Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
Tissue Eng Part A. 2010 Jun;16(6):1937-48. doi: 10.1089/ten.TEA.2009.0706.
Although human umbilical cord mesenchymal stromal cells (hUCMSCs) have been shown to differentiate along an osteogenic lineage in monolayer culture, the potential of these cells has seldom before been investigated in three-dimensional scaffolds for bone tissue engineering applications. In this 6-week study, we observed osteogenic differentiation of hUCMSCs on polyglycolic acid (PGA) nonwoven mesh scaffolds, and compared seeding densities for potential use in bone tissue engineering. Cells were seeded into PGA meshes with densities of 5, 25, or 50 x 10(6) cells/mL scaffold and then cultured in osteogenic medium. Cell proliferation, osteogenic differentiation, and matrix formation were evaluated at weeks 0, 3, and 6. Osteogenic differentiation was observed based on positive alkaline phosphatase activity and an increase of collagen production and calcium incorporation into the extracellular matrix, which increased with higher cell density. During differentiation, runt-related transcription factor (RUNX2), type I collagen (CI), and osteocalcin (OCN) gene expression levels were also increased. In conclusion, exposed to osteogenic signals, hUCMSCs differentiated along an osteogenic lineage as determined by expression of osteogenic markers and matrix formation, and increasing the density of hUCMSCs seeded onto three-dimensional PGA scaffolds led to better osteogenic differentiation.
尽管人脐带间充质干细胞(hUCMSCs)在单层培养中已被证明可沿着成骨谱系分化,但这些细胞在三维支架中的潜力在骨组织工程应用中很少被研究过。在这项 6 周的研究中,我们观察到 hUCMSCs 在聚乙醇酸(PGA)无纺网支架上的成骨分化,并比较了用于骨组织工程的潜在种子密度。细胞以 5、25 或 50 x 10(6)细胞/支架的密度接种到 PGA 网中,然后在成骨培养基中培养。在第 0、3 和 6 周评估细胞增殖、成骨分化和基质形成。成骨分化是基于碱性磷酸酶活性阳性和胶原产量增加以及细胞外基质中钙掺入的增加来观察到的,细胞密度越高,增加越明显。在分化过程中,RUNX2、I 型胶原(CI)和骨钙素(OCN)基因表达水平也增加。总之,在成骨信号的作用下,hUCMSCs 通过表达成骨标志物和基质形成沿着成骨谱系分化,并且增加接种到三维 PGA 支架上的 hUCMSCs 的密度可导致更好的成骨分化。