Visible-light-responsive reduced graphene oxide/g-C3N4/TiO2 composite nanocoating for photoelectric stimulation of neuronal and osteoblastic differentiation.

Restoration of nerve supply in newly formed bone is critical for bone defect repair. However, nerve regeneration is often overlooked when designing bone repair biomaterials. In this study, employing graphitic carbon nitride (g-C3N4) as a visible-light-driven photocatalyst and reduced graphene oxide (rGO) as a conductive interface, an rGO/g-C3N4/TiO2 (rGO/CN/TO) ternary nanocoating with photoelectric conversion ability was fabricated on a Ti-based orthopedic implant for photoelectric stimulation of both bone and nerve repair. Compared with g-C3N4/TiO2 (CN/TO) and TiO2 nanocoatings, the ternary nanocoating exhibited stronger visible-light absorption as well as higher transient photocurrent density and open circuit potential under blue LED exposure. The improved photo-electrochemical properties of the ternary nanocoating were attributed to the enhanced separation of photogenerated carriers at the heterointerface. For the tested nanocoatings, introducing blue LED light irradiation enhanced MC3T3-E1 osteoblastic differentiation and neurite outgrowth of PC12 cells. Among them, the rGO/CN/TO nanocoating exerted the greatest enhancement. In a coculture system, PC12 cells on the ternary nanocoating released a higher amount of neurotransmitter calcitonin gene-related peptide (CGRP) under light irradiation, which in turn significantly enhanced osteoblastic differentiation. The results may provide a prospective approach for targeting nerve regeneration to stimulate osteogenesis when designing bone repair biomaterials.

Zinc-α2-Glycoprotein Knockout Influenced Genes Expression Profile in Adipose Tissue and Decreased the Lipid Mobilizing After Dexamethasone Treatment in Mice.

Zinc-α2-glycoprotein (ZAG), as an adipokine, plays an important role in lipid metabolism. However, its influence on whole gene expression profile in adipose tissue is not known. Under stress condition, how ZAG affects the lipid metabolism is also unclear. Therefore, in this study ZAG systemic knockout (KO) mice were used as a model to reveal the genes expression profile in visceral fat tissues of ZAG KO mice and wild-type mice by genome-wide microarray screening. Then dexamethasone (DEX) was used to explore the effect of ZAG deletion on body fat metabolism under stress. Our results showed that 179 genes were differentially expressed more than 1.5 times between ZAG KO mice and wild type mice, of which 26 genes were upregulated dramatically and 153 genes were significantly downregulated. Under DEX simulated stress, ZAG systemic knockout in vivo resulted in a markedly decrease of triglycerides (TG) and nonesterified fatty acid (NEFA) content in in plasma. Similarly, for lipid catabolism, ZAG KO led to a significant increase of phosphorylated HSL (p-HSL) protein and a rising tendency of adipose triglyceride lipase (ATGL) protein relative to those of the DEX group. For lipid anabolism, fatty acid synthase (FAS) and adiponectin protein expression in visceral fat rose notably in ZAG KO mice after DEX treatment. In conclusion, ZAG knockout can affect the gene expression profile of adipose tissue, reduce elevated TG and NEFA levels in plasma, and alter lipid metabolism under DEX treatment. These findings provide new insights into the mechanism of lipid metabolic disorders in response to stress.

Improved osteogenesis of boron incorporated calcium silicate coatings via immunomodulatory effects.

Osteoimmunology has revealed the importance of a favorable immune response for successful biomaterial-mediated osteogenesis. Boron-incorporated calcium silicate (Ca11 Si4 B2 O22 , B-CS) coating has been reported as a potential candidate for improving osteogenesis in orthopedic applications in vitro. However, relatively little is known about its effects on the immune response and subsequent osteogenesis. In this work, the immunomodulatory properties of the B-CS coating and its specific mechanism of action were explored. We found that the B-CS coating decreased M1 polarization and converted macrophages to the M2 phenotype via restraining the toll-like receptor signaling pathway, thus inducing a significant reduction in pro-inflammatory cytokines and an increase in anti-inflammatory cytokines. Moreover, the B-CS coating inhibited osteoclastogenesis and osteoclastic activities by downregulating osteoclastogenic genes and inhibiting the RANKL/RANK system. BMP2 and VEGF were also significantly upregulated by macrophages and bone mesenchymal stem cells, leading to activation of the BMP2 signaling pathway and subsequent upregulation of osteogenesis-associated genes, finally promoting osteogenic differentiation. These findings show that the B-CS coating could be a promising coating material for hip and knee implants. Furthermore, incorporation of the element boron into bioceramic coatings could be a good strategy in the design of bone biomaterials with beneficial immune responses. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 12-24, 2019.

Incorporation of Cerium Oxide into Hydroxyapatite Coating Protects Bone Marrow Stromal Cells Against H2O2-Induced Inhibition of Osteogenic Differentiation.

Oxidative stress exerts a key influence in osteoporosis in part by inhibiting osteogenic differentiation of bone marrow stromal cells (BMSCs). With their unique antioxidant properties and reported biocompatibility, cerium oxide (CeO2) ceramics exhibit promising potential for the treatment of osteoporosis resulting from oxidative stress. In this study, protective effects of CeO2-incorporated hydroxyapatite coatings (HA-10Ce and HA-30Ce) on the viability and osteogenic differentiation of H2O2-treated BMSCs were examined. CeO2-incorporated HA coatings enhanced cell viability and attenuated cell apoptosis caused by H2O2. An increase in CeO2 content in HA coatings better alleviated H2O2-induced inhibition of osteogenic differentiation by increasing alkaline phosphatase (ALP) activity, calcium deposition activity, and mRNA expression levels of osteogenesis markers runt-related transcription factor 2 (Runx2), ALP, and osteocalcin (OCN) in BMSCs. Furthermore, the H2O2-induced decrease of gene and protein expressions of ß-catenin and cyclin D1 in the Wnt/ß-catenin signaling pathway was successfully rescued by the CeO2 incorporated HA coatings. Besides, the decreased expression of receptor activator of nuclear factor kappa-B ligand (RANKL) and the increased ratio of osteoprotegerin (OPG)/RANKL in BMSCs on the CeO2-modified coatings was observed, indicating the inhibition of osteoclastogenesis. The above results were mediated by the antioxidant properties of CeO2. The CeO2-incorporated HA coatings reversed the decreased superoxide dismutase (SOD) activity, reduced reactive oxygen species (ROS) generation, and suppressed the malondiadehyde (MDA) formation. The findings suggested that CeO2-modified HA coatings may be promising coating materials for osteoporotic bone regeneration.

Incorporation of cerium oxide into hydroxyapatite coating regulates osteogenic activity of mesenchymal stem cell and macrophage polarization.

Biomedical coatings for orthopedic implants should facilitate osseointegration and mitigate implant-induced inflammatory reactions. Cerium oxide (CeO2) ceramics possess anti-oxidative properties and can be used to decrease mediators of inflammation, which makes them attractive for biomedical applications. In our work, two kinds of CeO2 incorporated hydroxyapatite coatings (HA-10Ce and HA-30Ce) were prepared via plasma spraying technique and the effects of CeO2 addition on the responses of bone mesenchymal stem cells (BMSCs) and RAW264.7 macrophages were investigated. An increase in CeO2 content in the HA coatings resulted in better osteogenic behaviors of BMSCs in terms of cell proliferation, alkaline phosphatase (ALP) activity and mineralized nodule formation. RT-PCR and western blot analysis suggested that the incorporation of CeO2 may promote the osteogenic differentiation of BMSCs through the Smad-dependent BMP signaling pathway, which activated Runx2 expression and subsequently enhanced the expression of ALP and OCN. The expression profiles of macrophages cultured on the CeO2 modified coating revealed a tendency toward a M2 phenotype, because of an upregulation of M2 surface markers (CD163 and CD206), anti-inflammatory cytokines (TNF-α and IL-6) and osteoblastogenesis-related genes (BMP2 and TGF-ß1) as well as a downregulation of M1 surface markers (CCR7 and CD11c), proinflammatory cytokines (IL-10 and IL-1ra) and reactive oxygen species production. The results suggested the regulation of BMSCs behaviors and macrophage-mediated responses at the coating's surface were associated with CeO2 incorporation. The incorporation of CeO2 in HA coatings can be a valuable strategy to promote osteogenic responses and reduce inflammatory reactions.

[Evaluation of masticatory efficiency of lingualized occlusal complete denture].

PURPOSE: To compare the masticatory efficiency of lingualized occlusal complete denture with that of semi-anatomical occlusal complete denture. METHODS: Sixty cases with flat or depressed residual ridges were selected from 2009 to 2011. After randomly divided into two groups, the patients were treated with lingualized and semi-anatomical occlusal complete dentures, respectively. A comparative study of masticatory efficiency was carried out on patients wearing lingualized occlusal complete dentures with those wearing semi-anatomical occlusal complete dentures in different period (1st, 2nd, 3rd, 6th month) after wearing. SAS6.16 software package was used for data analysis. RESULTS: Once masticatory time or masticatory times were fixed, there was no significant difference between lingualized occlusal dentures and semi-anatomical occlusal complete dentures (P>0.05). CONCLUSION: Lingualized occlusal complete denture can achieve good masticatory efficiency for edentulous patients with flat or depressed residual ridges.

[Comparative adaptation of crowns of selective laser melting and wax-lost-casting method].

OBJECTIVE: To investigate the marginal adaptation of crowns fabricated by selective laser melting (SLM) and wax-lost-casting method, so as to provide an experimental basis for clinic. METHODS: Co-Cr alloy full crown were fabricated by SLM and wax-lost-casting for 24 samples in each group. All crowns were cemented with zinc phosphate cement and cut along longitudinal axis by line cutting machine. The gap between crown tissue surface and die was measured by 6-point measuring method with scanning electron microscope (SEM). The marginal adaptation of crowns fabricated by SLM and wax-lost-casting were compared statistically. RESULTS: The gap between SLM crowns were (36.51 ± 2.94), (49.36 ± 3.31), (56.48 ± 3.35), (42.20 ± 3.60) µm, and wax-lost-casting crowns were (68.86 ± 5.41), (58.86 ± 6.10), (70.62 ± 5.79), (69.90 ± 6.00) µm. There were significant difference between two groups (P < 0.05). CONCLUSIONS: Co-Cr alloy full crown fabricated by wax-lost-casting method and SLM method provide acceptable marginal adaptation in clinic, and the marginal adaptation of SLM is better than that of wax-lost-casting method.

[Study of the appearance difference of lower complete denture between functional and anatomic impression techniques].

PURPOSE: To compare the difference in oblique external ridge, oblique internal ridge and alveolar process crest of lower complete denture base made through functional impression and anatomic impression techniques. METHODS: Fifteen patients were chosen to treat with two kinds of complete dentures through functional impression and anatomic impression technique respectively. 3D laser scanner was used to scan the three-dimensional model of the denture base and the differences of the surface structural between two techniques in alveolar process crest, external and internal oblique ridges were analyzed, using paired t test with SPSS 12.0 software package. RESULTS: Between the two techniques, there were significant differences in the areas of internal and external oblique ridge(P<0.01); there was no significant difference in the main support areas(P>0.05). CONCLUSIONS: The results explain why there is less tenderness when functional impression technique is applied. The differences measured also indicate that sufficient buffering should be made in external and internal oblique ridge areas in clinic.

[The stress distribution of mandibular alveolar mucosa under functional impressive complete denture studied by 3-D finite element analysis].

PURPOSE: To analyze the stress distribution and displacement of the mandibular alveolar mucosa under functional impressive complete denture. METHODS: CT images were processed by Mimics and Geomagic studio, and the solid models and 3-D finite element models were established by Unigraphics NX. 3-D finite element analysis (3-D FEA) was used to study the stress distribution and displacement on the mandibular alveolar mucosa under functional impressive complete denture and anatomic impressive complete denture. RESULTS: The stress distribution on the mandibular alveolar mucosa under two types of complete denture were consistent, and arranged from large to small order as follow: alveolar process crest of molar area, alveolar process crest of anterior area, external oblique ridge and internal oblique ridge. At alveolar process crest and internal oblique ridge, the stress and Z-axis displacement of functional impressive complete denture were less than those of anatomic impressive complete denture. CONCLUSIONS: During masticatory period, the stress distribution on the mandibular alveolar mucosa under functional impressive complete denture is more even and more comprehensive than that under anatomic one.

An in vitro investigation of the mechanical-chemical and biological properties of calcium phosphate/calcium silicate/bismutite cement for dental pulp capping.

The properties of new calcium phosphate/calcium silicate/bismutite (CPCSBi) cement were compared with those of calcium hydroxide (CH) and Dycal cements in dental pulp-capping applications. CPCSBi is composed of hydroxyapatite, tetracalcium phosphate, bismutite, and calcium silicate, which was analyzed by SEM, FTIR, and XRD. The results of ion release from CPCSBi showed that the concentrations of Bi(3+), Ca(2+), PO4(2-), and Si(4+) increased with time in deionized water solutions. The setting time of CPCSBi and Dycal was 13 min 50 s and 2 min 25 s, respectively. There were no statistical differences in compressive strength and solubility between CPCSBi and Dycal (p > 0.05). The pH of CPCSBi (10.9) was lower than that of CH (11.6) and Dycal (12.5) after immersion for 24 h. Only slight cytotoxicity appeared for CPCSBi, whereas both CH and Dycal produced moderate discoloration and lysis. In antimicrobial tests against Sm, Av, La, and Sa, the antimicrobial potency of the CPCSBi was approximately 5-10 times greater than that of Dycal and CH groups. The dissoluble dentin matrix components (DDMCs) extracted from CPCSBi exposed to dentin powder demonstrated increased expression of dentin sialophosphoprotein (DSPP) and soteocalcin (OCN) dramatically in human pulp cells by RT-PCR. These results suggest that CPCSBi will be a good candidate for use as a dental pulp-capping agent in future.

Evidence of chemisorption of maleic acid to enamel and hydroxyapatite.

Maleic acid has been used as an etchant or non-rinse conditioner in adhesive dentistry. However, the inherent mechanisms of the interaction of maleic acid with hydroxyapatite/enamel have never been fully elucidated. The purpose of this study was to provide evidence for the chemisorption of maleic acid onto hydroxyapatite/enamel, and to identify the reaction products obtained following the interaction of maleic acid with hydroxyapatite. Hydroxyapatite particles were dissolved in a 15% (w/v) aqueous solution of maleic acid (pH = 0.98). Half of the solution was dried to obtain a desiccated mixture. This mixture, hydroxyapatite, maleic acid and self-prepared calcium maleate were analysed by X-ray diffraction (XRD). Acetone was added to the other half of the solution to obtain a precipitate. This precipitate, hydroxyapatite, maleic acid, unetched enamel and maleic acid-etched enamel were analysed by X-ray photoelectron spectroscopy (XPS). The precipitate was also analysed by (1)H NMR. A new binding energy, indicating carboxylate groups, was detected by XPS on the precipitate and maleic acid-etched enamel surface. XRD data indicated the formation of calcium maleate and calcium hydrogen phosphate after the reaction. NMR data revealed that one carboxylic group of maleic acid reacted with hydroxyapatite. Hence, maleic acid can chemisorb to hydroxyapatite and enamel via ionic interactions.