Cellular mechanics, a major regulating factor of cellular architecture and biological functions, responds to intrinsic stresses and extrinsic forces exerted by other cells and the extracellular matrix in the microenvironment. Cellular mechanics also acts as a fundamental mediator in complicated immune responses, such as cell migration, immune cell activation, and pathogen clearance. The principle of atomic force microscopy (AFM) and its three running modes are introduced for the mechanical characterization of living cells. The peak force tapping mode provides the most delicate and desirable virtues to collect high-resolution images of morphology and force curves. For a concrete description of AFM capabilities, three AFM applications are discussed. These applications include the dynamic progress of a neutrophil-extracellular-trap release by neutrophils, the immunological functions of macrophages, and the membrane pore formation mediated by perforin, streptolysin O, gasdermin D, or membrane attack complex.
Microscopy, Atomic Force ; Neutrophils

OBJECTIVES: This study investigated the effects of a hydrofluoric acid (HA; solution of hydrogen fluoride [HF] in water)-based smart etching (SE) solution at an elevated temperature on yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) ceramics in terms of bond strength and morphological changes.MATERIALS AND METHODS: Eighty sintered Y-TZP specimens were prepared for shear bond strength (SBS) testing. The bonding surface of the Y-TZP specimens was treated with 37% phosphoric acid etching at 20°C–25°C, 4% HA etching at 20°C–25°C, or HA-based SE at 70°C–80°C. In all groups, zirconia primers were applied to the bonding surface of Y-TZP. For each group, 2 types of resin cement (with or without methacryloyloxydecyl dihydrogen phosphate [MDP]) were used. SBS testing was performed. Topographic changes of the etched Y-TZP surface were analyzed using scanning electron microscopy and atomic force microscopy. The results were analyzed and compared using 2-way analysis of variance.RESULTS: Regardless of the type of resin cement, the highest bond strength was measured in the SE group, with significant differences compared to the other groups (p < 0.05). In all groups, MDP-containing resin cement yielded significantly higher bond strength values than MDP-free resin cement (p < 0.05). It was also shown that the Y-TZP surface was etched by the SE solution, causing a large change in the surface topography.CONCLUSIONS: Bond strength significantly improved when a heated HA-based SE solution was applied to the Y-TZP surface, and the etched Y-TZP surface was more irregular and had higher surface roughness.
Ceramics ; Hot Temperature ; Hydrofluoric Acid ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Resin Cements

PURPOSE: The purpose of this study was to evaluate the effects of various protocols and systems for finishing and polishing monolithic zirconia on surface topography, phase transformation, and bacterial adhesion. MATERIALS AND METHODS: Three hundred monolithic zirconia specimens were fabricated and then treated with three finishing and polishing systems (Jota [JO], Meisinger [ME], and Edenta [ED]) using four surface treatment protocols: coarse finishing alone (C); coarse finishing and medium polishing (CM); coarse finishing and fine polishing (CF); and coarse finishing, medium polishing, and fine polishing (CMF). Surface roughness, crystal phase transformation, and bacterial adhesion were evaluated using atomic force microscopy, X-ray diffraction, and streptococcal biofilm formation assay, respectively. One-way and two-way analysis of variance with Tukey post hoc tests were used to analyze the results (α=.05). RESULTS: In this study, the surface treatment protocols and systems had significant effects on the resulting roughness. The CMF protocol produced the lowest roughness values, followed by CM and CF. Use of the JO system produced the lowest roughness values and the smallest biofilm mass, while the ME system produced the smallest partial transformation ratio. The ED group exhibited the highest roughness values, biofilm mass, and partial transformation ratio. CONCLUSION: Stepwise surface treatment of monolithic zirconia, combined with careful polishing system selection, is essential to obtaining optimal microstructural and biological surface results.
Bacterial Adhesion ; Biofilms ; Clinical Protocols ; Dental Polishing ; Microscopy, Atomic Force ; X-Ray Diffraction

OBJECTIVE: The aim of this study was to analyze the surface composition, roughness, and relative friction of metal clips from various ceramic self-ligating brackets. METHODS: Six kinds of brackets were examined. The control group (mC) consisted of interactive metal self-ligating brackets while the experimental group (CC, EC, MA, QK, and WA) consisted of interactive ceramic self-ligating brackets. Atomic force microscopy-lateral force microscopy and scanning electron microscopy-energy-dispersive X-ray spectroscopy were used to analyze the surface of each bracket clip. RESULTS: All the clips in the experimental groups were coated with rhodium except for the QK clip. The results showed that the QK clip had the lowest average roughness on the outer surface, followed by the MA, EC, WA, and CC clips. However, the CC clip had the lowest average roughness on the inner surface, followed by the QK, WA, MA, and EC clips. The QK clip also had the lowest relative friction on the outer surface, followed by the MA, EC, CC, and WA clips. Likewise, the CC clip had the lowest relative friction on the inner surface, followed by the QK, WA, MA, and EC clips. CONCLUSIONS: The surface roughness and relative friction of the rhodium-coated clips were generally higher than those of the uncoated clips.
Ceramics* ; Friction ; Microscopy, Atomic Force ; Rhodium ; Spectrum Analysis

BACKGROUND: Osteolytic metastasis is a common destructive form of metastasis, in which there is an increased bone resorption but impaired bone formation. It is hypothesized that the changed mechanical properties of tumor affected bone cells could inhibit its mechanosensing, thus contributing to differences in bone remodeling. METHODS: Here, atomic force microscopy indentation on primary bone cells exposed to 50% conditioned medium from Walker 256 (W) carcinoma cell line or its adaptive tumor (T) cells was carried out. Nitric oxide levels of bone cells were monitored in response to low-magnitude, high-frequency (LMHF) vibrations. RESULTS: A stronger sustained inhibitive effect on bone cell viability and differentiation by T cells as compared to that of its cell line was demonstrated. This could be attributed to the higher levels of transforming growth factor-β1 (TGF-β1) in the T-conditioned medium as compared to W-conditioned medium. Bone cell elastic moduli in W and T-groups were found to decrease significantly by 61.0% and 69.6%, respectively compared to control and corresponded to filamentous actin changes. Nitric oxide responses were significantly inhibited in T-conditioned group but not in W-conditioned group. CONCLUSIONS: It implied that a change in cell mechanical properties is not sufficient as an indicator of change in mechanosensing ability. Moreover, inhibition of phosphoinositide 3-kinase/Akt downstream signaling pathway of TGF-β1 alleviated the inhibition effects on mechanosensing in T-conditioned cells, further suggesting that growth factors such as TGF-β could be good therapeutic targets for osteoblast treatment.
Actins ; Bone Neoplasms ; Bone Remodeling ; Bone Resorption ; Cell Line ; Cell Survival ; Culture Media, Conditioned ; Elastic Modulus ; Intercellular Signaling Peptides and Proteins ; Microscopy, Atomic Force ; Neoplasm Metastasis ; Nitric Oxide ; Osteoblasts ; Osteogenesis ; Sensitivity Training Groups ; T-Lymphocytes ; Vibration ; Walkers

OBJECTIVETo study the effects of Astragalus polysaccharide (APS), the primary effective component of the Chinese herb medicine Astragalus membranaceus (frequently used for its anti-hepatic fibrosis effects), on nanoscale mechanical properties of liver sinusoidal endothelial cells (SECs).

METHODSUsing endothelial cell medium as the control, 5 experimental groups were established utilizing different concentrations of APS, i.e. 12.5, 25, 50, 100, and 200 μg/mL. By using atomic force microscopy along with a microcantilever modified with a silicon dioxide microsphere as powerful tools, the value of Young's modulus in each group was calculated. SAS 9.1 software was applied to analyze the values of Young's modulus at the pressed depth of 300 nm. Environmental scanning electron microscopy was performed to observe the surface microtopography of the SECs.

RESULTSThe value of Young's modulus in each APS experimental group was significantly greater than that of the control group: as APS concentration increased, the value of Young's modulus presented as an increasing trend. The difference between the low-concentration (12.5 and 25 μg/mL) and high-concentration (200 μg/mL) groups was statistically significant (P<0.05), but no significant differences were observed between moderateconcentration (50 and 100 μg/mL) groups versus low- or high-concentration groups (P>0.05). Surface topography demonstrated that APS was capable of increasing the total area of fenestrae.

CONCLUSIONSThe values of Young's modulus increased along with increasing concentrations of APS, suggesting that the stiffness of SECs increases gradually as a function of APS concentration. The observed changes in SEC mechanical properties may provide a new avenue for mechanistic research of anti-hepatic fibrosis treatments in Chinese medicine.

Animals ; Astragalus Plant ; chemistry ; Biomechanical Phenomena ; drug effects ; Elastic Modulus ; Endothelial Cells ; cytology ; ultrastructure ; Liver ; cytology ; Microscopy, Atomic Force ; Microspheres ; Nanotechnology ; Polysaccharides ; pharmacology ; Rats ; Silicon Dioxide ; chemistry ; Surface Properties

OBJECTIVES: To examine the surface topography of intact WaveOne (WO; Dentsply Sirona Endodontics) and WaveOne Gold (WOG; Dentsply Sirona Endodontics) nickel-titanium rotary files and to evaluate the presence of alterations to the surface topography after root canal preparations of severely curved root canals in molar teeth. MATERIALS AND METHODS: Forty-eight severely curved canals of extracted molar teeth were divided into 2 groups (n = 24/each group). In group 1, the canals were prepared using WO and in group 2, the canals were prepared using WOG files. After the preparation of 3 root canals, instruments were subjected to atomic force microscopy analysis. Average roughness and root mean square values were chosen to investigate the surface features of endodontic files. The data was analyzed using one-way analysis of variance and post hoc Tamhane's tests at 5% significant level. RESULTS: The surface roughness values of WO and WOG files significantly changed after use in root canals (p < 0.05). The used WOG files exhibited higher surface roughness change when compared with the used WO files (p < 0.05). CONCLUSIONS: Using WO and WOG Primary files in 3 root canals affected the surface topography of the files. After being used in root canals, the WOG files showed a higher level of surface porosity value than the WO files.
Dental Pulp Cavity ; Microscopy, Atomic Force ; Molar ; Porosity ; Root Canal Preparation ; Tooth

PURPOSE: The purpose of this in vitro study is to examine the effects of a nano-structured alumina coating on the adhesion between resin cements and zirconia ceramics using a four-point bending test. MATERIALS AND METHODS: 100 pairs of zirconium bar specimens were prepared with dimensions of 25 mm × 2 mm × 5 mm and cementation surfaces of 5 mm × 2 mm. The samples were divided into 5 groups of 20 pairs each. The groups are as follows: Group I (C) – Control with no surface modification, Group II (APA) – airborne-particle-abrasion with 110 µm high-purity aluminum oxide (Al2O3) particles, Group III (ROC) – airborne-particle-abrasion with 110 µm silica modified aluminum oxide (Al₂O₃ + SiO2) particles, Group IV (TCS) – tribochemical silica coated with Al2O3 particles, and Group V (AlC) – nano alumina coating. The surface modifications were assessed on two samples selected from each group by atomic force microscopy and scanning electron microscopy. The samples were cemented with two different self-adhesive resin cements. The bending bond strength was evaluated by mechanical testing. RESULTS: According to the ANOVA results, surface treatments, different cement types, and their interactions were statistically significant (P < .05). The highest flexural bond strengths were obtained in nanostructured alumina coated zirconia surfaces (50.4 MPa) and the lowest values were obtained in the control group (12.00 MPa), both of which were cemented using a self-adhesive resin cement. CONCLUSION: The surface modifications tested in the current study affected the surface roughness and flexural bond strength of zirconia. The nano alumina coating method significantly increased the flexural bond strength of zirconia ceramics.
Aluminum Oxide ; Cementation ; Ceramics ; In Vitro Techniques ; Methods ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Resin Cements ; Silicon Dioxide ; Zirconium

The aim of this study was to investigate the synergistic effect of cold atmospheric plasma (CAP) treatment and RGD peptide coating for enhancing cellular attachment and proliferation over titanium (Ti) surfaces. The surface structure of CAP-treated and RGD peptide-coated Ti discs were characterized by contact angle goniometer and atomic force microscopy. The effect of such surface modification on human bone marrow derived mesenchymal stem cells (hMSCs) adhesion and proliferation was assessed by cell proliferation and DNA content assays. Besides, hMSCs' adhesion and morphology on surface modified Ti discs were observed via fluorescent and scanning electron microscopy. RGD peptide coating following CAP treatment significantly enhanced cellular adhesion and proliferation among untreated, CAP-treated and RGD peptide-coated Ti discs. The treatment of Ti surfaces with CAP may contribute to improved RGD peptide coating, which enables increased cellular integrations with the Ti surfaces.
Bone Marrow ; Cell Proliferation ; DNA ; Humans ; Mesenchymal Stromal Cells ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Plasma ; Plasma Gases ; Titanium

BACKGROUND: The mechanical deformability of cancer cells has attracted particular attention as an emerging biomarker for the prediction of anti-cancer drug sensitivity. Nevertheless, it has not been possible to establish a general rubric for the identification of drug susceptibility in breast cancer cells from a mechanical perspective. In the present study, we investigated the mechanical alteration associated with resistance to adjuvant therapy in breast cancer cells. METHODS: We performed an ‘atomic force microscopy (AFM)-based nanomechanical study’ on ‘drug-sensitive (MCF-7)’ and ‘drug-resistant (MCF-7/ADR)’ breast cancer cells. We also conducted cell viability tests to evaluate the difference in doxorubicin responsiveness between two breast cancer cell lines. We carried out a wound closure experiment to investigate the motility changes associated with chemotherapeutic resistance. To elucidate the changes in molecular alteration that accompany chemotherapeutic resistance, we investigated the expression of vinculin and integrin-linked kinase-1–which are proteins involved in substrate adhesion and the actin cytoskeleton–using Western blotting analysis. RESULTS: A MTT assay confirmed that the dose-dependent efficacy of doxorubicin was reduced in MCF-7/ADR cells compared to that in MCF-7 cells. The wound assay revealed enhanced two-dimensional motility in the MCF-7/ADR cells. The AFM mechanical assay showed evidence that the drug-resistant breast cancer cells exhibited a significant decrease in mechanical deformability compared to their drug-sensitive counterparts. The mechanical alteration in the MCF-7/ADR cells was accompanied by upregulated vinculin expression. CONCLUSIONS: The obtained results manifestly showed that the altered mechanical signatures–including mechanical deformability and motility–were closely related with drug resistance in the breast cancer cells. We believe that this investigation has improved our understanding of the chemotherapeutic susceptibility of breast cancer cells.
Actins ; Biophysics ; Blotting, Western ; Breast Neoplasms ; Breast ; Cell Line ; Cell Survival ; Doxorubicin ; Drug Resistance ; Drug Resistance, Multiple ; Elastic Modulus ; MCF-7 Cells ; Microscopy, Atomic Force ; Vinculin ; Wounds and Injuries