Treating Titanium Particle-Induced Inflammation with Genetically Modified NF-κB Sensing IL-4 Secreting or Preconditioned Mesenchymal Stem Cells in Vitro
Treating Titanium Particle-Induced Inflammation with Genetically Modified NF-κB Sensing IL-4 Secreting or Preconditioned Mesenchymal Stem Cells in Vitro
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DOI：
10.1021/acsbiomaterials.9b00560
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发表时间：
2019-06-01
影响因子：
5.8
通讯作者：
Goodman, Stuart B.
中科院分区：
文献类型：
--
作者：
Kohno, Yusuke;Lin, Tzuhua;Goodman, Stuart B.
钛和钛基合金广泛用于骨科植入物。全关节置换术是非常成功的，然而，由植入物衍生的生物材料碎片引起的异物反应和慢性炎症仍然是未解决的问题。无菌性松动伴随磨损碎屑诱导的骨溶解（骨丢失）是晚期失败和翻修手术的最常见原因之一。间充质干细胞（MSC）和IL-4可能是可能的治疗策略，因为它们的免疫调节特性。我们在钛颗粒诱导的假体周围组织炎症的创新细胞共培养模型中研究了基于MSC的新型治疗对免疫调节和成骨分化的有效性。从Balb/c小鼠的骨髓中收集MSC和巨噬细胞。将MSC和巨噬细胞（代表假体周围组织的内源性细胞）接种在24孔transwell板的底部威尔斯孔上。我们产生了基因修饰的NF-κ B感应IL-4分泌MSC（炎症反应MSC）和经脂多糖和TNF-α预处理的MSC以进一步增强其免疫调节功能。将这些修饰的MSC（代表植入假体周围组织的外源性治疗细胞）接种在transwell板的上室上。然后将这些共培养物暴露于钛颗粒7天。在第7天，NF-κ B敏感IL-4分泌MSC显示出强免疫调节（显著降低TNF-α并诱导Argl表达）并促进早期骨生成（显著诱导Runx 2、ALP和β-连环蛋白以及降低Smurf 2表达）。分泌IL-4的MSC也早在第3天就降低了TNF-α蛋白的分泌，并在第7天增加了IL-1 ra蛋白的分泌，这表明对颗粒诱导的炎症具有有效的免疫调节作用。在这个短期实验中，预处理的MSC没有显示出显著的免疫调节作用，但是在第7天ALP和β-连环蛋白表达被显著诱导。我们的研究结果表明，基因修饰的IL-4分泌间充质干细胞和预处理的间充质干细胞有可能优化骨再生的炎症条件下，包括假体周围骨溶解。
Titanium and titanium-based alloys are widely used in orthopaedic implants. Total joint replacement is very successful; however, the foreign body response and chronic inflammation caused by implant-derived biomaterial debris still remain as unsolved issues. Aseptic loosening accompanied by wear debris-induced osteolysis (bone loss) is one of the most frequent' causes for late failure and revision surgery. Mesenchymal stem cells (MSCs) and IL-4 may be possible treatment strategies because of their immunomodulatory properties. We investigated the efficacy of novel MSC-based treatments on immunomodulation and osteogenic differentiation in an innovative cell coculture model of titanium particle- induced inflammation in the periprosthetic tissues. MSCs and macrophages were collected from the bone marrow of Balb/c mice. Both MSCs and macrophages (representing endogenous cells at the periprosthetic tissue) were seeded on the bottom wells of the 24-well transwell plates. We generated genetically modified NF-kappa B sensing IL-4 secreting MSCs (inflammatory responsive MSCs) and MSCs preconditioned by lipopolysaccharide and TNF-alpha to further enhance their immunomodulatory function. These modified MSCs (representing exogenous therapeutic cells implanted to the periprosthetic tissue) were seeded on the upper chambers of the transwell plates. These cocultures were then exposed to titanium particles for 7 days. NF-kappa B sensing IL-4 secreting MSCs showed strong immunomodulation (significantly reduced TNF-alpha and induced Argl expression) and promoted early osteogenesis (significantly induced Runx2, ALP, and beta-catenin as well as reduced Smurf2 expression) at day 7. IL-4 secreting MSCs also decreased TNF-alpha protein secretion as early as day 3 and increased IL-1ra protein secretion at day 7, suggesting efficacious immunomodulation of particle-induced inflammation. Preconditioned MSCs did not show significant immunomodulation in this short-term experiment, but ALP and beta-catenin expression were significantly induced at day 7. Our results suggest that genetically modified IL-4 secreting MSCs and preconditioned MSCs have the potential to optimize bone regeneration in inflammatory conditions including periprosthetic osteolysis.