Scaffold loaded LPS-hUCMSC-sEVs promote Osteo/odontogenic differentiation and angiogenic potential of hDPSCs

IF 2.5 4区生物学 Q1 ANATOMY & MORPHOLOGY Tissue & cell Pub Date : 2024-12-01 Epub Date: 2024-08-30 DOI:10.1016/j.tice.2024.102549

Jingjie Zeng , Huidan Deng , Quanjie Li , Jingyi Kang , Yu Wu

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Abstract

Objective

The formation of dentin-pulp complex determines the success of vital pulp therapy. Human umbilical cord mesenchymal stem cell-derived small extracellular vesicles (hUCMSC-sEVs) appeared to have stronger effect in anti-inflammatory and promoting the proliferation and migration of human dental pulp stem cells (hDPSCs). Moreover, Lipopolysaccharides (LPS) pretreatment can enhance the rapeutic potency of extracellular vesicles. LPS pretreatment hUCMSC-sEVs have the potential to regenerate the dentin-pulp complex by recruiting hDPSCs. This paper aims to develop collagen sponge/self-assembling peptide nanofiber scaffold (CS/SAPNS) composite scaffold loaded with LPS pretreatment hUCMSC-sEVs (CS/SAPNS-sEVs), and assess the release characteristics of hUCMSC-sEVs and the effect of this composite scaffold on osteo/odontogenic differentiation and angiogenic potential in hDPSCs.

Methods

LPS pretreatment hUCMSC-sEVs (LPS-hUCMSC-sEVs) were mixed with self-assembling peptide hydrogel and loaded onto collagen sponge to obtain the CS/SAPNS-sEVs. BCA assay, nanoparticle analysis, transmission electron microscopy and laser confocal microscopy were used to investigate the characteristics of LPS-hUCMSC-sEVs loaded on CS/SAPNS. Osteo/odontogenic differentiation ability of hDPSCs were analyzed by ALP stainning, alizarin red staining. RT-PCR and Western blot analysis were performed to confirm the levels of osteo/odontogenic factors and angiogenic factors, and the involvement of NF-κB pathway was verified by immunocytochemical staining and Western blot analysis.

Results

CS/SAPNS could control LPS-hUCMSC-sEVs release for 7 days and keep their structural integrity. CS/SAPNS-sEVs promoted deposition of calcified nodules and expression of osteogenic/odontogenic and angiogenic factors in hDPSCs. On the contrary, inhibition of the NF-κB pathway down-regulated the expression of CS/SAPNS-sEVs-regulated osteo/odontogenic and angiogenic factors.

Conclusion

CS/SNAPS could be used as scaffold for LPS-hUCMSC-sEVs, and CS/SAPNS-sEVs may promote osteo/odontogenic differentiation and enhance the angiogenic potential of hDPSCs through activating the NF-κB pathway.

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负载 LPS-hUCMSC-sEV 的支架可促进 hDPSCs 的骨/牙源性分化和血管生成潜能

目的牙本质-牙髓复合体的形成决定了牙髓治疗的成败。人脐带间充质干细胞衍生的细胞外小泡（hUCMSC-sEVs）似乎具有更强的抗炎、促进牙髓干细胞（hDPSCs）增殖和迁移的作用。此外，脂多糖（LPS）预处理可增强细胞外囊泡的抗炎作用。经 LPS 预处理的 hUCMSC-sEVs 有可能通过招募 hDPSCs 来再生牙本质-牙髓复合体。本文旨在开发负载 LPS 预处理 hUCMSC-sEVs 的海绵胶原/自组装肽纳米纤维支架（CS/SAPNS）复合支架（CS/SAPNS-sEVs），并评估 hUCMSC-sEVs 的释放特性以及该复合支架对 hDPSCs 成骨/成髓分化和血管生成潜能的影响。方法将经 LPS 预处理的 hUCMSC-sEVs（LPS-hUCMSC-sEVs）与自组装肽水凝胶混合并负载到海绵胶原上，得到 CS/SAPNS-sEVs。利用 BCA 检测、纳米颗粒分析、透射电子显微镜和激光共聚焦显微镜研究了负载在 CS/SAPNS 上的 LPS-hUCMSC-sEVs 的特性。通过 ALP 染色和茜素红染色分析了 hDPSCs 的骨/牙源性分化能力。结果CS/SAPNS可控制LPS-hUCMSC-sEVs释放7天，并保持其结构的完整性。CS/SAPNS-sEVs 促进了 hDPSCs 中钙化结节的沉积以及成骨/牙生成因子和血管生成因子的表达。结论CS/SNAPS 可用作 LPS-hUCMSC-sEVs 的支架，CS/SAPNS-sEVs 可通过激活 NF-κB 通路促进 hDPSCs 的成骨/成牙分化并增强其血管生成潜能。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.