长期生物库牙髓干细胞保留血管生成潜力的血管化组织工程-实验室研究。

IF 7.1 1区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE
Shuntaro Yamada, Katerina Holomkova, Åshild Johansen, Masoumeh Jahani Kadousaraei, Niyaz Al-Sharabi, Francesco Torelli, Pierfrancesco Pagella, Ana Angelova Volponi, Hiroshi Egusa, Inge Fristad, Kamal Mustafa
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引用次数: 0

摘要

目的:本研究旨在评估人类牙髓干细胞(DPSCs)在长期生物银行(7-8年)后是否保留其促血管生成特性,并可用于工程血管化组织,解决其在再生牙科临床转化的潜力。方法:从青少年供体中冷冻保存的DPSCs从生物银行中回收,并以染色体完整性,MSC免疫表型和多能性为特征。在体外促血管生成条件下,通过RT-qPCR阵列、流式细胞术和高通量免疫分型分析基因和蛋白的表达。通过体外成管、体外CAM植入试验、器官芯片灌注模型和临床级GelMA水凝胶长期培养(45天),评估功能血管生成能力,有无huvec。结果:生物银行的DPSCs保留了MSC身份和多谱系分化潜力。促血管生成/内皮调节增强了血管生成/内皮基因(PECAM1, VEGFR2, NRP1, ACE)的表达,但大多数细胞保持周细胞样表型。在CAM模型中,初始和内皮条件的DPSCs(即分别为naiveDPSCs和endoDPSCs)均显著增强血管长入。在器官芯片系统中,幼稚的dpscs与HUVECs形成可灌注的脉管系统,并分化为血管周围细胞类型。最值得注意的是,在长时间的刺激后,endoDPSCs成功地在GelMA水凝胶中生成了含有CD31(+)和αSMA(+)细胞的血管化组织。结论:长期生物储存的DPSCs保持其血管生成潜力,并在延长内皮诱导后,可以在体外3D培养模型中独立生成血管化组织。这是第一份证明全面的促血管生成特性和使用生物库DPSCs进行血管化组织工程的可行性的报告,强调了它们在未来再生治疗中的强大临床适用性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Long-Term Biobanked Dental Pulp Stem Cells Retain Angiogenic Potential for Vascularised Tissue Engineering-Laboratory Investigation.

Aim: This study aimed to evaluate whether human dental pulp stem cells (DPSCs), after long-term biobanking (7-8 years), retain their pro-angiogenic properties and can be used to engineer vascularised tissues, addressing their potential for clinical translation in regenerative dentistry.

Methodology: Cryopreserved DPSCs from adolescent donors were recovered from biobanking and characterised for chromosomal integrity, MSC immunophenotype and multipotency. After conditioning in pro-angiogenic conditions in vitro, gene and protein expression were analysed by RT-qPCR array, flow cytometry and high-throughput immunophenotyping. Functional angiogenic capacity was assessed via in vitro tube formation, ex ovo CAM implantation assay, organ-on-chip perfusion model and long-term culture (45 days) in clinical-grade GelMA hydrogels, with and without HUVECs.

Results: Biobanked DPSCs retained MSC identity and multi-lineage differentiation potential. Pro-angiogenic/endothelial conditioning enhanced the expression of angiogenic/endothelial genes (PECAM1, VEGFR2, NRP1, ACE), yet most cells maintained a pericyte-like phenotype. Both naive and endothelial-conditioned DPSCs (i.e., naiveDPSCs and endoDPSCs, respectively) significantly enhanced vascular ingrowth in the CAM model. In the organ-on-chip system, naiveDPSCs formed perfusable vasculature with HUVECs and differentiated into perivascular cell types. Most notably, endoDPSCs alone successfully generated vascularised tissue with both CD31(+) and αSMA(+) cells present in GelMA hydrogels after prolonged stimulation.

Conclusion: Long-term biobanked DPSCs preserve their angiogenic potential and, following extended endothelial induction, can independently generate vascularised tissue in 3D in vitro culture models. This is the first report demonstrating the comprehensive pro-angiogenic characterisation and the feasibility of using biobanked DPSCs for vascularised tissue engineering, highlighting their strong clinical applicability for future regenerative therapies.

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来源期刊
International endodontic journal
International endodontic journal 医学-牙科与口腔外科
CiteScore
10.20
自引率
28.00%
发文量
195
审稿时长
4-8 weeks
期刊介绍: The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.
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