Heinz-Dieter Müller1, Barbara Cvikl2, Klara Janjić3, Sylvia Nürnberger4, Andreas Moritz3, Reinhard Gruber5, Hermann Agis6. 1. Department of Prosthodontics, Medical University of Vienna, Vienna, Austria; Department of Preventive, Restorative and Pediatric Dentistry, University of Bern, Bern, Switzerland; Austrian Cluster for Tissue Regeneration, Vienna, Austria. 2. Department of Preventive, Restorative and Pediatric Dentistry, University of Bern, Bern, Switzerland; Austrian Cluster for Tissue Regeneration, Vienna, Austria; Department of Conservative Dentistry and Periodontology, Medical University of Vienna, Vienna, Austria. 3. Austrian Cluster for Tissue Regeneration, Vienna, Austria; Department of Conservative Dentistry and Periodontology, Medical University of Vienna, Vienna, Austria. 4. Austrian Cluster for Tissue Regeneration, Vienna, Austria; Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria; Department of Trauma Surgery, Medical University of Vienna, Vienna, Austria. 5. Austrian Cluster for Tissue Regeneration, Vienna, Austria; Department of Oral Biology, Medical University of Vienna, Vienna, Austria; Laboratory for Oral Cell Biology, University of Bern, Bern, Switzerland. 6. Austrian Cluster for Tissue Regeneration, Vienna, Austria; Department of Conservative Dentistry and Periodontology, Medical University of Vienna, Vienna, Austria. Electronic address: hermann.agis@meduniwien.ac.at.
Abstract
INTRODUCTION: Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. METHODS: In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. RESULTS: In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. CONCLUSIONS: Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization.
INTRODUCTION: Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. METHODS: In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. RESULTS: In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. CONCLUSIONS: Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization.
Authors: Klara Janjić; Alwina Schellner; Alexander Engenhart; Kurt Kernstock; Barbara Schädl; Andreas Moritz; Hermann Agis Journal: J Periodontal Res Date: 2019-03-20 Impact factor: 4.419