Fabrication of Interleukin-4 Encapsulated Bioactive Microdroplets for Regulating Inflammation and Promoting Osteogenesis.
Fabrication of Interleukin-4 Encapsulated Bioactive Microdroplets for Regulating Inflammation and Promoting Osteogenesis.
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DOI：
10.2147/ijn.s397359
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发表时间：
2023
影响因子：
8
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中科院分区：
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尽管骨具有固有的再生能力，但大型骨缺损的再生仍然是骨科外科面临的主要临床挑战。M2表型巨噬细胞或M2巨噬细胞诱导剂的治疗策略已被广泛用于促进组织重塑。本研究制备了包被生物活性分子白介素4(IL4)的超声响应型生物活性微滴(MDs-IL4)，以调节巨噬细胞极化并增强人骨髓间充质干细胞(HBMSCs)的成骨分化。采用四甲基偶氮唑盐比色法、死活染色、鬼臼糖苷/DAPI双重染色评价材料的体外生物相容性。采用H&E染色评价材料的体内生物相容性。进一步通过内毒素刺激诱导炎性巨噬细胞，以模拟促炎状态。通过巨噬细胞表型标志物基因表达、促炎细胞因子水平、细胞形态分析、免疫荧光染色等检测MDS-IL4的免疫调节作用，并进一步研究hBMSCs通过巨噬细胞和hBMSCs相互作用的免疫-成骨效应。生物活性MDS-IL4支架在RAW 264.7巨噬细胞和人骨髓间充质干细胞中表现出良好的细胞相容性。结果证实，具有生物活性的MDS-IL4支架可以减少炎症表型巨噬细胞，表现为形态特征的改变，促炎标志物基因表达的降低，M2表型标志物基因的增加，以及促炎性细胞因子的分泌。此外，我们的结果表明，具有生物活性的MDS-IL4可以通过其潜在的免疫调节特性显著促进hBMSCs的成骨分化。我们的结果表明，生物活性MDS-IL4支架可以作为其他促成骨分子的新型载体系统，因此在骨组织再生方面具有潜在的应用前景。
Despite the inherent regenerative ability of bone, large bone defect regeneration remains a major clinical challenge for orthopedic surgery. Therapeutic strategies medicated by M2 phenotypic macrophages or M2 macrophage inducer have been widely used to promote tissue remodeling. In this study, ultrasound-responsive bioactive microdroplets (MDs) encapsulated with bioactive molecule interleukin-4 (IL4, hereafter designated MDs-IL4) were fabricated to regulate macrophage polarization and potentiate the osteogenic differentiation of human mesenchymal stem cells (hBMSCs). The MTT assay, live and dead staining, and phalloidin/DAPI dual staining were used to evaluate biocompatibility in vitro. H&E staining was used to evaluate biocompatibility in vivo. Inflammatory macrophages were further induced via lipopolysaccharide (LPS) stimulation to mimic the pro-inflammatory condition. The immunoregulatory role of the MDs-IL4 was tested via macrophage phenotypic marker gene expression, pro-inflammatory cytokine level, cell morphological analysis, and immunofluorescence staining, etc. The immune-osteogenic response of hBMSCs via macrophages and hBMSCs interactions was further investigated in vitro. The bioactive MDs-IL4 scaffold showed good cytocompatibility in RAW 264.7 macrophages and hBMSCs. The results confirmed that the bioactive MDs-IL4 scaffold could reduce inflammatory phenotypic macrophages, as evidenced by changing in morphological features, reduction in pro-inflammatory marker gene expression, increase of M2 phenotypic marker genes, and inhibition of pro-inflammatory cytokine secretion. Additionally, our results indicate that the bioactive MDs-IL4 could significantly enhance the osteogenic differentiation of hBMSCs via its potential immunomodulatory properties. Our results demonstrate that the bioactive MDs-IL4 scaffold could be used as novel carrier system for other pro-osteogenic molecules, thus having potential applications in bone tissue regeneration.