In vitro studies on the differentiation potential of human periodontal ligament-derived progenitor cells for developing functional periodontal ligament 3D cell culture models: A systematic review of methods and approaches

Background: The successful translation of periodontal regeneration therapies from laboratory to clinic relies on physiologically relevant preclinical models. 3D periodontal ligament (PDL) cell culture is a transformative technology in preclinical dental research, which serves as a link between laboratory experiments and human trials. PDL plays a central role in tooth anchorage, alveolar bone stability, and orthodontic tooth movement (OTM) due to the self-renewal ability of PDL-derived progenitor cells. Its regenerative capacity is often compromised by periodontitis or mechanical stress. PDL-derived progenitor cells are characterized by their self-renewal and multilineage differentiation. They are the primary drivers of tissue repair. However, the complexity of the PDL role in tooth regeneration necessitates advanced 3D modelling to accurately simulate in vivo responses. This review aims to summarize all previously described 3D PDL cell culture models and systematically analyze them with respect to their structure, cell composition, expression of PDL marker genes, and applications. Methods : PubMed, Scopus, Web of Science (WoS) and Google Scholar databases were searched for studies published from 2000 to 2026 by two researchers. Study selection, data extraction, and risk of bias assessment were performed independently by three reviewers following PICO criteria. Only studies published in English were included. Results: Fifty-eight in vitro studies of initially 1095 papers met our inclusion criteria and were included in this systematic review. These in vitro studies demonstrated that PDL-derived progenitor cells can differentiate into specialized dental cells within different 3D cell models, as verified by lineages specific markers and histological examination of their structure and mineralization. The use of 3D cell models in various functional applications was also analyzed. Conclusions: Under certain conditions, 3D PDL cell models can mimic native PDL tissue, making them a promising option for regenerative dentistry and for studying OTM, bone remodeling, and other functional aspects. However, the review results confirmed substantial heterogeneity in experimental conditions for 3D PDL cell culture manufacturing, complicating the comparability of PDL models and rendering a quantitative analysis impossible. More studies on this subject, along with consistency in experimental approaches, are needed to ensure comparability and quality control.