Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, Seoul, Republic of Korea.
BK21 Plus Future Biopharmaceutical Human Resources Training and Research Team, Hanyang University, Seoul, Republic of Korea.
Tissue Eng Part A. 2020 Dec;26(23-24):1243-1258. doi: 10.1089/ten.tea.2020.0015. Epub 2020 Apr 13.
Guided bone regeneration refers to the process in which bone defects could be regenerated by facilitated healing through the use of membranes, potentially with the delivery of osteoinductive molecules, however, the regeneration often failed due to inflammation during bone formation. In this study, we developed a membrane immobilized with lactoferrin to modulate both bone regeneration and inflammatory responses. Lactoferrin was immobilized on electrospun nanofibers (LF50) by exploiting an adhesive polydopamine coating method. When human adipose-derived stem cells (hADSCs) were seeded onto the nanofibers, the LF50 significantly increased the osteogenic differentiation. For example, the gene expression of osteopontin was 6.9 ± 2.3 times greater in the cells on LF50 than the cells on unmodified nanofibers without lactoferrin. In addition, the gene expression of tumor necrosis factor-alpha () of the macrophage cell line (RAW264.7) cultured on the LF50 was 0.3 ± 0.1 times reduced, indicating the lactoferrin was able to reduce inflammatory response. With implantation of nanofibers on mouse calvarial defects, the LF50 resulted in 60.9% ± 4.5% of new bone formation, which was six times greater than the results of other groups. Furthermore, when the fibers were implanted onto the mouse subcutaneous model challenged with lipopolysaccharide and interferon-γ, the area of inflammatory tissue was significantly reduced in the LF50 implanted group as 0.6 ± 0.1 mm as compared with the control group (1.1 ± 0.1 mm). Taken together, the lactoferrin immobilization onto the nanofiber by polydopamine chemistry may be an effective delivery method for improving bone regeneration while regulating the inflammation. Impact statement critical-sized bone reconstruction remains challenging due to the severe inflammation, which would be an unavoidable problem during surgical process. Therefore, the present study aims to develop a guided nanofibrous membrane immobilized with lactoferrin, which has dual functions with osteoinduction and anti-inflammation. The lactoferrin-immobilized fibers demonstrated significantly enhanced osteogenic differentiation of adipose-derived stem cells as well as decreased polarization of macrophage to M1 with relatively reduced amount than that reported from previous reports. We also found that the membrane improved bone regeneration and decreased inflammatory tissue formation. Taken together, this system would be a new platform for successful bone regeneration.
引导骨再生是指通过使用膜来促进骨缺损的愈合，从而实现骨再生的过程，这可能需要递送成骨诱导分子，然而，由于骨形成过程中的炎症，再生往往会失败。在这项研究中，我们开发了一种用乳铁蛋白固定的膜，以调节骨再生和炎症反应。乳铁蛋白通过利用粘性聚多巴胺涂层方法固定在静电纺纳米纤维上（LF50）。当人脂肪源性干细胞（hADSCs）接种在纳米纤维上时，LF50 显著促进了成骨分化。例如，在 LF50 上的细胞中骨桥蛋白的基因表达是未修饰纳米纤维上无乳铁蛋白的细胞的 6.9±2.3 倍。此外，在 LF50 上培养的巨噬细胞系（RAW264.7）的肿瘤坏死因子-α（TNF-α）的基因表达减少了 0.3±0.1 倍，表明乳铁蛋白能够减少炎症反应。在小鼠颅骨缺损上植入纳米纤维后，LF50 导致 60.9%±4.5%的新骨形成，是其他组的六倍。此外，当纤维植入脂多糖和干扰素-γ挑战的小鼠皮下模型时，LF50 植入组的炎症组织面积明显减少，为 0.6±0.1mm，而对照组为 1.1±0.1mm。总之，通过聚多巴胺化学将乳铁蛋白固定在纳米纤维上可能是一种有效的递送方法，可在改善骨再生的同时调节炎症。
由于严重的炎症，临界尺寸的骨重建仍然具有挑战性，这将是手术过程中不可避免的问题。因此，本研究旨在开发一种固定有乳铁蛋白的引导性纳米纤维膜，该膜具有成骨诱导和抗炎的双重功能。固定有乳铁蛋白的纤维表现出明显增强的脂肪源性干细胞成骨分化，以及相对减少的巨噬细胞向 M1 极化的程度，比以前的报告报道的要少。我们还发现该膜改善了骨再生并减少了炎症组织的形成。总的来说，该系统将成为成功骨再生的新平台。
