ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of tumor necrosis factor-α (TNF-α) on osteogenic differentiation and Notch signaling pathway of periodontal ligament stem cells (PDLSCs) and to investigate the regulatory role of Notch signaling pathway on the osteogenic differentiation of PDLSCs under the influence of TNF-α.</p><p><b>METHODS</b>PDLSCs were obtained through enzyme digestion and tissue block method. The expression levels of stem cell surface markers CD105, CD90, CD146, CD45, and CD31 were detected by fluorescence activated cell sorter (FACS). PDLSCs were divided into experimental (10 ng·mL⁻¹ TNF-α) and control groups (0 ng·mL⁻¹ TNF-α). The proliferation ability of PDLSCs was detected using cell counting kit-8 (CCK-8). The effect of TNF-α on the osteogenic ability of PDLSCs were tested by measuring alkaline phosphatase (ALP) activity and conducting alizarin red staining and quantitative real-time polymerase chain reaction (PCR). We tested Notch signal pathway receptors Notch1, Notch2, ligand JAG1, JGA2, and downstream gene Hes-1. Changes in DLL1 expression were detected by quantitative real-time PCR.</p><p><b>RESULTS</b>FACS profiling showed that PDLSCs were strongly positive for CD105, CD90, and CD146 but negative for CD45 and CD31. CCK-8 results showed that TNF-α could promote the proliferation of PDLSCs (P<0.05). ALP activity in the experimental group was lower than that in the control group (P<0.05). Alizarin red staining showed that the experimental group had decreased mineralized nodules as compared with the control group. Quantitative real-time PCR results showed that the mRNA expression of osteogenic marker genes cementum attachment protein (CAP), osteopontin (OPN), and Runt-related transcription factor 2 (Runx2) significantly decreased in the experimental group as compared with those in the control group (P<0.05). The expression levels of Notch1, Notch2, JAG1, JGA2 and Hes-1 were significantly decreased (P<0.05), whereas those of Notch3 and DLL1 were increased in Notch signaling pathway-related molecules (P<0.05).</p><p><b>CONCLUSIONS</b>TNF-α can promote PDLSCs proliferation and inhibit bone differentiation and Notch signaling pathway expression, indicating that the Notch signaling pathway regulates PDLSCs osteogenic differentiation.</p>
ABSTRACT
Bone tissue engineering is a scientific field devoted to the development of materials that can repair or replace human bone tissue with biological and engineering methods. The stent, which provides structural support and adhesion sites for cell and tissue growth, is one of the key elements in tissue engineering. The scaffold may comprise metal, polymer, and ceramic biomaterial. The polymer scaffold is widely used due to its biocompatibility, biodegradability, and mechanical stability. Chitosan, as a natural polymer, is derived from chitin and has played a particularly important role in bone tissue engineering over the past two decades. In recent years, chitosan composites and their application in bone tissue engineering have received considerable attention due to their small foreign body reaction, excellent antibacterial properties, plasticity, suitability for inward cell growth, and bone conduction. This review will discuss the biocompatibility and osteogenesis research in vivo and in vitro of several common chitosan composites in bone tissue engineering.