Characteristics and biological responses of selective laser melted Ti6Al4V modified by micro-arc oxidation.

Background/purpose: Additive manufacturing (AM) technology, such as selective laser melting (SLM), has been used to fabricate medical devices of Ti-6wt.% Al-4wt.%V (Ti6Al4V) alloys in dentistry. Strontium (Sr) has been shown to have the potential to treat osteoporosis. The aim of this study was to investigate the physicochemical and biological properties of strontium-containing coatings on selective laser melted Ti6Al4V (SLM-Ti6Al4V) substrate. Materials and methods: The disk of Ti6Al4V was prepared by SLM method. The strontium-containing coatings were prepared by micro-arc oxidation (MAO) in aqueous electrolytes. The surface topography, chemical composition, and phase of strontium-containing MAO (SrMAO) coatings were performed by scanning electron microscope (SEM), energy dispersive X-ray spectrometer (EDS), and thin film X-ray diffraction (TF-XRD), respectively. The apatite-forming ability of the MAO coatings was conducted in simulating body fluid (SBF), and the cell proliferation was determined by methylthiazoletetrazolium (MTT) assay. Results: The microstructure of SLM-Ti6Al4V displays acicular α-phase organization. The TF-XRD results indicated that the phase of SrMAO coating was anatase, rutile, and titanium. The calcium, phosphorus, and strontium were detected in the coatings by EDS. Using the SEM, the surface morphology of SrMAO coatings exhibited a uniform 3D porous structure. The SrMAO coatings could induce a bone-like apatite layer after immersion in SBF, and presented significantly higher cell proliferation than untreated specimens in in-vitro experiments. Conclusion: All findings in this study indicate that SrMAO coatings formed on SLM-Ti6Al4V surfaces exhibit a benefit on biological responses and thereby are suitable for biomedical applications.

The Uniform Distribution of Hydroxyapatite in a Polyurethane Foam-Based Scaffold (PU/HAp) to Enhance Bone Repair in a Calvarial Defect Model.

Polyurethane (PU) is a promising material for addressing challenges in bone grafting. This study was designed to enhance the bone grafting capabilities of PU by integrating hydroxyapatite (HAp), which is known for its osteoconductive and osteoinductive potential. Moreover, a uniform distribution of HAp in the porous structure of PU increased the effectiveness of bone grafts. PEG/APTES-modified scaffolds were prepared through self-foaming reactions. A uniform pore structure was generated during the spontaneous foaming reaction, and HAp was uniformly distributed in the PU structure (PU15HAp and PU30HAp) during foaming. Compared with the PU scaffolds, the HAp-modified PU scaffolds exhibited significantly greater protein absorption. Importantly, the effect of the HAp-modified PU scaffold on bone repair was tested in a rat calvarial defect model. The microstructure of the newly formed bone was analyzed with microcomputed tomography (µ-CT). Bone regeneration at the defect site was significantly greater in the HAp-modified PU scaffold group than in the PU group. This innovative HAp-modified PU scaffold improves current bone graft materials, providing a promising avenue for improved bone regeneration.

Predicting root fracture after root canal treatment and crown installation using deep learning.

Background/purpose: Vertical root fracture (VRF) is a prevalent reason for tooth extraction following root canal treatment and even after crown placement. Predicting fractures is challenging due to multifactorial nature. The current study aimed to predict the likelihood of fracture following root canal treatment and crown placement by developing a deep learning (DL) model. Materials and methods: DL techniques were employed to analyze a dataset comprising 145 clinical cases consisting of 97 fractured teeth and 48 non-fractured teeth. This dataset spanned a five-year period and encompassed cases involving root canal therapy and crown installation. The analysis identified several root fracture-related parameters, which were incorporated into the DL system. The dataset consisted of 17 features presented in a mixed-type tabular format. Results: The deep neural network (DNN) model surpassed the support vector machine (SVM) model with a higher accuracy (80.7 % vs. 71.7 %) and F1-score value (0.857 vs. 0.817) for predicting root fracture. Furthermore, in determining root fracture occurrence, it was observed that 17 significant characteristics in the DNN model outperformed the 7 features by 11.7 % in accuracy and 10 % in F1-score. Conclusion: DL shows promise in predicting root fracture post root canal therapy and prosthesis, and it may have the potential to aid clinicians in assessing fracture risk and improving decision-making.

A Multifunctional Polyethylene Glycol/Triethoxysilane-Modified Polyurethane Foam Dressing with High Absorbency and Antiadhesion Properties Promotes Diabetic Wound Healing.

The delayed healing of chronic wounds, such as diabetic foot ulcers (DFUs), is a clinical problem. Few dressings can promote wound healing by satisfying the demands of chronic wound exudate management and tissue granulation. Therefore, the aim of this study was to prepare a high-absorption polyurethane (PU) foam dressing modified by polyethylene glycol (PEG) and triethoxysilane (APTES) to promote wound healing. PEG-modified (PUE) and PEG/APTES-modified (PUESi) dressings were prepared by self-foaming reactions. Gauze and PolyMem were used as controls. Next, Fourier transform-infrared spectroscopy, thermomechanical analyses, scanning electron microscopy and tensile strength, water absorption, anti-protein absorption, surface dryness and biocompatibility tests were performed for in vitro characterization. Wound healing effects were further investigated in nondiabetic (non-DM) and diabetes mellitus (DM) rat models. The PUE and PUESi groups exhibited better physicochemical properties than the gauze and PolyMem groups. Moreover, PUESi dressing showed better anti-adhesion properties and absorption capacity with deformation. Furthermore, the PUESi dressing shortened the inflammatory phase and enhanced collagen deposition in both the non-DM and DM animal models. To conclude, the PUESi dressing not only was fabricated with a simple and effective strategy but also enhanced wound healing via micronegative-pressure generation by its high absorption compacity with deformation.

Development of a flexible film made of polyvinyl alcohol with chitosan based thermosensitive hydrogel.

Background/purpose: A challenge that arises with periodontal regeneration surgery has been associated with the future development of periodontal regeneration membrane to prevent gingiva and fibroblasts invade the wound and allow alveolar bone successfully regenerated. Materials and methods: Chitosan (CS) has the advantages of non-toxicity, biodegradation, biocompatibility, and has been widely used in wound dressings. A flexible film was made using polyvinyl alcohol (PVA) blending CS based thermosensitive hydrogel. Results: The proposed 2% PVA/CS hydrogel has the highest swelling ratio about 720% after 60 min incubation and keeps its area after 10 min incubation for surgery suture. The elastic modulus of 0%, 1%, 2%, and 4% PVA/CS hydrogel were 7.75 ± 1.96, 0.91 ± 0.16, 0.75 ± 0.21, and 0.37 ± 0.06 MPa, respectively. The maximum strain of 2% PVA/CS hydrogel was 101.00 ± 28.03 (%). After 8 weeks biodegradation, the remain weight of 2% PVA/CS hydrogel was 71.36 ± 0.79 (%). Conclusion: In vitro cytotoxicity tests were performed and demonstrated PVA/CS hydrogel significantly improving cell proliferation. This study realized a promising flexible film for periodontal regeneration membrane that can prevent the rapid growth of fibroblasts to invade the wound and be used for periodontal regeneration surgery.

The biological responses of osteoblasts on titanium: Effect of oxygen level and surface roughness.

BACKGROUND/PURPOSE: Due to the general application of in vitro test, cell culture is generally selected to evaluate the cytocompatibility of devices and materials. The choice of test condition should depend on the probable site and clinical application. The oxygen content of human body could be estimated around 5%∼12%, and the oxygen level of healing bone fracture range from 0.8%∼3.8%%. However, materials for bone implant are traditionally evaluated under laboratory normoxia condition (21% O2) in vitro. The aim was to study the effect of oxygen level on osteoblast upon high stiffness titanium with different roughness. METHODS: After sandblasted and acid-etched (SLA) process, we create titanium surfaces with four different roughness. The differentiation and proliferation of MC3T3-E1 osteoblast cultured on SLA-treated specimens were evaluated in designed chamber with oxygen level of 1%, 5%, 10%, 21%. RESULTS: By scanning electron microscopy, all samples had sub-micro pit inside the micro-holes upon SLA-treated Ti disk surface. The decrease of oxygen level from 21% to 5% promoted osteoblast growth of SLA-treated specimens, but 1% O2 delayed cell proliferation. The surface roughness of specimens influenced osteoblast cell differentiation. The differentiation and proliferation ability of the cells upon SLA-treated specimens is proportional to oxygen level. CONCLUSION: Our results demonstrated that 5% O2 will easily discriminate osteoblasts responses on different SLA-treated specimens. These results suggest that hypoxia (5% O2) environment is better model for biological evaluation of bone-related materials.

Effect of thermocycling-induced stress on properties of orthodontic NiTi wires.

Background/purpose: In orthodontic applications, NiTi wires are under continuous bending stress and exposed to fluctuations in temperature over long durations. The sensitivity of NiTi to temperature can have a considerable influence on its mechanical properties. This study investigated the effects of deflected NiTi wire, presented in stress-induced (detwinned) martensite microstructure, combined with thermal cycle on the microstructure and mechanical properties. Materials and methods: We tested four types of as-received orthodontic NiTi: (1) Nitinol Classic (3 M Unitek), (2) Sentalloy (Tomy), (3) 27 °C CuNiTi (Ormco) and (4) 40 °C CuNiTi (Ormco). Each group of specimens was subjected to three different testing conditions: (1) temperature fluctuations (5000 cycles) between 5 and 55 °C, (2) continuous three-point bending force and (3) combination of thermal cycling and bending stress. Results: The specimens that underwent thermocycling as well as loading exhibited a substantial narrowing in stress hysteresis, which may be attributed to crystallinity lower than that of as-received NiTi wires. Reduced crystallinity can manifest in a number of imperfections, such as dislocations and internal stress, as well as a less-organized structure. Micro X-ray diffraction (XRD) analysis revealed the existence of martensite phase in Sentalloy wires subject to thermal and stress conditions. Under loading conditions, stress-induced martensite of NiTi wires exposed to temperature fluctuations of 5-55 °C also induced cyclic changes in bending stress. In a simulated intra-oral environment, the stability of austenite↔martensite transformation decreased. Conclusion: This study determined that bending stress in conjunction with repeated temperature fluctuations can greatly affect the microstructure and mechanical properties of NiTi wires.

Novel system in vitro of classifying oral carcinogenesis based on feature extraction for gray-level co-occurrence matrix using scanned laser pico projector.

Oral cancer is among the top 10 causes of death due to cancer worldwide. The prognosis for oral cancer patients is not good, with a 5-year survival rate of only 50%. Earlier and more precise classification will help clinicians make a diagnosis and patients survive. With the advancement of technology, computer-aided detection methods are used to help clinicians form therapy strategies. Gray-level co-occurrence matrix (GLCM) feature extraction of images describing the spatial distribution of gray levels is widely used in medical imaging analysis. Scanned laser pico projector (SLPP) has advantages such as high intensity, directivity, coherence, and mono-color with low bandwidth. In this study, GLCM feature extraction and SLPP reflex images were combined to make a small, non-staining, noninvasive classification system. According to the various image characteristics in oral carcinogenesis, SLPP reflex images better define the borders and three-dimensional structures and provide effective GLCM features such as contrast, energy, and homogeneity to classify carcinogenesis in dysplastic oral keratinocyte (DOK) and normal oral keratinocyte (NOK) cells. Moreover, it also reliably classifies highly metastatic (HSC-3) and tongue cancer (CAL-27) cells. A promising computer-aided classification system for oral cancer was developed to build a reliable intraoral examination system for in situ computer-aided diagnosis in normal clinics.

Biopolymers Hybrid Particles Used in Dentistry.

This literature review provides an overview of the fabrication and application of biopolymer hybrid particles in dentistry. A total of 95 articles have been included in this review. In the review paper, the common inorganic particles and biopolymers used in dentistry are discussed in general, and detailed examples of inorganic particles (i.e., hydroxyapatite, calcium phosphate, and bioactive glass) and biopolymers such as collagen, gelatin, and chitosan have been drawn from the scientific literature and practical work. Among the included studies, calcium phosphate including hydroxyapatite is the most widely applied for inorganic particles used in dentistry, but bioactive glass is more applicable and multifunctional than hydroxyapatite and is currently used in clinical practice. Today, biopolymer hybrid particles are receiving more attention as novel materials for several applications in dentistry, such as drug delivery systems, bone repair, and periodontal regeneration surgery. The literature published on the biopolymer gel-assisted synthesis of inorganic particles for dentistry is somewhat limited, and therefore, this article focuses on reviewing and discussing the biopolymer hybrid particles used in dentistry.

Hydrothermal-aging-induced lattice distortion in yttria-stabilized zirconia using EBSD technique.

The destabilization problem is of importance in the application of yttria-stabilized zirconia (YSZ) bio-ceramic in the oral environment due to phase transformation between tetragonal to monoclinic. Thus, in this study, the lattice distortion induced by hydrothermal aging in yttria-stabilized zirconia (YSZ) was investigated, in which YSZ specimens were subjected to hydrothermal-aging treatment for 0-48 h. The Kikuchi-band based method was employed to calculate the lattice distortion after phase transformation and the results from EBSD were compared with these obtained by X-ray diffraction (XRD). Both measurement methods showed a similar tendency of the lattice change in the a- and c- axes. EBSD results showed that the strain rates were 7.98 % and -5.03 % at the a- and c-axes, respectively. A significant decrease in the c/a ratio from 1.429 to 1.257 for the tetragonal matrix after 48 h aging is observed, which indicated Kikuchi-band based method using EBSD technique can successfully determine the local strain in tetragonal matrices.

OmpR coordinates the expression of virulence factors of Enterohemorrhagic Escherichia coli in the alimentary tract of Caenorhabditis elegans.

Enterohemorrhagic Escherichia coli (EHEC), an enteropathogen that colonizes in the intestine, causes severe diarrhea and hemorrhagic colitis in humans by the expression of the type III secretion system (T3SS) and Shiga-like toxins (Stxs). However, how EHEC can sense and respond to the changes in the alimentary tract and coordinate the expression of these virulence genes remains elusive. The T3SS-related genes are known to be regulated by the locus of enterocyte effacement (LEE)-encoded regulators, such as Ler, as well as non-LEE-encoded regulators in response to different environmental cues. Herein, we report that OmpR, which participates in the adaptation of E. coli to osmolarity and pH alterations, is required for EHEC infection in Caenorhabditis elegans. OmpR protein was able to directly bind to the promoters of ler and stx1 (Shiga-like toxin 1) and regulate the expression of T3SS and Stx1, respectively, at the transcriptional level. Moreover, we demonstrated that the expression of ler in EHEC is in response to the intestinal environment and is regulated by OmpR in C. elegans. Taken together, we reveal that OmpR is an important regulator of EHEC which coordinates the expression of virulence factors during gastrointestinal infection in vivo.

Host CDK-1 and formin mediate microvillar effacement induced by enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) induces changes to the intestinal cell cytoskeleton and formation of attaching and effacing lesions, characterized by the effacement of microvilli and then formation of actin pedestals to which the bacteria are tightly attached. Here, we use a Caenorhabditis elegans model of EHEC infection to show that microvillar effacement is mediated by a signalling pathway including mitotic cyclin-dependent kinase 1 (CDK1) and diaphanous-related formin 1 (CYK1). Similar observations are also made using EHEC-infected human intestinal cells in vitro. Our results support the use of C. elegans as a host model for studying attaching and effacing lesions in vivo, and reveal that the CDK1-formin signal axis is necessary for EHEC-induced microvillar effacement.

Nano/Micro Hierarchical Bioceramic Coatings for Bone Implant Surface Treatments.

Bone implants with surface modifications that promote the physiological activities of osteoblasts are the first step for osseointegration in bone repair. Hydroxyapatite is the main inorganic component in mammal bones and teeth, and nanoscaled hydroxyapatite promotes the adhesion of osteoblastic cells. In this study, we created a nano/micro hierarchical structure using micro-arc oxidation coatings and hydrothermal treatments at 150 °C, 175 °C, and 200 °C for 2, 6, 12, and 24 h. After undergoing hydrothermal treatment for 24 h, CaTiO3 began forming regular-shaped crystals at the surface at 175 °C. In order to decrease the CaTiO3 formations and increase the apatite fabrication, a shorter time of hydrothermal treatment was required at 175 °C. There was still surface damage on samples treated for 6 h at 175 °C; however, the nano/micro hierarchical structures were formed in 2 h at 175 °C. The normalized alkaline phosphatase (ALP) activities of the MC3T3-E1 cells with micro-arc oxidation (MAO) coatings and nano/micro hierarchical bioceramics coatings were 4.51 ± 0.26 and 7.36 ± 0.51 µmol p-NP/mg protein (*** P value of <0.001), respectively. The MC3T3-E1 cells with coatings showed highly statistically significant results in terms of the ALP activity. This proposed nano/micro hierarchical structure promoted cell proliferation and osteogenic differentiation of the osteoblast MC3T3-E1 cells. This study realized a promising nano system for osseointegration via bone implant surface treatments, which can promote the physiological activities of osteoblasts.

Comparative investigation into effects of ZrO2 and Al2O3 addition in fluorapatite laser-clad composite coatings on Ti6Al4V alloy.

Composite coatings consisting of fluorapatite mixed with 20 wt% yttria (3 mol%) stabilized cubic phase zirconia (c-ZrO2, 3Y-TZP) or 20 wt% alumina (α-Al2O3) were deposited on Ti6Al4V substrates using a Nd:YAG laser cladding system. The interface morphology, phase composition, micro-hardness and biological properties of the two coatings were examined and compared. The results showed that the fluorapatite/Al2O3 specimen underwent a greater inter-diffusion at the interface between the coating layer and the transition layer than the fluorapatite/ZrO2 specimen. During the cladding process, the ZrO2 and Al2O3 components of the coating were completely decomposed or underwent phase transformation. In addition, the fluorapatite was partially decomposed. For both specimens, the coating layers contained fluorapatite, CaF2 and CaTiO3 phases. The coating layer of the fluorapatite/ZrO2 specimen additionally contained TTCP, CaO, CaZrO3 and m-ZrO2 (monoclinic phase), while that of the fluorapatite/Al2O3 specimen contained ß-TCP, CaAl2O4 and θ-Al2O3. The average micro-hardness of the fluorapatite/ZrO2 coating layer (1300 HV) was approximately 200 HV higher than that of the fluorapatite/Al2O3 coating layer (1100 HV). Both specimens generated dense bone-like apatite following immersion in simulated body fluid for 3 days. In other words, both specimens had a good in vitro bioactivity. However, the fluorapatite/ZrO2 specimen showed a better initial attachment and spread of osteoblast-like osteosarcoma MG63 cells than the fluorapatite/Al2O3 specimen in in vitro biocompatibility tests performed for 24 h.

The effect of plasma treatment on the osseointegration of rough titanium implant: A histo-morphometric study in rabbits.

ABSTRACT BACKGROUND/PURPOSE: The surface properties, such as hydrophilicity and functional OH groups, play an important role in bone fixation in vivo. In our previous study, the plasma treatments of large grit and acid etching (SLA) method produce functional OH groups on the rough surface. There is no report in discussing the integration between basic Ti-OH groups and bone-to-implant contact (BIC). The aim of this study was to evaluate the effect of the functional OH groups on the rough surface both in vitro and in vivo. MATERIALS AND METHODS: Functional hydroxyl groups were produced on a SLA-treated surface. The surface topography, roughness, wettability, and chemical composition were examined using various techniques. Twenty-four implants were inserted into the proximal tibia of four New Zealand white rabbits. The biological responses were measured in terms of histomorphometric analysis 4 and 8 weeks post-implantation. RESULTS: The surface morphology and roughness were similar among all groups. However, the concentration of OH groups and hydrophilicity were found increased in the plasma treatment. The cell morphology in RF-plasma treated groups had more polygonal type and higher expression of actin and vinculin. The bone-to-implant contact (BIC) ratios of RF-200W were significantly higher than other groups (P < 0.05). The relationship between basic OH groups and BIC showed linear correspondence. CONCLUSION: The Ti-OH groups introduced on the rough surface by plasma treatments can trigger cell adhesion which further initiate new bone apposition. We propose that RF-plasma treatment can help to enhance bone healing at 4 and 8 weeks.

Hydrothermal treatment and butylphosphonic acid derived self-assembled monolayers for improving the surface chemistry and corrosion resistance of AZ61 magnesium alloy.

The hydrothermal treatment followed by a self-assembled monolayer (SAM) of 1-butylphosphonic acid through the tethering by aggregation and growth (T-BAG) method was employed to produce protective surface coatings on the Mg-6Al-1Zn alloy (AZ61) for reducing the degradation rate in physiological environments. Potentiodynamic polarization measurements revealed that the organic self-assembled monolayer and Mg(OH)2 coating can further enhance the surface chemical stability and corrosion resistance of Mg alloys. SAM-treated Mg(OH)2 coatings can be served as a more passive surface layer as a result of their much higher charge transfer resistance and the presence of Warburg impedance in electrochemical impedance spectroscopy measurement.

Surface Modification of Direct-Current and Radio-Frequency Oxygen Plasma Treatments Enhance Cell Biocompatibility.

The sand-blasting and acid etching (SLA) method can fabricate a rough topography for mechanical fixation and long-term stability of titanium implant, but can not achieve early bone healing. This study used two kinds of plasma treatments (Direct-Current and Radio-Frequency plasma) to modify the SLA-treated surface. The modification of plasma treatments creates respective power range and different content functional OH groups. The results show that the plasma treatments do not change the micron scale topography, and plasma-treated specimens presented super hydrophilicity. The X-ray photoelectron spectroscopy (XPS)-examined result showed that the functional OH content of the RF plasma-treated group was higher than the control (SLA) and DC treatment groups. The biological responses (protein adsorption, cell attachment, cell proliferation, and differentiation) promoted after plasma treatments, and the cell responses, have correlated to the total content of amphoteric OH groups. The experimental results indicated that plasma treatments can create functional OH groups on SLA-treated specimens, and the RF plasma-treated SLA implant thus has potential for achievement of bone healing in early stage of implantation.

A Disease Model of Muscle Necrosis Caused by Aeromonas dhakensis Infection in Caenorhabditis elegans.

A variety of bacterial infections cause muscle necrosis in humans. Caenorhabditis elegans has epidermis and bands of muscle that resemble soft-tissue structures in mammals and humans. Here, we developed a muscle necrosis model caused by Aeromonas dhakensis infection in C. elegans. Our data showed that A. dhakensis infected and killed C. elegans rapidly. Characteristic muscle damage in C. elegans induced by A. dhakensis was demonstrated in vivo. Relative expression levels of host necrosis-associated genes, asp-3, asp-4, and crt-1 increased significantly after A. dhakensis infection. The RNAi sensitive NL2099 rrf-3 (pk1426) worms with knockdown of necrosis genes of crt-1 and asp-4 by RNAi showed prolonged survival after A. dhakensis infection. Specifically knockdown of crt-1 and asp-4 by RNAi in WM118 worms, which restricted RNAi only to the muscle cells, conferred significant resistance to A. dhakensis infection. In contrast, the severity of muscle damage and toxicity produced by the A. dhakensis hemolysin-deletion mutant is attenuated. In another example, shiga-like toxin-producing enterohemorrhagic E. coli (EHEC) known to elicit toxicity to C. elegans with concomitant enteropathogenicty, did not cause muscle necrosis as A. dhakensis did. Taken together, these results show that Aeromonas infection induces muscle necrosis and rapid death of infected C. elegans, which are similar to muscle necrosis in humans, and then validate the value of the C. elegans model with A. dhakensis infection in studying Aeromonas pathogenicity.