A Modified Glycosaminoglycan, GM-0111, Inhibits Molecular Signaling Involved in Periodontitis.
A Modified Glycosaminoglycan, GM-0111, Inhibits Molecular Signaling Involved in Periodontitis.
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DOI：
10.1371/journal.pone.0157310
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Lee WY
中科院分区：
文献类型：
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作者：
Savage JR;Pulsipher A;Rao NV;Kennedy TP;Prestwich GD;Ryan ME;Lee WY
牙周炎的特征是微生物感染、炎症、组织破坏和牙槽骨基质的加速损失。针对疾病的这些多个阶段的治疗提供了治疗或预防牙周炎的方法。某些糖胺聚糖（GAGs）可以阻断多种炎症介质并抑制细菌生长，这表明这些糖胺聚糖可用作牙周炎的治疗药物。我们研究了合成的GAG GM-0111对与牙周炎相关的各种分子事件的影响：牙龈卟啉单胞菌（P. gingivalis）和伴放线菌聚集菌（A.本发明涉及与牙周炎相关的致病菌（放线菌共生菌（actinomycetemcomitans））;在小鼠巨噬细胞RAW 264.7细胞和异源表达的HEK 293细胞中通过TLR 2和TLR 4激活促炎信号传导;在培养的小鼠前破骨细胞中破骨细胞形成和骨基质再吸收。(1)GM-0111对牙龈卟啉单胞菌和A.放线菌共生菌，即使在1%（w/v）的溶液。GM-0111在所有浓度下对牙龈卟啉单胞菌的抗菌作用均强于透明质酸（HA）或木糖醇，对A.放线菌共生菌在≥2%（w/v）的溶液中。我们还观察到GM-0111抑制牙龈卟啉单胞菌的生物膜形成，并且这些作用比HA强得多。(2)GM-0111抑制巨噬细胞和HEK 293细胞中TLR介导的促炎细胞信号传导，对TLR 2的选择性高于TLR 4（IC 50分别为1-10 ng/mL vs. > 100 μg/mL）。(3)GM-0111阻断RANKL诱导的破骨细胞形成（低至300 ng/mL）和骨基质吸收。虽然GM-0111显示出与RANKL的高亲和力结合，但其不干扰RANKL/RANK/NF-κB信号传导，表明GM-0111通过RANKL-RANK非依赖性机制抑制破骨细胞形成。我们报告说，GM-0111抑制牙周炎中涉及的多种分子事件，从早期促炎TLR信号传导到骨丢失后期激活的通路。
Periodontitis is characterized by microbial infection, inflammation, tissue breakdown, and accelerated loss of alveolar bone matrix. Treatment targeting these multiple stages of the disease provides ways to treat or prevent periodontitis. Certain glycosaminoglycans (GAGs) block multiple inflammatory mediators as well as suppress bacterial growth, suggesting that these GAGs may be exploited as a therapeutic for periodontitis. We investigated the effects of a synthetic GAG, GM-0111, on various molecular events associated with periodontitis: growth of Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) pathogenic bacteria associated with periodontitis; activation of pro-inflammatory signaling through TLR2 and TLR4 in mouse macrophage RAW 264.7 cells and heterologously expressed HEK 293 cells; osteoclast formation and bone matrix resorption in cultured mouse pre-osteoclasts. (1) GM-0111 suppressed the growth of P. gingivalis and A. actinomycetemcomitans even at 1% (w/v) solution. The antibacterial effects of GM-0111 were stronger than hyaluronic acid (HA) or xylitol in P. gingivalis at all concentrations and comparable to xylitol in A. actinomycetemcomitans at ≥2% (w/v) solution. We also observed that GM-0111 suppressed biofilm formation of P. gingivalis and these effects were much stronger than HA. (2) GM-0111 inhibited TLR-mediated pro-inflammatory cellular signaling both in macrophage and HEK 293 cells with higher selectivity for TLR2 than TLR4 (IC50 of 1–10 ng/mL vs. > 100 μg/mL, respectively). (3) GM-0111 blocked RANKL-induced osteoclast formation (as low as 300 ng/mL) and bone matrix resorption. While GM-0111 showed high affinity binding to RANKL, it did not interfere with RANKL/RANK/NF-κB signaling, suggesting that GM-0111 inhibits osteoclast formation by a RANKL-RANK-independent mechanism. We report that GM-0111 inhibits multiple molecular events involved in periodontitis, spanning from the early pro-inflammatory TLR signaling, to pathways activated at the later stage component of bone loss.