Abstract
Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale. Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage. To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes. The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including "anatomical structure development" and "cell adhesion." Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process "anatomical structure development," Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.
Highlights
Periodontitis is a polymicrobial, infection-induced inflammatory disease in the periodontium characterized by connective attachment loss and alveolar bone destruction
Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. This comprehensive transcriptomic assessment of human periodontal dental ligament (PDL) progenitor cells revealed that expression of transcripts related to the biological process “anatomical structure development,” Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype
The PDL-CD105+-enriched population exhibited a high proportion of cells that expressed mesenchymal stem cell (MSC)-related markers (Figure 1A)
Summary
Periodontitis is a polymicrobial, infection-induced inflammatory disease in the periodontium characterized by connective attachment loss and alveolar bone destruction. Epidemiological studies indicate that periodontitis is still a globally prevalent disease. This periodontal disease may lead to functionally compromised dentition, which affects the quality of life of many subjects.[1] In the last decade, several attempts have been made to regenerate the tissues impaired due to periodontitis, including bone replacement grafts, guided tissue regeneration, enamel matrix derivative, and combined therapy.[2] these clinical approaches have not shown complete and predictable regeneration of periodontal tissues, namely cementum, periodontal ligament (PDL), and alveolar bone.[2,3] emerging regenerative approaches based on a biological rationale have been proposed to achieve improved clinical outcomes, such as enamel matrix derivative (EMD), recombinant human plateletderived growth factor-BB (rhPDGF-BB)/beta tricalcium phosphate (b-TCP), and synthetic peptide-binding protein P-15/anorganic bovine bone matrix.[2,3]
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