多肽水凝胶对支架材料生物相容性和骨引导性影响的研究
发布时间：2018-08-03 19:11
【摘要】：本研究制备了一种新型的复合支架材料，通过多肽水凝胶对纳米羟基磷灰石/胶原（nano-Hydroxyapatite/Collagen, nHAC）进行表面修饰，未复合水凝胶的nHAC为对照组，探讨其对大鼠脂肪干细胞（Rat adipose-derived stemcells，rADSCs）黏附增殖和骨向分化能力以及体内骨再生能力的影响。本实验将rADSCs接种到两种支架材料上，使用扫描电子显微镜（Field emission scanningelectron microscope, FESEM）和共聚焦显微镜（Confocal laser scanningmicroscope, CLSM）进行表征，以CCK-8检测细胞毒性， DNA荧光比色法检测细胞增殖情况。以碱性磷酸酶（ALP）活性为早期成骨分化指标，并通过实时定量PCR检测成骨标志基因的表达，比较rADSCs的骨向分化情况。将支架植入大鼠颅顶骨缺损模型，在植入4周和6周后处死大鼠，观测体内骨再生情况。结果显示多肽水凝胶成纳米纤维多孔网状结构，孔径约300μm，在nHAC表面形成涂层，，rADSCs在多肽水凝胶复合支架材料（cnHAC）上黏附、铺展的更好。CCK-8及DNA荧光比色结果显示cnHAC能促进rADSCs在支架上的增殖，两组间差异有统计学意义。ALP和实时定量PCR结果表明cnHAC能显著促进rADSCs的骨向分化。大鼠颅顶骨缺损实验结果显示cnHAC能在早期有效地促进大鼠新骨形成。本研究结果表明cnHAC与nHAC相比，能显著促进细胞的黏附增殖、骨向分化及体内骨再生能力。用多肽水凝胶对nHAC进行表面修饰可作为一种简单有效的改性方式，提高支架材料的生物相容性和骨引导性。
[Abstract]:In this study, a new composite scaffold material was prepared. The nano-hydroxyapatite / collagen (nHAC) was modified by polypeptide hydrogel, and the nHAC without hydrogel was used as control group. Objective: to investigate the effects of ASCs on the adhesion, proliferation, bone differentiation and bone regeneration of rat adipose stem cells (Rat adipose-derived stem cells). RADSCs was inoculated into two scaffolds and characterized by scanning electron microscope (Field emission scanningelectron microscope, FESEM) and confocal microscope (Confocal laser scanningmicroscope, CLSM). Cytotoxicity was detected by CCK-8 and cell proliferation was detected by DNA fluorescence colorimetry. The activity of alkaline phosphatase (ALP) was used as the early osteogenic differentiation index, and the expression of osteogenic marker gene was detected by real-time quantitative PCR to compare the bone differentiation of rADSCs. The rat model of cranio-parietal bone defect was implanted with scaffold. The rats were killed at 4 and 6 weeks after implantation and bone regeneration in vivo was observed. The results showed that polypeptide hydrogels formed a porous network of nanofibers with pore size of about 300 渭 m, and a coating was formed on the surface of nHAC to adhere to the polypeptide hydrogel composite scaffold (cnHAC). The results of CCK-8 and DNA fluorescence colorimetry showed that cnHAC could promote the proliferation of rADSCs on the scaffold. The difference between the two groups was statistically significant. ALP and real-time quantitative PCR showed that cnHAC could significantly promote the bone differentiation of rADSCs. The experimental results of cranio-parietal bone defect in rats showed that cnHAC could effectively promote the formation of new bone in early stage. The results showed that cnHAC could significantly promote cell adhesion proliferation, bone differentiation and bone regeneration compared with nHAC. The surface modification of nHAC by polypeptide hydrogel can be used as a simple and effective way to improve the biocompatibility and bone guidance of scaffolds.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R783.1
[Abstract]:In this study, a new composite scaffold material was prepared. The nano-hydroxyapatite / collagen (nHAC) was modified by polypeptide hydrogel, and the nHAC without hydrogel was used as control group. Objective: to investigate the effects of ASCs on the adhesion, proliferation, bone differentiation and bone regeneration of rat adipose stem cells (Rat adipose-derived stem cells). RADSCs was inoculated into two scaffolds and characterized by scanning electron microscope (Field emission scanningelectron microscope, FESEM) and confocal microscope (Confocal laser scanningmicroscope, CLSM). Cytotoxicity was detected by CCK-8 and cell proliferation was detected by DNA fluorescence colorimetry. The activity of alkaline phosphatase (ALP) was used as the early osteogenic differentiation index, and the expression of osteogenic marker gene was detected by real-time quantitative PCR to compare the bone differentiation of rADSCs. The rat model of cranio-parietal bone defect was implanted with scaffold. The rats were killed at 4 and 6 weeks after implantation and bone regeneration in vivo was observed. The results showed that polypeptide hydrogels formed a porous network of nanofibers with pore size of about 300 渭 m, and a coating was formed on the surface of nHAC to adhere to the polypeptide hydrogel composite scaffold (cnHAC). The results of CCK-8 and DNA fluorescence colorimetry showed that cnHAC could promote the proliferation of rADSCs on the scaffold. The difference between the two groups was statistically significant. ALP and real-time quantitative PCR showed that cnHAC could significantly promote the bone differentiation of rADSCs. The experimental results of cranio-parietal bone defect in rats showed that cnHAC could effectively promote the formation of new bone in early stage. The results showed that cnHAC could significantly promote cell adhesion proliferation, bone differentiation and bone regeneration compared with nHAC. The surface modification of nHAC by polypeptide hydrogel can be used as a simple and effective way to improve the biocompatibility and bone guidance of scaffolds.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R783.1
【参考文献】
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1 嵇伟平;韩培;蒋W
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