Youjin Cheong

Ultrastructural investigation of intact orbital implant surfaces using atomic force microscopy

This study examined the surface nanostructures of three orbital implants: nonporous poly(methyl methacrylate) (PMMA), porous aluminum oxide and porous polyethylene. The morphological characteristics of the orbital implants surfaces were observed by atomic force microscopy (AFM). The AFM topography, phase shift and deflection images of the intact implant samples were obtained. The surface of the nonporous PMMA implant showed severe scratches and debris. The surface of the aluminum oxide implant showed a porous structure with varying densities and sizes. The PMMA implant showed nodule nanostructures, 215.56 ± 52.34 nm in size, and the aluminum oxide implant showed crystal structures, 730.22 ± 341.02 nm in size. The nonporous PMMA implant showed the lowest roughness compared with other implant biomaterials, followed by the porous aluminum oxide implant. The porous polyethylene implant showed the highest roughness and severe surface irregularities. Overall, the surface roughness of orbital implants might be associated with the rate of complications and cell adhesion.

Changes in ultrastructure and properties of bracket slots after orthodontic treatment with bicuspid extraction.

This study examined the effects of an orthodontic treatment using a bicuspid extraction on the surface roughness and mechanical properties of stainless steel (SS) brackets adjacent to the extraction space. Four experimental groups were employed; groups 1 and 2 used the Archist(®) SS brackets before and after the extraction treatment, respectively, and groups 3 and 4 used the Victory(®) SS brackets before and after the extraction treatment, respectively. The slot surfaces of the bracket were scanned in air at a resolution of 512 × 512 pixels with a scan speed of 0.8 line/s. The visco-elasticity of the bracket slot was determined from the force-distance curves of atomic force microscopy. The orthodontic treatment with bicuspid extraction led to a significant increase (p<0.0001) in surface roughness in both groups. In particular, the Archist(®) SS brackets showed more changes than the Victory(®) SS brackets (p<0.0005). However, there was no significant difference in properties of the Victory(®) and Archist(®) brackets between before and after treatment. This suggests that the orthodontic treatment with bicuspid extraction is more responsible for the changes in surface roughness than the properties of the brackets.

Effect of fluoride pretreatment on primary and permanent tooth surfaces by acid‐etching

This study observed the effect of fluoride application on a 37% phosphoric acid etching for 20 s of the enamel surfaces of primary and permanent teeth based on a clinical protocol employed in dental hospitals, through atomic force microscopy and scanning electron microscopy. Enamel samples were prepared from 84 exfoliated and noncarious teeth. Primary (groups 1-4) and permanent (groups 5-8) tooth samples were assigned randomly to one of eight groups based on the timing of acid-etching with 37% phosphoric acid after an acidulated phosphate fluoride (APF) treatment. Groups 1 and 5 received no fluoride application. Groups 2-4 and 6-8 were pretreated with fluoride and received acid-etching 2 weeks later (groups 2 and 6), 1 week later (groups 3 and 7), and immediately (groups 4 and 8). The acid-etching process led to a significant increase in roughness (p<0.0001), and the APF treatment led to a decrease in primary and permanent tooth surface roughness (p<0.005). An acid-etching procedure 2 weeks after performing an APF pretreatment might be recommended to obtain the maximum enamel adhesion of a resin composite.

Label-free and quantitative evaluation of cytotoxicity based on surface nanostructure and biophysical property of cells utilizing AFM.

In this study, the four commonly used cytotoxicity assays and the mechanical properties as evaluated by atomic force microscopy (AFM) were compared in a cellular system. A cytotoxicity assay is the first and most essential test to evaluate biocompatibility of various toxic substances. Many of the cytotoxicity methods require complicated and labor-intensive process, as well as introduce experimental error. In addition, these methods cannot provide instantaneous and quantitative cell viability information. AFM has become an exciting analytical tool in medical, biological, and biophysical research due to its unique abilities. AFM-based force-distance curve measurements precisely measure the changes in the biophysical properties of the cell. Therefore, we observed the morphological changes and mechanical property changes in L929 cells following sodium lauryl sulfate (SLS) treatment utilizing AFM. AFM imaging showed that the toxic effects of SLS changed not only the spindle-like shape of L929 cells into a round shape, but also made a rough cell surface. As the concentration of SLS was increased, the surface roughness of L929 cell was increased, and stiffness decreased. We confirmed that inhibition of proliferation clearly increased with increases in SLS concentration based on results from MTT, WST, neutral red uptake, and LIVE/DEAD viability/cytotoxicity assays. The estimated IC₅₀ value by AFM analysis was similar to those of other conventional assays and was included within the 95% confidence interval range. We suggest that an AFM quantitative analysis of the morphological and biophysical changes in cells can be utilized as a new method for evaluating cytotoxicity.

Evaluation of aneurysm-associated wall shear stress related to morphological variations of circle of Willis using a microfluidic device

Although microfluidic systems have been important tools in analytical chemistry, life sciences, and medical research, their application was rather limited for drug-screening and biosensors. Here, we described a microfluidic device consisting of a multilayer micro-channel system that represented the hemodynamic cerebral vascular system. We analyzed wall shear stresses related to aneurysm formation in the circle of Willis (CoW) and their morphological variations using this system. This device was controlled by pneumatic valves, which occluded various major arteries by closing the associated channels. The hemodynamic analysis indicated that higher degrees of shear stress occurred in an anterior communicating artery (ACoA), particularly in the hypoplastic region of the posterior communicating artery (PCoA) and the P1 segment. Furthermore, occlusion of a common carotid artery (CCA) or a middle cerebral artery (MCA) increased the shear stress, whereas occlusion of a vertebral artery (VA) decreased the shear stress. These results indicate that the morphological variation of the CoW may affect aneurysm formation resulting from increased wall shear stress. Therefore, the technique described in this paper provides a novel method to investigate the hemodynamics of complex cerebral vascular systems not accessible from previous clinical studies.

Structural and biomechanical effects of photooxidative collagen cross-linking with photosensitizer riboflavin and 370 nm UVA light on human corneoscleral tissues.

This study quantitatively investigated the immediate effects of a photooxidative collagen cross-linking treatment with photosensitizer riboflavin (RF) and 370 nm UVA light in in vitro human corneoscleral collagen fibrils using histology, thickness, scanning electron microscopy, and atomic force microscopy analyses. Twenty 8 x 2 mm corneoscleral strips were dissected sagittally from donor tissue using a scalpel. Four parameters were investigated, including the density, thickness, adhesion force, and stiffness of corneoscleral tissues before and after the collagen cross-linking treatment. The RFUVA-catalyzed collagen cross-linking treatment led to an increase in the density of both corneal (8%) and scleral (23%) stromal collagens. However, there was no difference in corneoscleral thickness. Furthermore, RFUVA-catalyzed collagen cross-linking treatment led to an increased biomechanical response of corneosclera: 25 and 8% increases in corneoscleral stiffness, and 24 and 22% increases in corneoscleral adhesion force. The collagen cross-linking treatment through RF-sensitized photoreaction may cause structural and biomechanical changes in the collagen fibril network of the cornea and the sclera. This is due to narrowing of the interfibrillar spacing and the stromal edema.

AFM Study for Morphological Characteristics and Biomechanical Properties of Human Cataract Anterior Lens Capsules

The aim of this study was to quantitatively investigate the morphologies (surface roughness) and biomechanical properties (Youngs modulus) of human anterior lens capsules (ALCs) for noncataract and cataract groups using atomic force microscopy. Eight human ALCs obtained during phacoemulsification from patients with senile cataracts (72 ± 13 years) were investigated in both the hydrated and dehydrated conditions. The cataract group showed clearly the proliferated lens epithelial cells (LECs) with a monomorphic cell structure, a diameter of 12.54 ± 4.31 μm, and a height of 0.23 ± 0.04 μm, whereas the control group showed no LECs. A substantial amount of false-positive calcification was observed caused by the deposition of remnants of dried salt solution. Cataract group showed significantly higher surface roughness (382.06 nm, p ≤ 0.001) than control group in the anterior side of ALCs, whereas cataract group showed significantly lower surface roughness (353.79 nm, p ≤ 0.001) than control group in their posterior side. Cataract group showed significantly higher Youngs modulus (69.52 kPa, p ≤ 0.001) compared to the control group, regardless of the ALC side. Therefore, it is significant that this study provides a new method to examine the nanostructural characteristic and biomechanical property of human ALCs through a nanometer-scale resolution microscopy technique.

Potential effects of tooth‐brushing on human dentin wear following exposure to acidic soft drinks

This study used scanning electron microscopy and atomic force microscopy to examine the short‐term potential effects of brushing time and the start‐time of tooth‐brushing after demineralization on primary dentin wear in vitro. Thirty‐six noncarious primary central incisors were assigned to 12 experimental groups. Exposure to cola drinks was used to initiate the demineralization process. Three brushing times (5, 15 and 30 s) and four start‐times of brushing (0, 30, 60 and 120 min) after an erosive attack were used for the abrasion process. Tooth‐brushing the softened dentin surface led to increases in the open tubular fraction and microstructural changes on the dentin surface. Brushing immediately after exposure to cola resulted in the greatest irreversible dentin loss, whereas brushing 60 or 120 min after pretreatment resulted in the least irreversible dentin loss. However, brushing time had no effect on the irreversible loss of dentin wear. Based on these experimental results, tooth‐brushing should be performed at least 60 min after consuming a cola drink to achieve the desired tooth cleaning and avoid the introduction of surface lesions on dentin.

Short-term effect of cryotherapy on human scleral tissue by atomic force microscopy.

This study investigated the inflammatory effect of cryotherapy application on collagen matrix network in human infant sclera. Donor scleral tissues taken from three infant patients divided into five groups: control group, sham-treated group, and three cryotreated groups. In the cryotherapy groups, the sclera was treated for 5 s, 10 s, and 20 s with -80°C freezing by a cryosurgical system. The cryotreated reactions were examined using double histological analysis with hematoxylin-eosin and Massons trichrome, and atomic force microscopy analysis to quantify the diameter and D-banding of collagen fibrils. The infant scleral tissues treated with cryotherapy showed a significantly increased collagen density associated with inflammatory response (p < 0.05), increased fibril diameter (p < 0.005) compared to the scleral tissues in the control group. The results directly suggest that the cryotherapy affects the morphology of scleral collagen.

Ultrastructural effect of self-ligating bracket materials on stainless steel and superelastic niTi wire surfaces

Frictional interactions between wires and brackets reduce the efficacy in orthodontic treatments. Self‐ligating brackets (SLBs) are now more often used due to lower frictional forces when compared with conventional‐ligating brackets. In this study, scanning electron microscopy and atomic force microscopy were used to examine the microstructural effects of stainless steel and ceramic SLBs on the surface roughness of stainless steel and superelastic NiTi wires both after in vivo clinical orthodontic treatment as well as in in vitro three‐point bending experiments. A combination of two wires—0.019 in. × 0.025 in. stainless steel wires and 0.016 in. superelastic NiTi wires—and two SLBs—both passive‐type stainless steel SLBs and active‐type ceramic SLBs—was applied for 4 months (bicuspid‐extraction) in an in vivo setting and for 1 month in an in vitro setting (200 g loads). After the SLB treatments, all wires exhibited severe scratches secondary to frictional interactions with the brackets. When used with the stainless steel SLBs (Damon 3MX®), the surfaces of 0.019 in. × 0.025 in. stainless steel (P < 0.0001) and 0.016 in. superelastic NiTi wires (P < 0.05) were significantly smoother than when used with the ceramic SLBs (Clippy‐C®). Such results suggest that orthodontic treatments with stainless steel SLBs are more effective than with ceramic SLBs. Microsc. Res. Tech. 75:1076–1083, 2012.