Sutherland Amanda J, Detamore Michael S
Bioengineering Graduate Program, University of Kansas, Lawrence, 66045, Kansas, USA.
Department of Chemical and Petroleum Engineering, University of Kansas, 4149 Learned Hall, 1530 W 15th St., Lawrence, 66045, Kansas, USA.
Macromol Biosci. 2015 Jul;15(7):979-89. doi: 10.1002/mabi.201400472. Epub 2015 Mar 27.
The aim of this study was to fabricate mechanically functional microsphere-based scaffolds containing decellularized cartilage (DCC), with the hypothesis that this approach would induce chondrogenesis of rat bone marrow-derived mesenchymal stem cells (rBMSCs) in vitro. The DCC was derived from porcine articular cartilage and decellularized using a combination of physical and chemical methods. Four types of scaffolds were fabricated: poly(d,l-lactic-co-glycolic acid) (PLGA) only (negative control), TGF-β-encapsulated (positive control), PLGA surface coated with DCC, and DCC-encapsulated. These scaffolds were seeded with rBMSCs and cultured up to 6 weeks. The compressive modulus of the DCC-coated scaffolds prior to cell seeding was significantly lower than all other scaffold types. Gene expression was comparable between DCC-encapsulated and TGF-β-encapsulated groups. Notably, DCC-encapsulated scaffolds contained 70% higher glycosaminoglyan (GAG) content and 85% more hydroxyproline compared to the TGF-β group at week 3 (with baseline levels subtracted out from acellular DCC scaffolds). Certainly, bioactivity was demonstrated in eliciting a biosynthetic response from the cells with DCC, although true demonstration of chondrogenesis remained elusive under the prescribed conditions. Encapsulation of DCC appeared to lead to improved cell performance relative to coating with DCC, although this finding may be a dose-dependent observation. Overall, DCC introduced via microsphere-based scaffolds appears to be promising as a bioactive approach to cartilage regeneration, although additional studies will be required to conclusively demonstrate chondroinductivity.
本研究的目的是制备含有脱细胞软骨(DCC)的具有机械功能的微球基支架,假设该方法将在体外诱导大鼠骨髓间充质干细胞(rBMSCs)的软骨形成。DCC来源于猪关节软骨,并通过物理和化学方法相结合进行脱细胞处理。制备了四种类型的支架:仅聚(d,l-乳酸-共-乙醇酸)(PLGA)(阴性对照)、包封转化生长因子-β(TGF-β)的(阳性对照)、表面涂覆DCC的PLGA以及包封DCC的。将这些支架接种rBMSCs并培养长达6周。细胞接种前,DCC涂层支架的压缩模量显著低于所有其他类型的支架。DCC包封组和TGF-β包封组之间的基因表达相当。值得注意的是,在第3周时,与TGF-β组相比,DCC包封的支架中糖胺聚糖(GAG)含量高70%,羟脯氨酸多85%(从无细胞DCC支架中减去基线水平)。当然,DCC能够引发细胞的生物合成反应,证明了其生物活性,尽管在规定条件下仍难以真正证明软骨形成。相对于DCC涂层,DCC包封似乎导致细胞性能得到改善,尽管这一发现可能是剂量依赖性观察结果。总体而言,通过微球基支架引入的DCC作为软骨再生的生物活性方法似乎很有前景,尽管需要进一步的研究来最终证明其软骨诱导性。