Abstract
Periodontal disease is the main reason for tooth loss in adults. Tissue engineering and regenerative medicine are advanced technologies used to manage soft and hard tissue defects caused by periodontal disease. We developed a transforming growth factor-β3/chitosan sponge (TGF-β3/CS) to repair periodontal soft and hard tissue defects. We investigated the proliferation and osteogenic differentiation behaviors of primary human periodontal ligament stem cells (hPDLSCs) to determine the bioactivity and potential application of TGF-β3 in periodontal disease. We employed calcein-AM/propidium iodide (PI) double labeling or cell membranes (CM)-Dil labeling coupled with fluorescence microscopy to trace the survival and function of cells after implantation in vitro and in vivo. The mineralization of osteogenically differentiated hPDLSCs was confirmed by measuring alkaline phosphatase (ALP) activity and calcium content. The levels of COL I, ALP, TGF-βRI, TGF-βRII, and Pp38/t-p38 were assessed by western blotting to explore the mechanism of bone repair prompted by TGF-β3. When hPDLSCs were implanted with various concentrations of TGF-β3/CS (62.5–500 ng/mL), ALP activity was the highest in the TGF-β3 (250 ng/mL) group after 7 d (p < 0.05 vs. control). The calcium content in each group was increased significantly after 21 and 28 d (p < 0.001 vs. control). The optimal result was achieved by the TGF-β3 (500 ng/mL) group. These results showed that TGF-β3/CS promotes osteogenic differentiation of hPDLSCs, which may involve the p38 mitogen-activated protein kinase (MAPK) signaling pathway. TGF-β3/CS has the potential for application in the repair of incomplete alveolar bone defects.
Highlights
Periodontal disease is a chronic inflammatory condition that affects the supporting tissues around teeth, resulting in periodontal tissue breakdown or tooth loss in severe cases
DLSCs were cultured on Transforming growth factor-β3 (TGF-β3)/CS for 3 d, human periodontal ligament stem cells (hPDLSCs) grew well
GF-β3, we observed that hPDLSCs grew and proliferated on CS
Summary
Periodontal disease is a chronic inflammatory condition that affects the supporting tissues around teeth, resulting in periodontal tissue breakdown or tooth loss in severe cases. Repairing periodontal support tissues, such as alveolar bone, is indispensable for the treatment of periodontal disease. With the increasing popularity of dental implant surgery, the lack of bone mass in patients with periodontitis has limited the need for implant surgery, and the need to repair alveolar bone defects is increasing [2,3]. The current clinical techniques mainly used for the treatment of alveolar bone defects [4] are bone grafting and guided bone regeneration (GBR). Periodontal tissue engineering technology, which is characterized by the use of stem cells, bioactive molecules (e.g., growth factors), and scaffold materials as the three basic elements, has provided a new solution to reconstruct alveolar bone defects
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