RESUMEN
AIM: To evaluate the role of biomimetic pulp scaffolds derived from the extracellular matrix derived of stem cells from human exfoliated deciduous teeth (SHED-ECM-PS) in promoting pulp-dentine complex regeneration. METHODOLOGY: SHED-ECM-PS was prepared through cell aggregation and decellularization techniques. Histological and immunofluorescence analyses, scanning electron microscopy, and DNA quantification assays were used to characterize the SHED-ECM-PS. Additionally, a tooth slice implantation model was established to evaluate the effects of SHED-ECM-PS on regeneration of the pulp-dentine complex in vivo. Extraction medium for SHED-ECM-PS was prepared, and its effect on bone marrow mesenchymal stem cells (BMMSCs) was assessed in vitro. Cell counting kit-8 and Ki-67 staining assays were performed to determine cell proliferation. The rate of apoptosis was evaluated by flow cytometry. Wound healing and transwell assays were conducted to evaluate cell migration. Alizarin red S staining was performed to examine mineralized nodule formation. Western blotting was used to detect the expression of osteogenic and odontogenic markers. The results were analysed using an independent two-tailed Student's t-test. p < .05 was considered statistically significant. RESULTS: SHED-ECM-PS was successfully constructed, exhibiting a striped dental pulp-like shape devoid of nuclear structures or DNA components, and rich in fibronectin, collagen I, DMP1 and DSPP. Notably, SHED-ECM-PS showed no impact on the proliferation or apoptosis of BMMSCs. Histological analysis revealed that dental pulp fibroblasts formed an interwoven mesh in the root canal, and angiogenesis was observed in the SHED-ECM-PS group. Moreover, a continuous, newly formed tubular dentine layer with polarized odontoblast-like cells was observed along the inner wall of the root canal. SHED-ECM-PS promoted the migration, polar alignment and mineralized nodule formation of BMMSCs and specifically elevated the expression levels of odontogenic markers, but not osteogenic markers, compared with the control group in vitro. CONCLUSION: SHED-ECM-PS exhibited no cytotoxicity and promoted pulp-dentine complex regeneration in vivo as well as cell migration and odontogenic differentiation of BMMSCs in vitro. These findings provide evidence that SHED-ECM-PS, as a novel biological scaffold, has the potential to improve the outcomes of REPs.
Asunto(s)
Proliferación Celular , Pulpa Dental , Dentina , Matriz Extracelular , Regeneración , Andamios del Tejido , Diente Primario , Humanos , Diente Primario/citología , Pulpa Dental/citología , Células Madre , Células Madre Mesenquimatosas , Movimiento Celular , Animales , Microscopía Electrónica de Rastreo , Odontogénesis , Diferenciación Celular , Células CultivadasRESUMEN
OBJECTIVES: To investigate in vitro effects of a nanoparticle bioceramic material, iRoot BP Plus, on stem cells from apical papilla (SCAP) and in vivo capacity to induce pulp-dentin complex formation. MATERIALS AND METHODS: The sealing ability of iRoot BP Plus was measured via scanning electron microscopy (SEM). SCAP were isolated and treated in vitro by iRoot BP Plus conditioned medium, with mineral trioxide aggregate (MTA) conditioned medium and regular medium used as controls, respectively. Cell proliferation was assessed by BrdU labeling and MTT assay and cell migration was evaluated with wound healing and transwell assays. Osteo/odontogenic potential was evaluated by Alizarin red S staining and qPCR. Pulp-dentin complex formation in vivo was assessed by a tooth slice subcutaneous implantation model. RESULTS: iRoot BP Plus was more tightly bonded with the dentin. There was no difference in SCAP proliferation between iRoot BP Plus and control groups (P > 0.05). iRoot BP Plus had a greater capacity to elevated cell migration (P < 0.05) and osteo/odontogenic marker expression and mineralization nodule formation of SCAP compared with MTA groups (P < 0.05). Furthermore, the new continuous dentine layer and pulp-like tissue was observed in the iRoot BP Plus group in vivo. CONCLUSIONS: iRoot BP Plus showed excellent sealing ability, promoted the migration and osteo/odontogenesis of SCAP and induced pulp-dentin complex formation without affecting the cell proliferation, which indicated iRoot BP Plus was a promising coronal sealing material in REPs. CLINICAL RELEVANCE: The coronal sealing materials play crucial roles for the outcomes of REPs. This study showed that iRoot BP Plus has good coronal sealing and promote pulp-dentin complex formation compared with MTA, providing experimental evidences for the clinical application of iRoot BP Plus as a promising coronal seal material in REPs.
Asunto(s)
Endodoncia Regenerativa , Humanos , Medios de Cultivo Condicionados/farmacología , Diferenciación Celular , Pulpa Dental , Silicatos/farmacología , Proliferación Celular , Óxidos/farmacología , Compuestos de Calcio/farmacología , Combinación de Medicamentos , Compuestos de Aluminio/farmacologíaRESUMEN
The rapid development of the internet has brought about a comprehensive transformation in human life. However, the challenges of cybersecurity are becoming increasingly severe, necessitating the implementation of effective security mechanisms. Cybersecurity situational awareness can effectively assess the network status, facilitating the formulation of better cybersecurity defense strategies. However, due to the low accuracy of existing situational assessment methods, situational assessment remains a challenge. In this study, a new situational assessment method, MSWOA-BiGRU, combining optimization algorithms and temporal neural networks, was proposed. Firstly, a scientific indicator system proposed in this research is used to calculate the values of each indicator. Then, the Analytic Hierarchy Process is used to derive the actual situation values, which serve as labels. Taking into account the temporal nature of network traffic, the BiGRU model is utilized for cybersecurity situational assessment. After integrating time-related features and network traffic characteristics, the situational assessment value is obtained. During the evaluation process, a whale optimization algorithm (MSWOA) improved with a mix of strategies proposed in this study was employed to optimize the model. The performance of the proposed MSWOA-BiGRU model was evaluated on publicly available real network security datasets. Experimental results indicate that compared to traditional optimization algorithms, the optimization performance of MSWOA has seen significant enhancement. Furthermore, MSWOA-BiGRU demonstrates superior performance in cybersecurity situational assessment compared to existing evaluation methods.