Computational analysis of curcumin-mediated alleviation of inflammation in periodontitis patients with experimental validation in mice.

AIM: Using network pharmacology and experimental validation to explore the therapeutic efficacy and mechanism of curcumin (Cur) in periodontitis treatment. MATERIALS AND METHODS: Network pharmacology was utilized to predict target gene interactions of Cur-Periodontitis. Molecular docking was used to investigate the binding affinity of Cur for the predicted targets. A mouse model with ligature-induced periodontitis (LIP) was used to verify the therapeutic effect of Cur. Microcomputed tomography (micro-CT) was used to evaluate alveolar bone resorption, while western blotting, haematoxylin-eosin staining and immunohistochemistry were used to analyse the change in immunopathology. SYTOX Green staining was used to assess the in vitro effect of Cur in a mouse bone marrow-isolated neutrophil model exposed to lipopolysaccharide. RESULTS: Network pharmacology identified 114 potential target genes. Enrichment analysis showed that Cur can modulate the production of neutrophil extracellular traps (NETs). Molecular docking experiments suggested that Cur effectively binds to neutrophil elastase (ELANE), peptidylarginine deiminase 4 (PAD4) and cathepsin G, three enzymes involved in NETs. In LIP mice, Cur alleviated alveolar bone resorption and reduced the expression of ELANE and PAD4 in a time-dependent but dose-independent manner. Cur can directly inhibit NET formation in the cell model. CONCLUSIONS: Our research suggested that Cur may alleviate experimental periodontitis by inhibiting NET formation.

Quercetin promotes autophagy to alleviate cigarette smoke-related periodontitis.

BACKGROUND AND OBJECTIVES: Cigarette smoking has been reported as an independent risk factor for periodontitis. Tobacco toxins affect periodontal tissue not only locally but also systemically, leading to the deterioration and recurrence of periodontitis. However, the mechanism of cigarette smoke-related periodontitis (CSRP) is unclear and thus lacks targeted treatment strategies. Quercetin, a plant-derived polyphenolic flavonoid, has been reported to have therapeutic effects on periodontitis due to its documented antioxidant activity. This study aimed to evaluate the effects of quercetin on CSRP and elucidated the underlying mechanism. METHODS: The cigarette smoke-related ligature-induced periodontitis mouse model was established by intraperitoneal injection of cigarette smoke extract (CSE) and silk ligation of bilateral maxillary second molars. Quercetin was adopted by gavage as a therapeutic strategy. Micro-computed tomography was used to evaluate the alveolar bone resorption. Immunohistochemistry detected the oxidative stress and autophagy markers in vivo. Cell viability was determined by Cell Counting Kit-8, and oxidative stress levels were tested by 2,7-dichlorodihydrofluorescein diacetate probe and lipid peroxidation malondialdehyde assay kit. Alkaline phosphatase and alizarin red staining were used to determine osteogenic differentiation. Network pharmacology analysis, molecular docking, and western blot were utilized to elucidate the underlying molecular mechanism. RESULTS: Alveolar bone resorption was exacerbated and oxidative stress products were accumulated during CSE exposure in vivo. Oxidative stress damage induced by CSE caused inhibition of osteogenic differentiation in vitro. Quercetin effectively protected the osteogenic differentiation of human periodontal ligament cells (hPDLCs) and periodontal tissue by upregulating the expression of Beclin-1 thus to promote autophagy and reduce oxidative stress damage. CONCLUSION: Our results established a role of oxidative stress damage and autophagy dysfunction in the mechanism of CSE-induced destruction of periodontal tissue and hPDLCs, and provided a potential application value of quercetin to ameliorate CSRP.

Electroacupuncture regulates macrophage, neutrophil, and oral microbiota to alleviate alveolar bone loss and inflammation in experimental ligature-induced periodontitis.

AIM: Electroacupuncture (EA) regulates distant body physiology through somatic sensory autonomic reflexes, balances the microbiome, and can promote the release of immune cells into bloodstream, thereby inhibiting severe systemic inflammation. This makes it possible to use EA as an integrated treatment for periodontitis. MATERIALS AND METHODS: In this study, EA was applied to the ST36 acupoints in a ligature-induced periodontitis (LIP) mouse model. Then the effects of EA on periodontal myeloid cells, cytokines, and the microbiome were comprehensively analysed using flow cytometry, quantitative Polymerase Chain Reaction (PCR), and 16 S sequencing. RESULTS: Results demonstrated that EA could significantly relieve periodontal bone resorption. EA also suppressed the infiltration of macrophages and neutrophils, reduced gene expression of the pro-inflammatory cytokines IL-1ß, IL-6, IL-17 and TNF-α, and increased expression of the anti-inflammatory factors IL-4 and IL-10 in periodontal tissues. Moreover, composition of the periodontal microbiome was regulated by EA, finding that complex of microbiota, including supragingival Veillonella, subgingival Streptococcus, and subgingival Erysipelatoclostridium, were significantly reduced. Meanwhile, nitrate and nitrate-related activities of subgingival microbiota were reversed. Network analysis revealed close relationships among Veillonella, Streptococcus, and Bacteroides. CONCLUSIONS: Our study indicates that EA can effectively alleviate inflammation and bone resorption in LIP mice, potentially via the regulation of myeloid cells, cytokines, and periodontal microbiome.

Predicting the risk of dental implant loss using deep learning.

AIM: To investigate the feasibility of predicting dental implant loss risk with deep learning (DL) based on preoperative cone-beam computed tomography. MATERIALS AND METHODS: Six hundred and three patients who underwent implant surgery (279 high-risk patients who did and 324 low-risk patients who did not experience implant loss within 5 years) between January 2012 and January 2020 were enrolled. Three models, a logistic regression clinical model (CM) based on clinical features, a DL model based on radiography features, and an integrated model (IM) developed by combining CM with DL, were developed to predict the 5-year implant loss risk. The area under the receiver operating characteristic curve (AUC) was used to evaluate the model performance. Time to implant loss was considered for both groups, and Kaplan-Meier curves were created and compared by the log-rank test. RESULTS: The IM exhibited the best performance in predicting implant loss risk (AUC = 0.90, 95% confidence interval [CI] 0.84-0.95), followed by the DL model (AUC = 0.87, 95% CI 0.80-0.92) and the CM (AUC = 0.72, 95% CI 0.63-0.79). CONCLUSIONS: Our study offers preliminary evidence that both the DL model and the IM performed well in predicting implant fate within 5 years and thus may greatly facilitate implant practitioners in assessing preoperative risks.

Nanotube-decorated hierarchical tantalum scaffold promoted early osseointegration.

This study aimed to fabricate a hierarchical tantalum scaffold mimicking natural bone structure to enhance osseointegration. Porous tantalum scaffolds (p-Ta) with microgradients were fabricated by selective laser melting according to a computer-aided design model. Electrochemical anodization produced nanotubes on the p-Ta surface (p-Ta-nt). SEM verified the construction of a unique nanostructure on p-Ta-nt. Contact angle and protein adsorption measurements demonstrated that p-Ta-nt have enhanced hydrophilicity and protein absorption. MC3T3-E1 preosteoblasts showed increased filamentous pseudopods and comparable cell proliferation when cultured on p-Ta-nt. Osteogenic marker gene (Osterix, Runx2, COL-I) transcription was significantly upregulated in MC3T3-E1 cells cultured on p-Ta-nt after 7 days. After implantation into the femurs of New Zealand white rabbits for 2 weeks, histological examination found improved early osseointegration in the p-Ta-nt group. This study showed that a hierarchical tantalum structure could enhance early osteogenic effects in vitro and in vivo.

Mitochondrial Calcium Ion Nanogluttons Alleviate Periodontitis via Controlling mPTPs.

The mitochondrial permeability transition (mPT) directly affects mitochondrial function in macrophages. Under inflammatory conditions, mitochondrial calcium ion (mitoCa2+ ) overload triggers the persistent opening of mPT pores (mPTPs), further aggravating Ca2+ overload and increasing reactive oxygen species (ROS) to form an adverse cycle. However, there are currently no effective drugs targeting mPTPs to confine or unload excess Ca2+ . It is novelly demonstrated that the initiation of periodontitis and the activation of proinflammatory macrophages depend on the persistent overopening of mPTPs, which is mainly triggered by mitoCa2+ overload and facilitates further mitochondrial ROS leakage into the cytoplasm. To solve the above problems, mitochondrial-targeted "nanogluttons" with PEG-TPP conjugated to the surface of PAMAM and BAPTA-AM encapsulated in the core are designed. These nanogluttons can efficiently "glut" Ca2+ around and inside mitochondria to effectively control the sustained opening of mPTPs. As a result, the nanogluttons significantly inhibit the inflammatory activation of macrophages. Further studies also unexpectedly reveal that the alleviation of local periodontal inflammation in mice is accompanied by diminished osteoclast activity and reduced bone loss. This provides a promising strategy for mitochondria-targeted intervention in inflammatory bone loss in periodontitis and can be extended to treat other chronic inflammatory diseases associated with mitoCa2+ overload.

Three-Dimensional Matrix Stiffness Activates the Piezo1-AMPK-Autophagy Axis to Regulate the Cellular Osteogenic Differentiation.

Extracellular matrix (ECM) stiffness is a key stimulus affecting cellular differentiation, and osteoblasts are also in a three-dimensional (3D) stiff environment during the formation of bone tissues. However, it remains unclear how cells perceive matrix mechanical stiffness stimuli and translate them into intracellular signals to affect differentiation. Here, for the first time, we constructed a 3D culture environment by GelMA hydrogels with different amino substitution degrees and found that Piezo1 expression was significantly stimulated by the stiff matrix with high substitution; meanwhile, the expressions of osteogenic markers OSX, RUNX2, and ALP were also observably improved. Moreover, knockdown of Piezo1 in the stiff matrix revealed significant reduction of the abovementioned osteogenic markers. In addition, in this 3D biomimetic ECM, we also observed that Piezo1 can be activated by the static mechanical conditions of the stiff matrix, leading to the increase of the intracellular calcium content and accompanied with a continuous change in cellular energy levels as ATP was consumed during cellular differentiation. More surprisingly, we found that in the 3D stiff matrix, intracellular calcium as a second messenger promoted the activation of the AMP-activated protein kinase (AMPK) and unc-51-like autophagy-activated kinase 1 (ULK1) axis and modestly modulated the level of autophagy, bringing it more similar to differentiated osteoblasts, with increased ATP energy metabolism consumption. Our study innovatively clarifies the regulatory role of the mechanosensitive ion channel Piezo1 in a static mechanical environment on cellular differentiation and verifies the activation of the AMPK-ULK1 axis in the cellular ATP energy metabolism and autophagy level. Collectively, our research develops the understanding of the interaction mechanisms of biomimetic extracellular matrix biomaterials and cells from a novel perspective and provides a theoretical basis for bone regeneration biomaterials design and application.

Natural Plant Tissue with Bioinspired Nano Amyloid and Hydroxyapatite as Green Scaffolds for Bone Regeneration.

Bone defects have been increasingly prevalent around the globe and traditional bone substitutes are constantly limited by low abundance and biosafety due to their animal-based resources. Plant-based scaffolds are currently studied as a green candidate but the bioinertia of cellulose to mammalian cells leads to uncertain bone regeneration. Inspired by the cross-kingdom adhesion of plants and bacteria, this work proposes a concept of a novel plant bone substitute, involving coating decellularized plant with nano amyloids and nano hydroxyapatites, to bridge the plant scaffold and animal tissue regeneration. Natural microporosity of plants can guide alignment of mammalian cells into various organ-like structures. Taking advantage of the bioactive nano amyloids, the scaffolds drastically promote cell adhesion, viability, and proliferation. The enhanced bio-affinity is elucidated as positively charged nano amyloids and serum deposition on the nanostructure. Nano-hydroxyapatite crystals deposited on amyloid further prompt osteogenic differentiation of pre-osteoblasts. In vivo experiments prove successful trabeculae regeneration in the scaffold. Such a hierarchical design leverages the dedicated microstructure of natural plants and high bioactivity of nano amyloid/hydroxyapatite coatings, and addresses the abundant resource of bone substitutes. Not limited to their current application, plant materials functionalized with nano amyloid/hydroxyapatite coatings allow many cross-kingdom tissue engineering and biomedical applications.

Tumor-specific design of PEGylated gadolinium-based nanoscale particles: Facile synthesis, characterization, and improved magnetic resonance imaging of metastasis lung cancer.

Herein we report the synthesis and characterization of the antifouling Gadolinium oxide (Gd2O3) nanoparticles (NPs) modified with PEG with improved biocompatibility for MR imaging purposes. In this report, using the solvothermal decomposition of Gadolinium (III) in the presence of Na3cit, monitored by surface modification with PEG and L-Cys. The synthesized nanoparticles were confirmed by the TEM, DLS and UV-Visible spectroscopy. The morphological results show normal distance across of the flawless Gd2O3-PEG-Cys-NPs show 7.9 ±â€¯0.4 nm, discretely, with a thin size exchange. This infers the surface adjustment does not obviously alteration the center size of the Gd2O3-NPs when contrasted with the perfect sodium citrate-balanced out Gd2O3-NPs. The Gd2O3-PEG-L-Cys-NPs are highly stable at room temperature, water dispersible and importantly less cytotoxic at high concentration of the NPs. The T1-weighted MR phantasm readings evidentially displayed that the formed PEG coated Gd2O3-PEG and Gd2O3-PEG-Cys-NPs with and without Cys may be performed as the promising T1-weighted MR imaging. The NPs displays no signs of toxicity against the human blood, which represents the biocompatibility for the human medicine applications. The Gd2O3-PEG-Cys-NPs shows relatively, high r1 acceptable cytocompatibility, target specific cancer cells and activate the dual mode MR imaging of lung metastasis cancer model in vitro. The development of versatile zwitterion functionalized Gd2O3 may be promising as an active nanoparticle probe for improved multi-model of MR imaging agents for various cancer diseases.

Nanotopography on titanium promotes osteogenesis via autophagy-mediated signaling between YAP and ß-catenin.

Nanostructured titanium implants are recognized for inducing osteogenesis, but the cell signal transductions related to topography are not fully understood. Implant topography is associated with the functionality of osteogenic transcription factors directed by ß-catenin in the nucleus, and autophagic flux in the cytoplasm; YAP (Yes-associated protein) is implicated in the destruction of ß-catenin in the cytoplasm and is susceptible to autophagic flux. This study investigated whether surface topography of the titanium implant modulates autophagy-lysosome degradation of cytoplasmic YAP. Titanium surfaces were modified with smooth, micro, or nanotopographies. Compared with the smooth and micro surfaces, nanotopography was associated with higher ß-catenin nuclear translocation, osteogenic differentiation, and autophagy, and less cytoplasmic YAP. Blockade of the autophagy-lysosome pathway resulted in YAP retention in MC3T3-E1 cells. Cytoplasmic YAP restricted ß-catenin nuclear translocation. In the nano surface group, ß-catenin accumulation in the nucleus and expression of osteogenesis genes was improved. However, in the absence of cell-cell (confluent) contact, manipulation of YAP and ß-catenin localization associated with topography-induced autophagy was lost. In summary, the osteogenesis observed in response to titanium implants with nanotopography involves a signaling link between YAP and ß-catenin. STATEMENT OF SIGNIFICANCE: Titanium with rough topographical surfaces is extensively applied in orthopedic and dental clinics. However, the cellular response to topographies that promotes osteogenesis and underlying mechanisms are not fully understood. In this study, we modified titanium surfaces to produce smooth, micro, or nano topographies. Experiments indicated that the nanotopography induced a stronger autophagic response, leading to degraded cytoplasmic YAP. With the lower levels of YAP, ß-catenin transported and accumulated in the nucleus to activate TCF/LEF transcription factors, resulting in stronger osteogenesis. Additionally, cell-cell contact was essential in the autophagy-mediated signaling link between YAP and ß-catenin. Consequently, our investigation revealed a novel signal transduction in nanotopography-regulated osteogenesis, and supports the modification of biomaterial surfaces to maximize osseointegration.

Nano-hydroxyapatite mineralized silk fibroin porous scaffold for tooth extraction site preservation.

OBJECTIVE: To fabricate a novel nano-hydroxyapatite mineralized silk fibroin (MSF) scaffold in order to diminish the resorption of alveolar ridge and accelerate new bone formation within tooth sockets. Also, to investigate the biocompatibility and osteogenic ability of the MSF in vitro, and the effect of site preservation of the MSF graft in post-extractive sockets in vivo. METHODS: SEM, EDX, FTIR and XRD were used to analyze the mineral crystals deposited on the silk fibroin (SF) surface. Pre-osteoblasts (MC3T3-E1) were seeded on SF and MSF scaffolds. Cell viability, distribution and differentiation were examined using a live-dead assay, histological analysis and Alizarin Red S staining. Furthermore, prepared grafts (SF or MSF scaffold) were implanted into the maxillary right first molar sockets of Sprague Dawley rats for 6 weeks and newly formed bone tissue was analyzed by micro-CT and histological examination. RESULTS: The SEM, EDX, FTIR and XRD analysis demonstrated that granulate nano-hydroxyapatite (nHA) crystals were uniformly distributed on the SF scaffold. In addition, the MSF hydrophilicity measured by water contact angle and swelling ratio was superior to plain SF scaffold. The effect of nHA inorganic crystals on osteogenic differentiation of MC3T3-E1 cells indicated the MSF scaffolds improved osteogenesis. Furthermore, MSF grafts induced more bone formation and reduced the height of alveolar bone resorption after tooth extraction. SIGNIFICANCE: The MSF scaffold partially simulated the structure and composition of natural bone matrix. It induced osteogenic differentiation of MC3T3-E1 cells in vitro, and also promoted new bone regeneration in tooth extraction sockets in vivo, indicating it is a biomaterial with great potential for tooth extraction site preservation.