RESUMEN
Severe bone defects that extend beyond a critical size do not heal on their own, increasing the risk of complications and leading to poor outcomes for patients. Healing is a highly coordinated and complex process in which immune cells have an important function making the design and preparation of biomaterials with immunomodulatory functions an important new therapeutic strategy. 1,25-dihydroxyvitamin D3 (VD3) is crucial for bone metabolism and immune regulation. For post-defect bone regeneration, we developed a drug delivery system (DDS) based on chitosan (CS) and nanoparticles (NPs) to sustain the release effect of VD3 and desirable biological characteristics. The hydrogel system was physically characterized and confirmed to have good mechanical strength, degradation rate, and drug release rate. In vitro experiments showed that the cells had good biological activity when the hydrogel was co-cultured with MC3T3-E1 and RAW264.7. The high expression of ARG-1 and low expression of iNOS in macrophages confirmed that VD3-NPs/CS-GP hydrogel transformed lipopolysaccharide-induced M1 macrophages into M2 macrophages. Alkaline phosphatase and alizarin red staining showed that VD3-NPs/CS-GP hydrogel promoted osteogenic differentiation under inflammatory conditions. In conclusion, VD3-NPs/CS-GP hydrogel with synergistic anti-inflammatory and pro-osteogenic differentiation effects may serve as a potential immunomodulatory biomaterial for bone repair and regeneration in cases of bone defects.
Asunto(s)
Quitosano , Nanopartículas , Humanos , Hidrogeles/farmacología , Quitosano/farmacología , Osteogénesis , Regeneración Ósea , Materiales Biocompatibles/farmacología , Diferenciación CelularRESUMEN
The CUE domain-containing 2 (CUEDC2) protein plays critical roles in many biological processes, such as the cell cycle, inflammation, and tumorigenesis. However, whether CUEDC2 is involved in osteoblast differentiation and plays a role in bone regeneration remains unknown. This study investigated the role of CUEDC2 in osteogenesis and its underlying molecular mechanisms. We found that CUEDC2 is expressed in bone tissues. The expression of CUEDC2 decreased during bone development and BMP2-induced osteoblast differentiation. The overexpression of CUEDC2 suppressed the osteogenic differentiation of precursor cells, while the knockdown of CUEDC2 showed the opposite effect. In vivo studies showed that the overexpression of CUEDC2 decreased bone parameters (bone volume, bone area, and bone mineral density) during ectopic bone formation, whereas its knockdown increased bone volume and the reconstruction percentage of critical-size calvarial defects. We found that CUEDC2 affects STAT3 activation by regulating SOCS3 protein stability. Treatment with a chemical inhibitor of STAT3 abolished the promoting effect of CUEDC2 silencing on osteoblast differentiation. Together, we suggest that CUEDC2 functions as a key regulator of osteoblast differentiation and bone formation by targeting the SOCS3-STAT3 pathway. CUEDC2 manipulation could serve as a therapeutic strategy for controlling bone disease and regeneration.
Asunto(s)
Diferenciación Celular , Osteoblastos/metabolismo , Osteogénesis , Proteínas Represoras/metabolismo , Factor de Transcripción STAT3/metabolismo , Cráneo/metabolismo , Proteína 3 Supresora de la Señalización de Citocinas/metabolismo , Células 3T3 , Animales , Regulación del Desarrollo de la Expresión Génica , Masculino , Ratones , Ratones Endogámicos C57BL , Osteoblastos/patología , Fosforilación , Estabilidad Proteica , Proteínas Represoras/genética , Transducción de Señal , Cráneo/patología , Cráneo/cirugíaRESUMEN
The peroxisome proliferator-activated receptor (PPAR)-α agonist fenofibrate is used as a lipid-lowering agent to reduce cholesterol and triglyceride in blood. In this study, we investigated whether fenofibrate affects osteoblast differentiation of osteogenic precursor cells. Quantitative real-time PCR and alkaline phosphatase (ALP) staining assays revealed that fenofibrate can enhance the osteoblast differentiation of C3H10T1/2 and MC3T3-E1 cells. In contrast with fenofibrate, the PPARγ agonist rosiglitazone decreased or did not affect the expression of osteogenic genes in these cells. Fenofibrate dose- and time-dependently increased PPARα expression, and concomitantly increased the expression of bone morphogenetic protein 2 (BMP2). Knockdown of PPARα abolished fenofibrate-induced BMP2 expression, activity of the BMP2 promoter gene, and calcium deposition. The chromatin immunoprecipitation assay demonstrated that fenofibrate increased BMP2 expression by inducing direct binding of PPARα to the BMP2 promoter region. Taken together, we suggest that fenofibrate has a stimulatory effect on osteoblast differentiation via the elevation of PPARα levels and the PPARα-mediated BMP2 expression. Our findings provide fenofibrate as a useful agent for controlling hypercholesterolemic patients with osteoporosis.