Gingival Stem Cell-Conditioned Media and Low-Level Laser Therapy Enhance Periodontal Ligament Stem Cells Function by Upregulating Wnt and TGF-β Pathway Components: An In Vitro Study

IF 2.2 Q3 DENTISTRY, ORAL SURGERY & MEDICINE

Clinical and Experimental Dental Research Pub Date : 2025-05-26 DOI:10.1002/cre2.70151

Mohammed Y. Aljabri, Yaser A. Alhazmi, Salah A. Elsayyad, Shehabeldin M. Saber, Mohamed Shamel

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Abstract

Objectives

This study investigated the synergistic effects of gingival stem cell conditioned media (GSC-CM) and low-level laser therapy (LLLT) on stimulating PDLSCs, explicitly focusing on the molecular basis of enhancing the Wnt and TGF-β signaling pathways.

Material and Methods

PDLSCs were isolated and GSC-CM were prepared and characterized. Five groups were utilized: control, osteogenic induction medium (positive control), GSC-CM, LLLT (980 nm, 1.5 J/cm²), and a combination of GSC-CM with LLLT. Cell viability was assessed using the MTT assay on Days 1, 2, and 3. Osteogenic differentiation of PDLSCs was assessed using mineralization assays employing Alizarin Red staining, and molecular analyses of osteogenic markers (RUNX2, ALP, OCN) and signaling-related genes (CTNNβ1, TGFβ1) conducted via RT‒PCR.

Results

The findings revealed that the dual application of GSC-CM and LLLT significantly enhanced cell viability and osteogenic differentiation of PDLSCc compared to the effects of the individual treatment modalities. Moreover, the study revealed elevated expression levels of osteogenic markers across all the experimental groups, specially the combination group, which showed the highest levels. Significantly, the combination treatment group displayed superior outcomes in this regard. The results also suggest potential activation of Wnt/β-catenin and TGF-β signaling components.

Conclusions

This study highlights the significant effects of using GSC-CM and LLLT to improve PDLSC survival and osteogenic differentiation.

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牙龈干细胞条件介质和低水平激光治疗通过上调Wnt和TGF-β通路成分增强牙周韧带干细胞功能的体外研究

目的研究牙龈干细胞条件培养基（GSC-CM）和低水平激光治疗（LLLT）对PDLSCs刺激的协同作用，明确关注Wnt和TGF-β信号通路增强的分子基础。材料与方法分离PDLSCs，制备GSC-CM并对其进行表征。采用对照、成骨诱导培养基（阳性对照）、GSC-CM、LLLT （980 nm, 1.5 J/cm²）、GSC-CM与LLLT联合使用五组。在第1、2和3天使用MTT法评估细胞活力。采用茜素红染色矿化法评估PDLSCs的成骨分化，并通过RT-PCR对成骨标志物（RUNX2、ALP、OCN）和信号相关基因（CTNNβ1、TGFβ1）进行分子分析。结果与单独治疗相比，GSC-CM和LLLT的双重应用显著提高了PDLSCc的细胞活力和成骨分化。此外，研究显示，在所有实验组中，成骨标志物的表达水平均有所升高，特别是联合组，表达水平最高。值得注意的是，联合治疗组在这方面表现出更好的结果。结果还提示Wnt/β-catenin和TGF-β信号成分的潜在激活。结论GSC-CM和LLLT对改善PDLSC存活和成骨分化有显著作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical and Experimental Dental Research DENTISTRY, ORAL SURGERY & MEDICINE-

CiteScore

3.30

自引率

5.60%

发文量

165

审稿时长

26 weeks

期刊介绍： Clinical and Experimental Dental Research aims to provide open access peer-reviewed publications of high scientific quality representing original clinical, diagnostic or experimental work within all disciplines and fields of oral medicine and dentistry. The scope of Clinical and Experimental Dental Research comprises original research material on the anatomy, physiology and pathology of oro-facial, oro-pharyngeal and maxillofacial tissues, and functions and dysfunctions within the stomatognathic system, and the epidemiology, aetiology, prevention, diagnosis, prognosis and therapy of diseases and conditions that have an effect on the homeostasis of the mouth, jaws, and closely associated structures, as well as the healing and regeneration and the clinical aspects of replacement of hard and soft tissues with biomaterials, and the rehabilitation of stomatognathic functions. Studies that bring new knowledge on how to advance health on the individual or public health levels, including interactions between oral and general health and ill-health are welcome.