The effect of Azacitidine and Trichostatin A on impaired osteogenic differentiation capacity of bone marrow and periodontal ligament derived-mesenchymal stem cells
The effect of Azacitidine and Trichostatin A on impaired osteogenic differentiation capacity of bone marrow and periodontal ligament derived-mesenchymal stem cells
Mahshid Hodjat,1,*Mahdi Gholami,2Mohammad Sadegh Ahmad Akhoundi ,3
1. Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences (TUMS), Tehran, Iran 2. Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran 3. Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences (TUMS), Tehran, Iran
Introduction: Diabetes Mellitus is an increasingly prevalent metabolic disease in the 21st century that is associated with long-term malfunction of different organs. Alteration of bone metabolism is the common complication of hyperglycemic exposure leading to an increased risk of osteoporosis-related fractures. Teeth surrounding alveolar bone could highly be affected by diabetes, undergoes gradual deterioration. In this project, we aimed to investigate the effect of epigenetic modifiers drugs on the osteogenic differentiation capacity of mesenchymal stem cell derived from diabetic rat bone marrow (BM) and tooth periodontal ligament tissue (PDL).
Methods: 8 wistar rats were included in the study and divided into two groups; Group-I: rats injected intraperitoneally with streptozotocin 60 mg/kg; Group-II: Control group injected with normal saline. The mandibles as well as femurs were dissected after three months. The cells were isolated and cultured from BM and PDL. The MSCs were characterized for surface markers using flow cytometry. The osteogenic capacity of MSCs-derived BM and PDL were assessed in the presence of epidrugs; Trichostatin-A (TSA) and Azacitidine (5-AZA) using Alizarin Red staining, ALP activity test and real-time PCR for gene expression analysis.
Results: The isolated cells were positive for CD105 and CD90 while there was low detectable cells with CD45+. The results showed reduced mineralization capacity of diabetic and normal rat BM and PDL MSCs in the presence of osteogenic medium containing high glucose. Adding Trichostatin A or 5-Azacytidine increased mineralization in MSC derived from diabetic rat BM and PDL. Adding TSA or 5-Aza also increased beta-catenin and GSK3ß the member of wnt signaling.
Conclusion: Hyperglycemic condition affects negatively the osteogenic potential of MSCs derived from bone marrow and periodontal ligament. Our results demonstrate that suppression of DNA methylation and histone deacetylase could alleviate the impaired osteogenic differentiation capacity of diabetic MSCs. Our data indicate a role for the canonical Wnt signaling pathway in diabetogenic conditions. The data further implicated a role of epigenetic therapy in the treatment of osteogenesis complications of patients suffering from diabetic alveolar bone loss and diabetes-related bone diseases.