Effects of the controlled-released TGF-β1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold
Effects of the controlled-released TGF-β1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold
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DOI：
10.1016/j.biomaterials.2003.10.057
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发表时间：
2004-08-01
期刊：
影响因子：
14
通讯作者：
Lee, MC
中科院分区：
文献类型：
--
作者：
Lee, JE;Kim, KE;Lee, MC
本研究的目的是：(1)构建三维胶原/壳聚糖/糖胺聚糖（GAG）支架，并结合负载转化生长因子- β 1 （tgf - β 1）的壳聚糖微球，(2)评价释放的tgf - β 1对支架中兔软骨细胞成软骨潜能的影响。利用乳剂交联法将tgf - β 1负载到壳聚糖微球中。用酶联免疫吸附试验（ELISA）监测tgf - β 1的控释7天。在硫酸软骨素（CS）的存在下，采用冷冻干燥技术制备了含有胶原和壳聚糖的多孔支架，并用1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDC）交联。将tgf - β 1微球包埋在支架中，浓度为10 ng tgf - β 1/scaffold，然后将软骨细胞植入支架中，体外培养3周。tgf - β 1微球掺入支架的增殖率和糖胺聚糖（GAG）产量均显著高于未掺入微球的对照支架。tgf - β 1微球支架细胞外基质红素O染色和II型胶原免疫组化染色均升高。这些结果表明，tgf - β 1微球加入到支架中时，具有促进软骨形成的潜力。(C) 2003 Elsevier Ltd.版权所有。
The objectives of this study were (1) to develop a three-dimensional collagen/chitosan/glycosaminoglycan (GAG) scaffold in combination with transforming growth factor-beta1 (TGF-beta1)-loaded chitosan microspheres, and (2) to evaluate the effect of released TGF-beta1 on the chondrogenic potential of rabbit chondrocytes in such scaffolds.TGF-beta1 was loaded into chitosan microspheres using an emulsion-crosslinking method. The controlled release of TGF-beta1, as measured by enzyme-linked immunosorbent assay (ELISA), was monitored for 7 days. The porous scaffolds containing collagen and chitosan were fabricated by using a freeze drying technique and crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC) in the presence of chondroitin sulfate (CS), as a GAG component. The TGF-beta1 microspheres were encapsulated into the scaffold at a concentration of 10 ng TGF-beta1/scaffold and then chondrocytes were seeded in the scaffold and incubated in vitro for 3 weeks. Both proliferation rate and glycosaminoglycan (GAG) production were significantly higher in the TGF-beta1 microsphere-incorporated scaffolds than in the control scaffolds without microspheres. Extracellular matrix staining by Safranin O and imimmohistochemistry for type II collagen were elevated in the scaffold with TGF-beta1 microspheres. These results suggest that TGF-beta1 microspheres when incorporated into a scaffold have the potential to enhance cartilage formation. (C) 2003 Elsevier Ltd. All rights reserved.