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.

Electromagnetic interference effect of dental equipment on cardiac implantable electrical devices: A systematic review.

BACKGROUND: The electromagnet interference (EMI) effect resulting from using dental equipment near cardiovascular implantable electronic devices (CIEDs) is controversial based on in vitro and in vivo studies. We aimed to summarize the available evidence to investigate the safety of using dental equipment on patients with CIEDs. METHODS: An electronic search was performed in PubMed, Embase, MEDLINE Ovid, and the Cochrane Library for relevant studies published between January 2000 and May 2020. The search strategy centered on terms related to dental devices and CIEDs. Two independent reviewers determined the final inclusion of the studies in the systematic review. The EMI effect was summarized based on different dental instruments detected in in vitro or in vivo studies. RESULTS: The primary search identified 84 articles, and 18 studies were finally included in this systematic review after exclusions. Most in vitro studies (n = 12) reported background noise or severe EMI affecting CIED function at a close distance from the lead tip or at a high sensitivity setting of CIEDs. In in vivo studies (n = 6), EMI that altered CIED function was not detected at clinical distance and sensitivity settings. The summary, based on electronic apex locators, ultrasonic devices, and electric pulp testers, demonstrated the compatibility of these common dental devices with CIEDs. CONCLUSIONS: This systematic review indicates that most dental instruments can be used safely in routine dental practice. The EMI effect of dental equipment depends on the exposure distance and lead-related parameters of the CIEDs.

From Bone Remodeling to Wound Healing: An miR-146a-5p-Loaded Nanocarrier Targets Endothelial Cells to Promote Angiogenesis.

Wound healing is a complex challenge that demands urgent attention in the clinical realm. Efficient angiogenesis is a pivotal factor in promoting wound healing. microRNA-146a (miR-146a) inhibitor has angiogenic potential in the periodontal ligament. However, free microRNAs (miRNAs) are poorly delivered into cells due to their limited tissue specificity and low intracellular delivery efficiency. To address this hurdle, we developed a nanocarrier for targeted delivery of the miR-146a inhibitor into endothelial cells. It is composed of a polyethylenimine (PEI)-modified mesoporous silica nanoparticle (MSN) core and a pentapeptide (YIGSR) layer that recognizes endothelial cells. In vitro, we defined that the miR-146a inhibitor and adiponectin (ADP) can modulate angiogenesis and the remodeling of periodontal tissues by activating the ERK and Akt signaling pathways. Then, we confirm the specificity of YIGSR to endothelial cells, and importantly, the nanocarrier effectively delivers the miR-146a inhibitor into endothelial cells, promoting angiogenesis. In a C57 mouse skin wound model, the miR-146a inhibitor is successfully delivered into endothelial cells at the wound site using the nanocarrier, resulting in the formation of new blood vessels with strong CD31 expression. Additionally, no significant differences are found in the expression levels of inflammatory markers interleukin-6 and tumor necrosis factor-α. This outcome not only brings new strategies for angiogenesis but also exhibits broader implications for bone remodeling and wound healing. The breakthrough holds significance for future research and clinical interventions.

The role of WTAP in regulating macrophage-mediated osteoimmune responses and tissue regeneration in periodontitis.

Periodontitis, delineated by the destruction of structures that support teeth, is predominantly propelled by intricate immune responses. Immunomodulatory treatments offer considerable promise for the management of this ailment; however, the modulation of the periodontal immune microenvironment to facilitate tissue regeneration presents a substantial biomedical challenge. Herein, our study investigates the role of Wilms' tumor 1-associating protein (WTAP), a critical m6A methyltransferase, in the immunomodulation of periodontitis and assesses its viability as a therapeutic target. We observed heightened expression of WTAP in macrophages extracted from gingival tissues impacted by periodontitis, with a strong association with M1 polarization. Via loss-of-function experiments, we demonstrated that diminishing WTAP expression precipitates a transition from M1 to M2 macrophage phenotypes amidst inflammatory conditions, thus improving the periodontal immune landscape. Further, RNA sequencing and indirect co-culture assays indicated that suppressing of WTAP expression modulates osteoimmune responses and enhances the osteogenic differentiation of bone marrow stromal cells. The local deployment of adeno-associated virus-shWTAP in murine models of periodontitis robustly validated the therapeutic promise of targeting WTAP in this disease. Collectively, our findings highlight the crucial role of WTAP in orchestrating macrophage-mediated osteoimmune responses and tissue regeneration in periodontitis, proposing novel avenues for immunotherapeutic interventions in its treatment.

Young Exosome Bio-Nanoparticles Restore Aging-Impaired Tendon Stem/Progenitor Cell Function and Reparative Capacity.

Aging impairs tendon stem/progenitor cell function and tendon homeostasis, however, effective treatments for aging-induced tendon diseases are lacking. Exosomes are naturally derived nanoparticles that contain bioactive molecules, and therefore, have attracted great interest in tissue engineering and regenerative medicine. In this study, it is shown that young exosomes secreted by stem cells from human exfoliated deciduous teeth (SHED-Exos) possess abundant anti-aging signals. These young bio-nanoparticles can alleviate the aging phenotypes of aged tendon stem/progenitor cells (AT-SCs) and maintain their tenogenic capacity. Mechanistically, SHED-Exos modulate histone methylation and inhibit nuclear factor-κB to reverse AT-SC aging. In a naturally aging mouse model, systemic administration of SHED-Exo bio-nanoparticles retards tendon degeneration. Interestingly, local delivery of SHED-Exos-loaded microspheres confers anti-aging phenotypes, including reduced senescent cells and decreased ectopic bone formation, thereby functionally and structurally rescuing endogenous tendon regeneration and repair capacity in aged rats. Overall, SHED-Exos, as natural bioactive nanoparticles, have promising translational and therapeutic potential for aging-related diseases.

Induction of Cartilage Regeneration by Nanoparticles Loaded with Dentin Matrix Extracted Proteins.

Due to the limited self-repair capacity of articular cartilage, tissue engineering has good application prospects for cartilage regeneration. Dentin contains several key growth factors involved in cartilage regeneration. However, it remains unknown whether dentin matrix extracted proteins (DMEP) can be utilized as a complex growth factor mixture to induce cartilage regeneration. In this work, we extracted DMEP from human dentin and improved the content and activity of chondrogenic-related growth factors in DMEP by alkaline conditioning. Afterward, mesoporous silica nanoparticles (MSNs) with particular physical and chemical properties were composed to selectively load and sustain the release of proteins in DMEP. MSN-DMEP promoted chondrogenic differentiation of rat bone marrow-derived mesenchymal stem cells with fewer growth factors than exogenously added transforming growth factor-ß1 (TGF-ß1). Therefore, MSN-DMEP may serve as a promising candidate for cartilage regeneration as an alternative to expensive synthetic growth factors. Impact statement Several growth factors embedded in dentin matrix could be involved in cartilage regeneration. This article reports that alkaline conditioning could improve the content and activity of chondrogenic-related growth factors in dentin matrix extracted proteins (DMEP). Mesoporous silica nanoparticles (MSNs) with particular physical and chemical properties performed well in loading and sustained releasing of proteins in DMEP. In vitro and in vivo studies suggest that MSN-DMEP could be a promising candidate for cartilage regeneration as an alternative to expensive synthetic growth factors.

Assessment of microbiota diversity in dental unit waterline contamination.

BACKGROUND: Dental unit waterlines (DUWLs) provide water for handpieces, air/water syringes, and mouth-rinse water outlets. DUWL contamination can negatively affect the operating environment and public health. Therefore, it is important to elucidate the bacterial concentrations and microbial composition in the DUWLs from different dental specialties. METHODS: We collected 350 5-mL dental water samples (from high-speed handpieces, air/water syringes, and mouth-rinse water outlets) from 60 dental chair units (DCUs) at a dental hospital to determine the bacterial concentrations by culture methods. Meanwhile, to investigate the diversity and community structure of microbe in the DUWLs, 17 high-quality DNA from 60 250-mL air/water syringe water samples, which were collected from the same 60 DCUs, were analyzed using 16S rDNA high-throughput sequencing. RESULTS: The median bacterial concentration was 166 (31.5, 672.5) CFU/mL and the range was 0-3,816,000 CFU/mL. Only 42.6% of the water samples had bacterial concentrations below 100 CFU/mL. The Kruskal-Wallis H-test revealed that the water samples from three dental specialties had significantly different bacterial concentrations (H = 27.441, P < 0.01). High-throughput sequencing results showed significant differences in bacterial community structure between periodontics and the other two dental specialties. In the samples from three dental specialties, 508 OTUs were detected, with 160, 182 and 176 OTUs unique to the periodontics, endodontics and prosthodontics specialties, respectively. Linear discriminant analysis (LDA) effect size (LEfSe) suggested that Hydrocarboniphaga, Zoogloea, Aquabacterium, and Hydrogenophaga were enriched in the periodontics specialty; Acinetobacter, Geothrix, and Desulfovibrio were enriched in the prosthodontics specialty; and Alistipes, Clostridium XIVa, and Serratia were enriched in the endodontics specialty. Seven potentially human-pathogenic genera (Pseudomonas, Acinetobacter, Sphingomonas, Ochrobactrum, Rhizobium, Brevundimonas, and Methylobacterium) with relative abundance exceeding 1% were also detected in the DUWLs. CONCLUSIONS: The bacterial concentrations and microbial composition were influenced by different dental specialties, so a validated disinfection protocol should be used to control DUWL contamination in different dental specialties.

Electrophoretic deposition of novel semi-permeable coatings on 3D-printed Ti-Nb alloy meshes for guided alveolar bone regeneration.

OBJECTIVE: Guided bone regeneration (GBR) techniques use barrier membranes to augment the alveolar ridge for the site-specific growth of bone defects. However, current approaches using cast metal substructures exhibit poor adaptation to the surgical site and increased risk of infection. This study aimed to fabricate multi-functional coatings with 3D-printed porous titanium-niobium (Ti-Nb) alloy meshes to maintain space, prevent the ingrowth of fibroblasts and inhibit the colonization of bacteria for GBR. METHODS: Ti-Nb alloy meshes were prepared by selective laser melting (SLM) and used as substrates for novel surface coatings. Porous chitosan (CS)/ gelatin (G)/ doxycycline (Dox) coatings were formed on the meshes using electrophoretic deposition (EPD) and freeze-drying. The process of EPD was characterized through Fourier transform infrared spectroscopy (FT-IR), zeta potential, and particle size analysis. The cytotoxicity of the coatings was evaluated through the culture of osteoblasts and immunostaining. The antibacterial activity of the coatings was tested using inhibition zone tests against Staphylococcus aureus (S. aureus) and scanning electron microscope (SEM). The inhibition of fibroblasts infiltration and nutrients transfer properties were analyzed using immunostaining and permeability tests. RESULTS: High yield strength (567.5 ± 3.5 MPa) and low elastic modulus (65.5 ± 0.2 GPa) were achieved in Ti-Nb alloy bulk samples. The data of zeta potential, FT-IR and SEM indicated that porous spongy coatings were chemically bonded following EPD. In vitro analysis of CSGDox1 (containing Dox at 1 mg·mL-1) coating revealed its antibacterial effect and biocompatibility. Moreover, the CSGDox1 coating was proved to be effective for preventing the ingrowth of fibroblasts, whilst allowing the infiltration of nutrients. SIGNIFICANCE: This study verified that the EPD of CSGDox coatings on the 3D-printed Ti-Nb meshes can maintain space, provide antibiotic release whilst maintaining a barrier against soft-tissue growth, which is essential for the success of GBR treatment.

Alkaline activation of endogenous latent TGFß1 by an injectable hydrogel directs cell homing for in situ complex tissue regeneration.

Utilization of the body's regenerative potential for tissue repair is known as in situ tissue regeneration. However, the use of exogenous growth factors requires delicate control of the dose and delivery strategies and may be accompanied by safety, efficacy and cost concerns. In this study, we developed, for the first time, a biomaterial-based strategy to activate endogenous transforming growth factor beta 1 (TGFß1) under alkaline conditions for effective in situ tissue regeneration. We demonstrated that alkaline-activated TGFß1 from blood serum, bone marrow fluids and soaking solutions of meniscus and tooth dentin was capable of increasing cell recruitment and early differentiation, implying its broad practicability. Furthermore, we engineered an injectable hydrogel (MS-Gel) consisting of gelatin microspheres for loading strong alkaline substances and a modified gelatin matrix for hydrogel click crosslinking. In vitro models showed that alkaline MS-Gel controllably and sustainably activated endogenous TGFß1 from tooth dentin for robust bone marrow stem cell migration. More importantly, infusion of in vivo porcine prepared root canals with alkaline MS-Gel promoted significant pulp-dentin regeneration with neurovascular stroma and mineralized tissue by endogenous proliferative cells. Therefore, this work offers a new bench-to-beside translation strategy using biomaterial-activated endogenous biomolecules to achieve in situ tissue regeneration without the need for cell or protein delivery.

A simple layer-stacking technique to generate biomolecular and mechanical gradients in photocrosslinkable hydrogels.

Physicochemical and biological gradients are desirable features for hydrogels to enhance their relevance to biological environments for three-dimensional (3D) cell culture. Therefore, simple and efficient techniques to generate chemical, physical and biological gradients within hydrogels are highly desirable. This work demonstrates a technique to generate biomolecular and mechanical gradients in photocrosslinkable hydrogels by stacking and crosslinking prehydrogel solution in a layer by layer manner. Partial crosslinking of the hydrogel allows mixing of prehydrogel solution with the previous hydrogel layer, which makes a smooth gradient profile, rather than discrete layers. This technique enables the generation of concentration gradients of bovine serum albumin in both gelatin methacryloyl (GelMA) and poly(ethylene glycol) diacrylate hydrogels, as well as mechanical gradients across a hydrogel containing varying gel concentrations. Fluorescence microscopy, mechanical testing, and scanning electron microscopy show that the gradient profiles can be controlled by changing both the volume and concentration of each layer as well as intensity of UV exposure. GelMA hydrogel gradients with different Young's moduli were successfully used to culture human fibroblasts. The fibroblasts migrated along the gradient axis and showed different morphologies. In general, the proposed technique provides a rapid and simple approach to design and fabricate 3D hydrogel gradients for in vitro biological studies and potentially for in vivo tissue engineering applications.

An approach to prepare polyethylenimine functionalized silica-based spheres with small size for siRNA delivery.

A novel approach has been developed to prepare polyethylenimine functionalized hybrid silica spheres with a diameter of ∼10 nm, which show excellent delivery efficiency of siRNA into osteosarcoma cancer cells and human colon cancer cells with a significant cell inhibition comparable to commercial agents.

Nanoparticles mimicking viral surface topography for enhanced cellular delivery.

Novel silica nanoparticles mimicking virus surface topography are prepared. It is demonstrated that increases in nanoscale surface roughness promote both binding of biomolecules and cellular uptake; thus, the cellular delivery efficiency is significantly increased (scale bars 20 µm).