Transforming Growth Factor-β3/Chitosan Sponge (TGF-β3/CS) Facilitates Osteogenic Differentiation of Human Periodontal Ligament Stem Cells

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.

Chitosan-Based Coacervate Polymers for Propolis Encapsulation: Release and Cytotoxicity Studies

“…However, the CS-P experimental group exhibited a statistically significant difference with respect to the control group (p < 0.0001), which presented lower cell viability. This result is in accordance with previous studies, which observed an increase of viable cells in different linage cells when chitosan was used as a biomaterial base [63][64][65]. The rates of cell viability exhibited by both groups make them suitable for biomedical use according to the International Organization for Standardization (ISO) and the national standards for Biological Evaluation of Medical Devices, ISO 10993-5 [66].…”

Enhanced Osteogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells by a Hybrid Hydroxylapatite/Collagen Scaffold

“…It is reported that Sp7 is a transcription factor for osteoblast differentiation, whereas Runx2 is known as a downstream gene ( Xin et al, 2019 ). BMP2 pathway is known to up-regulate Sp7 expression through two distinct transcription factors such as Runx2 and Msx2 during osteoblast differentiation ( Li et al, 2019 ; Tang et al, 2019 ). Accumulating data proved that BMP2 is involved in bone formation, bone remodeling, bone development, and osteoblast differentiation through hBMSC osteogenic differentiation ( Biswas et al, 2018 ; Wu et al, 2018 ).…”

“…TGF-β3 induces endochondral bone formation and completes bone remodeling. The signal transduction mediated by TGF-β 3 in osteogenic differentiation and bone regeneration specifically occurs through both canonical SMAD -dependent pathways with TGF-β3 ligands, receptors, and SMADs ( Li et al, 2019 ). Indeed, TGFβ3 gene and SMAD3 transcription factor up-regulation could induce bone regeneration by amplifying hBMSCs recruitment at the damaged site ( Deng et al, 2017 ).…”

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“…It is reported that Sp7 is a transcription factor for osteoblast differentiation, whereas Runx2 is known as a downstream gene (Xin et al, 2019). BMP2 pathway is known to upregulate Sp7 expression through two distinct transcription factors such as Runx2 and Msx2 during osteoblast differentiation (Li et al, 2019;Tang et al, 2019). Accumulating data proved that BMP2 is involved in bone formation, bone remodeling, bone development, and osteoblast differentiation through hBMSC osteogenic differentiation (Biswas et al, 2018;Wu et al, 2018).…”

“…TGF-β3 induces endochondral bone formation and completes bone remodeling. The signal transduction mediated by TGF-β 3 in osteogenic differentiation and bone regeneration specifically occurs through both canonical SMAD-dependent pathways with TGF-β3 ligands, receptors, and SMADs (Li et al, 2019). Indeed, TGFβ3 gene and SMAD3 transcription factor up-regulation could induce bone regeneration by amplifying hBMSCs recruitment at the damaged site (Deng et al, 2017).…”