Reactive Oxygen Species and Folate Receptor-Targeted Nanophotosensitizers Composed of Folic Acid-Conjugated and Poly(ethylene glycol)-Chlorin e6 Tetramer Having Diselenide Linkages for Targeted Photodynamic Treatment of Cancer Cells.

Folic acid-conjugated nanophotosensitizers composed of folic acid (FA), poly(ethylene glycol) (PEG) and chlorin e6 (Ce6) tetramer were synthesized using diselenide linkages for reactive oxygen species (ROS)- and folate receptor-specific delivery of photosensitizers. Ce6 was conjugated with 3-[3-(2-carboxyethoxy)-2,2-bis(2-carboxyethoxymethyl)propoxy]propanoic acid (tetra acid, or TA) to make Ce6 tetramer via selenocystamine linkages (TA-sese-Ce6 conjugates). In the carboxylic acid end group of the TA-sese-Ce6 conjugates, FA-PEG was attached again using selenocystamine linkages to make FA-PEG/TA-sese-Ce6 conjugates (abbreviated as FAPEGtaCe6 conjugates). Nanophotosensitizers were fabricated by a dialysis procedure. In the morphological observations, they showed spherical shapes with small diameters of less than 200 nm. Stability of the aqueous FAPEGtaCe6 nanophotosensitizer solution was maintained (i.e., their particle sizes were not significantly changed until 7 days later). When H2O2 was added to the nanophotosensitizer solution, the particle size distribution was changed from a monomodal pattern to a multimodal pattern. In addition, the fluorescence intensity and Ce6 release rate from the nanophotosensitizers were also increased by the addition of H2O2. These results indicated that the nanophotosensitizers had ROS-sensitive properties. In an in vitro cell culture study, an FAPEGtaCe6 nanophotosensitizer treatment against cancer cells increased the Ce6 uptake ratio, ROS generation and light-irradiated cytotoxicity (phototoxicity) compared with Ce6 alone against various cancer cells. When the folic acid was pretreated to block the folate receptors of the Y79 cells and KB cells (folate receptor-overexpressing cells), the intracellular Ce6 uptake, ROS generation and thereby phototoxicity were decreased, while the MCF-7 cells did not significantly respond to blocking of the folate receptors. These results indicated that they could be delivered by a folate receptor-mediated pathway. Furthermore, an in vivo pulmonary metastasis model using Y79 cells showed folate receptor-specific delivery of FAPEGtaCe6 nanophotosensitizers. When folic acid was pre-administered, the fluorescence intensity of the lungs was significantly decreased, indicating that the FAPEGtaCe6 nanophotosensitizers had folate receptor specificity in vitro and in vivo. We suggest that FAPEGtaCe6 nanophotosensitizers are promising candidates for a targeted photodynamic therapy (PDT) approach against cancer cells.

Reconstruction of Mandibular Defects With Bone Marrow-Derived Stem Cells in Odontogenic Myxoma.

Odontogenic myxoma is a benign tumor, mostly located in the mandible. It shows locally aggressive behavior and requires surgical removal. Common treatment options for reconstructing the bone defects are immediate or delayed autologous bone graft or free flap. In this article, the authors present the successful reconstruction with autogenous bone graft and autologous human bone marrow mesenchymal stem, followed by distraction osteogenesis, dental implant placement and prosthodontic restoration in the mandibular defect.

Combinatorial Effect of Cold Atmosphere Plasma (CAP) and the Anticancer Drug Cisplatin on Oral Squamous Cell Cancer Therapy.

Cold atmospheric plasma (CAP) has been extensively investigated in the local treatment of cancer due to its potential of reactive oxygen species (ROS) generation in biological systems. In this study, we examined the synergistic effect of combination of CAP and cisplatin-mediated chemotherapy of oral squamous cell carcinoma (OSCC) in vitro. SCC-15 OSCC cells and human gingival fibroblasts (HGF-1) cells were treated with cisplatin, and then, the cells were irradiated with CAP. Following this, viability and apoptosis behavior of the cells were investigated. The viability of SCC-15 cells was inhibited by cisplatin with a dose-dependent manner and CAP treatment time. HGF-1 cells also showed decreased viability by treatment with cisplatin and CAP. Combination of 1 µM cisplatin plus 3 min of CAP treatment or 3 µM cisplatin plus 1 min of CAP treatment showed a synergistic anticancer effect with appropriate cytotoxicity against normal cells. ROS generation and dead cell staining were also increased by the increase in CAP treatment time. Furthermore, tumor-suppressor proteins and apoptosis-related enzymes also increased according to the treatment time of CAP. We showed the synergistic effect of cisplatin and CAP treatment against SCC-15 cells with low cytotoxicity against normal cells.

Cell Proliferation on Macro/Nano Surface Structure and Collagen Immobilization of 3D Polycaprolactone Scaffolds.

In this study, 3D polycaprolactone (PCL) scaffolds were fabricated by 3D printing technique. The macro/nano morphology of, 3D PCL scaffolds surface was etched with oxygen plasma. Acrylic acid (AA) plasma-polymerization was performed to functionalize the macro/nano surface with carboxyl groups and then collagen was immobilized with plasma-polymerized 3D PCL scaffolds. After O2 plasma and AA plasma-polymerization, contact angles were decreased. The FE-SEM and AFM results showed that O2 plasma is increased the surface roughness. The MTT assay results showed that proliferation of the M3CT3-E1 cells increased on the oxygen plasma treated and collagen immobilized 3D PCL scaffolds.

O2/Ar Plasma Treatment for Enhancing the Biocompatibility of Hydroxyapatite Nanopowder and Polycaprolactone Composite Film.

In this study we performed O2/Ar plasma treatment to remove the polycaprolactone on hydroxyapatite nanopowder and polycaprolactone (HAp-NP/PCL) composite film. After plasma ashing, the HAp-NP was exposed on the composite film. The 25 wt% HAp-NP/PCL treated with plasma showed the hydrophilic surface property with reducing the aging effect. The MTT and ALP results indicated that the plasma etching increased the biocompatibility of HAp-NP/PCL composite film. The present simple plasma etching technique can be applicable in a development of biomaterials.

Characteristics of Plasma Treated Electrospun Polycaprolactone (PCL) Nanofiber Scaffold for Bone Tissue Engineering.

Polycaprolactone (PCL) nanofibers (PCL-NF) with uniform fibrous structure were fabricated by electrospinning. However, PCL-NF has hydrophobic surface, lacks functional groups and hence it is not a good substrate for cell adhesion. To improve the cell adhesion, PCL-NF surfaces were modified by low pressure RF discharge plasma treatment using monomer such as acrylic acid or oxygen gas. The plasma treated PCL-NFs improved the wettability and cell proliferation.

Oxygen and Nitrogen Gas Plasma Surface Treatments on Titanium for Initial Adhesion and Proliferation of Preosteoblast Cells.

In this study, titanium (Ti) surface was treated through the use of oxygen and nitrogen plasma for improving the initial adhesion and proliferation of preosteoblast cells. The plasma treated Ti surface demonstrated hydrophilic surface property and promoted initial adhesion and proliferation of preosteoblast cells.

Plasma Polymerization of 1,2-Diaminocyclohexane for Covalent Bonding of Bone Morphogenic Protein-2 on Titanium Surface.

Among the bone morphogenic protein (BMP) family members, BMP-2 is a potent osteoinductive factor that plays key roles during bone formation. In this study, plasma polymerization of 1,2-diaminocyclohexane (DACH) was performed to immobilize the BMP-2 on Ti surface. The plasma polymerization of DACH was carried out at a discharge power of 60 W and 100 W under a pressure of 10 mTorr for 90 sec. The BMP-2 was successfully immobilized on the DACH plasma treated Ti surface. The BMP-2 immobilized Ti surface showed the excellent cell differentiation. The results indicate that the DACH plasma polymerized Ti surface has a potential for immobilization of biomolecules in bone tissue engineering.

In Vitro Antibacterial Activity of Nano Silver Ion Substituted Poly Acrylic Acid Films on Titanium by Plasma Polymerization.

Antibacterial activity of oral pathogens such as Streptococcus mutans, Streptococcus sobrinus when silver ion immobilized on commercially pure (CP) titanium (Ti) surface was investigated in this study. Plasma-polymerized acrylic acid to have carboxyl group was deposited on CP-Ti surface and then ion-exchanged with Ag+ ions in 0.1 N AgNO3. In anti-adherent experiment, antibacterial activity was tested using broth culture methods. The biofilm formation assay was performed using semi-defined biofilm medium with sucrose. The silver coated CP-Ti completely inhibited the growth of S. mutans and S. sobrinus. In addition, the biofilm formation was significantly inhibited in silver-coated CP-Ti group.

Secretory Leukocyte Protease Inhibitor (SLPI) Increases Focal Adhesion in MC3T3 Osteoblast on Titanium Surface.

An appropriate interaction between implanted materials and the surrounding tissue is essential for successful implantation. Titanium (Ti) and some of its alloys have been used in dentistry and orthopedics as a substitutive material for hard tissue, such as teeth or natural bone. Nevertheless, metal ions released from titanium and alloy implants have adverse biological effects on biological tissues or cells. Secretory leukocyte protease inhibitor (SLPI) promotes cell migration, proliferation and wound healing. FAK and ERK1/2 signaling regulate cell adhesion and proliferation for cell survival. This study evaluated the potential of SLPI as a molecule to increase the cell adhesion on the Ti surface. Compared with the untreated cells, SLPI increased the adhesion of MC3T3-E1 cells to Ti discs, formation of actin stress fibers, paxillin expression and the phosphorylation of FAK. Moreover, SLPI enhanced the level of Grb2 and Ras expression and ERK1/2 phosphorylation in the MC3T3-E1 cells on Ti discs. These results suggest that SLPI can increase the interaction between the implanted Ti material and surrounding bone in orthodontic and dental surgery, making an effective nanomolecule for successful implantation.

Preparation of O2 Plasma Treated Polycaprolactone/Nano TiO2 Composites and In Vitro Bioactivity.

Polycaprolactone (PCL)/TiO2 composite films (PTCFs) were prepared by a solvent casting method at various concentrations of TiO2 (1, 3, 5, and 10 wt%) and then treated using oxygen plasma. The hydrophilicity of the oxygen plasma treated PTCFs increased as the treatment time was increased, due to the oxygen induced production of polar species at the surface of the PTCFs. In vitro bioactivities of the composite films were examined by immersion in simulated body fluid for up to 7 days. It was found that the oxygen plasma treatment significantly influenced the in vitro bioactivity of the PTCFs.

660 nm Red LED Induces Secretory Leukocyte Protease Inhibitor (SLPI) in Lipopolysaccharide-Stimulated RAW264.7 Cell.

SLPI acts as a modulator of the innate immune responses of macrophages, neutrophils and odontoblasts, and LPS-inducible anti-inflammatory cytokine to suppress the production of pro-inflammatory products by macrophages. Many studies have revealed the effects of light emitting diodes (LEDs) on the tissue repair and inflammatory responses. Although the anti-inflammatory mechanisms of irradiation with LEDs in gingival fibroblasts are known, the effects of 660 nm red LEDs on the inflammation remain unclear. Moreover, there is no report regarding the molecular mechanism for the relationship between SLPI and biological effects of LEDs. The effects of 660 nm red LEDs on inflammation with SLPI were investigated by examining the effects of 660 nm LED on the SLPI expression of RAW264.7 cells after LPS stimulation. This paper reports that the 660 nm red LED induced SLPI expression or reduced the LPS response, and inhibited NF-κB activation directly, leading to the suppression of pro-inflammatory cytokines, such as TNF-α and IL-1ß, suggesting that it might be a useful wavelength LED for inflammation therapy.

Up-and-Coming Mandibular Reconstruction Technique With Autologous Human Bone Marrow Stem Cells and Iliac Bone Graft in Patients With Large Bony Defect.

Large bony defects followed by resection of the mandible need to be reconstructed by various surgical techniques such as the fibular flap. In this article, we report the case of mandibular reconstruction with autologous human bone marrow mesenchymal stem cells and autogenous bone graft, followed by placement of dental implants and prosthodontic treatment in a patient who has been failed to reconstruct mandibular bone defect after resection of mandible.

Drug release control and anti-inflammatory effect of biodegradable polymer surface modified by gas phase chemical functional reaction.

The plasma technique has been widely used to modify the surfaces of materials. The purpose of this study was to evaluate the probability of controlling the prednisolone delivery velocity on a polylactic acid (PLA) surface modified by plasma surface treatment. Surface modification of PLA was performed at a low-pressure radio frequency under conditions of 100 W power, 50 mTorr chamber pressure, 100-200 sccm of flow rate, and Ar, O2, and CH4gases. The plasma surface-modified PLA was characterized using scanning emission microscope, x-ray photoelectron spectroscopy (XPS), and contact angle measurements.In vitroevaluations were performed to determine cellular response, drug release behavior, and anti-inflammatory effects. The PLA surface morphology was changed to a porous structure (with a depth of approximately 100 µm) and the surface roughness was also significantly increased. The XPS results demonstrated higher oxygenized carbon contents than those in the non-treated PLA group. The prednisolone holding capacity increased and the release was relatively prolonged in the surface-modified PLA group compared to that in the non-treated PLA group. In addition, cell migration and proliferation significantly increased after PLA treatment alone. The activity of cytokines such as cyclooxygenase-2 (COX-2), tumor necrosis factor-a (TNF-α), interleukin (IL-1ß), and IL-6 were considerably reduced in the plasma-treated and prednisolone holding group. Taken together, surface-modified PLA by plasma can provide an alternative approach to conventional physicochemical approaches for sustained anti-inflammatory drug release.

Osteoblastic cell response and bone formation of phosphate ion coated on plasma polymerized Ti surface.

This study examined the bone formation ability and cell response on a phosphate (PO3(4-)) ion exchanged amine plasma polymerized titanium (Ti) surface. The enhanced bone-like apatite (hydroxyapatite, HAp)-forming ability was attributed to the PO3(4-) ion exchanged amine plasma polymerized Ti (P/NH2/Ti) surface, which was formed by the reduction of PO3(4-) ions. PO3(4-) ions promote HAp nucleation and growth on Ti in SBF, and PO3(4-) ions improve the crystallinity of the HAp deposited layer. The cell viability tests revealed significantly greater cell viability on the P/NH2/Ti surfaces than on the other surfaces.

DMP1 and IFITM5 Regulate Osteogenic Differentiation of MC3T3-E1 on PEO-Treated Ti-6Al-4V-Ca2+/Pi surface.

Despite numerous studies on various surface modifications on titanium and its alloys, it remains unclear what kind of titanium-based surface modifications are capable of controlling cell activity. This study aimed to understand the mechanism at the cellular and molecular levels and investigate the in vitro response of osteoblastic MC3T3-E1 cultured on the Ti-6Al-4V surface modified by plasma electrolytic oxidation (PEO) treatment. A Ti-6Al-4V surface was prepared by PEO at 180, 280, and 380 V for 3 or 10 min in an electrolyte containing Ca2+/Pi ions. Our results showed that PEO-treated Ti-6Al-4V-Ca2+/Pi surfaces enhanced the cell attachment and differentiation of MC3T3-E1 compared to the untreated Ti-6Al-4V control but did not affect cytotoxicity as shown by cell proliferation and cell death. Interestingly, on the Ti-6Al-4V-Ca2+/Pi surface treated by PEO at 280 V for 3 or 10 min, MC3T3-E1 showed a higher initial adhesion and mineralization. In addition, the alkaline phosphatase (ALP) activity significantly increased in MC3T3-E1 on the PEO-treated Ti-6Al-4V-Ca2+/Pi (280 V for 3 or 10 min). In RNA-seq analysis, the expression of dentin matrix protein 1 (DMP1), sortilin 1 (Sort1), signal-induced proliferation-associated 1 like 2 (SIPA1L2), and interferon-induced transmembrane protein 5 (IFITM5) was induced during the osteogenic differentiation of MC3T3-E1 on the PEO-treated Ti-6Al-4V-Ca2+/Pi. DMP1 and IFITM5 silencing decreased the expression of bone differentiation-related mRNAs and proteins and ALP activity in MC3T3-E1. These results suggest that the PEO-treated Ti-6Al-4V-Ca2+/Pi surface induces osteoblast differentiation by regulating the expression of DMP1 and IFITM5. Therefore, surface microstructure modification through PEO coatings with Ca2+/Pi ions could be used as a valuable method to improve biocompatibility properties of titanium alloys.

Formation of nano-phase hydroxyapatite film on TiO2 nano-network.

Nano- and micro-phase HA film formed on TiO2 nano-network surface by simple electrochemical treatment. The range of lateral pore size of the network specimen was about 10-120 nm on Ti surface by anodized in 5 M NaOH solution at 0.3 A for 10 min. Nano-network TiO2 surface were formed by this anodization step which acted as templates and anchorage for growth of the HA during subsequent pulsed electrochemical deposition process at 85 degrees C. The phase and morphologies of deposits HA were influenced by the electrolyte concentration. The nano needle-like precipitates formed under low SBF concentration were identified to be HA crystals orientated parallel to the c-axis direction. Increasing electrolyte concentration, needle-like deposits transferred to the plate-like and micro plate like precipitates in the case of high SBF concentration.

Development and structure of a novel barrier membrane composed of drug-loaded poly(lactic-co-glycolic acid) particles for guided bone regeneration.

A novel barrier membrane composed of poly(lactic-co-glycolic acid) particles loaded with dexamethasone (DEX) as a bioactive molecule was produced via a modified nanoprecipitation method without any mixing. The particle membranes had a bilayer structure: one side was smooth and had a compact surface that was connected to larger particles, while the opposite side was rough, porous and connected to smaller particles. Additionally, a cross-section of the particle membrane had a porous structure with nano and micro sized irregular pores. Process optimization revealed that NaCl concentration in the water phase, with acetone as solvent and water as a non-solvent, played critical roles in determining the properties of the particle membranes, such as DEX encapsulation efficiency, thickness and surface morphologies of the particle membranes. A novel barrier membrane containing DEX using polymer particle drug capture technology has been successfully developed.

Corrosion behavior of nanotubular oxide on the Ti-29Nb-xZr alloy.

This work reports the corrosion behavior of nanotubular oxide layers on Ti-29Nb-xZr alloys with different compositions by anodization in 1 M H3PO4 + 0.8 wt% NaF. Depending on the alloy composition, nanoporous or highly ordered nanotube structures can be formed. The nanotube oxides on the Ti-29Nb-xZr alloy showed lower corrosion current density compared to non-treated sample.

Study in bactericidal properties of chlorhexidine grafting on the modified titanium.

Chlorhexidine (CHX) is widely used in a number of dental applications to reduce inflammation as well as swelling of gums and gum bleeding. We investigate anti-adherent effect of CHX grafted titanium on oral pathogens such as S. mutans and S. aureus respectively. CHX grafted titanium surfaces show good anti-adherent properties.