Sheng-Wei Feng

Biological surface modification of titanium surfaces using glow discharge plasma

To improve the biological activity of titanium, by using of glow discharge plasma (GDP), albumin-grafted titanium disk have been implemented and carefully studied. Titanium disks were pre-treated with GDP in an environment filled with argon and allylamine gas. Glutaraldehyde was used as a cross-linking agent for albumin grafting. Then, the surface of the albumin-grafted titanium was examined using scanning electron microscopy and X-ray photoelectron spectroscopy. In addition, the static water contact angles of the albumin-grafted titanium disks were measured using goniometry. To observe the effects of albumin adsorption on cell behavior, MG-63 osteoblast-like cells were cultured on the surface-modified titanium disks. Blood coagulation resistance of the modified titanium was monitored and compared to the control titanium disks. The results demonstrated that MG-63 osteoblast-like cells cultured on the albumin-grafted titanium disks expressed better-differentiated morphology compare to cells grown on the control disks. Furthermore, albumin-grafting treatment significantly improved the surface wettability of the titanium disks and resulted in a significantly negative effect on thrombus formation. Based on these results, it was believed that the GDP can potentially improve the biofunctionality of titanium surfaces.

Slow freezing coupled static magnetic field exposure enhances cryopreservative efficiency--a study on human erythrocytes.

The aim of this study was to assess the cryoprotective effect of static magnetic fields (SMFs) on human erythrocytes during the slow cooling procedure. Human erythrocytes suspended in 20% glycerol were slowly frozen with a 0.4-T or 0.8-T SMF and then moved to a −80°C freezer for 24 hr. The changes in survival rate, morphology, and metabolites of the thawed erythrocytes were examined. To understand possible cryoprotective mechanisms of SMF, membrane fluidity and dehydration stability of SMF-exposed erythrocytes were tested. For each test, sham-exposed erythrocytes were used as controls. Our results showed that freezing coupled with 0.4-T or 0.8-T SMFs significantly increased the relative survival ratios of the frozen-thawed erythrocytes by 10% and 20% (p<0.001), respectively. The SMFs had no effect on erythrocyte morphology and metabolite levels. However, membrane fluidity of the samples exposed to 0.8-T SMF decreased significantly (p<0.05) in the hydrophobic regions. For the dehydration stability experiments, the samples exposed to 0.8-T SMF exhibited significantly lower (p<0.05) hemolysis. These results demonstrate that a 0.8-T SMF decreases membrane fluidity and enhances erythrocyte membrane stability to resist dehydration damage caused by slow cooling procedures.

Influence of a static magnetic field on the slow freezing of human erythrocytes.

Abstract Purpose: The aim of this study was to test whether or not a strong static magnetic field (SMF) had a positive effect on the survival rate of frozen erythrocytes. Materials and methods: Human erythrocytes were slow freezing at a rate of −1°C/min, to a final temperature of −20°C. During the freezing process, the cells were simultaneously exposed to an SMF with a magnetic induction of 0.2 or 0.4 T. After the cells were thawed, the survival rate, morphology, and function of the thawed erythrocytes were evaluated. Furthermore, tests of membrane fluidity were performed to assess the effect of the SMF on the cell membrane. Results: The slow freezing process coupled with an SMF increased the survival rate of frozen erythrocytes, without any negative effect on the cell morphology or function. The increases in relative survival rates of frozen erythrocytes were 5.7% and 9.1% when the cells were frozen in 0.2 T and 0.4 T groups, respectively. In addition, the 0.4 T group significantly increased the membrane rigidity of the erythrocytes. Conclusions: Slow freezing coupled with a strong SMF produced positive effects on the survival rate of thawed erythrocytes, without changing their normal function.

Development and Testing of X-Ray Imaging-Enhanced Poly-L-Lactide Bone Screws.

Nanosized iron oxide particles exhibit osteogenic and radiopaque properties. Thus, iron oxide (Fe3O4) nanoparticles were incorporated into a biodegradable polymer (poly-L-lactic acid, PLLA) to fabricate a composite bone screw. This multifunctional, 3D printable bone screw was detectable on X-ray examination. In this study, mechanical tests including three-point bending and ultimate tensile strength were conducted to evaluate the optimal ratio of iron oxide nanoparticles in the PLLA composite. Both injection molding and 3D printing techniques were used to fabricate the PLLA bone screws with and without the iron oxide nanoparticles. The fabricated screws were implanted into the femoral condyles of New Zealand White rabbits. Bone blocks containing the PLLA screws were resected 2 and 4 weeks after surgery. Histologic examination of the surrounding bone and the radiopacity of the iron-oxide-containing PLLA screws were evaluated. Our results indicated that addition of iron oxide nanoparticles at 30% significantly decreased the ultimate tensile stress properties of the PLLA screws. The screws with 20% iron oxide exhibited strong radiopacity compared to the screws fabricated without the iron oxide nanoparticles. Four weeks after surgery, the average bone volume of the iron oxide PLLA composite screws was significantly greater than that of PLLA screws without iron oxide. These findings suggested that biodegradable and X-ray detectable PLLA bone screws can be produced by incorporation of 20% iron oxide nanoparticles. Furthermore, these screws had significantly greater osteogenic capability than the PLLA screws without iron oxide.

Er:YAG Laser-Roughened Enamel Promotes Osteoblastic Differentiation

OBJECTIVE The aim of this study was to test whether Er:YAG laser-etched enamel of human teeth could act as a biologically active scaffold for tissue regeneration. BACKGROUND DATA Hydroxylapatite (HA) with rough surface created by acid etching treatment has been used as a scaffold for tissue engineering. However, whether tooth HA can be a scaffold for osteoblastic cell seeding is still unclear. MATERIALS AND METHODS Enamel samples from human teeth were pretreated with an Er:YAG laser to create a rough surface. Then the surface of the laser-treated enamel was examined using a surface roughness profilometer and a scanning electron microscope. In addition, static water contact angles of the Er:YAG laser-treated enamel samples were measured using goniometry. To observe the effects of cell behavior on an Er:YAG laser-roughened enamel surface, we cultured MG63 osteoblast-like cells on the surface-modified enamel samples. Alkaline phosphatase activity, a marker of cell proliferation and differentiation, was monitored and compared with that in untreated control and acid-etched enamel samples. RESULTS Er:YAG laser treatment significantly improved the surface roughness of the enamel samples. Furthermore, MG63 osteoblast-like cells cultured on the Er:YAG laser-roughened enamel surface expressed more alkaline phosphatase activity and exhibited greater degrees of cellular differentiation than did cells that had been cultured on untreated enamel samples. CONCLUSIONS These results demonstrate that Er:YAG laser-roughened enamel promotes osteoblastic differentiation. This finding suggests that Er:YAG laser-roughened enamel surfaces can potentially serve as a scaffold for tissue engineering.

In Vitro Analysis of Fibronectin-Modified Titanium Surfaces

Background Glow discharge plasma (GDP) procedure is an effective method for grafting various proteins, including albumin, type I collagen, and fibronectin, onto a titanium surface. However, the behavior and impact of titanium (Ti) surface modification is yet to be unraveled. Purpose The purpose of this study is to evaluate and analyze the biological properties of fibronectin-grafted Ti surfaces treated by GDP. Materials and Methods Grade II Ti discs were initially cleaned and autoclaved to obtain original specimens. Subsequently, the specimens were GDP treated and grafted with fibronectin to form Ar-GDP (Argon GDP treatment only) and GDP-fib (fibronectin coating following GDP treatment) groups. Blood coagulation test and MG-63 cell culture were performed to evaluate the biological effects on the specimen. Results There was no significant difference between Ar-GDP and GDP-fib groups in blood compatibility analysis. While in the MTT test, cellular proliferation was benefited from the presence of fibronectin coating. The numbers of cells on Ar-GDP and GDP-fib specimens were greater than those in the original specimens after 24 h of culturing. Conclusions GDP treatment combined with fibronectin grafting favored MG-63 cell adhesion, migration, and proliferation on titanium surfaces, which could be attributed to the improved surface properties.

Development and biocompatibility tests of electrospun poly-l-lactide nanofibrous membranes incorporating oleic acid-coated Fe3O4

Abstract The aim of this study was to develop an electrospun poly-l-lactide (PLLA) nanofibrous membrane incorporating oleic acid-coated Fe3O4. The Fe3O4 nanoparticles were prepared using the chemical co-precipitation method, and particle diameters were analyzed using transmission electron microscopy. After mixing the oleic acid-coated Fe3O4 nanoparticles and PLLA, a membrane with nanofibers was manufactured using the electrospinning technique. Our results showed that Fe3O4 nanoparticle diameters fabricated in this study were concentrated at 2–8 nm (84.2%). After magnetizing, there exists an approximately linear relationship between magnetic flux density and membrane thickness (R2=0.7, p<0.05). NIH-3T3 fibroblast cells cultured on the magnetized Fe3O4/PLLA nanomembranes exhibited a more spreading and attached phenotype. These results can serve as a reference for future advanced studies.

A novel HA/β-TCP-collagen composite enhanced new bone formation for dental extraction socket preservation in beagle dogs

Past studies in humans have demonstrated horizontal and vertical bone loss after six months following tooth extraction. Many biomaterials have been developed to preserve bone volume after tooth extraction. Type I collagen serves as an excellent delivery system for growth factors and promotes angiogenesis. Calcium phosphate ceramics have also been investigated because their mineral chemistry resembles human bone. The aim of this study was to compare the performance of a novel bioresorbable purified fibrillar collagen and hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ceramic composite versus collagen alone and a bovine xenograft-collagen composite in beagles. Collagen plugs, bovine graft-collagen composite and HA/β-TCP-collagen composite were implanted into the left and right first, second and third mandibular premolars, and the fourth molar was left empty for natural healing. In total, 20 male beagle dogs were used, and quantitative and histological analyses of the extraction ridge was done. The smallest width reduction was 19.09% ± 8.81% with the HA/β-TCP-collagen composite at Week 8, accompanied by new bone formation at Weeks 4 and 8. The HA/β-TCP-collagen composite performed well, as a new osteoconductive and biomimetic composite biomaterial, for socket bone preservation after tooth extraction.

A Novel Porcine Graft for Regeneration of Bone Defects

Bone regeneration procedures require alternative graft biomaterials to those for autogenous bone. Therefore, we developed a novel porcine graft using particle sizes of 250–500 μm and 500–1000 μm in rabbit calvarial bone defects and compared the graft properties with those of commercial hydroxyapatite (HA)/beta-tricalcium phosphate (β-TCP) over eight weeks. Surgery was performed in 20 adult male New Zealand white rabbits. During a standardized surgical procedure, four calvarial critical-size defects of 5 mm diameter and 3 mm depth were prepared. The defects were filled with HA/β-TCP, 250–500 μm or 500–1000 μm porcine graft, and control defects were not filled. The animals were grouped for sacrifice at 1, 2, 4, and 8 weeks post-surgery. Subsequently, sample blocks were prepared for micro-computed tomography (micro-CT) scanning and histological sectioning. Similar bone formations were observed in all three treatment groups, although the 250–500 μm porcine graft performed slightly better. Rabbit calvarial bone tissue positively responded to porcine grafts and commercial HA/β-TCP, structural analyses showed similar crystallinity and porosity of the porcine and HA/β-TCP grafts, which facilitated bone formation through osteoconduction. These porcine grafts can be considered as graft substitutes, although further development is required for clinical applications.

A novel porcine collagen GTR membrane for treatment of Class II molar furcation involvement

Abstract Guided tissue regeneration (GTR) in the management of the periodontal furcation invasion is effective. Recent studies showed significant clinical improvement with different materials. The purpose of this study is to provide clinical evaluations of the bone repair potential of GTR combined with a novel porcine collagen membrane in the treatment of human molar Class II furcations. Fifty-one patients, 33 males and 18 females, with no systemic disease and a mean age of 49.1 years, participated in this study after phase I periodontal therapy. Probing depth, attachment loss and radiographic examination were recorded and analyzed before and after GTR therapy with collagen membrane. In the soft tissue measurement, the periodontal pocket (PD) depth was 5.283±0.283 mm before the surgery and improved to 2.631±0.147 mm at 9 months after surgery (p<0.05). Attachment loss also showed similar results. The attachment level was 5.547±0.3 mm before the surgery, and 2.911±0.188 mm at 9 months after surgery (p<0.05). According to the measurements the attachment gain was 2.636±0.168 mm after GTR surgery with porcine collagen membrane. According to this study, the porcine collagen membrane can perform with exceptional results in attachment level gain for the Class II periodontal furcation involvement.