laminin nanofibers, random-oriented PHBV nanofibers and random-oriented PHBV/Laminin nanofibers. In the vitro study, four kinds of nanofibers were fabricated for bone regeneration were investigated. The BMSCs cultured on the flat films were tested by CCK-8 kits and osteogenic markers. Cell proliferation tested with CCK-8 assay indicated that the BMSCs attached and proliferated more favorably on random-oriented PHBV nanofibrous meshes without HA. After one, two and four weeks of cell seeding, osteogenic markers including alkaline phosphate (ALP), osteocalcin (OCN), and mineralized matrix deposits were detected, respectively. The results indicated that the introduction of HA could induce the BMSCs to differentiate into osteoblasts. As similar, the Schwann cells seeded on four kinds of nanofibers fabricated for nerve regeneration were investigated through CCK-8 kits test and specific markers test. The results showed that the attachment and proliferation of Schwann cells on the PHBV meshes was good. SEM showed the BMSCs and Schwann cells migrated and elongated along the same diection on the aligned meshes, BMSCs and Schwann cells migrated and elongated randomly on the random-oriented meshes. In vivo study, the PHBV nanofibers were fabricated into 3D scaffolds and then the scaffolds were implanted into the rabbit 1.5cm radius defect for bone regeneration. The outcome of repairing the defect were tested through gross observation, X-ray, histological examination, SEM, CT three dimensional reconstruction and biomechanical test. The results showed that the four kinds nanoibrous scaffolds could heal the bone defect 16 weeks after the operation, the random-oriented PHBV/HA nanofibrous scaffolds had the best osteogenic capacity among the four kinds nanoibrous scaffolds. The results in vivo and vitro study indicated that the HA loaded on the PHBV nanofibers could improve the bone regeneration capacity of the nanofibers. The random-oriented nanofibers may affect bone regeneration in a certain extent. The PHBV nanofibers were fabricated into nerve guide conduits and then the conduits were implanted into the SD rats sciactic nerve defect for repairing the 12mm nerve gap. The outcome of nerve regeneration were tested through gross observation, sciatic functional test, histological examination, and electrophysiological test. The results showed that the four kinds nanoibrous scaffolds could repair the sciatic nerve gap 8 weeks after the operation, the aligned PHBV/Laminin nanofibrous conduits and the aligned PHBV nanofibrous conduits could repair the nerve defect better than the random-oriented nanofibrous conduits, the Laminin did not affect the nerve regeneration capacity of the nanofibers both in vivo and vitro.
学术讨论
主办单位时间地点报告人报告主题
东南大学生物科学与医学工程学院 2011.10.24 逸夫科技馆南三楼会议室 张晓峰 干细胞修复神经损伤的研究
东南大学生物科学与医学工程学院 2011.12.5 逸夫科技馆南三楼会议室 张晓峰 干细胞修复坐骨神经损伤
东南大学生物科学与医学工程学院 2012.1.4 逸夫科技馆南三楼会议室 张晓峰 坐骨神经损伤模型及术后检测指标
东南大学生物科学与医学工程学院 2012.5.7 逸夫科技馆南三楼会议室 张晓峰 大鼠坐骨神经损伤模型的建立及材料的植入
东南大学生物科学与医学工程学院 2012.6.4 逸夫科技馆南三楼会议室 张晓峰 大鼠坐骨神经修复后步态测试研究
东南大学生物科学与医学工程学院 2012.10.8 逸夫科技馆南三楼会议室 张晓峰 大鼠坐骨神经修复后的观察
东南大学生物科学与医学工程学院 2012.11.5 逸夫科技馆南三楼会议室 张晓峰 新生坐骨神经的组织学观察
东南大学生物科学与医学工程学院 2013.4.8 逸夫科技馆南三楼会议室 张晓峰 神经导管新进展
     
学术会议
会议名称时间地点本人报告本人报告题目
全国生物与医学纳米技术博士生学术论坛 2012.7.12 苏州
全国生物与医学纳米技术博士生学术论坛 2013.7.25 苏州 静电纺丝制备的PHBV/HA纳米纤维材料修复兔桡骨缺损的实验研究
东南大学校庆报告会 2017.5.4 南京 PHBV材料对于修复骨组织及神经组织的实验研究
微流控芯片与组织工程研讨会 2017.6.10 南京 PHBV Nanofibrous Nerve Conduits Promote Peripheral Nerve Regeneration
     
代表作
论文名称
Laminin-modified and aligned PHBV/PEO nanofibrous nerve conduits promote peripheral nerve regenerati
 
答辩委员会组成信息
姓名职称导师类别工作单位是否主席备注
陆祖宏 正高 教授 博导 东南大学
王连生 正高 教授 博导 南京医科大学
陈立娟 正高 教授 博导 东南大学
柴人杰 正高 教授 博导 东南大学
肖守军 正高 博导 南京大学
      
答辩秘书信息
姓名职称工作单位备注
孙博 副高 副教授 东南大学