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Interaction of bone cells with biomimetic hydroxyapatite gelatin nanocomposites towards developing bone tissue engineering.

机译:骨细胞与仿生羟基磷灰石明胶纳米复合材料之间的相互作用促进了骨组织工程的发展。

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摘要

Significance. Bone tissue engineering provides an alternative to restore the complex morphologic features and functions of bone during repair of large bone defects. The biomaterials investigated in bone tissue engineering to date have their inherent disadvantages. An ideal scaffold biomaterial remains to be developed. Objectives. A novel biomimetic hydroxyapatite/gelatin nanocomposite (HAP/GEL) was developed to be a potential scaffold material. The objective is; therefore, to investigate the interaction between bone cells and the HAP/GEL in vitro and in vivo. This dissertation lays the groundwork and provides the protocols for the future development of this HAP/GEL nanocomposite as a scaffold for bone tissue engineering. Materials and methods. The human fetal osteoblastic cell line was used to assess cell morphology, cell spreading, cell growth and cell differentiation on HAP/GEL in vitro by using techniques of confocal laser scanning microscopy, PicoGreen DNA assay, and real-time PCR. The drill-hole defects were created in rat femurs to examine the biocompatibility, osteoconductivity, and biodegradation of HAP/GEL by using the techniques of fluorochrome labeling, histology, histomorphometry, and immunohstochemistry. Results. In vitro: Cells adhered on HAP/GEL appeared more elongated spindle shape but exhibited same degree of cell spreading compared to cells adhered on hydroxyapatite (HA) or glass. The number of cells grown on the HAP/GEL or HA increased over time, but at a slower rate than that increased on glass. The gene expression of alkaline phosphatase, bone sialoprotein, osteopontin, or osteocalcin was similar for cells grown on HAP/GEL, HA, or glass. Type I collagen expression was lower for cells grown on HAP/GEL. In vivo. Osteoblastic activity and new bone formation occurred on the surface of HAP/GEL. There was a trend that HAP/GEL may enhance bone formation in defect walls at two weeks postopertiavely. The osteoconductivity of HAP/GEL increased over time during eight weeks of healing period. Foreign body reaction was found at the early stage. Osteoclasts were observed on the surface of HAP/GEL. Conclusions. HAP/GEL provides a satisfactory surface for cell spreading, cell growth, and cell differentiation in vitro. HAP/GEL supports the recruitment of the osteogenic cells and osteogenesis in vivo. Osteoclasts may participate in the biodegradation of HAP/GEL.
机译:意义。骨组织工程学提供了一种在修复大骨缺损时恢复复杂的骨骼形态和功能的方法。迄今为止,在骨组织工程中研究的生物材料具有其固有的缺点。理想的支架生物材料仍有待开发。目标。一种新型的仿生羟基磷灰石/明胶纳米复合材料(HAP / GEL)被开发为一种潜在的支架材料。目的是;因此,在体外和体内研究骨细胞与HAP / GEL之间的相互作用。本论文奠定了基础,并为该HAP / GEL纳米复合材料作为骨组织工程支架的未来发展提供了协议。材料和方法。通过共聚焦激光扫描显微镜技术,PicoGreen DNA检测和实时PCR技术,将人类胎儿成骨细胞系用于体外评估HAP / GEL上的细胞形态,细胞扩散,细胞生长和细胞分化。通过使用荧光染料标记,组织学,组织形态学和免疫组织化学技术,在大鼠股骨中创建钻孔缺陷,以检查HAP / GEL的生物相容性,骨传导性和生物降解性。结果。体外:与粘附在羟基磷灰石(HA)或玻璃上的细胞相比,粘附在HAP / GEL上的细胞表现出更细长的纺锤形,但表现出相同程度的细胞扩散。随着时间的推移,在HAP / GEL或HA上生长的细胞数量会增加,但其速度要比玻璃上的速度慢。对于在HAP / GEL,HA或玻璃上生长的细胞,碱性磷酸酶,骨唾液蛋白,骨桥蛋白或骨钙素的基因表达相似。对于在HAP / GEL上生长的细胞,I型胶原蛋白表达较低。体内。 HAP / GEL表面发生了成骨细胞活性和新骨形成。手术后两周,HAP / GEL可能会增强缺损壁的骨形成。在愈合期的八周内,HAP / GEL的骨电导率随时间增加。早期发现异物反应。在HAP / GEL表面观察到破骨细胞。结论。 HAP / GEL为体外细胞扩散,细胞生长和细胞分化提供了令人满意的表面。 HAP / GEL支持体内成骨细胞的募集和成骨。破骨细胞可能参与HAP / GEL的生物降解。

著录项

  • 作者

    Wu, Ying-Lien.;

  • 作者单位

    University of Minnesota.;

  • 授予单位 University of Minnesota.;
  • 学科 Health Sciences Dentistry.;Engineering Biomedical.
  • 学位 Ph.D.
  • 年度 2007
  • 页码 227 p.
  • 总页数 227
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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