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. 2024 Aug 2.
doi: 10.1007/s00056-024-00541-2. Online ahead of print.

TNF reduces osteogenic cell fate in PDL cells at transcriptional and functional levels without alteration of periodontal proliferative capacity

Isabel Knaup  1 , Rafael Kramann  2 , Martha-Julia Sasula  3 , Paula Mack  4 , Rogério Bastos Craveiro  4 , Christian Niederau  4 , Franziska Coenen  4 , Sabine Neuss  5   6 , Joachim Jankowski  7 , Michael Wolf  4
Affiliations

TNF reduces osteogenic cell fate in PDL cells at transcriptional and functional levels without alteration of periodontal proliferative capacity

Isabel Knaup et al. J Orofac Orthop. .

Abstract
in English, German

Aims: To investigate the effect of tumor necrosis factor (TNF) on the growth of human periodontal ligament (PDL) cells, their osteogenic differentiation and modulation of their matrix secretion in vitro.

Methods: The influence of 10 ng/ml TNF on proliferation and metabolic activity of PDL cells was analyzed by cell counting (DAPI [4',6-diamidino-2-phenylindole] staining) and the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. In addition, cells were cultured under control conditions and osteogenic conditions (media containing 10 mM β-glycerophosphate). Quantitative expression analysis of genes encoding the osteogenic markers alkaline phosphatase (ALP), collagen type I alpha 1 chain (COL1A1), osteoprotegerin (OPG), and osteopontin (OPN) was performed after 7 and 14 days of cultivation. Calcium deposits were stained with alizarin red.

Results: Our studies showed that 10 ng/ml TNF did not affect the survival and metabolic activity of PDL cells. Quantitative expression analysis revealed that long-term cultures with TNF impaired osteogenic cell fate at early and late developmental stages. Furthermore, TNF significantly reduced matrix secretion in PDL cells.

Conclusion: The present data confirm TNF as a regulatory factor of proinflammatory remodeling that influences the differentiation behavior but not the metabolism and cell proliferation of the periodontium. Therefore, TNF represents an interesting target for the regulation of orthodontic remodeling processes in the periodontium.

Zusammenfassung: ZIELE: In-vitro-Untersuchung des Einflusses von Tumornekrosefaktor (TNF) auf das Wachstum humaner PDL(Parodontalligament)-Zellen, ihre osteogene Differenzierung und die Modulation ihrer Matrixsekretion.

Methoden: Der Einfluss von 10 ng/ml TNF auf Proliferation und metabolische Aktivität der PDL-Zellen wurde anhand der Zellzählung (DAPI [4ʼ,6-Diamidino-2-Phenylindol]-Färbung) und des MTS (3-(4,5-Dimethylthiazol-2-yl)-5-(3-Carboxymethoxyphenyl)-2-(4-Sulfophenyl)-2H-Tetrazolium) Assays analysiert. Darüber hinaus wurden die Zellen unter Kontrollbedingungen und osteogenen Bedingungen (Medien mit 10 mM β‑Glycerophosphat) kultiviert. Eine quantitative Expressionsanalyse von Genen, die für die osteogenen Marker alkalische Phosphatase (ALP), Kollagen-Typ-I-Alpha-1-Kette (COL1A1), Osteoprotegerin (OPG) und Osteopontin (OPN) kodieren, erfolgte nach 7 und 14 Tagen Kultivierung. Kalziumablagerungen wurden mit Alizarinrot angefärbt.

Ergebnisse: Unsere Untersuchungen zeigten, dass 10 ng/ml TNF keinen Einfluss auf das Überleben und die Stoffwechselaktivität von PDL-Zellen hatte. Quantitative Expressionsanalysen ergaben, dass Langzeitkulturen mit TNF das osteogene Zellverhalten in frühen und späten Entwicklungsstadien beeinträchtigten. Darüber hinaus ist die Matrixsekretion von TNF-stimulierten PDL-Zellen signifikant reduziert.

Schlussfolgerungen: Die vorliegenden Daten bestätigen TNF als regulatorischen Faktor des proinflammatorischen Remodelings, welcher das Differenzierungsverhalten, aber nicht den Metabolismus und die Zellproliferation des Parodonts beeinflusst. TNF stellt somit ein interessantes Target zur Modulation kieferorthopädischer Umbauprozesse im Zahnhalteapparat dar.

Keywords: Orthodontic tooth movement; Osteogenic differentiation; PDL fibroblasts; Periodontal ligament; Tumor necrosis factor.

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