Ganapathy Amudha, Chen Yinghua, Bakthavachalam Velavan, George Anne
Brodie Tooth Development Genetics & Regenerative Medicine Research Laboratory, Department of Oral Biology, University of Illinois at Chicago, Chicago, IL, United States.
Front Cell Dev Biol. 2025 Aug 8;13:1630129. doi: 10.3389/fcell.2025.1630129. eCollection 2025.
Dental pulp stem cells (DPSCs), can differentiate into endothelial cells (ECs), offering a promising strategy for generation of new blood vessels which is crucial for tissue repair and regeneration. Many studies have focused on optimizing conditions for differentiating DPSCs into ECs and subsequent validation of the vasculogenic potential of newly generated ECs . Previously, we demonstrated the ability of the HUVEC ECM scaffold along with DMP1 stimulation would drive endothelial-specific lineage of DPSCs.
In this study, DMP1-treated DPSCs were cultured on HUVEC ECM for 7 days and sorted using angiogenic-specific markers CD31 and CD144. The cells were separated into a positive fraction (CD31/CD144) and a negative fraction (CD31/CD144). To assess if ECs transformed from DMP1 stimulated DPSCs maintain their endothelial properties over time, we cultured both the positive CD31/CD144 and negative CD31/CD144 fractions along with unstimulated DPSCs and assessed their angiogenic characteristics by gene expression analysis, functional properties using a tubule formation assay and subcutaneous implantation model.
The findings of this study indicate that the CD31/CD144 fraction, retains both the phenotypic and functional characteristics of ECs, in contrast to the CD31/CD144 fraction. Furthermore, analysis of the sorted ECs using the subcutaneous implantation model exhibited neovascularization along with the expression of vasculogenic markers. Overall, DPSC-derived ECs obtained by stimulation with DMP1 and cultured on HUVEC-ECM function as typical vascular ECs. This strategy, could be exploited for the development of vasculogeneis and as a therapeutic potential for tissue repair and regeneration.
牙髓干细胞(DPSCs)可分化为内皮细胞(ECs),为生成新血管提供了一种有前景的策略,这对组织修复和再生至关重要。许多研究集中于优化将DPSCs分化为ECs的条件以及随后对新生成ECs血管生成潜力的验证。此前,我们证明了人脐静脉内皮细胞(HUVEC)细胞外基质(ECM)支架与牙本质基质蛋白1(DMP1)刺激相结合能够驱动DPSCs向内皮细胞特异性谱系分化。
在本研究中,用DMP1处理的DPSCs在HUVEC ECM上培养7天,并使用血管生成特异性标志物CD31和CD144进行分选。细胞被分为阳性组分(CD31/CD144)和阴性组分(CD31/CD144)。为了评估从DMP1刺激的DPSCs转化而来的ECs是否随时间维持其内皮特性,我们将CD31/CD144阳性组分和CD31/CD144阴性组分与未刺激的DPSCs一起培养,并通过基因表达分析、使用小管形成试验的功能特性以及皮下植入模型评估它们的血管生成特征。
本研究结果表明,与CD31/CD144阴性组分相比,CD31/CD144阳性组分保留了ECs的表型和功能特征。此外,使用皮下植入模型对分选的ECs进行分析显示有新血管形成以及血管生成标志物的表达。总体而言,通过DMP1刺激获得并在HUVEC-ECM上培养的DPSC来源的ECs具有典型血管ECs的功能。这种策略可用于血管生成的开发以及作为组织修复和再生的治疗潜力。