Metabolomic signatures distinguish extracellular vesicles from osteoclasts and odontoclasts.
Metabolomic signatures distinguish extracellular vesicles from osteoclasts and odontoclasts.
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DOI：
10.1111/ocr.12658
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发表时间：
2023-11
影响因子：
3.1
通讯作者：
Holliday, L. Shannon
中科院分区：
文献类型：
--
作者：
Rody Jr, Wellington J.;Reuter, Nathan G.;Brooks, Shannen E.;Hammadi, Lina I.;Martin, Macey L.;Cagmat, Joy G.;Garrett, Timothy J.;Holliday, L. Shannon
病理性牙根吸收和牙槽骨丢失通常只有在不可逆性损伤后才能发现。牙龈沟液或唾液中的生物标记物可以提供一种早期检测的手段；然而，事实证明，这样的生物标记物难以捉摸。我们假设，多组学方法可能为牙根吸收和牙槽骨丢失提供可靠的诊断信号。以前，我们发现来自破骨细胞和破牙细胞的细胞外小泡(EV)的蛋白质组成不同。在这项研究中，我们研究了来自破骨细胞、破牙细胞和破骨细胞(非吸收破骨细胞)的EV的代谢组。在重组RANKL和CSF-1存在下，在牙本质、骨或塑料上培养小鼠造血祖细胞，以触发沿碎裂线的分化。第7天，细胞固定，确认碎裂细胞的分化状态和吸收状态。第7天从条件培养液中分离出EVS，并通过纳米颗粒跟踪和电子显微镜进行表征，以确保质量。使用带有Dionex UHPLC和自动进样器的Thermo Q-Exactive Orbitrap质谱仪进行全球代谢组谱分析。我们在碎屑EV中鉴定出978种代谢物。其中，79个是可变相互依赖参数得分大于或等于2的潜在生物标志物。与破骨细胞相比，已知的代谢产物胞苷、异胞嘧啶、胸腺嘧啶、琥珀酸和瓜氨酸在来自破牙细胞的EVS中被发现有统计学上更高的水平。我们的结论是，在破牙细胞EV中发现的大量代谢物不同于破骨细胞EV中的代谢物，因此是牙根吸收和牙周组织破坏的潜在生物标志物。
Pathological dental root resorption and alveolar bone loss are often detected only after irreversible damage. Biomarkers in the gingival crevicular fluid or saliva could provide a means for early detection; however, such biomarkers have proven elusive. We hypothesize that a multiomic approach might yield reliable diagnostic signatures for root resorption and alveolar bone loss. Previously, we showed that extracellular vesicles (EVs) from osteoclasts and odontoclasts differ in their protein composition. In this study, we investigated the metabolome of EVs from osteoclasts, odontoclasts and clasts (non-resorbing clastic cells). Mouse haematopoietic precursors were cultured on dentine, bone or plastic, in the presence of recombinant RANKL and CSF-1 to trigger differentiation along the clastic line. On Day 7, the cells were fixed and the differentiation state and resorptive status of the clastic cells were confirmed. EVs were isolated from the conditioned media on Day 7 and characterized by nanoparticle tracking and electron microscopy to ensure quality. Global metabolomic profiling was performed using a Thermo Q-Exactive Orbitrap mass spectrometer with a Dionex UHPLC and autosampler. We identified 978 metabolites in clastic EVs. Of those, 79 are potential biomarkers with Variable Interdependent Parameters scores of 2 or greater. Known metabolites cytidine, isocytosine, thymine, succinate and citrulline were found at statistically higher levels in EVs from odontoclasts compared with osteoclasts. We conclude that numerous metabolites found in odontoclast EVs differ from those in osteoclast EVs, and thus represent potential biomarkers for root resorption and periodontal tissue destruction.
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DOI：
10.1007/978-1-0716-0239-3_3
发表时间：
2020
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Du X;Smirnov A;Pluskal T;Jia W;Sumner S
通讯作者：
Sumner S
影响因子：
3.7
作者：
Chamberlain, Casey A.;Hatch, Marguerite;Garrett, Timothy J.
通讯作者：
Garrett, Timothy J.
影响因子：
1.9
作者：
Harokopakis-Hajishengallis, Evlambia
通讯作者：
Harokopakis-Hajishengallis, Evlambia
影响因子：
3.1
作者：
Holliday LS;McHugh KP;Zuo J;Aguirre JI;Neubert JK;Rody WJ Jr
通讯作者：
Rody WJ Jr
质膜受体参与破骨细胞与骨骼非细胞成分的结合和反应。
影响因子：
5.6
作者：
Karanth DS;Martin ML;Holliday LS
通讯作者：
Holliday LS