Ultra-Sensitive and Selective Electrochemical Bio-Fluid Biopsy for Oral Cancer Screening.

The early diagnosis of recurrence and metastasis is critically important for decreasing the morbidity and mortality associated with oral cancers. Although liquid biopsy methods hold great promise that provide a successive "time-slice" profile of primary and metastatic oral cancer, the development of non-invasive, rapid, simple, and cost-effective liquid biopsy techniques remains challenging. In this study, an ultrasensitive and selective electrochemical liquid biopsy is developed for oral cancer screening based on tracking trace amounts of cancer biomarker by functionalized asymmetric nano-channels. Detection via antigen-antibody reactions is assayed by evaluating changes in ionic current. Upon the recognition of cancer biomarker antigens in bio-fluids, the inner wall of nano-channel immobilized with the corresponding antibodies undergoes molecular conformation transformation and surface physicochemical changes, which significantly regulate the ion transport through the nano-channel and help achieve sensitivity with a detection limit of 10-12 g mL-1 . Furthermore, owing to the specificity of the monoclonal antibody for the antigen, the nano-channel exhibits high selectivity for the biomarker than for structurally similar biological molecules present in bio-fluids. The effectiveness of this technique is confirmed through the diagnosis of clinical cases of oral squamous cell carcinoma. This study presents a novel diagnostic tool for oral cancer detection in bio-fluids.

An Amorphous Peri-Implant Ligament with Combined Osteointegration and Energy-Dissipation.

Progress toward developing metal implants as permanent hard-tissue substitutes requires both osteointegration to achieve load-bearing support, and energy-dissipation to prevent overload-induced bone resorption. However, in existing implants these two properties can only be achieved separately. Optimized by natural evolution, tooth-periodontal-ligaments with fiber-bundle structures can efficiently orchestrate load-bearing and energy dissipation, which make tooth-bone complexes survive extremely high occlusion loads (>300 N) for prolonged lifetimes. Here, a bioinspired peri-implant ligament with simultaneously enhanced osteointegration and energy-dissipation is presented, which is based on the periodontium-mimetic architecture of a polymer-infiltrated, amorphous, titania nanotube array. The artificial ligament not only provides exceptional osteoinductivity owing to its nanotopography and beneficial ingredients, but also produces periodontium-similar energy dissipation due to the complexity of the force transmission modes and interface sliding. The ligament increases bone-implant contact by more than 18% and simultaneously reduces the effective stress transfer from implant to peri-implant bone by ≈30% as compared to titanium implants, which as far as is known has not previously been achieved. It is anticipated that the concept of an artificial ligament will open new possibilities for developing high-performance implanted materials with increased lifespans.

The Dynamic Counterbalance of RAC1-YAP/OB-Cadherin Coordinates Tissue Spreading with Stem Cell Fate Patterning.

Tissue spreading represents a key morphogenetic feature of embryonic development and regenerative medicine. However, how molecular signaling orchestrates the spreading dynamics and cell fate commitment of multicellular tissue remains poorly understood. Here, it is demonstrated that the dynamic counterbalance between RAC1-YAP and OB-cadherin plays a key role in coordinating heterogeneous spreading dynamics with distinct cell fate patterning during collective spreading. The spatiotemporal evolution of individual stem cells in spheroids during collective spreading is mapped. Time-lapse cell migratory trajectory analysis combined with in situ cellular biomechanics detection reveal heterogeneous patterns of collective spreading characteristics, where the cells at the periphery are faster, stiffer, and directional compared to those in the center of the spheroid. Single-cell sequencing shows that the divergent spreading result in distinct cell fate patterning, where differentiation, proliferation, and metabolism are enhanced in peripheral cells. Molecular analysis demonstrates that the increased expression of RAC1-YAP rather than OB-cadherin facilitated cell spreading and induced differentiation, and vice versa. The in vivo wound healing experiment confirms the functional role of RAC1-YAP signaling in tissue spreading. These findings shed light on the mechanism of tissue morphogenesis in the progression of development and provide a practical strategy for desirable regenerative therapies.

Enamel Repair with Amorphous Ceramics.

Developing high-performance materials in physiological conditions to clinically repair stiff tissue for long lifespan remains a great challenge. Here, an enamel repair strategy is reported by efficiently growing a biocompatible ZrO2 ceramic layer on defective enamel through controllable hydrolysis of Zr4+ in oral-tolerable conditions. Detailed analysis of the grown layer indicates that the grown ZrO2 ceramic is amorphous without grain boundary and dislocation, which endows the repaired enamel with natural enamel comparable mechanical performance (modulus ≈82.5 GPa and hardness ≈5.2 GPa). Besides, the strong chemical connection between unsaturated coordinated Zr4+ in amorphous structure and PO4 3- greatly strengthen the crystalline-amorphous interface of the repaired enamel to endure the long-time mastication damage. Moreover, these ZrO2 ceramics provide hydrophilic, electronegative, and smooth surfaces to resist the adhesion and proliferation of cariogenic bacteria. The hybrid amorphous-crystalline interface design with advantages in biomechanical compatibility would promote the evolution of a variety of cutting-edge functional materials for medical and engineering application.

Endodontic management of the maxillary first molars with two root canals: A case report and review of the literature.

BACKGROUND: The complex anatomy of the maxillary first molars has always been a major challenge for complete root canal treatment in endodontic therapy. Here, we present two cases of maxillary first molars, each with only two root canals, which have been rarely reported. We also perform a literature review of maxillary first molar anatomy. CASE SUMMARY: The two patients were referred to the hospital after 1) finding a cavity in their tooth with a color change and, 2) a toothache during mastication, respectively. Both of these cases were diagnosed as apical periodontitis by X-ray imaging and cone beam computed tomography (CBCT). Non-surgical endodontic therapy was performed with the assistance of a dental operating microscope (DOM). CBCT showed rare but accurate images of both patients, each with two root canals and two roots in their maxillary first molars. Both roots were located in the buccal in the palatal direction, and each root had only one clear root canal. In addition, each maxillary first molar in both patients was symmetrical to that on the opposing side with only two separate root canals. Non-surgical endodontic therapy was performed with the assistance of a DOM. Finally, the teeth were restored using composite resin and the patients were satisfied with the results. CONCLUSION: Making full use of CBCT and DOM would contribute to helping dentists make correct diagnoses and successfully treat teeth with rare root canal morphologies.

Nanogels of carboxymethyl chitosan and lysozyme encapsulated amorphous calcium phosphate to occlude dentinal tubules.

This study aimed to develop of a rapid and effective method to occlude dentinal tubules using carboxymethyl chitosan and lysozyme (CMC/LYZ) nanogels with encapsulated amorphous calcium phosphate (ACP) based on the transformation of ACP to HAP. In this work, CMC/LYZ was used to stabilize ACP and form CMC/LYZ-ACP nanogels, and then the nanogel-encapsulated ACP was applied to exposed dentinal tubule surfaces. The morphology of the nanogels was examined by transmission electron microscopy (TEM). Distribution and quantity of elements in CMC/LYZ-ACP nanogels were determined by element mapping and energy dispersive X-Ray spectroscopy (EDX). Scanning electron microscopy (SEM) images, XRD measurements and nanoindentation were applied to check the efficacy of tubular occlusion. TEM revealed that CMC/LYZ-ACP nanogels were spherical dense gel particles with size approximately 50-500 nm. Element mapping and EDX indicated that C, N, O, Ca, P, and S in the microspheres are thoroughly represented. SEM images shows that the thickness of the coating layer was approximately 1-2 µm and the depth to which the mineralized substance enters the dentinal tubule was approximately 4-8 µm. XRD measurements and nanoindentation indicated that the occluding mineralized substance observed were similar to nature dentin. CMC can form spherical dense nanogels loaded with ACP under the participation of lysozyme. The CMC/LYZ-ACP nanogels could increase the dentinal tubule occluding effectiveness. These results indicated that finding and developing novel nanomaterials of CMC/LYZ-ACP would be an effective strategy for the treatment of dentin hypersensitivity.

Rapid biomimetic remineralization of the demineralized enamel surface using nano-particles of amorphous calcium phosphate guided by chimaeric peptides.

OBJECTIVE: The objective of this study was to develop a rapid and effective method to remineralize human carious-like enamel using chimaeric peptide-mediated nanocomplexes of carboxymethyl chitosan/amorphous calcium phosphate (CMC/ACP), mimicking the mineralizing pattern of the oriented assembly of ACP guided by amelogenin in the biomineralization of enamel. METHODS: CMC/ACP nanocomplex solution was first synthesized through the successive addition of carboxymethyl chitosan, calcium chloride, and dipotassium phosphate into distilled water. ACP nanoparticles were degraded by 1% NaClO from CMC/ACP nanocomplexes. The morphology of the particles at different periods was tested by transmission electron microscopy (TEM). The chimaeric peptides were added to guide the arrangement of ACP nanoparticles and to bind ACP nanoparticles to the demineralized enamel surface specifically. X-ray diffraction (XRD)/scanning electron microscope (SEM)/confocal laser scanning microscopy (CLSM)/nano-indentation tests were applied to check the remineralization effects. RESULTS: CMC/ACP nanocomplexes were obtained and could be kept without precipitation for a long time. After the degradation of NaClO and guidance of chimaeric peptides, ACP nanoparticles were arranged into oriented arrays before transforming into crystals, and the enamel-like crystals were tightly bound onto the demineralized surface. The newly formed enamel-like crystals were nearly well-organized and equipped with strong mechanical properties.

Oriented and Ordered Biomimetic Remineralization of the Surface of Demineralized Dental Enamel Using HAP@ACP Nanoparticles Guided by Glycine.

Achieving oriented and ordered remineralization on the surface of demineralized dental enamel, thereby restoring the satisfactory mechanical properties approaching those of sound enamel, is still a challenge for dentists. To mimic the natural biomineralization approach for enamel remineralization, the biological process of enamel development proteins, such as amelogenin, was simulated in this study. In this work, carboxymethyl chitosan (CMC) conjugated with alendronate (ALN) was applied to stabilize amorphous calcium phosphate (ACP) to form CMC/ACP nanoparticles. Sodium hypochlorite (NaClO) functioned as the protease which decompose amelogenin in vivo to degrade the CMC-ALN matrix and generate HAP@ACP core-shell nanoparticles. Finally, when guided by 10 mM glycine (Gly), HAP@ACP nanoparticles can arrange orderly and subsequently transform from an amorphous phase to well-ordered rod-like apatite crystals to achieve oriented and ordered biomimetic remineralization on acid-etched enamel surfaces. This biomimetic remineralization process is achieved through the oriented attachment (OA) of nanoparticles based on non-classical crystallization theory. These results indicate that finding and developing analogues of natural proteins such as amelogenin involved in the biomineralization by natural macromolecular polymers and imitating the process of biomineralization would be an effective strategy for enamel remineralization. Furthermore, this method represents a promising method for the management of early caries in minimal invasive dentistry (MID).