El-Amin Saadiq F, Botchwey Edward, Tuli Richard, Kofron Michelle D, Mesfin Addisu, Sethuraman Swaminathan, Tuan Rocky S, Laurencin Cato T
Department of Orthopaedic Surgery, University of Virginia, 400 Ray C. Hunt Drive, Charlottesville, Virginia 22903, USA.
J Biomed Mater Res A. 2006 Mar 1;76(3):439-49. doi: 10.1002/jbm.a.30411.
We performed a detailed examination of the isolation, characterization, and growth of human osteoblast cells derived from trabecular bone. We further examined the morphology, phenotypic gene expression, mineralization,and growth of these human osteoblasts on polyester polymers used for musculoskeletal tissue engineering. Polylactic-co-glycolic acid [PLAGA (85:15, 50:50, 75:25)], and poly-lactic acid (L-PLA, D,L-PLA) were examined. The osteoblastic expression of key phenotypic markers osteocalcin, alkaline phosphatase, collagen, and bone sialoprotein at 4 and 8 weeks was examined. Reverse transcription-polymerase chain reaction studies revealed that trabecular-derived osteoblasts were positive for all markers evaluated with higher levels expressed over long-term culture. These cells also revealed mineralization and maturation as evidenced by energy dispersive X-ray analysis and scanning electron microscopy. Growth studies on PLAGA at 50:50,75:25, and 85:15 ratios and PLA in the L and DL isoforms revealed that human osteoblasts actively grew, with significantly higher cell numbers attached to scaffolds composed of PLAGA 50:50 in the short term and PLAGA 85:15 in the long term compared with PLA (p < 0.05). We believe human cell adhesion among these polymeric materials may be dependent on differences in cellular integrin expression and extracellular matrix protein elaboration.
我们对源自松质骨的人成骨细胞的分离、特性鉴定及生长进行了详细研究。我们进一步研究了这些人成骨细胞在用于肌肉骨骼组织工程的聚酯聚合物上的形态、表型基因表达、矿化及生长情况。对聚乳酸-乙醇酸共聚物[PLAGA(85:15、50:50、75:25)]和聚乳酸(L-PLA、D,L-PLA)进行了检测。检测了4周和8周时关键表型标志物骨钙素、碱性磷酸酶、胶原蛋白和骨唾液蛋白的成骨细胞表达情况。逆转录-聚合酶链反应研究显示,源自松质骨的成骨细胞对所有评估标志物均呈阳性,且在长期培养中表达水平更高。能量色散X射线分析和扫描电子显微镜结果表明,这些细胞也呈现出矿化和成熟现象。对比例为50:50、75:25和85:15的PLAGA以及L型和DL型PLA上的生长研究显示,人成骨细胞能够活跃生长,与PLA相比,短期内附着在由50:50的PLAGA组成的支架上、长期内附着在85:15的PLAGA组成的支架上的细胞数量显著更多(p<0.05)。我们认为,这些聚合材料之间的人细胞黏附可能取决于细胞整合素表达和细胞外基质蛋白分泌的差异。