气电纺纳米羟基磷灰石/聚羟基丁酸酯复合纤维支架对大鼠骨髓基质细胞成骨分化的影响  

Effect of gas-jet/electrospun nanosized hydroxyapatite/Poly (3-hydroxybutyrate) ultrafine-fibrous scaffolds on the osteogenic differentiation of rat bone marrow-derived mescenchymal stem cells

作　　者：管东华[1] 林映荷[2] 宋光保[1] 陈治清[2] 

机构地区：[1]广东省口腔医院.南方医科大学附属口腔医院修复科,广东省广州市510280 [2]四川大学华西口腔医学院,四川省成都市610041

出　　处：《中国组织工程研究与临床康复》2009年第47期9277-9281,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省医学科学技术研究基金资助(B2009029);四川省2008年科技计划项目(2008SG0020;2008JY0024-2)~~

摘　　要：背景:利用静电纺丝技术制备的纤维支架材料具有类似于细胞外间质的形态和结构,且其独特的生产工艺可以很便捷地将功能性纳米颗粒复合入高分子纤维内,在制备组织工程支架方面具有独特的优势。目的:创新性地将物理共混法与气流-高压静电纺丝技术相结合,仿生构建纳米羟基磷灰石/聚羟基丁酸酯复合纳米纤维支架材料,评价其作为骨组织工程支架在体外的生物活性。设计、时间及地点:细胞学体外实验,于2008-03/2009-04在四川大学口腔疾病研究国家重点实验室完成。材料:按照预定参数进行气流-高压静电纺丝,分别制备气电纺纯聚羟基丁酸酯纤维支架及含质量分数为10%纳米羟基磷灰石的纳米羟基磷灰石/聚羟基丁酸酯复合纤维支架。方法:将大鼠骨髓基质细胞接种于纳米羟基磷灰石/聚羟基丁酸酯纳米纤维支架后进行体外培养(实验组),以接种于气电纺纯聚羟基丁酸酯纳米纤维支架为对照组,接种于细胞培养板为空白对照组。主要观察指标:通过RT-PCR检测连续培养14d后细胞成骨分化标志物碱性磷酸酶、Ⅰ型胶原和骨钙素的mRNA表达。结果:各组均能检测到3种成骨标志物碱性磷酸酶、Ⅰ型胶原和骨钙素的mRNA表达。通过QuantityOne软件分析计算目的条带吸光度值与内参β-actin条带吸光度值的比值,可以发现3种目的基因在实验组具有最高表达水平,其次为对照组,空白对照组的表达明显最弱。结论:气电纺纳米羟基磷灰石/聚羟基丁酸酯复合纤维支架可促进大鼠骨髓基质细胞成骨分化,证实其在体外具有优良的生物活性。BACKGROUND： In order to mimic the structure of natural bone, which is a biocomposite of hydroxyapatite mineral crystals arranged in an organic collagen matrix, introducing bioactive ceramics into polymer matrices become an attractive strategy for development of bone tissue scaffolds materials. Besides the selection of scaffolds materials, the structure of bone tissue engineering scaffolds also is critical. Many techniques have been developed to fabricate scaffolds. Among them, electrospinning has attracted much attention due to its consistency in producing fibers with submicron diameters and convenience in filling functional phase into the matrix phase. OBJECTIVE： To evaluate the bioactivity of gas-jet/electrospun nanosized hydroxyapatite （nHAP）/poly （3-hydroxybutyrate） （PHB） scaffolds for being used as bone tissue engineering scaffolds. DESIGN, TIME AND SETTING： The in vitro cytology experiment was performed at the State Key Laboratoty of Oral Diseases, Sichuan University between March 2008 and April 2009. MATEIALS： ①The scaffold materials samples （2.4？2.4cm2） were sterilized by high pressure stream and positioned at the bottom of individual 6-well tissue culture plates. ②Passage two isolated bone marrow-derived mescenchymal stem cells （BMSCs） were seeded at a density of 4？105 cells/sample on pretreated samples. Cells were grown in 2 ml culture medium and incubated at 37℃ and 5% CO2 for the duration of the experiment. The medium was changed every 2 days. Methods： The osteogenic differentiation of the BMSCscultured on the scaffolds materials were accessed using reverse transcription-polymerase chain reaction （RT–PCR）. MAIN OUTCOME MEASURES： mRNA transcript expression of bone-related markers, including alkaline phosphatase （ALP）, collagen type I （COL- I）, and osteocalcin （OC）, were quantified utilizing RT–PCR on days 14. RESULTS： After 14 days, while these bone-related markers were expressed in three groups, they had higher transcript levels in the ce

关 键 词：高压静电纺丝 纳米羟基磷灰石 聚羟基丁酸酯 骨组织工程 骨髓基质细胞 

分 类 号：R318[医药卫生—生物医学工程]