Autophagy regulates age-related delayed jawbone regeneration and decreased osteoblast osteogenesis by degrading FABP3.

The regenerative ability of limb bones after injury decreases during aging, but whether a similar phenomenon occurs in jawbones and whether autophagy plays a role in this process remain unclear. Through retrospective analysis of clinical data and studies on a mouse model of jawbone defects, we confirmed the presence of delayed or impaired bone regeneration in the jawbones of old individuals and mice. Subsequently, osteoblasts (OBs) derived from mouse jawbones were isolated, showing reduced osteogenesis in senescent osteoblasts (S-OBs). We observed a reduction in autophagy within both aged jawbones and S-OBs. Additionally, pharmacological inhibition of autophagy in normal OBs (N-OBs) led to cell aging and decreased osteogenesis, while autophagic activation reversed the aging phenotype of S-OBs. The activator rapamycin (RAPA) increased the autophagy level and bone regeneration in aged jawbones. Finally, we found that fatty acid-binding protein 3 (FABP3) was degraded by autolysosomes through its interaction with sequestosome 1 (P62/SQSTM1). Autophagy inhibition within senescent jawbones and S-OBs led to the excessive accumulation of FABP3, and FABP3 knockdown partially rescued the decreased osteogenesis in S-OBs and alleviated age-related compromised jawbone regeneration. In summary, we confirmed that autophagy inhibition plays an important role in delaying bone regeneration in aging jawbones. Autophagic activation or FABP3 knockdown can partially rescue the osteogenesis of S-OBs and the regeneration of aging jawbones, providing insight into jawbone aging.

Suppression of the NLRP3 Inflammasome through Activation of the Transient Receptor Potential Channel Melastatin 2 Promotes Osteogenesis in Tooth Extraction Sockets of Periodontitis.

This study explored the role of transient receptor potential channel melastatin 2 (TRPM2)-mediated activation of NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome in osteogenesis during healing of tooth extraction sockets. Tooth extraction socket tissue samples were collected from patients with or without periodontitis. In a TRPM2 knockout mouse model of socket healing, mice with or without periodontitis and their wild-type littermates were used for comparing the socket healing phenotypes. Micro-computed tomography imaging, three-dimensional reconstruction of the sockets, and hematoxylin and eosin staining for histopathologic analysis were performed. Immunofluorescence, immunohistochemistry, and Western blot analysis were used for evaluation of protein expression; the mRNA levels were evaluated by quantitative RT-PCR. Osteogenic, chondrogenic, and adipogenic differentiation potential of human bone marrow mesenchymal stem cells (BMMSCs) was evaluated. Calcium deposition was evaluated using Alizarin Red S staining. NLRP3 and CASP1 were up-regulated in tooth sockets of periodontitis patients. NLRP3 knockdown promoted the osteogenic differentiation of maxillary BMMSCs under inflammatory conditions. TRPM2 was up-regulated in the tooth extraction socket tissue of periodontitis. Inhibiting TRPM2 expression mitigated the NLRP3 inflammasome and its deleterious effect on osteogenesis. Activation of the TRPM2 ion channel regulated osteogenesis of BMMSCs under inflammatory conditions via Ca2+ influx, the mitochondrial dynamics, and pyroptosis. Targeting the TRPM2/Ca2+/NLRP3 axis could be beneficial in the healing process of the tooth extraction sockets of patients with periodontitis.

Genome wide analysis of dexamethasone stimulated mineralization in human dental pulp cells by RNA sequencing.

Human dental pulp cells (hDPCs) contain mesenchymal stem cells and are therefore indispensible for reparative dentin formation. Here, we present a pilot study of transcriptomic profiles of mineralized hDPCs isolated from sound human maxillary third molars. We observed altered gene expression of hDPCs between control (dexamethasone free) and experimental (dexamethasone 1 nm) groups. Differential expression analysis revealed up-regulation of several inflammation and mineralization-related genes in the experimental group. After a Gene Ontology analysis for predicting genes involved in biological process, cellular component and molecular function, we found enrichment of genes related to protein binding. Based on the results of Kyoto Encylopedia of Genes and Genomes pathway analysis, it is suggested up-regulated genes in mineralized hDPCs were mostly enriched in the mitogen-activated protein kinase (MAPK) signaling pathway, fluid shear stress and the atherosclerosis signaling pathway, etc. Importantly, Gene Set Enrichment Analysis revealed dexamethasone was positively related to the Janus kinase/signal transducer and activator of transcription, MAPK and Notch signaling pathway. Moreover, it was suggested that dexamethasone regulates signaling pathway in pluripotency of stem cells. Collectively, our work highlights transcriptome level gene regulation and intercellular interactions in mineralized hDPCs. The database produced in the present study paves the way for further investigations looking to explore genes that are involved in dental pulp cells mineralization.

Optimal therapy for concomitant EGFR and TP53 mutated non-small cell lung cancer: a real-world study.

BACKGROUND: Non-small cell cancer (NSCLC) patients with concomitant epidermal growth factor receptor (EGFR) and TP53 mutations have a poor prognosis with the treatment of tyrosine kinase inhibitors (TKIs), and may benefit from a combination regimen preferentially. The present study aims to compare the benefits of EGFR-TKIs and its combination with antiangiogenic drugs or chemotherapy in patients with NSCLC harboring EGFR and TP53 co-mutation in a real-life setting. METHODS: This retrospective analysis included 124 patients with advanced NSCLC having concomitant EGFR and TP53 mutations, who underwent next-generation sequencing prior to treatment. Patients were classified into the EGFR-TKI group and combination therapy group. The primary end point of this study was progression-free survival (PFS). The Kaplan-Meier (KM) curve was drawn to analyze PFS, and the differences between the groups were compared using the logarithmic rank test. Univariate and multivariate cox regression analysis was performed on the risk factors associated with survival. RESULTS: The combination group included 72 patients who received the regimen of EGFR-TKIs combined with antiangiogenic drugs or chemotherapy, while the EGFR-TKI monotherapy group included 52 patients treated with TKI only. The median PFS was significantly longer in the combination group than in the EGFR-TKI group (18.0 months; 95% confidence interval [CI]: 12.1-23.9 vs. 7.0 months; 95% CI: 6.1-7.9; p < 0.001) with greater PFS benefit in TP53 exon 4 or 7 mutations subgroup. Subgroup analysis showed a similar trend. The median duration of response was significantly longer in the combination group than in the EGFR-TKI group. Patients with 19 deletions or L858R mutations both achieved a significant PFS benefit with combination therapy versus EGFR-TKI alone. CONCLUSION: Combination therapy had a higher efficacy than EGFR-TKI alone for patients with NSCLC having concomitant EGFR and TP53 mutations. Future prospective clinical trials are needed to determine the role of combination therapy for this patient population.

Enhanced osteogenesis by addition of cancellous bone chips at xenogenic bone augmentation: In vitro and in vivo experiments.

OBJECTIVES: To investigate and compare the influence of deproteinized bovine bone mineral (DBBM) combined with autologous cortical (CorBC) or cancellous bone chips (CanBC) as bone grafts on guided bone regeneration (GBR) in vivo and in vitro. MATERIALS AND METHODS: Defects were created in the mandibular buccal alveolar ridges in dogs and randomly filled with 3 groups of bone grafts: DBBM, DBBM + CorBC, or DBBM + CanBC. Osteogenesis was evaluated by sequential fluorescent labeling and histological analysis. Moreover, rat bilateral calvaria defects were randomly grafted with DBBM, DBBM + CorBC, or DBBM + CanBC. A blank group was included as control. Defect healing was assessed by histological staining, micro-CT, and quantitative polymerase chain reaction. In vitro migration, proliferation, and osteogenic differentiation assays were performed by stimulating rat bone marrow mesenchymal stem cells (rBMSCs) with cortical (CorBCM) or cancellous bone conditioned medium (CanBCM) to unveil the cellular mechanism. RESULTS: In the canine model, the augmented sites of DBBM + CanBC exhibited higher mineralized tissue proportion than the other two groups (DBBM: 0.61 ± 0.03 versus DBBM + CorBC: 0.69 ± 0.07 versus DBBM + CanBC: 0.86 ± 0.06; p < .05). In the rat model, the BV/TV value of DBBM + CanBC (0.51 ± 0.01) was higher than those of DBBM + CorBC (0.41 ± 0.02), DBBM (0.31 ± 0.01), and Control (0.10 ± 0.01; p < .01). Further radiological, histological and transcriptional results showed similar trends. In vitro experiments revealed that CorBCM and especially CanBCM could enhance rBMSCs migration, proliferation, and osteogenic differentiation. CONCLUSION: In vivo and in vitro experiments verified favorable synergistic effect of mixing autologous bone chips with DBBM on osteogenesis. Furthermore, CanBC presented more powerful osteogenic effect than CorBC.

LAMA5-inspired adhesive dodecapeptide facilitates efficient dentine regeneration: An in vitro and in vivo study.

AIM: The primary goal of this study was to investigate the potential effects of A5G81 in inducing reparative dentine (RD) formation both in vitro and in vivo. METHODOLOGY: Cell adhesion was observed by crystal violet staining and quantified by Sodium Dodecyl Sulphate (SDS) extraction. Cell proliferation was investigated using Cell Counting Kit-8 (CCK-8) assay. Spreading of cytoskeleton was visualized using immunofluorescence staining. Protein expression level of Akt signalling pathway was compared in a human Akt pathway phosphorylation array. Genes that were up or downregulated by A5G81 were identified by RNA sequencing. The mRNA expression of odontoblastic markers was detected by quantitative real-time polymerase chain reaction (qPCR). Moreover, mineralization of human dental pulp cells (hDPCs) was visualized by alizarin red staining and quantified using cetylpyridinium chloride (CPC). A direct pulp-capping model was established in SD rats and the RD formation at 2 weeks after operation was observed using HE staining. RESULTS: A5G81 (optimal coating concentration: 0.5 mg/mL) promoted hDPCs adhesion and proliferation to a level that was similar to Type I collagen (COL-1). Meanwhile, A5G81 activated Akt signalling pathway, albeit to a lesser extent than COL-1. An inhibition test indicated that A5G81 induced hDPCs adhesion by activating PI3K pathway. A5G81 induced the expression of ECM remodelling genes and odontoblastic genes, which were demonstrated by RNA-seq and qPCR, respectively. In addition, A5G81 efficiently accelerated the mineralization of hDPCs in both immobilized and soluble forms, a property that makes it more applicable in dental clinic. Finally, the pulp-capping study in rats suggested that use of A5G81 could successfully induce the formation of RD within 2 weeks. CONCLUSION: Coating of A5G81 to non-tissue culture-treated polystyrene facilitates spreading, proliferation and differentiation of hDPCs, resulting in rapid RD formation in artificially exposed pulp.

Triggering receptor expressed on myeloid cells-2 stimulates osteoclast differentiation and bone loss in periodontitis.

OBJECTIVE: To investigate the expression of triggering receptor expressed on myeloid cells 2 (TREM-2) in the healthy and diseased tissue, including gingivitis or periodontitis, and then to assess whether it has an impact on the development of periodontitis. METHODS AND MATERIALS: The gingival tissues from healthy controls, gingivitis, and periodontitis underwent hematoxylin-eosin and immunohistochemical staining, and the association of TREM-2 expression or TREM-2+ cell counts with clinical parameters was assessed. An anti-TREM-2 antibody was used to block the osteoclastogenesis in vitro and during the experimental periodontitis by injection into the gingiva. The relative gene expression of TREM-2 in different gingival tissues was analyzed by quantitative PCR. RESULTS: In the gingival tissues of periodontitis, TREM-2 expression and TREM-2+ cell counts were significantly higher than those of gingivitis and healthy controls (p<0.05). In the group of periodontitis showing moderate signs, the gingival tissues displayed significantly lower TREM-2 expression, in contrast with the group with advanced periodontal symptoms (p < 0.05). Consistently, blocking TREM-2 significantly decreased osteoclast formation both in vitro and in vivo (p < 0.05). CONCLUSION: Increased TREM-2 expression and TREM-2+ cells were positively associated with the development of periodontitis. Osteoclast differentiation and stimulating alveolar bone loss were partly relied on TREM-2, which could be a target to be blocked for attenuating osteoclastogenesis in periodontitits.

Parenchymal and stromal tissue regeneration of tooth organ by pivotal signals reinstated in decellularized matrix.

Cells are transplanted to regenerate an organs' parenchyma, but how transplanted parenchymal cells induce stromal regeneration is elusive. Despite the common use of a decellularized matrix, little is known as to the pivotal signals that must be restored for tissue or organ regeneration. We report that Alx3, a developmentally important gene, orchestrated adult parenchymal and stromal regeneration by directly transactivating Wnt3a and vascular endothelial growth factor. In contrast to the modest parenchyma formed by native adult progenitors, Alx3-restored cells in decellularized scaffolds not only produced vascularized stroma that involved vascular endothelial growth factor signalling, but also parenchymal dentin via the Wnt/ß-catenin pathway. In an orthotopic large-animal model following parenchyma and stroma ablation, Wnt3a-recruited endogenous cells regenerated neurovascular stroma and differentiated into parenchymal odontoblast-like cells that extended the processes into newly formed dentin with a structure-mechanical equivalency to native dentin. Thus, the Alx3-Wnt3a axis enables postnatal progenitors with a modest innate regenerative capacity to regenerate adult tissues. Depleted signals in the decellularized matrix may be reinstated by a developmentally pivotal gene or corresponding protein.

Glycosylation of DMP1 maintains cranial sutures in mice.

Craniosynostosis, a severe craniofacial developmental disease, can only be treated with surgery currently. Recent studies have shown that proteoglycans are involved in the suture development. For the bone matrix protein, dentin matrix protein 1 (DMP1), glycosylation on the N-terminal of it could generate a functional proteoglycan form of DMP1 during osteogenesis. We identified that the proteoglycan form of DMP1 (DMP1-PG) is highly expressed in mineralisation front of suture. But, the potential role of DMP1-PG in suture fusion remain unclear. To investigate the role of DMP1-PG in cranial suture fusion and craniofacial bone development. By using a DMP1 glycosylation site mutation mouse model, DMP1-S89G mice, we compared the suture development in it with control mice. We compared the suture phenotypes, bone formation rate, expression levels of bone formation markers in vivo between DMP1-S89G mice and wild-type mice. Meanwhile, cell culture and organ culture were performed to detect the differences in cell differentiation and suture fusion in vitro. Finally, chondroitin sulphate (CHS), as functional component of DMP1-PG, was employed to test whether it could delay the premature suture fusion and the abnormal differentiation of bone mesenchymal stem cells (BMSCs) of DMP1-PG mice. DMP1-S89G mice had premature closure of suture and shorter skull size. Lack of DMP1-PG accelerated bone formation in cranial suture. DMP1-PG maintained the essential stemness of BMSCs in suture through blocking the premature differentiation of BMSCs to osteoblasts. Finally, chondroitin sulphate, a major component of DMP1-PG, successfully delayed the premature suture fusion by organ culture of skull in vitro. DMP1-PG could inhibit premature fusion of cranial suture and maintain the suture through regulating the osteogenic differentiation of BMSCs.

Rat sinus mucosa- and periosteum-derived exosomes accelerate osteogenesis.

Guided bone regeneration (GBR) is commonly used for alveolar bone augmentation. The paracrine mechanism in the field of bone tissue engineering has been emphasized in recent years and exosomes are considered to have the potential of promoting osteogenesis. We aimed to study the influence of sinus mucosa and periosteum on bone regeneration through paracrine stimulation, especially via exosomes, and compare the differences between them. Here, we report that conditioned medium (CM) from sinus mucosa-derived cells (SMCs) and periosteum-derived cells (PCs) and the isolated exosomes enhanced the proliferation, migration and osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs) in vitro. A rat model of femoral bone defects was used to demonstrate that the exosomes derived from SMCs (SMC-Exos) and PCs (PC-Exos) can accelerate bone formation in vivo. Furthermore, we present a preliminary discussion of the possible functional components involved in the effects of SMC-Exos and PC-Exos on bone regeneration. In conclusion, these results demonstrated that the sinus mucosa and periosteum can accelerate osteogenesis through paracrine effects and the exosomes play important roles in this process.

Adrenaline inhibits osteogenesis via repressing miR-21 expression.

Sympathetic signaling is involved in bone homeostasis; however, the cellular and molecular mechanisms remain unknown. In this study, we found that the psychological stress mediator adrenaline inhibited osteogenic differentiation of human bone marrow-derived stem cells (hMSC) by reducing microRNA-21 (miR-21) expression. Briefly, adrenaline significantly inhibited the osteogenic differentiation of hMSCs, as observed with both Alizarin red staining and maker gene expression (RUNX2, OSX, OCN, and OPN). During this process, miR-21 was suppressed by adrenaline via inhibition of histone acetylation, as verified by H3K9Ac chromatin immunoprecipitation (ChIP) assay. MiR-21 was confirmed to promote hMSC osteogenic differentiation, and overexpression of miR-21 reversed the impeditive effect of adrenaline on hMSC osteogenic differentiation. Our results demonstrate that down-regulation of miR-21 is responsible for the adrenaline-mediated inhibition of hMSC osteogenic differentiation. These findings indicate a regulation of bone metabolism by psychological stress and also provide a molecular basis for psychological stress-associated bone diseases.

A comparative study of the effect of Bio-Oss® in combination with concentrated growth factors or bone marrow-derived mesenchymal stem cells in canine sinus grafting.

BACKGROUND: To compare the effects of Bio-Oss® in combination with concentrated growth factors (CGFs) and bone marrow-derived mesenchymal stem cells (BMSCs) on bone regeneration for maxillary sinus floor augmentation in beagle dogs. METHODS: Six beagle dogs received bilateral maxillary sinus floor augmentation. Venous blood drawn from dogs was collected and centrifuged to obtain CGFs. BMSCs derived from canine bone marrow were cultured using density gradient centrifugation. The suspension of BMSCs was added onto Bio-Oss® granules at a density of 2 × 106 cells/ml, and the BMSCs/Bio-Oss® constructs were incubated for an additional 4 h before use. Twelve sinuses were grafted with a mixture of CGFs/Bio-Oss® , BMSCs/Bio-Oss® construct, or Bio-Oss® alone. Six months later, the bone formation of bilateral sinuses was evaluated by Micro-CT, microhardness test, histological examination, and histomorphometry. RESULTS: No adverse effect was found in these dogs. The dome-shaped augmentation protruded into the sinus cavity. Micro-CT revealed that there was significant difference in BV/TV but not in Tb. N, between groups A, B, and C. The extent of microhardness in groups A and B was significantly higher than in group C. The proportion of newly formed bone in groups A and B showed significant difference when compared to group C (P ≤ 0.01). The amount of residual grafts in groups A and B was significantly lower than in group C. CONCLUSIONS: Grafting with Bio-Oss® in combination with CGFs can increase new bone formation more efficiently than using Bio-Oss® alone in a canine model.

S100A4 upregulation suppresses tissue ossification and enhances matrix degradation in experimental periodontitis models.

AIM: S100A4, also known as fibroblast-specific protein 1 or metastasin 1, is not only highly expressed in growth-stimulated cultured cells and metastatic tumor cells, but also in the periodontal ligament. The aim of this study was to investigate the roles of S100A4 in the pathogenesis of periodontitis and its regulatory mechanisms in inflammatory milieu. METHODS: Experimental periodontitis was induced in rats by submarginal silk ligatures. TRAP activity and S100A4 expression in periodontal ligaments were examined using immunohistochemistry and immunofluorescence methods. IL-1ß-treated human periodontal ligament cells (hPDLCs) were used as in vitro model of experimental periodontitis. S100A4 mRNA and protein were assessed using qRT-PCR and Western blot, respectively. hPDLCs were transfected with either S100A4 overexpression plasmids or shRNAs plasmids. The mineralization in hPDLCs was evaluated with a 12-d osteogenic induction assay, and the expression of ALP, OCN, MMP-2 and MMP-13 was analyzed by qRT-PCR. RESULTS: In the periodontal ligaments of rats with experimental periodontitis, TRAP activity and S100A4 protein staining were considerably more intense compared with those in the control rats. Treatment of hPDLCs with IL-1ß (10, 50 and 100 ng/mL) dose-dependently increased the mRNA and protein levels of S100A4. Transfection with shRNAs markedly increased mineralized nodule formation and the osteogenic-related markers ALP and OCN levels in hPDLCs, whereas the overexpression of S100A4 significantly reduced mineralized nodule formation, and increased the matrix degradation enzymes MMP-2 and MMP-13 levels in hPDLCs. CONCLUSION: S100A4 is upregulated in the experimental rat periodontitis and in IL-1ß-treated hPDLCs, where S100A4 suppresses osteogenic differentiation and enhances matrix degradation. Thus, S100A4 is a potential target for the treatment of periodontitis.

Comparison of tissue-engineered bone from different stem cell sources for maxillary sinus floor augmentation: a study in a canine model.

PURPOSE: To compare the potential of tissue-engineered bone derived from different stem cell sources for canine maxillary sinus augmentation. MATERIALS AND METHODS: Bilateral maxillary sinus floor augmentations were performed in 6 beagles and were randomly repaired with 3 graft types: Bio-Oss granules alone (n = 4; group A), a complex of osteoblasts derived from bone marrow mesenchymal stem cells (BMMSCs) and Bio-Oss (n = 4; group B), and a complex of osteoblasts derived from periodontal ligament stem cells (PDLSCs) and Bio-Oss (n = 4; group C). After 12 weeks, fluorescent labeling, maxillofacial computed tomography, scanning electron microscopy, and histologic and histomorphometric analyses were used to evaluate new bone deposition, mineralization, and remodeling in the augmented area. RESULTS: The osteogenic capacity was greater in groups B and C than in group A. The level tended to be higher in group C than in group B; however, the difference was not statistically significant. CONCLUSIONS: Seeding of PDLSCs or BMMSCs onto Bio-Oss can promote bone formation and mineralization and maintain the maximum volume of the augmented maxillary sinus. These tissue-engineered bone complexes might be a good option for augmentation of the maxillary sinus in edentulous patients.

The detection of Gper1 as an important gene promoting jawbone regeneration in the context of estrogen deficiency.

Numerous studies have demonstrated that estrogen deficiency inhibit the proliferation and differentiation of pre-osteoblasts in skeleton by affecting osteogenic signaling, lead to decreased bone mass and impaired regeneration. To explore the mechanisms maintaining bone regeneration under estrogen deficiency, we randomly selected 1102 clinical cases, in which female patients aged between 18 and 75 have underwent tooth extraction in Stomatological Hospital of Tongji University, there is little difference in the healing effect of extraction defects, suggesting that to some extent, the regeneration of jawbone is insensitive to the decreased estrogen level. To illuminate the mechanisms promoting jawbone regeneration under estrogen deficiency, a tooth extraction defect model was established in the maxilla of female rats who underwent ovariectomy (OVX) or sham surgery, and jawbone marrow stromal cells (BMSCs) were isolated for single-cell sequencing. Further quantitative PCR, RNA interference, alizarin red staining, immunohistochemistry and western blotting experiments demonstrated that in the context of ovariectomy, maxillary defects promoted G protein-coupled estrogen receptor 1 (Gper1) expression, stimulate downstream cAMP/PKA/pCREB signaling, and facilitate cell proliferation, and thus provided sufficient progenitors for osteogenesis and enhanced the regeneration capacity of the jawbone. Correspondingly, the heterozygous deletion of the Gper1 gene attenuated the phosphorylation of CREB, led to decreased cell proliferation, and impaired the restoration of maxillary defects. This study demonstrates the importance of Gper1 in maintaining jawbone regeneration, especially in the context of estrogen deficiency.

Paired-Related Homeobox 1-Positive Cells Are Needed for Osseointegration.

PURPOSE: To explore the contribution of paired-related homeobox 1-positive cells to the implant-induced osseointegration process in adult alveolar bone and the potential underlying mechanisms. MATERIALS AND METHODS: Cre recombinase-induced lineage tracing and cell ablation were conducted in a murine dental implant model. Scratch and transwell assays were used to assess MC3T3-E1 cell migration after paired-related homeobox 1 overexpression. Single-cell RNA sequencing were applied to identify potential genes involved in pairedrelated homeobox 1-positive cells-driven osteogenesis. RESULTS: Paired-related homeobox 1- positive cells were observed to accumulate in the peri-implant area in a time-dependent manner. The number of these cells were found to reach its maximum on day 14. Osseointegration in mice were noticeably impaired after ablation of paired-related homeobox 1-positive cells. Further, it was discovered that paired-related homeobox 1 promotes MC3T3- E1 cell migration, a process which is indispensable for sound healing of peri-implant tissue. Finally, Semaphorin 3C was detected exclusively and abundantly expressed by paired-related homeobox 1-positive cells. Knockdown of semaphorin 3C in paired-related homeobox 1- positive cells significantly weakened their osteogenic potential. CONCLUSION: Our data suggest that paired-related homeobox 1-positive cells contribute to the osseointegration process under stress stimulation and semaphorin 3C may play a critical role in paired-related homeobox 1- positive cell-driven osteogenesis. Paired-related homeobox 1 could significantly promote MC3T3-E1 cell migration.

Revealing the solid solution tendency of tungsten ions in phases of Ordinary Portland cement clinker: A study based on experiments and DFT calculations.

The collaborative utilization of solid waste through cement kiln represents a highly effective approach in the current era for harnessing solid waste resources. In this paper, density functional theory simulations is used to predict the substitution tendency of tungsten (W) in Ordinary Portland cement (OPC) clinker. By employing experimental design, X-ray diffraction testing, and element distribution spectrum analysis, the doping preference of W ions in OPC clinker was comprehensively investigated. The findings demonstrate that a minor fraction of WO3 firstly infiltrates C4AF through the substitution of Fe atoms, whereas the majority of WO3 infiltrates C3S and C2S secondly by substituting Si atoms, with negligible infiltration observed in C3A finally. The substitution of Fe with W exhibits a lower formation energy compared to other ions, thereby indicating its preference for the formation of solid solutions in C4AF. This preference is primarily determined by the overlapping distribution of WO and FeO bond order-bond length and their similar electron contributions in spatial distribution. However, it should be noted that the newly formed WO bond has weaker strength than the FeO bond, which may explain the limited solubility of W in C4AF. The in-depth investigation of these fundamental issues is expected to offer an effective approach for enhancing solubility of W in OPC clinker through increasing content of C4AF and silicate minerals, thereby providing valuable guidance for synthesizing OPC clinker using W-bearing solid wastes.

Study on activation of fluorogypsum by sodium sulfate and sodium nitrite.

Given the issues related to poor hydration activity, long setting time and low early strength of industrial by-product fluorogypsum (FG), the composite modifiers (Na2SO4 and NaNO2) were utilized to enhance its reactivity. The investigation of the mechanism involved the utilization of contemporary analytical methods, including X-ray diffraction (XRD), 1H low-field nuclear magnetic resonance (NMR), and Scanning electron microscope and Energy Dispersive Spectrometer (SEM-EDS). The results demonstrated that the incorporation of modifiers significantly enhanced both the hydration rate and activity of fluorogypsum. The optimum concentration of the composite modifier was found to be 1.5 wt% Na2SO4 and 0.5 wt% NaNO2. The addition of modifiers (1.5 wt% Na2SO4 and 0.5 wt% NaNO2) significantly shortens the setting time of FG paste, reducing it by approximately 500 min compared to the control sample. After 28 days of curing, the flexural strength and compressive strength of the fluorogypsum sample containing modifiers (1.5 wt% Na2SO4 and 0.5 wt% NaNO2) increased by 55.5 % (reaching 4.2 MPa) and 31.5 % (reaching 37.6 MPa), respectively. The modifiers facilitate the transformation from anhydrite (CaSO4, AH) to dihydrate gypsum (CaSO4·2H2O, DH). Both NaNO2 and Na2SO4 alter the growth rates of different crystal axes during DH crystal growth, transforming them into prismatic and needle-shaped DH. The prismatic and needle-shaped DH crystals were arranged in layers, resulting in a compact structure with low hole content and few pores, which led to increased density of the hardened paste and higher strength. The current study provides evidence that the inclusion of composite modifiers greatly improves the activity of FG, making it more efficient in the field of building materials.

Leptin receptor (+) stromal cells respond to periodontitis and attenuate alveolar bone repair via CCRL2-mediated Wnt inhibition.

The impaired bone healing in tooth extraction sockets due to periodontitis presents a major obstacle to restoring oral health. The mechanisms regulating the osteogenic capacity of jawbone-derived stromal cells in the periodontitis microenvironment remain elusive. Leptin receptor (LepR) expressing stromal cells, which largely overlap with Cxcl12-abundant reticular (CAR) cells in bone tissue, rapidly proliferate and differentiate into bone-forming cells during extraction socket healing to support alveolar bone repair. In this study, we identify that CCRL2 is significantly expressed and inhibits osteogenesis in LepR+/CAR cells of alveolar bones with periodontitis. The Ccrl2-KO mice exhibit significant improvements in bone healing in extraction sockets with periodontitis. Specifically, the binding of CCRL2 to SFRP1 on the surface of LepR+/CAR cells can amplify the suppressive effect of SFRP1 on Wnt signaling under inflammation, thus hindering the osteogenic differentiation of LepR+/CAR cells and resulting in poor bone healing in extraction sockets with periodontitis. Together, we clarify that the CCRL2 receptor of LepR+/CAR cells can respond to periodontitis and crosstalk with Wnt signaling to deteriorate extraction socket healing.

The impaired bone healing in tooth extraction sockets due to periodontitis presents a major obstacle to restoring oral health. Alterations in the cellular activity of LepR+/CAR cells, an essential stromal cell population for extraction socket healing, in the periodontitis microenvironment have yet to be determined. In this study, we identify that CCRL2, as a potent agent of inflammation-bone crosstalk, is significantly expressed and inhibits osteogenesis in LepR+/CAR cells of alveolar bones with periodontitis. Specifically, the binding of CCRL2 to SFRP1 on the surface of LepR+/CAR cells can amplify the suppressive effect of SFRP1 on the Wnt/ß-catenin signaling under inflammation, thus hindering the osteogenic differentiation of LepR+/CAR cells and resulting in poor bone healing in tooth extraction sockets with periodontitis.

Effects of Implant Diameter on Implant Stability and Osseointegration in the Early Stage in a Dog Model.

Purpose: To determine the optimal implant diameter under limited bone width by comparing the effects of implants with different diameters on implant stability, peri-implant bone stability, and osseointegration. In addition, to evaluate the reliability of resonance frequency analysis (RFA) in detecting osseointegration and marginal bone level (MBL). Materials and Methods: Mandibular premolars and first molars of seven beagle dogs were extracted. After 8 weeks, their mandibular models and radiographic information were collected to fabricate implant templates. Implant sites were randomly divided into three groups according to diameter: Ø3.3, Ø4.1, and Ø4.8 mm. Implant stability quotient (ISQ) measurement and radiographic evaluation were performed after surgery (baseline) and at 4, 8, and 12 weeks. Three dogs were euthanized at 4 weeks to observe osteogenesis and implant-tissue interface biology. Four dogs were euthanized at 12 weeks to observe osseointegration. Hard tissue sections were prepared to analyze osteogenesis (fluorescence double labeling) and osseointegration (methylene blue-acid fuchsin staining). Results: At baseline and at 4, 8, and 12 weeks, the ISQ values of Ø4.1- and Ø4.8-mm implants did not differ (P > .05), but both had higher values than the Ø3.3-mm implants (P < .05). The mean marginal bone resorption (MBR) associated with Ø3.3-, Ø4.1-, and Ø4.8-mm implants was 0.65 ± 0.58 mm, 0.37 ± 0.28 mm, and 0.73 ± 0.37 mm, respectively. The buccal MBR of Ø4.8-mm implants was significantly higher than that of Ø4.1-mm implants (P < .05). The bone-to-implant contact (BIC) percentage at 12 weeks did not differ for any group (P > .05). The correlation coefficients between the ISQ and MBL of the Ø3.3-, Ø4.1-, and Ø4.8-mm implants were -0.84 (P < .01), -0.90 (P < .001), and -0.93 (P < .001), respectively, while that between the ISQ and BIC was 0.15 (P > .05). Conclusions: During the early healing stage, the performance of Ø4.1- and Ø4.8-mm implants in terms of implant stability was better than that of Ø3.3-mm implants. Implant diameter may not influence BIC percentage. RFA can be used to evaluate implant stability and MBL but is not suitable to assess the degree of osseointegration.