Suliman Salwa, Mieszkowska Anna, Folkert Justyna, Rana Neha, Mohamed-Ahmed Samih, Fuoco Tiziana, Finne-Wistrand Anna, Dirscherl Kai, Jørgensen Bodil, Mustafa Kamal, Gurzawska-Comis Katarzyna
Centre of Translational Oral Research (TOR), Department of Clinical Dentistry, Faculty of Medicine, University of Bergen, Årstadveien 19, 5009, Bergen, Norway.
Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 31-007, Krakow, Poland.
Inflamm Regen. 2022 Apr 3;42(1):12. doi: 10.1186/s41232-022-00196-9.
Age-driven immune signals cause a state of chronic low-grade inflammation and in consequence affect bone healing and cause challenges for clinicians when repairing critical-sized bone defects in elderly patients.
Poly(L-lactide-co-ɛ-caprolactone) (PLCA) scaffolds are functionalized with plant-derived nanoparticles from potato, rhamnogalacturonan-I (RG-I), to investigate their ability to modulate inflammation in vitro in neutrophils and macrophages at gene and protein levels. The scaffolds' early and late host response at gene, protein and histological levels is tested in vivo in a subcutaneous rat model and their potential to promote bone regeneration in an aged rodent was tested in a critical-sized calvaria bone defect. Significant differences were tested using one-way ANOVA, followed by a multiple-comparison Tukey's test with a p value ≤ 0.05 considered significant.
Gene expressions revealed PLCA scaffold functionalized with plant-derived RG-I with a relatively higher amount of galactose than arabinose (potato dearabinated (PA)) to reduce the inflammatory state stimulated by bacterial LPS in neutrophils and macrophages in vitro. LPS-stimulated neutrophils show a significantly decreased intracellular accumulation of galectin-3 in the presence of PA functionalization compared to Control (unmodified PLCA scaffolds). The in vivo gene and protein expressions revealed comparable results to in vitro. The host response is modulated towards anti-inflammatory/ healing at early and late time points at gene and protein levels. A reduced foreign body reaction and fibrous capsule formation is observed when PLCA scaffolds functionalized with PA were implanted in vivo subcutaneously. PLCA scaffolds functionalized with PA modulated the cytokine and chemokine expressions in vivo during early and late inflammatory phases. PLCA scaffolds functionalized with PA implanted in calvaria defects of aged rats downregulating pro-inflammatory gene markers while promoting osteogenic markers after 2 weeks in vivo.
We have shown that PLCA scaffolds functionalized with plant-derived RG-I with a relatively higher amount of galactose play a role in the modulation of inflammatory responses both in vitro and in vivo subcutaneously and promote the initiation of bone formation in a critical-sized bone defect of an aged rodent. Our study addresses the increasing demand in bone tissue engineering for immunomodulatory 3D scaffolds that promote osteogenesis and modulate immune responses.
年龄驱动的免疫信号会引发慢性低度炎症状态，进而影响骨愈合，并给临床医生修复老年患者的临界尺寸骨缺损带来挑战。
聚（L-丙交酯-共-ε-己内酯）（PLCA）支架用来自马铃薯的植物源纳米颗粒鼠李糖半乳糖醛酸聚糖-I（RG-I）进行功能化处理，以研究其在基因和蛋白质水平上调节体外中性粒细胞和巨噬细胞炎症的能力。在皮下大鼠模型中对支架在基因、蛋白质和组织学水平上的早期和晚期宿主反应进行体内测试，并在临界尺寸的颅骨骨缺损中测试其促进老年啮齿动物骨再生的潜力。使用单向方差分析检验显著差异，随后进行多重比较的Tukey检验，p值≤0.05被认为具有显著性。
基因表达显示，用植物源RG-I功能化且半乳糖含量相对高于阿拉伯糖的PLCA支架（马铃薯脱阿拉伯糖（PA））可降低体外细菌脂多糖刺激中性粒细胞和巨噬细胞所引发的炎症状态。与对照组（未修饰的PLCA支架）相比，在PA功能化存在的情况下，脂多糖刺激的中性粒细胞中半乳糖凝集素-3的细胞内积累显著减少。体内基因和蛋白质表达显示出与体外相当的结果。在基因和蛋白质水平上，宿主反应在早期和晚期时间点均朝着抗炎/愈合方向调节。当用PA功能化的PLCA支架皮下植入体内时，观察到异物反应和纤维囊形成减少。用PA功能化的PLCA支架在体内早期和晚期炎症阶段调节细胞因子和趋化因子的表达。植入老年大鼠颅骨缺损中的用PA功能化的PLCA支架在体内2周后下调促炎基因标记物，同时促进成骨标记物。
我们已经表明，用植物源RG-I功能化且半乳糖含量相对较高的PLCA支架在体外和皮下体内均在炎症反应调节中发挥作用，并促进老年啮齿动物临界尺寸骨缺损中骨形成的启动。我们的研究满足了骨组织工程对促进成骨和调节免疫反应的免疫调节3D支架日益增长的需求。
