Abstract
Different structures and cell types of the periodontium respond to orthodontic tooth movement (OTM) individually. Cementoblasts (OC/CM) located in the immediate vicinity of the fibroblasts on the cement have found way to the centre of actual research. Here, we identify and validate possible reference genes for OC/CM cells by RT-qPCR with and without static compressive loading. We investigated the suitability of 3 reference genes in an in vitro model of cementoblast cells using four different algorithms (Normfinder, geNorm, comparative delta-Ct method and BestKeeper) under different confluences and time. Comparable to our previous publications about reference genes in OTM in rats and human periodontal ligament fibroblasts (hPDLF), Rpl22 in murine OC/CM cells appears as the least regulated gene so that it represents the most appropriate reference gene. Furthermore, unlike to the expression of our recommended reference genes, the expression of additionally investigated target genes changes with confluence and under loading compression. Based on our findings for future RT-qPCR analyses in OC/CM cells, Rpl22 or the combination Rpl22/Tbp should be favored as reference gene. According to our results, although many publications propose the use of Gapdh, it does not seem to be the most suitable approach.
Highlights
The periodontal ligament (PDL), a component of the periodontal apparatus, represents a microenvironment essential for anchoring teeth, fulfils functions of proprioception and buffering of acting forces and it enables periodontal remodeling allowing for orthodontic tooth movement
The individual role of different cell types involved in orthodontic tooth movement must be identified and understood in order to gain a better insight into the mechanisms of orthodontic tooth movement for research purposes
It has been shown that root resorption lacunas are regenerated with cementum in absence of the causal stimulus[10,11]
Summary
The periodontal ligament (PDL), a component of the periodontal apparatus, represents a microenvironment essential for anchoring teeth, fulfils functions of proprioception and buffering of acting forces and it enables periodontal remodeling allowing for orthodontic tooth movement. Cementum, produced by cementoblasts all lifelong, plays an important role in anchoring the tooth to the surrounding alveolar b one[6,7] It represents the key structure for attaching Sharpey’s fibers on the tooth side[8]. In hPDLF, hypoxia related VEGF mRNA upregulation is published[22] Considering these findings we already investigated the influence of hypoxia regarding to loading compression experiments with hPDLF with the result, that hypoxic effects appear to play a minor role in the regulation of o steoclastogenesis[23]. These findings need to be verified for cementoblasts. A systematic validation of reference genes with focus on the in vitro model of mechanical loading in cementoblasts is missing
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