Karina Gonzales Silvério, Gabriela Bessa Marconato Antunes, Bruno Cazotti Pereira, Francisco Naldo Gomes Filho, Nathalia Reiche Moreira, Renato Corrêa Viana Casarin, Enilson Antonio Sallum, Catharina Marques Sacramento
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引用次数: 0
摘要
了解调节骨/骨水泥分化的分子机制对于优化牙周再生的细胞策略至关重要。本研究采用多元统计模型，在体外研究人牙周韧带干细胞（hPDLSCs）中骨形态发生蛋白2 （bone morphogenetic protein 2, BMP2）与ASPN的相互作用及其对矿化电位的影响。在标准（SDM）和成骨/骨水泥（OM）条件下培养4个初级hPDLSC群体，其中2个具有高成骨/骨水泥潜力（HOP）， 2个具有低成骨/骨水泥潜力（LOP）。采用茜素红染色（AR-S）和碱性磷酸酶（ALP）活性检测矿化程度，采用实时荧光定量聚合酶链式反应（qRT-PCR）检测ASPN、BMP2、骨钙素（OCN）和矮子相关转录因子2 （RUNX2）的基因表达。使用Luminex系统测量BMP2蛋白水平。数据分析采用了RStudio中的主成分分析（PCA）、线性判别分析（LDA）、Pearson相关性、相关网络和多元回归模型。与LOP细胞相比，HOP细胞表现出更高的矿化和BMP2表达(p)。萨克拉门托，C.（2025）。补充信息：在线版本包含补充资料，可在10.1007/s10616-025-00810-x获得。
Modulatory roles of bone morphogenetic protein 2 and asporin in osteo/cementoblast differentiation potential of human periodontal ligament stem cells: a multivariate model analysis.
Understanding the molecular mechanisms regulating osteo/cementogenic differentiation is critical for optimizing cell-based strategies in periodontal regeneration. This study employed multivariate statistical modeling to investigate, in vitro, the interaction between bone morphogenetic protein 2 (BMP2) and asporin (ASPN) in human periodontal ligament stem cells (hPDLSCs) and its impact on mineralization potential. Four primary hPDLSC populations-two with high osteo/cementogenic potential (HOP) and two with low potential (LOP)-were cultured under standard (SDM) and osteo/cementogenic (OM) conditions. Mineralization was assessed using Alizarin Red Staining (AR-S) and alkaline phosphatase (ALP) activity, while gene expression of ASPN, BMP2, osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) was quantified by quantitative real-time polymerase chain reaction (qRT-PCR). BMP2 protein levels were measured using the Luminex system. Data analysis incorporated Principal Component Analysis (PCA), Linear Discriminant Analysis (LDA), Pearson correlations, correlation networks, and multivariate regression models in RStudio. HOP cells exhibited higher mineralization and BMP2 expression compared to LOP cells (p < 0.05). ASPN negatively correlated with BMP2 expression and mineralization, particularly in LOP cells (p < 0.01), reinforcing its inhibitory role. Multivariate modeling identified BMP2 as a key positive regulator of osteo/cementogenesis, while ASPN emerged as a significant inhibitory factor (p < 0.001). This study highlights the potential of multivariate models as powerful tools for uncovering molecular interactions and identifying novel therapeutic targets, paving the way for advancements in periodontal regeneration and cell-based therapies.
Graphical abstract: Created in BioRender. Sacramento, C. (2025).
Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00810-x.
期刊介绍：
The scope of the Journal includes:
1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products.
2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools.
3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research.
4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy.
5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.
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