The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, H2SO4 and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven (37℃). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of H₂SO₄ and temperature of acid-solution. Acid-treatment conditions (concentration of H₂SO₄, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with 90℃ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.
Calcium ; Ceramics ; Microscopy, Confocal ; Photoelectron Spectroscopy ; Spectrometry, X-Ray Emission ; Surface Properties ; Titanium ; Ultrasonics ; Water

BACKGROUND: Sand blasted titanium (Ti) is commonly used in designing endosseous dental implants due to its biocompatibility and ability to form bonds with bone tissues. However, titanium implants do not induce strong interactions with teeth bones. To increase strong interactions between Ti disk implants and teeth bones, the L-glutamic acid grafted hydroxyapatite nanorods (nHA) were immobilized on albumin modified Ti disk implants (Ti-Alb). METHODS: For modification of Ti disk implants by nHA, the L-glutamic acid grafted nHA was synthesized and then immobilized on albumin modified Ti disk implants. Fourier transformed infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscope; energy dispersive spectroscopy and confocal laser scanning microscopy were used to confirm the modification of Ti disk implants. The bioactivity of nHA-modified Ti disk implants was evaluated by seeding MC3T3-E1 cells on Ti-nHA implants. RESULTS: Characterization techniques have confirmed the successful modification of Ti disk implants by L-glutamic acid grafted nHA. The nHA-modified Ti disk implants have shown enhanced adhesion, proliferation and cytotoxicity of MC3T3-E1 cells in comparison to pristine Ti implants. CONCLUSION: The modification of Ti implants by L-glutamic acid grafted nHA has produced highly osteogenic Ti disk plants in comparison to pristine Ti disk implants due to the formation of bioactive surfaces by hydroxyapatite nano rods on Ti disk implants. Ti-nHA disk implants showed enhanced adhesion, proliferation, and MC3T3-E1 cells viability in comparison to pristine Ti disk implants. Thus nHA might be to be useful to enhance the osseointegration of Ti implants with teeth bones.
Bone and Bones ; Dental Implants* ; Durapatite* ; Fourier Analysis ; Glutamic Acid ; Microscopy, Confocal ; Nanotubes ; Osseointegration ; Photoelectron Spectroscopy ; Spectrum Analysis ; Titanium* ; Tooth ; Transplants

OBJECTIVE: Microbial aggregation around dental implants can lead to loss/loosening of the implants. This study was aimed at surface treating titanium microimplants with silver nanoparticles (AgNPs) to achieve antibacterial properties. METHODS: AgNP-modified titanium microimplants (Ti-nAg) were prepared using two methods. The first method involved coating the microimplants with regular AgNPs (Ti-AgNP) and the second involved coating them with a AgNP-coated biopolymer (Ti-BP-AgNP). The topologies, microstructures, and chemical compositions of the surfaces of the Ti-nAg were characterized by scanning electron microscopy (SEM) equipped with energy-dispersive spectrometer (EDS) and X-ray photoelectron spectroscopy (XPS). Disk diffusion tests using Streptococcus mutans, Streptococcus sanguinis, and Aggregatibacter actinomycetemcomitans were performed to test the antibacterial activity of the Ti-nAg microimplants. RESULTS: SEM revealed that only a meager amount of AgNPs was sparsely deposited on the Ti-AgNP surface with the first method, while a layer of AgNP-coated biopolymer extended along the Ti-BP-AgNP surface in the second method. The diameters of the coated nanoparticles were in the range of 10 to 30 nm. EDS revealed 1.05 atomic % of Ag on the surface of the Ti-AgNP and an astounding 21.2 atomic % on the surface of the Ti-BP-AgNP. XPS confirmed the metallic state of silver on the Ti-BP-AgNP surface. After 24 hours of incubation, clear zones of inhibition were seen around the Ti-BP-AgNP microimplants in all three test bacterial culture plates, whereas no antibacterial effect was observed with the Ti-AgNP microimplants. CONCLUSIONS: Titanium microimplants modified with Ti-BP-AgNP exhibit excellent antibacterial properties, making them a promising implantable biomaterial.
Aggregatibacter actinomycetemcomitans ; Biopolymers ; Dental Implants ; Diffusion ; Methods ; Microscopy, Electron, Scanning ; Nanoparticles* ; Photoelectron Spectroscopy ; Silver* ; Streptococcus ; Streptococcus mutans ; Titanium

Increasing bioactivity and mechanical properties of polymers to produce more suitable scaffold for tissue engineering is a recurrent goal in the development of new biomedical materials. In this study, collagen-functionalized poly (lactic acid), PLA, was obtained by means of a simple grafting route, and electrospun scaffolds were produced to grow cells in vitro; their bioactivity was compared with scaffolds made of physical blends of PLA and collagen. Grafting was verified via nuclear magnetic resonance, attenuated total reflection-Fourier transform infrared and X-ray photoelectron spectroscopy. The cell adhesion performance of the scaffolds was studied using macrophages. Elastic modulus (74.7 megapascals) and tensile strength (3.0 megapascals) of the scaffold made from PLA grafted with collagen were substantially higher than the scaffolds made from physical blends of collagen and PLA: 32 and 2.16 megapascals, respectively, implying a more resistant material because of the chemical bond of the polypeptide to PLA. Besides, the fibers had more uniform diameter without defects. Scaffolds made from PLA grafted with collagen presented four-fold increase in cell adhesion than those of PLA blended with collagen. Furthermore, cell spreading within the scaffolds occurred only when collagen-functionalized poly (lactic acid) was used. These results open a new option for the easy tailoring of nanofiber-based scaffolds in three dimensions for tissue engineering.
Cell Adhesion* ; Collagen* ; Elastic Modulus ; In Vitro Techniques ; Macrophages ; Magnetic Resonance Spectroscopy ; Photoelectron Spectroscopy ; Polymers ; Tensile Strength ; Tissue Engineering ; Transplants*

Secreted protein, acidic, cysteine-rich (SPARC)-related modular calcium binding 1 (SMOC1) has been implicated in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs). In this study, we found that a peptide (16 amino acids in length), which is located in the extracellular calcium (EC) binding domain of SMOC1, stimulated osteogenic differentiation of human BMSCs in vitro and calvarial bone regeneration in vivo. Treatment of BMSCs with SMOC1-EC peptide significantly stimulated their mineralization in a dose-dependent manner without changing their rate of proliferation. The expression of osteogenic differentiation marker genes, including type 1 collagen and osteocalcin, also increased in a dose-dependent manner. To examine the effect of the SMOC1-EC peptide on bone formation in vivo, the peptide was covalently immobilized onto hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) particles. X-ray photoelectron spectroscopy analysis showed that the peptide was successfully immobilized onto the surface of HA/β-TCP. Implantation of the SMOC1-EC peptide-immobilized HA/β-TCP particles into mouse calvarial defects and subsequent analyses using microcomputed tomography and histology showed significant bone regeneration compared with that of calvarial defects implanted with unmodified HA/β-TCP particles. Collectively, our data suggest that a peptide derived from the EC domain of SMOC1 induces osteogenic differentiation of human BMSCs in vitro and efficiently enhances bone regeneration in vivo.
Amino Acids ; Animals ; Bone Marrow ; Bone Regeneration ; Calcium ; Ceramics* ; Collagen Type I ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Mice ; Miners ; Osteocalcin ; Osteogenesis ; Photoelectron Spectroscopy ; Regeneration* ; X-Ray Microtomography

PURPOSE: This study aimed to investigate the efficacy of cleaning solutions on saliva-contaminated zirconia in comparison to air-abrasion in terms of resin bonding. MATERIALS AND METHODS: For saliva-contaminated airabraded zirconia, seven cleaning methods)-no contamination (NC), water-spray rinsing (WS), additional airabrasion (AA), and cleaning with four solutions (Ivoclean [IC]; 1.0 wt% sodium dodecyl sulfate [SDS], 1.0 wt% hydrogen peroxide [HP], and 1.0 wt% sodium hypochlorite [SHC])-were tested. The zirconia surfaces for each group were characterized using various analytical techniques. Three bonded resin (Panavia F 2.0) cylinders (bonding area: 4.5 mm2) were made on one zirconia disk specimen using the Ultradent jig method [four disks (12 cylinders)/group; a total of 28 disks]. After 5,000 thermocycling, all specimens were subjected to a shear bond strength test with a crosshead speed of 1.0 mm/minute. The fractured surfaces were observed using an optical and scanning electron microscope (SEM). RESULTS: Contact angle measurements showed that groups NC, AA, IC, and SHC had hydrophilic surfaces. The X-ray photoelectron spectroscopy (XPS) analysis showed similar elemental distributions between group AA and groups IC and SHC. Groups IC and SHC showed statistically similar bond strengths to groups NC and AA (P>.05), but not groups SDS and HP (P<.05). For groups WS, SDS, and HP, blister-like bubble formations were observed on the surfaces under SEM. CONCLUSION: Within the limitations of this in vitro study, some of the cleaning solutions (IC or SHC) were effective in removing saliva contamination and enhancing the resin bond strength.
Dental Bonding ; Hydrogen Peroxide ; Photoelectron Spectroscopy ; Saliva ; Sodium Dodecyl Sulfate ; Sodium Hypochlorite

PURPOSE: This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS: Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS: From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION: The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.
Alkaline Phosphatase ; Calcium ; Cell Adhesion ; Cell Proliferation ; Chemistry ; Durapatite ; Microscopy, Electron, Scanning ; Osteoblasts* ; Photoelectron Spectroscopy ; Pseudopodia

PURPOSE: The aim of this study was to evaluate the surface properties and in vitro bioactivity to osteoblasts of magnesium and magnesium-hydroxyapatite coated titanium. MATERIALS AND METHODS: Themagnesium (Mg) and magnesium-hydroxyapatite (Mg-HA) coatings on titanium (Ti) substrates were prepared by radio frequency (RF) and direct current (DC) magnetron sputtering.The samples were divided into non-coated smooth Ti (Ti-S group), Mg coatinggroup (Ti-Mg group), and Mg-HA coating group (Ti-MgHA group).The surface properties were evaluated using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The surface roughness was evaluated by atomic force microscopy (AFM). Cell adhesion, cell proliferation and alkaline phosphatase (ALP) activity were evaluated using MC3T3-E1 cells. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed. RESULTS: Cross-sectional SEM images showed that Mg and Mg-HA depositionson titanium substrates were performed successfully. The surface roughness appeared to be similaramong the three groups. Ti-MgHA and Ti-Mg group had improved cellular responses with regard to the proliferation, alkaline phosphatase (ALP) activity, and bone-associated markers, such as bone sialoprotein (BSP) and osteocalcin (OCN) mRNA compared to those of Ti-S group. However, the differences between Ti-Mg group and Ti-MgHA group were not significant, in spite of the tendency of higher proliferation, ALP activity and BSP expression in Ti-MgHA group. CONCLUSION: Mg and Mg-HAcoatings could stimulate the differentiation into osteoblastic MC3T3-E1 cells, potentially contributing to rapid osseointegration.
Alkaline Phosphatase ; Biocompatible Materials ; Calcium Phosphates ; Calcium* ; Cell Adhesion ; Cell Proliferation ; Coated Materials, Biocompatible ; Integrin-Binding Sialoprotein ; Magnesium* ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Osseointegration ; Osteoblasts* ; Osteocalcin ; Photoelectron Spectroscopy ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Messenger ; Surface Properties ; Titanium*

BACKGROUNDSurface modification by ion implantation-deposition is well established in materials science and can be an effective way to improve biocompatibility. The aim of this study is to evaluate the chemical composition of a modified titanium (Ti) surface after zinc (Zn) ion implantation and deposition and examine the effect of the modification on the formation of cellular focal adhesion plaques in vitro.

METHODScp-Ti discs were modified with Zn ion implantation and deposition via PIIID. The chemical composition of the surface modification was characterized by X-ray photoelectron spectroscopy (XPS). The formation of focal adhesion plaques on the modified Ti was investigated with human osteoblast-like MG-63 cells that were seeded onto the Ti surfaces and quantified by morphometric analysis under a confocal microscope.

RESULTSXPS data revealed that the modified Zn-Ti surface consisted of Ti, oxygen, Zn, and carbon. In addition, Gaussian fitting of the spectra indicated that the modified surface contained titanium dioxide and zinc oxide. After 6 hours of MG-63 cell culture, there were significantly more focal adhesion plaques on the modified surfaces than observed on the nonmodified Ti (P < 0.05).

CONCLUSIONZn ion implantation and deposition greatly improved the biocompatibility of Ti for the growth of MG-63 cells.

Cell Adhesion ; drug effects ; Cell Line ; Humans ; Osteoblasts ; cytology ; drug effects ; Photoelectron Spectroscopy ; Titanium ; pharmacology ; Zinc Oxide ; pharmacology

PURPOSE: The aim of this study was to evaluate the surface properties and biological response of an anodized titanium surface by cell proliferation and alkaline phosphatase activity analysis. METHODS: Commercial pure titanium (Ti) disks were prepared. The samples were divided into an untreated machined Ti group and anodized Ti group. The anodization of cp-Ti was formed using a constant voltage of 270 V for 60 seconds. The surface properties were evaluated using scanning electron microscopy, X-ray photoelectron spectroscopy, and an image analyzing microscope. The surface roughness was evaluated by atomic force microscopy and a profilometer. The contact angle and surface energy were analyzed. Cell adhesion, cell proliferation, and alkaline phosphatase activity were evaluated using mouse MC3T3-E1 cells. RESULTS: The anodized Ti group had a more porous and thicker layer on its surface. The surface roughness of the two groups measured by the profilometer showed no significant difference (P>0.001). The anodized Ti dioxide (TiO2) surface exhibited better corrosion resistance and showed a significantly lower contact angle than the machined Ti surface (P>0.001). Although there was no significant difference in the cell viability between the two groups (P>0.001), the anodized TiO2 surface showed significantly enhanced alkaline phosphatase activity (P<0.001). CONCLUSIONS: These results suggest that the surface modification of Ti by anodic oxidation improved the osteogenic response of the osteoblast cells.
Alkaline Phosphatase ; Animals ; Cell Adhesion ; Cell Proliferation ; Cell Survival ; Corrosion ; Dental Implants ; Durapatite ; Mice ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Osteoblasts ; Photoelectron Spectroscopy ; Surface Properties ; Titanium