Lithium-Modified TiO2 Surface by Anodization for Enhanced Protein Adsorption and Cell Adhesion.

Promoting osseointegration is an essential step in improving implant success rates. Lithium has gradually gained popularity for promoting alkaline phosphatase activity and osteogenic gene expression in osteoblasts. The incorporation of lithium into a titanium surface has been reported to change its surface charge, thereby enhancing its biocompatibility. In this study, we applied anodization as a novel approach to immobilizing Li on a titanium surface and evaluated the changes in its surface characteristics. The objective of this study was to determine the effect of Li treatment of titanium on typical proteins, such as albumin, laminin, and fibronectin, in terms of their adsorption level as well as on the attachment of osteoblast cells. Titanium disks were acid-etched by 66 wt % H2SO4 at 120 °C for 90 s and set as the control group. The etched samples were placed in contact with an anode, while a platinum bar served as the counter electrode. Both electrodes were mounted on a custom electrochemical cell filled with 1 M LiCl. The samples were anodized at constant voltages of 1, 3, and 9 V. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) results showed no significant differences in the topography. However, the ζ potentials of the 3 V group were higher than those of the control group at a physiological pH of 7.4. Interestingly, the adsorption level of the extracellular matrix protein was mostly enhanced on the 3 V-anodized surface. The number of attached cells on the Li-anodized surfaces increased. The localization of vinculin at the tips of the stretching cytoplasmic projections was observed more frequently in the osteoblasts on the 3 V-anodized surface. Although the optimal concentration or voltage for Li application should be investigated further, this study suggests that anodization could be an effective method to immobilize lithium ions on a titanium surface and that modifying the surface charge characteristics enables a direct protein-to-material interaction with enhanced biological adhesion.

A review in titanium-zirconium binary alloy for use in dental implants: Is there an ideal Ti-Zr composing ratio?

Binary titanium-zirconium alloys have been studied as promising alternatives for Ti implants. The commercial Ti-15Zr alloy (Roxolid, Straumann) has been the major subject of numerous binary Ti-Zr alloys-related studies and has gained wide recognition in laboratory studies and clinical practices. However, binary Ti-Zr alloys of other composition ratios are still being investigated by researchers. This review aims to provide information on the potential of binary Ti-Zr alloys other than Ti-15Zr as implant materials in terms of mechanical strengths, chemical or electrochemical corrosion resistance capabilities, and biological performances. In addition, in this review, the Ti-15Zr alloy is discussed only when compared with other binary Ti-Zr alloys. From the included 26 studies, it is confirmed that the mechanical, chemical, electrochemical, and biological properties of Ti-Zr alloys are related to the Ti and Zr composition ratio in the alloy, phase, manufacturing process, and surface treatment. Among the studied alloys, α-or α' phase-Ti-5 wt, 45 wt/30at, and 50 wt. %Zr exhibited relatively more promising results for further investigation. More research is necessary to evaluate the potential for future use of these materials for implants.

A Comparative Study on Fungal Diversity in Organic and Conventionally Cultivated Lemons During Accelerated Storage.

There has been a growing interest in organic farming as a countermeasure to the environmental burden caused by chemical pesticides. We analyzed and compared the fungal diversity of lemon fruits from organic and conventional cultivation by automated rRNA intergenic spacer analysis (ARISA), accompanied by isolation of cultured colonies and metagenomic analysis. Lemon peels were cut out and subjected to the analyses at purchase and after accelerated storage at 28 °C. The organic lemons did not decay even after 14 weeks, while most of the conventional lemons did decay. The fungal colony counts were not significantly different, although the number of fungal species together with the Shannon index, considering the abundance of each species, clearly showed more diversity in organic lemons than in conventional lemons (p = 0.011). Fusarium sp. (putative F. solani) accounted for as much as 90% of the relative abundance in the decayed conventional lemons. Metagenomic analysis also supported the lack of fungal diversity in conventional lemons. These results may suggest that organic cultivation maintains the diversity of native fungal flora in lemon fruit and could contribute to preventing decay during ambient storage.

Knowledge acquisition efficacy of a remote flipped classroom on learning about removable partial dentures.

PURPOSE: Coronavirus disease 2019 has forced the use of remote education worldwide. In 2020, the authors conducted a fully digitized, remote flipped (RF) classroom focusing on removable partial dentures. This study aimed to evaluate the learning outcomes of the RF classroom and compare these with those of the on-site classroom. METHODS: The fourth-year dental students in 2017-2019 attended on-site flipped classroom (OF; n=70) or on-site lecture classroom (OL; n=67), and those in 2020 (n=58) attended the RF classroom. Lecture videos and text were provided online in the OF and RF groups, while only the text was given to the OL group. Individual and group tests were conducted to assess knowledge acquisition one week after all the classes ended. A one-way analysis of variance following Tukey's test was performed to compare individual/group test scores among the OF, OL, and RF groups. Multilevel linear regression analysis was performed to identify the factors associated with an increase in each test score. RESULTS: The mean individual test scores in the RF and OF groups were significantly higher than in the OL group (P<0.01). The mean group test score in the RF group was significantly higher than in the OF and OL groups (P<0.001). The multilevel analysis identified remote and flipped classroom styles in the individual test, and the remote style in the group test, as significant effective factors in learning (P<0.05). CONCLUSIONS: These results suggest that the RF classroom is more effective than the on-site lecture in knowledge acquisition in undergraduate prosthodontic education.

Impaired dental implant osseointegration in rat with streptozotocin-induced diabetes.

OBJECTIVE: Few studies have reported on the impact of oxidative stress on the dental implant failure. The aim of this study was to investigate the impact of hyperglycemia-induced oxidative stress on dental implant osseointegration in diabetes mellitus (DM). METHODS: Acid-treated titanium implants were bilaterally placed in the maxillary alveolar ridge of streptozotocin-induced diabetic (DM group) and control rats after extraction of first molars. Histological analysis and micro-push-out test were performed 4 weeks after surgery. Oxidative stress and osteogenic markers in the surrounding bone were quantified by real-time polymerase chain reaction. In the in vitro study, rat bone marrow-derived mesenchymal stem cells (BMMSCs) were cultured on acid-treated titanium discs in a high-glucose (HG) or normal environment. Intracellular reactive oxygen species (ROS), cell proliferation, alkaline phosphatase (ALP) activity, and extracellular calcification were evaluated following antioxidant treatment with N-acetyl-L-cysteine (NAC). RESULTS: The implant survival rate was 92.9% and 75.0% in control and DM group, respectively. Bone-implant contact and push-out loads were significantly lower in the DM group. Expression of superoxide dismutase 1 at the mRNA level and on immunohistochemistry was significantly lower in the DM group. In vitro experiments revealed that the HG condition significantly increased ROS expression and suppressed the proliferation and extracellular calcification of BMMSCs, while NAC treatment significantly restored ROS expression, cell proliferation, and calcification. The ALP activity of both groups was not significantly different. CONCLUSION: In diabetes, high-glucose-induced oxidative stress downregulates proliferation and calcification of BMMSCs, impairing osseointegration and leading to implant failure.

A systematic review of digital removable partial dentures. Part I: Clinical evidence, digital impression, and maxillomandibular relationship record.

PURPOSE: This study comprehensively reviewed the current status of digital workflows in fabricating removable partial dentures (RPDs) using evidence from clinical trials and case reports. STUDY SELECTION: We performed a systematic review of the literature on the materials and fabrication of RPDs using digital technologies published in online databases from 1980 to 2020. We selected eligible articles from the search results, retrieved information on digital RPDs from these, and conducted a qualitative analysis. We report evidence from clinical papers and case reports, digital impression-taking methods, and maxillomandibular relationship (MMR) records. RESULTS: A case report electronically published in 2019 introduced a clasp-retained RPD fabricated via a full-digital workflow without a gypsum definitive cast. Computer-aided design and computer-aided manufacturing of double-crown-retained RPDs with nonmetal materials were described in some case reports. Intraoral scanners were used to obtain digital impressions and MMR records in the fabrication of digital RPDs, which have potential advantages for reducing the number of clinical appointments and simplifying laboratory procedures. Evidence from clinical trials is scarce; a randomized controlled trial reported higher patient satisfaction with digital clasp-retained RPDs than with conventional RPDs. CONCLUSIONS: Full-digital RPDs can be fabricated without a gypsum definitive cast. However, the indication for full-digital RPDs is limited to cases with Kennedy Class III/IV partially edentulous arches with several missing teeth. Challenges in digital impression-taking and MMR recording remain to be solved to extend these indications. More evidence from clinical trials is required to evaluate the efficacy and usefulness of digital RPDs.

A systematic review of digital removable partial dentures. Part II: CAD/CAM framework, artificial teeth, and denture base.

PURPOSE: This study comprehensively reviewed the current status of the digital workflow of removable partial dentures (RPDs) and summarized information about the fabrication methods and material properties of the dental framework, artificial teeth, and denture base. STUDY SELECTION: We performed a systematic review of the literature published in online databases from January 1980 to April 2020 regarding RPD fabrication and materials used in the related digital technology. We selected eligible articles, retrieved information regarding digital RPDs, and conducted qualitative/quantitative analyses. In this paper, the computer-aided design/computer-aided manufacturing (CAD/CAM) framework, artificial teeth, and denture base materials are reported. RESULTS: A variety of materials, such as cobalt-chromium alloy, titanium, zirconia, and polyether ether ketone, are used for dental CAD/CAM frameworks. The mechanical strength of the metal materials used for the CAD/CAM framework was superior to that of the cast framework. However, the fitness and surface roughness of the framework and clasp fabricated using a selective laser melting (SLM) method were not superior to those obtained via cast fabrication. Most material properties and the surface roughness of poly methyl methacrylate (PMMA) discs used for digital RPDs were superior to those of heat-cured PMMA. CONCLUSION: The use of a CAD/CAM framework and PMMA disc for digital RPDs offers numerous advantages over conventional RPDs. However, technical challenges regarding the accuracy and durability of adhesion between the framework and denture base remain to be solved. In digital fabrication, human technical factors influence the quality of the framework.

Surface properties and biocompatibility of sandblasted and acid-etched titanium-zirconium binary alloys with various compositions.

Ti-Zr alloys have been investigated as an alternative to commercially pure Ti (c.p.Ti). According to our previous studies on the mechanical properties of Ti-Zr alloys, a Zr proportion in the range of 30-50 mol% has competitive advantages over Ti-10Zr and c.p.Ti. The aim of this study is to evaluate the biological response to Ti-Zr alloys with different compositions and their surface characteristics. Alloy surfaces are modified by sandblasting and sulfuric acid etching. As a result, similar surface structures are observed for c.p.Ti, Ti-10Zr, and Ti-30Zr, whereas Ti-50Zr does not form a micro-rough structure by the same treatment process. No significant difference is found in the viability of cells on c.p.Ti, Ti-10Zr, and Ti-30Zr, whereas lower cell attachment levels are detected on Ti-50Zr. In summary, Ti-30Zr reliably forms a micro-rough structure, which provides one evidence for its application in a new dental implant material.

Ultraviolet Treatment of Titanium to Enhance Adhesion and Retention of Oral Mucosa Connective Tissue and Fibroblasts.

Peri-implantitis is an unsolved but critical problem with dental implants. It is postulated that creating a seal of gingival soft tissue around the implant neck is key to preventing peri-implantitis. The objective of this study was to determine the effect of UV surface treatment of titanium disks on the adhesion strength and retention time of oral connective tissues as well as on the adherence of mucosal fibroblasts. Titanium disks with a smooth machined surface were prepared and treated with UV light for 15 min. Keratinized mucosal tissue sections (3 × 3 mm) from rat palates were incubated for 24 h on the titanium disks. The adhered tissue sections were then mechanically detached by agitating the culture dishes. The tissue sections remained adherent for significantly longer (15.5 h) on the UV-treated disks than on the untreated control disks (7.5 h). A total of 94% of the tissue sections were adherent for 5 h or longer on the UV-treated disks, whereas only 50% of the sections remained on the control disks for 5 h. The adhesion strength of the tissue sections to the titanium disks, as measured by tensile testing, was six times greater after UV treatment. In the culture studies, mucosal fibroblasts extracted from rat palates were attached to titanium disks by incubating for 24, 48, or 96 h. The number of attached cells was consistently 15-30% greater on the UV-treated disks than on the control disks. The cells were then subjected to mechanical or chemical (trypsinization) detachment. After mechanical detachment, the residual cell rates on the UV-treated surfaces after 24 and 48 h of incubation were 35% and 25% higher, respectively, than those on the control surfaces. The remaining rate after chemical detachment was 74% on the control surface and 88% on the UV-treated surface for the cells cultured for 48 h. These trends were also confirmed in mouse embryonic fibroblasts, with an intense expression of vinculin, a focal adhesion protein, on the UV-treated disks even after detachment. The UV-treated titanium was superhydrophilic, whereas the control titanium was hydrophobic. X-ray photoelectron spectroscopy (XPS) chemical analysis revealed that the amount of carbon at the surface was significantly reduced after UV treatment, while the amount of TiOH molecules was increased. These ex vivo and in vitro results indicate that the UV treatment of titanium increases the adhesion and retention of oral mucosa connective tissue as a result of increased resistance of constituent fibroblasts against exogenous detachment, both mechanically and chemically, as well as UV-induced physicochemical changes of the titanium surface.

Multilevel factor analysis of flipped classroom in dental education: A 3-year randomized controlled trial.

PURPOSE: Previous studies have rarely attempted to test the confounding factors that may affect learning outcomes of the flipped classroom. The purpose of this study was to assess how flipped classrooms affect the acquisition of knowledge in clinical dental education based on multilevel factor analysis. METHOD: The authors conducted a 3-year (2017, 2018, and 2019) randomized controlled trial in a series of introductory prosthodontics courses in dental education. A total of 137 participants were randomly assigned to flipped classroom (n = 70, 51%) or lecture (n = 67, 49%) formats. The flipped group was instructed to self-learn knowledge-based content through online preparation materials, including videos and text, while the lecture group was given text only. Both groups were provided with the same study content and opportunities for different styles of learning. The session attendance rate and number of times the materials were accessed were monitored. Individual and team readiness assurance tests (IRAT/TRAT) were conducted to evaluate knowledge acquisition. A multilevel linear regression analysis was conducted on both instructional styles (flipped vs. lecture) as an intervention factor, and confounding factors that could affect the outcomes were implemented. RESULTS: The average number of online accesses was 2.5 times per session in the flipped group and 1.2 in the lecture group, with a significant difference (p < .05). The average IRAT score was significantly higher in the flipped than in the lecture group (effect size [ES] 0.58, p < .001). The number of online accesses was significantly and positively correlated with IRAT scores (0.6 [0.4, 0.8]). The instructional style was significantly and positively correlated with TRAT scores (coefficient [95% confidence interval]: 4.6 [2.0, 7.3]), but it was not correlated with IRAT (4.3 [-0.45, 9.0]). CONCLUSIONS: The flipped classroom was more effective than the lecture format regarding knowledge acquisition; however, the decisive factor was not the instructional style but the number of individual learning occasions. The employment of the flipped classroom was the decisive factor for team-based learning outcomes.

UV-Mediated Photofunctionalization of Indirect Restorative Materials Enhances Bonding to a Resin-Based Luting Agent.

PURPOSE: The potential of UV-mediated photofunctionalization to enhance the resin-based luting agent bonding performance to aged materials was investigated. METHODS: Sixty samples of each material were prepared. Yttria-stabilized zirconia (YZr) and Pd-Au alloy (Pd-Au) plates were fabricated and sandblasted. Lithium disilicate glass-ceramic (LDS) was CAD-CAM prepared and ground with #800 SiC paper. Half of the specimens were immersed in machine oil for 24 h to simulate the carbon adsorption. Then, all of the specimens (noncarbon- and carbon-adsorbed) were submitted to UV-mediated photofunctionalization with a 15 W UV-LED (265 nm, 300 mA, 7692 µW/cm2) for 0 (control groups), 5, and 15 min and subjected to contact angle (Ɵ) measurement and bonded using a resin cement (Panavia™ V5, Kuraray Noritake, Japan). The tensile bond strength (TBS) test was performed after 24 h. The Ɵ (°) and TBS (MPa) data were statistically analyzed using two-way ANOVA and Bonferroni correction tests (α = 0.05). RESULTS: In the carbon-adsorbed groups, UV-mediated photofunctionalization for 5 min significantly decreased Ɵ of all materials and increased TBS of YZr, and UV for 15 min significantly increased the TBS of LDS and Pd-Au. In noncarbon-adsorbed groups, UV-photofunctionalization did not significantly change the Ɵ or TBS except YZr specimens UV-photofunctionalized for 15 min. CONCLUSION: UV-mediated photofunctionalization might have removed the adsorbed hydrocarbon molecules from the materials' surfaces and enhanced bond strengths of Panavia™ V5 to YZr, LDS, and Pd-Au. Additionally, UV-mediated photofunctionalization improved the overall TBS of YZr. Further investigation on the optimum conditions of UV photofunctionalization on indirect restorative materials should be conducted.

Tumor contact length of prostate cancer determined by a three-dimensional method on multiparametric magnetic resonance imaging predicts extraprostatic extension and biochemical recurrence.

OBJECTIVE: To evaluate the clinical benefit of tumor contact length as a predictor of pathological extraprostatic extension and biochemical recurrence in patients undergoing prostatectomy. METHODS: A total of 91 patients who underwent 3T multiparametric magnetic resonance imaging before prostatectomy from April 2014 to July 2019 were included. A total of 94 prostate cancer foci were analyzed retrospectively. We evaluated maximum tumor contact length, which was determined to be the maximum value in the three-dimensional directions, as a predictor of pathological extraprostatic extension and biochemical recurrence. RESULTS: A total of 19 lesions (20.2%) had positive pathological extraprostatic extension. Areas under the curves showed maximum tumor contact length to be a significantly better parameter to predict pathological extraprostatic extension than the Prostate Imaging Reporting and Data System (P = 0.002), tumor maximal diameter (P = 0.001), prostate-specific antigen (P = 0.020), Gleason score (P < 0.001), and clinical T stage (P < 0.001). Multivariate analysis showed maximum tumor contact length (P = 0.003) to be an independent risk factor for predicting biochemical recurrence. We classified the patients using preoperative factors (prostate-specific antigen >10, Gleason score >3 + 4 and maximum tumor contact length >10 mm) into three groups: (i) high-risk group (patients having all factors); (ii) intermediate-risk group (patients having two of three factors); and (iii) low-risk group (patients having only one or none of the factors). Kaplan-Meier curves showed that the high-risk group had significantly worse biochemical recurrence than the intermediate-risk group (P = 0.042) and low-risk group (P < 0.001). CONCLUSIONS: Our findings suggest that maximum tumor contact length is an independent predictor of pathological extraprostatic extension and biochemical recurrence. A risk stratification system using prostate-specific antigen, Gleason score and maximum tumor contact length might be useful for preoperative assessment of prostate cancer patients.

[Outcomes of Laparoscopic Radical Prostatectomies by a Single Surgeon Alternating Operating Position].

The clinical outcome of laparoscopic radical prostatectomy (LRP) was retrospectively investigated taking into consideration the surgeon's position during the procedure. The study cohort included 184 consecutive patients who had undergone LRP performed by a single surgeon from February 2013 to July 2018. During the study period,the surgeon stood alternately on either the left or right side of the patient. The D'Amico risk classification was low,intermediate and high in 26 (14.1%),45 (24.5%) and 113 (61.4%) patients,respectively. Mean surgical duration was 203.5 minutes and mean estimated blood loss was 437.6 ml. Nerve sparing (NS) was implemented in 82 (44. 6%) patients. The mean period of having an indwelling urethral catheter was 5. 0 days. Perioperative Clavien-Dindo degree ≥IIIa complications occurred in three (1.6%) patients. Except for cases with presurgical hormonal treatment,surgical margins were positive in 41 (22.3%) patients,among whom 23 (17.4%) had pT2 disease. The 5-year biochemical recurrence-free survival rate was 81.4%,and 84.8% of patients regained urinary continence at 12 months after surgery. Where the surgeon stood during LRP was not associated with significant differences in any parameter. However,the margin positive rate was higher on the side away from where the surgeon stood than the side closer to the surgeon (70.7% vs 29.3%). In conclusion,the position of the surgeon during LRP does not influence the outcome.

Correction to: Stiripentol alleviates neuropathic pain in L5 spinal nerve-transected mice.

In the original publication of the article, the value "40-µm thickness" was incorrect in the legend of Figure 4. The correct value is 10-µm thickness.

Stiripentol alleviates neuropathic pain in L5 spinal nerve-transected mice.

PURPOSE: Antiepileptic drugs are used not only for the treatment of epilepsy but also for that of neuropathic pain. However, their action mechanisms have not always been well explained. Stiripentol, an effective antiepileptic drug indicated as a therapeutic for Dravet syndrome, was recently shown to act as an inhibitor of lactate dehydrogenase in astrocytes. In this present study, we examined the effect of stiripentol on neuropathic pain in L5 spinal nerve-transected mice. METHODS: We carried out behavioral tests using calibrated von Frey filaments and the immunohistochemistry of glial fibrillary acidic protein, an astrocyte marker, in L5 spinal nerve-transected mice after intrathecal administration of drugs. RESULTS: Like other anticonvulsants such as gabapentin and carbamazepine, stiripentol alleviated mechanical hyperalgesia induced by L5 spinal nerve transection in a dose-dependent manner, when intrathecally administered to mice 7, 14, and 28 days after L5 spinal nerve transection. Likewise, α-cyano-4-hydroxycinnamic acid, a broad inhibitor of monocarboxylate transporters, diminished mechanical hyperalgesia induced by L5 spinal nerve transection. Simultaneous administration of L-lactate negated the analgesic effect elicited by stiripentol, carbamazepine or α-cyano-4-hydroxycinnamic acid, but not that by gabapentin. None of the anticonvulsants affected the immunoreactivity of glial fibrillary acidic protein. CONCLUSIONS: This present study demonstrated that stiripentol was effective against neuropathic pain and suggested that the astrocyte-neuron lactate shuttle was involved in such pain.

Magnetic resonance imaging/transrectal ultrasonography fusion targeted prostate biopsy finds more significant prostate cancer in biopsy-naïve Japanese men compared with the standard biopsy.

OBJECTIVE: To assess the clinical benefits of magnetic resonance imaging/transrectal ultrasound fusion-targeted biopsy for biopsy-naïve Japanese men. METHODS: Between February 2017 and August 2018, 131 biopsy-naïve men who underwent targeted biopsy together with 10-core systematic biopsy at Hiroshima University Hospital were retrospectively investigated. Multiparametric magnetic resonance imaging findings were reported based on Prostate Imaging Reporting and Data System version 2. RESULTS: The overall cancer detection rates per patient were 69.5% in systematic biopsy + targeted biopsy cores, 61.1% in systematic biopsy cores and 61.1% in targeted biopsy cores. The detection rates for clinically significant prostate cancer were 43.5% in targeted biopsy cores and 35.9% in systematic biopsy cores (P = 0.04), whereas the detection rates for clinically insignificant prostate cancer were 17.6% and 25.2% respectively (P = 0.04). Lesions in the peripheral zone were diagnosed more with clinically significant prostate cancer (54.8% vs 20.7%, P < 0.001) and International Society of Urological Pathology grade (3.2 vs 2.7, P = 0.02) than that in the inner gland. Just 4.2% (3/71) of Prostate Imaging Reporting and Data System category 2 and 3 lesions in the middle or base of the inner gland were found to have clinically significant prostate cancer. The cancer detection rate per core was 42.3% in targeted biopsy cores, whereas it was 17.9% in systematic biopsy cores (P < 0.001). CONCLUSIONS: Targeted biopsy is able to improve the diagnostic accuracy of biopsy in detection of clinically significant prostate cancer by reducing the number of clinically insignificant prostate cancer detections compared with 10-core systematic biopsy in biopsy-naïve Japanese men. In addition, the present findings suggest that patients with Prostate Imaging Reporting and Data System category 2 or 3 lesions at the middle or base of the inner gland might avoid biopsies.

Cytoprotective Preconditioning of Osteoblast-Like Cells with N-Acetyl-L-Cysteine for Bone Regeneration in Cell Therapy.

Oxidative stress hinders tissue regeneration in cell therapy by inducing apoptosis and dysfunction in transplanted cells. N-acetyl-L-cysteine (NAC) reinforces cellular antioxidant capabilities by increasing a major cellular endogenous antioxidant molecule, glutathione, and promotes osteogenic differentiation. This study investigates the effects of pretreatment of osteoblast-like cells with NAC on oxidative stress-induced apoptosis and dysfunction and bone regeneration in local transplants. Rat femur bone marrow-derived osteoblast-like cells preincubated for 3 h with and without 5 mM NAC were cultured in a NAC-free osteogenic differentiation medium with continuous exposure to 50 µM hydrogen peroxide to induce oxidative stress. NAC preincubation prevented disruption of intracellular redox balance and alleviated apoptosis and negative impact on osteogenic differentiation, even under oxidative stress. Autologous osteoblast-like cells with and without NAC pretreatment in a collagen sponge vehicle were implanted in critical-size defects in rat femurs. In the third week, NAC-pretreated cells yielded complete defect closure with significantly matured lamellar bone tissue in contrast with poor bone healing by cells without pretreatment. Cell-tracking analysis demonstrated direct bone deposition by transplanted cells pretreated with NAC. Pretreatment of osteoblast-like cells with NAC enhances bone regeneration in local transplantation by preventing oxidative stress-induced apoptosis and dysfunction at the transplanted site.

The change of surface charge by lithium ion coating enhances protein adsorption on titanium.

Surface charge is one of the essential physicochemical properties of titanium surfaces for extracellular protein adsorption. Titanium surfaces are generally electronegatively charged at physiological pH. Typical cellular adhesive proteins and cell membranes are also negatively charged. Therefore, there are no direct electric interactions between proteins and titanium surfaces at physiological pH. The objective of this study was to determine how different electrical charges on titanium surfaces affect protein adsorption. Commercially pure grade-2 titanium disks, 19 mm in diameter and 1.5 mm in thickness, having acid-etched micro-roughed surfaces, were prepared. Electropositive charge was supplied by soaking in LiOH solution at concentrations of 0.05, 0.1, 0.25, 0.5, and 1.0 M. After LiOH treatment, X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) were performed. The zeta potential, isoelectric point, and wettability of titanium surfaces were measured. The adsorption levels of proteins, including albumin, laminin, and fibronectin, were evaluated. Osteoblastic cell attachment level was also determined. Incorporation of Li was detected in the oxide layer of titanium without surface morphological modification. The zeta potential was shifted up and the isoelectric point was increased from 3.94 to 5.63 by LiOH treatment. Long-term super-hydrophilicity was also obtained on Li-treated surfaces. The adsorption of albumin and laminin increased with increasing concentration of LiOH treatment solution, whereas fibronectin adsorption was highest upon treatment with 0.25 M. The osteoblastic cell attachment level was shown to be dependent on the amount of fibronectin adsorbed. In conclusion, LiOH treatment enhances biological adhesion on titanium with an increase in surface charge and hydrophilicity. This study suggests that modifying the surface charge provides a direct protein-to-materials interaction and the optimal application of Li should be investigated further.

Evaluation of corrosion resistance of implant-use Ti-Zr binary alloys with a range of compositions.

Although titanium-zirconium (Ti-Zr) alloy has been adopted for clinical applications, the ideal proportion of Zr in the alloy has not been identified. In this study, we investigated the biocompatibility of Ti-Zr alloy by evaluating its corrosion resistance to better understand whether there is an optimal range or value of Zr proportion in the alloy. We prepared pure Ti, Ti-30Zr, Ti-50Zr, Ti-70Zr, and pure Zr (mol% of Zr) samples and subjected them to anodic polarization and immersion tests in a lactic acid + sodium chloride (NaCl) solution and artificial saliva. We observed pitting corrosion in the Ti-70Zr and Zr after exposure to both solutions. After the immersion test, we found that pure Ti exhibited the greatest degree of dissolution in the lactic acid + NaCl solution, with the addition of Zr dramatically reducing Ti ion dissolution, with the reduction ultimately exceeding 90% in the case of the Ti-30Zr. Hence, although the localized corrosion resistance under severe conditions was compromised when the Zr content was more than 70%, metal ion release reduced owing to Zr addition and the corresponding formation of a stable passive layer. The results suggest that Ti-30Zr or a Zr proportion of less than 50% would offer an ideal level of corrosion resistance for clinical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 73-79, 2018.

Inverse response of osteoblasts and fibroblasts to growth on carbon-deposited titanium surfaces.

Titanium implant surfaces may serve as attachment substrates for various cell types. Since carbon adsorption on titanium is inevitable, this study examined the negative/positive biological reaction of osteoblasts and fibroblasts on carbon-deposited titanium surfaces. Osteogenic MC3T3-E1 and fibrogenic NIH/3T3 cells were separately cultured on titanium disks on which carbon deposition was experimentally regulated to achieve titanium/carbon ratios of 6.5, 0.02, 0.005, and 0. The initial attachment of cells demonstrated that the quantity of attached osteoblasts on Ti/C (0.005) surfaces was 20% lower than that on Ti/C (6.5) surfaces at 4 h of culture. A 40% reduction in cell attachment at 24 h transferring from Ti/C (6.5) to Ti/C (0.005) surfaces highlighted the negative effect of carbon deposition on osteoblast attachment. However, the initial attachment of fibroblasts, which depended on carbon deposition, increased, and the quantity of cells on Ti/C (0.005) surfaces was almost twice that on Ti/C (6.5) surfaces at 4 h of culture. The levels of common differentiation markers of collagen synthesis were also differentially carbon-dependent as total collagen deposition on Ti/C (0.005) decreased by > 30% compared to that on Ti/C (6.5) in osteoblasts after 7 days of culture. In contrast, collagen synthesis in fibroblasts markedly increased as was evident by the increase in carbon deposition. These inverse effects indicate that carbon deposition on a titanium surface would likely be a disadvantage for bone formation, but might represent an effective option for achieving better wound healing and soft tissue sealing around the surface of an implant-neck region. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1869-1877, 2018.