Tim4 deficiency reduces CD301b+ macrophage and aggravates periodontitis bone loss.

Periodontitis is a common chronic inflammatory disease that causes the periodontal bone destruction and may ultimately result in tooth loss. With the progression of periodontitis, the osteoimmunology microenvironment in periodontitis is damaged and leads to the formation of pathological alveolar bone resorption. CD301b+ macrophages are specific to the osteoimmunology microenvironment, and are emerging as vital booster for conducting bone regeneration. However, the key upstream targets of CD301b+ macrophages and their potential mechanism in periodontitis remain elusive. In this study, we concentrated on the role of Tim4, a latent upstream regulator of CD301b+ macrophages. We first demonstrated that the transcription level of Timd4 (gene name of Tim4) in CD301b+ macrophages was significantly upregulated compared to CD301b- macrophages via high-throughput RNA sequencing. Moreover, several Tim4-related functions such as apoptotic cell clearance, phagocytosis and engulfment were positively regulated by CD301b+ macrophages. The single-cell RNA sequencing analysis subsequently discovered that Cd301b and Timd4 were specifically co-expressed in macrophages. The following flow cytometric analysis indicated that Tim4 positive expression rates in total macrophages shared highly synchronized dynamic changes with the proportions of CD301b+ macrophages as periodontitis progressed. Furthermore, the deficiency of Tim4 in mice decreased CD301b+ macrophages and eventually magnified alveolar bone resorption in periodontitis. Additionally, Tim4 controlled the p38 MAPK signaling pathway to ultimately mediate CD301b+ macrophages phenotype. In a word, Tim4 might regulate CD301b+ macrophages through p38 MAPK signaling pathway in periodontitis, which provided new insights into periodontitis immunoregulation as well as help to develop innovative therapeutic targets and treatment strategies for periodontitis.

Extrafibrillarly Demineralized Dentin Matrix for Bone Regeneration.

Dentin is a natural extracellular matrix, but its availability in bone grafting and tissue engineering applications is underestimated due to a lack of proper treatment. In this study, the concept of extrafibrillar demineralization is introduced into the construction of dentin-derived biomaterials for bone regeneration for the first time. Calcium chelating agents with large molecular weights are used to selectively remove the extrafibrillar apatite minerals without disturbing the intrafibrillar minerals within dentin collagen, resulting in the formation of an extrafibrillarly demineralized dentin matrix (EDM). EDM with distinctive nanotopography and bone-like mechanical properties is found to significantly promote cell adhesion, migration, and osteogenic differentiation in vitro while enhancing in vivo bone healing of rat calvarial defects. The outstanding osteogenic performance of EDM is further confirmed to be related to the activation of the focal adhesion-cytoskeleton-nucleus mechanotransduction axis. Overall, this study shows that extrafibrillar demineralization of dentin has great potential to produce hierarchical collagen-based scaffolds for bone regeneration, and this facile top-down fabrication method brings about new ideas for the biomedical application of naturally derived bioactive materials.

Fabrication of an exosome-loaded thermosensitive chitin-based hydrogel for dental pulp regeneration.

Injective thermosensitive hydrogels are considered promising scaffolds to trigger dental pulp regeneration in devitalized human teeth. In this study, we developed a hydroxypropyl chitin (HPCH)/chitin whisker (CW) thermosensitive hydrogel with enhanced mechanical properties and biological activities. Exosomes can serve as biomimetic tools for tissue engineering, but the rapid clearance of unconjugated exosomes in vivo limits their therapeutic effects. To address this challenge, exosomes were isolated from human pulp stem cells (hDPSCs) and directly embedded into the HPCH/CW pre-gel to form an exosome-loaded hydrogel (HPCH/CW/Exo). The exosome-loaded thermosensitive hydrogel can be easily injected into an irregular endodontic space and gelated in situ. In vitro cell experiments proved that the delivery of exosomes significantly improved the ability of hydrogels to promote odontogenesis and angiogenesis. Meanwhile, in vivo animal experiments revealed the formation of new dental pulp-like tissues in an implanted tooth root model. Therefore, the proposed hydrogel provides a great potential alternative to traditional root canal therapy in dental clinics.

Evaluation of resveratrol-doped adhesive with advanced dentin bond durability.

OBJECTIVES: This paper aimed to evaluate the influence of resveratrol-doped adhesive on the durability and antibiofilm capability of dentin bonding. METHODS: Experimental adhesives were prepared by incorporating resveratrol into a universal adhesive at concentrations of 0 (control), 0.1, 1, and 10 mg/mL. The microtensile bond strength, fracture modes, and adhesive-dentin interface nanoleakage were assessed after 24 h of water storage, 10,000 times of thermocycling or 1-month of collagenase ageing. Relevant antibiofilm capability on Streptococcus mutans (S. mutans), in situ zymography, degree of conversion, and cytotoxicity of resveratrol-doped adhesives were also determined. RESULTS: Irrespective of thermocycled or collagenase ageing, the resveratrol-doped adhesive (1 mg/mL) maintained the bond strength and reduced the nanoleakage expression. Meanwhile, the inhibitory ability on endogenous protease activity and S. mutans biofilm formation with acceptable biocompatibility were obtained. CONCLUSIONS: This study suggested that the resveratrol-doped adhesive achieved effective improvement on dentin bond durability and secondary caries management. CLINICAL SIGNIFICANCE: The application of the resveratrol-doped adhesive indicates promising benefits to increase the lifetime of composite restorations.

Epigallocatechin-3-gallate/nanohydroxyapatite platform delivery approach to adhesive-dentin interface stability.

Current adhesive techniques allow clinicians to bond composite resin to dentin for esthetic restoration of defected tooth. However, a vulnerable adhesive-dentin interface remains clinically challenging resulting in frequent replacement of the restorations. The inappropriate management of exposed dentin plays a major role in jeopardizing the bond stability of the adhesive-dentin interface. To overcome this problem, this paper highlights an epigallocatechin-3-gallate/nanohydroxyapatite (EGCG/nHAp) platform (mesoporous silica-based) delivery approach to the adhesive-dentin interface and investigates its effectiveness on dentin bonding durability. Microtensile bond strength, interfacial nanoleakage, and in situ zymography were determined. The inhibition of Streptococcus mutans (S. mutans) biofilm formation along the adhesive-dentin interface was assessed by confocal-laser scanning microscopy, colony forming units counts, and field-emission scanning electron microscopy. Results revealed that applying the EGCG/nHAp delivery platform on exposed dentin could preserve the dentin bond strength and reduce interfacial nanoleakage after collagenase ageing; moreover, it could inactivate the activity of matrix metalloproteinase within the hybrid layer and inhibit the adhesion and biofilm formation of S. mutans. The proposed approach demonstrates great potential for stabilizing the adhesive-dentin interface to improve dentin bonding durability and prevent secondary caries progression, thereby indicating a promising strategy to prolong the service life of dental restorations.

Effects of resveratrol/ethanol pretreatment on dentin bonding durability.

To determine the effects of resveratrol/ethanol solution on the durability of resin-dentin bonding interfaces. Sixty-four non-caries third molars were randomly divided into four groups (n = 16) after sectioning, and then pretreated with one of the following concentrations of resveratrol/ethanol solutions: 0 (control group), 1, 10 and 20 mg/mL, followed by a universal adhesive and resin composites. All microtensile samples were divided into three subgroups: immediate group, collagenase ageing group and thermocycled group. The microtensile bond strength (MTBS), failure modes, interfacial nanoleakage and in situ zymography were measured, whereas the inhibitory effects of pretreated dentin slices on S. mutans biofilms were determined by confocal laser scanning microscopy and MTT assay. The results indicated that bonding strength was not only influenced by pretreatment factors (P < 0.05) but also ageing factors (P < 0.05). Regardless of collagenase ageing or thermocycling, the 10 mg/mL resveratrol/ethanol pretreatment group presented significantly higher (P < 0.05) MTBS and lower (P < 0.05) expression of nanoleakage than the control group, showed better inhibitory effect of matrix metalloproteinases and S. mutans activity with acceptable cytotoxicity. Meanwhile, cohesive failure in dentin decreased gradually with increasing resveratrol concentration. Therefore, the resveratrol/ethanol solution had the potential to serve as a versatile dentin primer, which can effectively improve dentin bonding durability and prevent secondary caries.

Chlorhexidine-encapsulated mesoporous silica-modified dentin adhesive.

OBJECTIVES: This work aims to explore the feasibility of chlorhexidine-encapsulated mesoporous silica (CHX@pMSN) as a modifier of a commercial dental adhesive via the evaluation of physicochemical properties and antibacterial capabilities of adhesive-dentin interface. METHODS: Therapeutic adhesives were developed in the present study by incorporating CHX@pMSN into a commercial adhesive at four mass fractions (0, 1, 5 and 10 wt.%). The antibacterial capability on Streptococcus mutans (S. mutans) biofilm, conversion degree, adhesive morphology, microtensile bond strength (MTBS) and nanoleakage expression were evaluated comprehensively. RESULTS: MTT and CLSM evaluation showed that CHX@pMSN-doped adhesive inhibits S. mutans biofilm growth, while CHX is released from the modified adhesive continuously. The incorporation of CHX@pMSN did not affect immediate bond strength at the concentration of 1% and 5% (P > 0.05). Moreover, these bonds were mainly preserved in 5% CHX@pMSN group after one month of collagenase ageing. Meanwhile, CHX@pMSN-doped adhesive groups exhibited similar nanoleakage distribution compared with the control. CONCLUSION: This study showed that the 5% CHX@pMSN-modified adhesive achieved balance amongst unaffected immediate bonding strength, well-preserved bonds against collagenase ageing and effective inhibition of S. mutans biofilm growth. CLINICAL SIGNIFICANCE: CHX@pMSN-modified dentin adhesive can potentially extend the service life of adhesive restoration in clinic.