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新型聚(L-乳酸)/透明质酸大孔杂化支架:细胞毒性的表征和评估。

Novel poly(L-lactic acid)/hyaluronic acid macroporous hybrid scaffolds: characterization and assessment of cytotoxicity.

机构信息

University Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, , Guimarães, Portugal.

出版信息

J Biomed Mater Res A. 2010 Sep 1;94(3):856-69. doi: 10.1002/jbm.a.32753.

DOI:10.1002/jbm.a.32753
PMID:20336752
Abstract

Poly(L-lactic acid), PLLA, a synthetic biodegradable polyester, is widely accepted in tissue engineering. Hyaluronic acid (HA), a natural polymer, exhibits an excellent biocompatibility, influences cell signaling, proliferation, and differentiation. In this study, HA crosslinking was performed by immersion of the polysaccharide in water-acetone mixtures containing glutaraldehyde (GA). The objective of this work is to produce PLLA scaffolds with the pores coated with HA, that could be beneficial for bone tissue engineering applications. PLLA tridimensional scaffolds were prepared by compression molding followed by salt leaching. After the scaffolds impregnation with soluble HA solutions of distinct concentration, a GA-crosslinking reaction followed by inactivation of the unreacted GA with glycine was carried out. An increase on surface roughness is shown by scanning electron microscopy (SEM) with the addition of HA. Toluidine blue staining indicates the present of stable crosslinked HA. An estimation of the HA original weight in the hybrid scaffolds was performed using thermal gravimetric analyses. FTIR-ATR and XPS confirmed the crosslinking reaction. Preliminary in vitro cell culture studies were carried out using a mouse lung fibroblast cell line (L929). SEM micrographs of L929 showed that cells adhered well, spread actively throughout all scaffolds, and grew favorably. A MTS test indicated that cells were viable when cultured onto the surface of all scaffolds, suggesting that the introduction of crosslinked HA did not increase the cytotoxicity of the hybrid scaffolds.

摘要

聚(L-乳酸),PLLA,一种合成的可生物降解聚酯,在组织工程中被广泛接受。透明质酸(HA),一种天然聚合物,表现出极好的生物相容性,影响细胞信号转导、增殖和分化。在这项研究中,通过将多糖浸入含有戊二醛(GA)的水-丙酮混合物中来进行 HA 交联。这项工作的目的是生产具有 HA 涂层的孔的 PLLA 支架,这可能有益于骨组织工程应用。通过压缩成型和盐浸提制备 PLLA 三维支架。在将可溶性 HA 溶液浸渍到支架中之后,进行 GA 交联反应,然后用甘氨酸使未反应的 GA 失活。扫描电子显微镜(SEM)显示,随着 HA 的添加,表面粗糙度增加。甲苯胺蓝染色表明存在稳定的交联 HA。使用热重分析对混合支架中 HA 的原始重量进行了估计。FTIR-ATR 和 XPS 证实了交联反应。使用小鼠肺成纤维细胞系(L929)进行了初步的体外细胞培养研究。L929 的 SEM 显微照片表明,细胞很好地粘附,在所有支架中积极扩散并生长良好。MTS 测试表明,当细胞在所有支架的表面上培养时,细胞是存活的,这表明交联 HA 的引入没有增加杂交支架的细胞毒性。

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