Abstract
Particulate autologous tooth roots are increasingly used for alveolar bone augmentation; however, the proteomic profile of acid dentin lysate and the respective cellular response have not been investigated. Here we show that TGF-β1 is among the 226 proteins of acid dentin lysate (ADL) prepared from porcine teeth. RNA sequencing identified 231 strongly regulated genes when gingival fibroblasts were exposed to ADL. Out of these genes, about one third required activation of the TGF-β receptor type I kinase including interleukin 11 (IL11) and NADPH oxidase 4 (NOX4). Reverse transcription-quantitative polymerase chain reaction and immunoassay confirmed the TGF-β-dependent expression of IL11 and NOX4. The activation of canonical TGF-β signaling by ADL was further confirmed by the phosphorylation of Smad3 and translocation of Smad2/3, using Western blot and immunofluorescence staining, respectively. Finally, we showed that TGF-β activity released from dentin by acid lysis adsorbs to titanium and collagen membranes. These findings suggest that dentin particles are a rich source of TGF-β causing a major response of gingival fibroblasts.
Highlights
Particulate autologous tooth roots are increasingly used for alveolar bone augmentation; the proteomic profile of acid dentin lysate and the respective cellular response have not been investigated
We report here the gene expression changes of gingival fibroblasts exposed to acid dentin lysate (ADL), similar to what we have done with acid bone lysate[13] and platelet-rich fibrin lysate[21]
In the present investigation we report that (i) TGF-β1 is among the proteomic profile of acid dentin lysate; (ii) which provokes a robust activation of genes including interleukin 11 (IL11) and NADPH oxidase 4 (NOX4) via the canonical TGF-β signaling in gingival fibroblasts; (iii) which is paralleled by the phosphorylation of Smad[3] and the nuclear translocation of Smad2/3
Summary
Particulate autologous tooth roots are increasingly used for alveolar bone augmentation; the proteomic profile of acid dentin lysate and the respective cellular response have not been investigated. As dentin is free of costs and accessible, the use of autogenous tooth roots has received increasing attention in alveolar bone augmentations The rationale of this clinical approach rests on the similarities between dentin and bone in terms of biological and structural features[10,11]. We report here the gene expression changes of gingival fibroblasts exposed to ADL, similar to what we have done with acid bone lysate[13] and platelet-rich fibrin lysate[21] Both studies identified interleukin 11 (IL11) and NADPH oxidase 4 (NOX4) to be among the most strongly regulated genes in gingival fibroblast cells requiring TGF-β receptor type I kinase[13,21], and showed. We determined the TGF-β activity of acid dentin lysate by using the established bioassay strategy that we followed for the analysis of acid bone lysate[13]
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