Department of Clinical Dentistry-Center for Clinical Dental Research, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
Tissue Eng Part A. 2011 Nov;17(21-22):2817-31. doi: 10.1089/ten.tea.2010.0660. Epub 2011 Sep 9.
One of the principal goals in tissue engineering is to produce scaffold materials that will guide cells to differentiate and regenerate functional replacement tissue at the site of injury. Poly(l-lactide-co-1,5-dioxepan-2-one) [Poly(LLA-co-DXO)], a potential scaffolding material for bone tissue engineering, has high hydrophilicity. Previous in vitro studies using human osteoblast-like cells (HOBs) demonstrated greater cytocompatibility and enhanced osteogenic differentiation when HOBs were seeded onto Poly(LLA-co-DXO) compared to Poly(l-lactide) [P(LLA)] scaffolds. The aim of the study was to identify the gene expression profiles of HOBs obtained from alveolar bone and grown on Poly(LLA-co-DXO) biodegradable polymer scaffolds compared to P(LLA) one. Illumina HumanWG-6 v3.0 Expression BeadChips were used for the gene expression analysis. Several genes were found as differentially expressed at 24 h and at 21 days. Expression of genes related to cell adhesion, cytoskeleton, antiapoptosis, proliferation, and bone mineralization was influenced by adding the monomer 1,5-dioxepan-2-one to the L-lactide. Genes related to three biological pathways involving Integrin, Notch, and Ras were found to be upregulated. For selected genes, results were confirmed by quantitative reverse transcriptase-polymerase chain reaction. Further, calcium content analysis revealed a significant enhancement of calcium deposition on both tested scaffolds. This observation was confirmed by Von Kossa and Alizarin Red S staining. Findings of this study are relevant to a better understanding of the molecular mechanisms underlying the behavior of HOBs in bone regenerative procedure.
组织工程的主要目标之一是生产支架材料,这些材料将引导细胞在损伤部位分化并再生功能性替代组织。聚(L-丙交酯-co-1,5-二氧杂环戊酮)[聚(LLA-co-DXO)]是一种有潜力的骨组织工程支架材料,具有较高的亲水性。先前使用人成骨样细胞(HOB)的体外研究表明,与聚(L-丙交酯)[P(LLA)]支架相比,HOB 接种到聚(LLA-co-DXO)上时具有更好的细胞相容性和增强的成骨分化。本研究的目的是鉴定从牙槽骨中获得的 HOB 的基因表达谱,并将其在聚(LLA-co-DXO)可生物降解聚合物支架上与 P(LLA)支架进行比较。Illumina HumanWG-6 v3.0 Expression BeadChips 用于基因表达分析。在 24 小时和 21 天时,发现了几个差异表达的基因。细胞黏附、细胞骨架、抗凋亡、增殖和骨矿化相关基因的表达受到添加 1,5-二氧杂环戊酮单体的影响。涉及整合素、Notch 和 Ras 的三个生物学途径的基因被发现上调。对于选定的基因,通过定量逆转录聚合酶链反应(qRT-PCR)对结果进行了验证。此外,钙含量分析显示两种测试支架上钙沉积的显著增强。Von Kossa 和茜素红 S 染色证实了这一观察结果。本研究的结果有助于更好地理解 HOB 在骨再生过程中的行为的分子机制。