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评估源自各种口腔来源的间充质干细胞条件培养基的血管生成潜力。

Assessment of angiogenic potential of mesenchymal stem cells derived conditioned medium from various oral sources.

作者信息

Shekatkar Madhura Rajendra, Kheur Supriya Mohit, Kharat Avinash Haribhau, Deshpande Shantanu Sanjeev, Sanap Avinash Purushottam, Kheur Mohit Gurunath, Bhonde Ramesh Ramchandra

机构信息

Department of Oral Pathology and Microbiology, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India.

Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India.

出版信息

J Clin Transl Res. 2022 Jul 25;8(4):323-338. eCollection 2022 Aug 29.

PMID:36090765
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9450500/
Abstract

BACKGROUND

Abnormal angiogenesis hamper blood vessel proliferation implicated in various biological processes. The current method available to clinically treat patients to enhance angiogenesis is administering the angiogenic growth factors. However, due to a lack of spatiotemporal control over the substantial release of these factors, numerous drawbacks are faced such as leaky vasculature. Hence, stem-cell-based therapeutic applications are running their race to evolve as potential targets for deranged angiogenesis. In clinical dentistry, adequate tissue vascularization is essential for successful endodontic therapies such as apexogenesis and apexification. Furthermore, wound healing of the extraction socket and tissue regeneration post-surgical phase of treatment including implant placement require angiogenesis as a foundation for the ultimate success of treatment. Mesenchymal stem cells (MSCs) secrete certain growth factors and cytokines in the culture medium during the proliferation. These factors and cytokines are responsible for various biological activities inside human body. Oral cavity-derived stem cells can secrete growth factors that enhance angiogenesis.

AIM

The aim of the study was to investigate the angiogenic potential of conditioned medium (CM) of MSCs derived from different oral sources.

METHODS

Oral tissues such as dental pulp of adult and deciduous teeth, gingiva, and buccal fat were used to isolate dental pulp MSCs (DPSCs), exfoliated deciduous teeth, gingival MSCs, and buccal fat derived MSCs. MSCs conditioned medium (CM) from passage four cells from all the sources were obtained at 48 h interval and growth factor analysis was performed using flow cytometry. To assess the functionality of the CM, Chick Yolk Sac Membrane (YSM) assay was performed.

RESULTS

CM obtained from DPSCs showed higher levels of vascular endothelial growth factor, fibroblast growth factor, and hepatocyte growth factor as evidenced by flow cytometry. Furthermore, DPSC-CM exhibited significantly higher pro-angiogenic potential when assessed in in-ovo YSM assay.

CONCLUSION

DPSCs so far seems to be the best source as compare to the rest of oral sources in promoting angiogenesis. A novel source of CM derived from buccal fat stem cells was used to assess angiogenic potential. Thus, the present study shows that CM derived from oral cavity-derived-MSCs has a dynamic and influential role in angiogenesis.

RELEVANCE FOR PATIENTS

CM derived from various oral sources of MSCs could be used along with existing therapies in medical practice where patients have compromised blood supply like in diabetes and in patients with debilitating disorders. In clinical dentistry, adequate tissue vascularization is essential for successful wound healing, grafting procedures, and endodontic therapies. DPSCs-CM shows better angiogenic potential in comparison with other oral sources of MSCs-CM. Our findings could be a turning point in the management of all surgical and regenerative procedures requiring increased angiogenesis.

摘要

背景

异常血管生成会阻碍参与各种生物过程的血管增殖。目前临床上用于治疗患者以促进血管生成的方法是给予血管生成生长因子。然而,由于缺乏对这些因子大量释放的时空控制,面临许多缺点,如血管渗漏。因此,基于干细胞的治疗应用正在竞相发展成为血管生成紊乱的潜在靶点。在临床牙科中,充足的组织血管化对于诸如根尖诱导成形术和根尖屏障术等成功的牙髓治疗至关重要。此外,拔牙窝的伤口愈合以及包括种植体植入在内的治疗后手术阶段的组织再生需要血管生成作为治疗最终成功的基础。间充质干细胞(MSCs)在增殖过程中在培养基中分泌某些生长因子和细胞因子。这些因子和细胞因子负责人体内的各种生物活性。口腔来源的干细胞可以分泌增强血管生成的生长因子。

目的

本研究的目的是研究不同口腔来源的间充质干细胞条件培养基(CM)的血管生成潜力。

方法

使用成人和乳牙的牙髓、牙龈和颊脂等口腔组织来分离牙髓间充质干细胞(DPSCs)、脱落乳牙、牙龈间充质干细胞和颊脂来源的间充质干细胞。每隔48小时从所有来源的第4代细胞中获得间充质干细胞条件培养基(CM),并使用流式细胞术进行生长因子分析。为了评估CM的功能,进行了鸡胚卵黄囊膜(YSM)试验。

结果

通过流式细胞术证明,从DPSCs获得的CM显示出更高水平的血管内皮生长因子、成纤维细胞生长因子和肝细胞生长因子。此外,在鸡胚卵黄囊膜试验中评估时,DPSC-CM表现出显著更高的促血管生成潜力。

结论

与其他口腔来源相比,迄今为止DPSCs似乎是促进血管生成的最佳来源。使用一种源自颊脂干细胞的新型CM来源来评估血管生成潜力。因此,本研究表明源自口腔来源间充质干细胞的CM在血管生成中具有动态且有影响的作用。

对患者的意义

源自各种口腔来源间充质干细胞的CM可与现有疗法一起用于医疗实践中血液供应受损的患者,如糖尿病患者和患有衰弱性疾病的患者。在临床牙科中,充足的组织血管化对于成功的伤口愈合、移植手术和牙髓治疗至关重要。与其他口腔来源的间充质干细胞CM相比,DPSCs-CM显示出更好的血管生成潜力。我们的发现可能是所有需要增加血管生成的手术和再生程序管理中的一个转折点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/4af9f83d57bd/jclintranslres-2022-8-4-323-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/171b0355f8ef/jclintranslres-2022-8-4-323-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/0cd4ea749d18/jclintranslres-2022-8-4-323-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/2bf6eefabc77/jclintranslres-2022-8-4-323-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/7859b1b08c0d/jclintranslres-2022-8-4-323-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/4af9f83d57bd/jclintranslres-2022-8-4-323-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/9c97b52b32f1/jclintranslres-2022-8-4-323-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/96e91e45b3d7/jclintranslres-2022-8-4-323-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/5096e972642f/jclintranslres-2022-8-4-323-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/3da0b20e10d8/jclintranslres-2022-8-4-323-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/171b0355f8ef/jclintranslres-2022-8-4-323-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/0cd4ea749d18/jclintranslres-2022-8-4-323-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/2bf6eefabc77/jclintranslres-2022-8-4-323-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/7859b1b08c0d/jclintranslres-2022-8-4-323-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f9b3/9450500/42eccd7cb03c/jclintranslres-2022-8-4-323-g009.jpg
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