Lee Jong Eun, Kim Ko Eun, Kwon Ick Chan, Ahn Hyun Jeong, Lee Sang-Hoon, Cho Hyunchul, Kim Hee Joong, Seong Sang Chul, Lee Myung Chul
Seoul National University College of Medicine, Orthopedic Surgery, 28 Yongon-dong Chongno-gu, Seoul 110-744, South Korea.
Biomaterials. 2004 Aug;25(18):4163-73. doi: 10.1016/j.biomaterials.2003.10.057.
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-beta 1)-loaded chitosan microspheres, and (2) to evaluate the effect of released TGF-beta 1 on the chondrogenic potential of rabbit chondrocytes in such scaffolds. TGF-beta 1 was loaded into chitosan microspheres using an emulsion-crosslinking method. The controlled release of TGF-beta 1, 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-beta 1 microspheres were encapsulated into the scaffold at a concentration of 10 ng TGF-beta 1/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-beta 1 microsphere-incorporated scaffolds than in the control scaffolds without microspheres. Extracellular matrix staining by Safranin O and immunohistochemistry for type II collagen were elevated in the scaffold with TGF-beta 1 microspheres. These results suggest that TGF-beta 1 microspheres when incorporated into a scaffold have the potential to enhance cartilage formation.
（1）开发一种三维胶原/壳聚糖/糖胺聚糖（GAG）支架，并结合负载转化生长因子-β1（TGF-β1）的壳聚糖微球；（2）评估释放的TGF-β1对兔软骨细胞在这种支架中软骨形成潜能的影响。采用乳化交联法将TGF-β1负载到壳聚糖微球中。通过酶联免疫吸附测定（ELISA）测量TGF-β1的控释情况，并监测7天。含有胶原和壳聚糖的多孔支架采用冷冻干燥技术制备，并在作为GAG成分的硫酸软骨素（CS）存在下，使用1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDC）进行交联。将TGF-β1微球以10 ng TGF-β1/支架的浓度包封到支架中，然后将软骨细胞接种到支架中并在体外培养3周。与不含微球的对照支架相比，含有TGF-β1微球的支架中的增殖率和糖胺聚糖（GAG）产量均显著更高。用番红O进行的细胞外基质染色和针对II型胶原的免疫组织化学在含有TGF-β1微球的支架中有所增强。这些结果表明，当TGF-β1微球掺入支架中时，具有增强软骨形成的潜力。
