Rutin promotes the formation and osteogenic differentiation of human periodontal ligament stem cell sheets in vitro

Cell sheet technology is a novel tissue engineering technology that has been rapidly developed in recent years. As a novel technology, cell sheet technology is expected to become one of the preferred methods for cell transplantation. The present study investigated the biological effects of rutin on the formation of periodontal ligament stem cell (PDLSC) sheets and their resultant osteogenic properties. The results of Cell Counting Kit-8 (CCK-8) assay demonstrated that a concentration of 1×10(−6) mol/l rutin promoted the proliferation of PDLSCs more effectively compared with other designed concentrations. Rutin-modified cell sheets could be induced by complete medium supplemented with 20 µg/ml vitamin C (VC) and 1×10(−6) mol/l rutin. Rutin-modified cell sheets appeared thicker and more compact compared with the VC-induced PDLSC sheets, demonstrating more layers of cells (3 or 4 layers), which secreted a richer extracellular matrix (ECM). Furthermore, the improved cell sheets exhibited varying degrees of increases in the mRNA and protein expression of collagen type I (COL1), alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2) and osteopontin (OPN). Combined treatment with VC and rutin promoted the formation of PDLSC sheets and enhanced the osteogenic differentiation potential of the cell sheets. Therefore, rutin-modified cell sheets of PDLSCs are expected to play an important role in the treatment of periodontal tissue regeneration by stem cells.