摘要
目的构建聚乳酸／多巴胺／骨形成发生蛋白-2（PLLA-PDA-BMP-2）控释支架并评价其成骨能力。方法构建PLLA-PDA-BMP-2支架，设立PLLA-PDA支架及PLLA支架为对照。以骨髓间充质干细胞（BMSCs）为种子细胞在支架上培养，采用扫描电镜（SEM）、X光电子能谱仪（XPS）、接触角测量仪、共聚焦显微镜观察支架表面结构及细胞生长状态。通过检测碱性磷酸酶（ALP）活性、茜素红染色、定量反转录聚合酶链反应（RT—PCR）评价支架上BMSCs的成骨分化能力。结果（1）支架表征：PLLA-PDA-BMP-2支架表面氮元素（N）含量为（8．7±0．7）％，高于PLLA-PDA支架[（3．9±0．4）％]与PLLA支架（0％），差异有统计学意义（P=0．001）；PLLA-PDA-BMP-2支架表面接触角为（17．6±2．2）°，低于PLLA-PDA支架[（31．4±1．8）°]与PLLA支架[（120．8±5．0）°]，差异有统计学意义（P=0．001）；（2）体外实验：PLLA-PDA-BMP-2支架上细胞增殖能力优于PLLA-PDA支架与PLLA支架，差异有统计学意义（P=0．003）；培养14d后，PLLA-PDA-BMP-2支架ALP活性为（136．5±7．1）μmol／mg，高于PLLA-PDA支架[（44．2±4．2）μmoL／mg]及PLLA支架[（24．4±3．6）μmol／mg]，差异有统计学意义（P=0．013）；PLLA-PDA-BMP-2支架COL-1和OCN基因表达明显升高，与PIJA-PDA支架及PLLA支架比较，差异有统计学意义（P=0．002）。结论PLLA-PDA-BMP-2支架能够长期缓释BMP-2多肽，有效促进BMSCs增殖及成骨分化。
Objective To synthesize poly dopamine （PDA） coated polylactic acid nanofiber （PLLA） scaffold immobilized with bone morphogenetic protein -2 （BMP -2 ） peptide and evaluate its os- teogenetic effects in vitro. Methods The PLLA - PDA - BMP - 2 peptide scaffold was set as experiment group. The PLLA -PDA and the PLLA scaffold were set as control group. The X -ray photoelectron spec- troscopy （XPS）, contact angle viewer, scanning electron microscope （SEM） and confocal microscopy were utilized to observe the surface morphology of scaffold and cell growth morphology respectively. The osteo- genie differentiation of the cell on the scaffolds were assessed by alkaline phosphatase （ALP） , alizarin red stain and reverse transeriptase- polymerase chain reaction （RT- PCR） examination respectively. Results For scaffold characterization, the percentage of N content increased from PLLA - PDA - BMP - 2 scaffold （8.7±0.7）% to thePLLA-PDAscaffold （3.9±0.4）% and to thePLLA scaffold 0% （P=0.001）. The water contact angle significantly decreased from the PLLA scaffold 120. 8 ± 5.0° to the PLLA -PDA scaffold （ 31.4± 1.8 ） °and to the PLLA scaffold （ 17.6 ± 2. 2 ） ° （ P = 0. 001 ）. In vitro experiment, the PLLA- PDA -BMP- 2 scaffold remarkably promted cell proliferation than the PLLA scaffold and the PLLA scaffold （ P = 0. 003 ）. The ALP activity obviously increased from the PLLA scaffold （ 24. 4 ± 3.6） μmol/mg to the PLLA - PDA scaffold （44. 2 ± 4. 2） pLmol/mg and to the PLLA - PDA - BMP - 2 peptide scaffold （136. 5±7.1 ） μmol/mg （P =0. 013）. The expression levels of COL- 1 and OCN in the PLLA - PDA - BMP - 2 peptide scaffold were higher than that in the PLLA - PDA scaffold and the PLLA scaffold （ P = 0. 002 ）. Conclusion PLLA - PDA - BMP - 2 peptide nanofiber scaffold can release BMP - 2 peptide, promote the proliferation and osteogenic differentiation of BMSCs, and show potential ap- plications in bone tissue engineering.
作者
刘勇
陈亮
张云庆
姜雪峰
Liu Yong;Chen Liang;Zhang Yunqing;Jiang Xuefeng(Department of Orthopedics Surgery,Affiliated Jiangyin Renmin Hospital of Dongnan University,Jiangyin 214400,China;Department of Orthopedics Surgery,the First Affiliated Hospital of Soochow University,Suzhou 215006,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第9期1713-1716,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81772312）
关键词
聚乳酸
多巴胺
骨形成发生蛋白-2
骨
u vPolylactic acid
Poly dopamine
Bone morphogenetic protein - 2
Bone
