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新型纺织壳聚糖支架促进成骨细胞的铺展、增殖和分化。

Novel textile chitosan scaffolds promote spreading, proliferation, and differentiation of osteoblasts.

作者信息

Heinemann Christiane, Heinemann Sascha, Bernhardt Anne, Worch Hartmut, Hanke Thomas

机构信息

Max Bergmann Center of Biomaterials and Institute of Materials Science, Dresden University of Technology, Budapester Str. 27, D-01069 Dresden, Germany.

出版信息

Biomacromolecules. 2008 Oct;9(10):2913-20. doi: 10.1021/bm800693d. Epub 2008 Sep 5.

Abstract

Two novel scaffold models made of chitosan fibers were designed, fabricated, and investigated. Raw chitosan fibers were either tightened between plastic rings or were processed into stand-alone scaffolds. Chitosan fiber scaffolds were further modified by coating with a thin layer of fibrillar collagen type I to biologize the surface. Cell culture experiments were carried out using murine osteoblast-like cells (7F2). Confocal laser scanning microscopy (cLSM) as well as scanning electron microscopy (SEM) revealed fast attachment and morphological adaptation of the cells on both the raw chitosan fibers and the collagen-coated scaffolds. Cells were cultivated for up to 4 weeks on the materials and proliferation as well as osteogenic differentiation was quantitatively analyzed in terms of lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) activity. We found a 14-16-fold increase of cell number and the typical pattern of ALP activity, whereas the collagen coating does not remarkably influence these parameters. The maintenance of osteogenic phenotype on the novel materials was furthermore confirmed by immunostaining of osteocalcin and study of matrix mineralization. The feature of the collagen-coated but also the raw chitosan fiber scaffolds to support the attachment, proliferation, and differentiation of osteoblast-like cells suggest a potential application of chitosan fibers and textile chitosan scaffolds for the tissue engineering of bone.

摘要

设计、制造并研究了两种由壳聚糖纤维制成的新型支架模型。未加工的壳聚糖纤维要么被紧束在塑料环之间,要么被加工成独立的支架。壳聚糖纤维支架通过涂覆一层薄薄的I型原纤维胶原蛋白进行进一步修饰,以使表面生物化。使用小鼠成骨样细胞(7F2)进行细胞培养实验。共聚焦激光扫描显微镜(cLSM)以及扫描电子显微镜(SEM)显示,细胞在未加工的壳聚糖纤维和胶原蛋白涂层支架上均能快速附着并发生形态适应。在材料上培养细胞长达4周,并根据乳酸脱氢酶(LDH)和碱性磷酸酶(ALP)活性对细胞增殖和成骨分化进行定量分析。我们发现细胞数量增加了14至16倍,且出现了典型的ALP活性模式,而胶原蛋白涂层对这些参数没有显著影响。通过骨钙素免疫染色和基质矿化研究,进一步证实了新型材料上成骨表型的维持。胶原蛋白涂层的以及未加工的壳聚糖纤维支架支持成骨样细胞附着、增殖和分化的特性表明,壳聚糖纤维和纺织壳聚糖支架在骨组织工程中具有潜在应用价值。

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