Gene delivery system of pDNA using the blood glycoprotein fetuin

Fetuin is a biocompatible plasma protein and strongly enhances phagocytosis of bacteria, DNA and apoptotic cells by peripheral blood cells such as monocytes, macrophages and dendritic cells. We developed a novel gene delivery system: ternary complexes constructed with pDNA, polyethylenimine (PEI) and fetuin. Without covalent binding, fetuin was able to coat pDNA–PEI complexes, and stable anionic nanoparticles formed at a weight ratio greater than 30. Optimised pDNA–PEI–fetuin complexes significantly decreased the cytotoxicity of pDNA–PEI complexes in the melanoma cell line B16F10. Furthermore, the pDNA–PEI–fetuin complexes had higher transgene efficiency compared to that of commercial lipofectin previously reported in B16F10 cells despite an anionic surface. The pDNA–PEI–fetuin complexes did not agglutinate with erythrocytes. The pDNA–PEI–fetuin complexes had high gene expression in the spleen after intravenous administration in mice. Thus, the pDNA–PEI–fetuin complexes were a useful in vivo gene delivery system with tropism for the spleen.     

A comparative study of unilateral dislocated mandibular condyle fractures in the rabbit.

PURPOSE: The purpose of this study was to clarify the influence of patients age and the degree of dislocation on the healing of mandibular condyle fractures. MATERIAL AND METHODS: Sixty-two Japanese white rabbits (30 adolescent and 30 adult ones) were used. They underwent unilateral condylar neck osteotomy. The proximal fragments were positioned differently. In group I animals, the fragments were reduced into the original position, in group II animals the small fragment was dislocated 55-90 degrees to the ramus. In group III animals the proximal fragment was dislocated 135 degrees to the ramus. Gross and microscopic assessment was performed 4, 6, 8, 12 and 24 weeks postoperatively. RESULTS: In adolescent animals, complete healing of the fracture was observed, irrespective of the degree of dislocation. In the adult animals, there was complete healing of the fractures in groups I and II. In group III animals, there was a decrease of ramus height accompanied by some fibro-cartilaginous changes. CONCLUSION: Even in rabbits condylar deformation has been found following heavily dislocated artificial condylar fractures in adult animals. This result will help to decide for or against surgical treatment of condylar fractures in humans.     

A two-year comparison of resin-based composite tunnel and Class II restorations in a randomized controlled trial

PURPOSE: To evaluate the performance of total tunnel restoration with resin-based composites compared to Class II resin-based composite restorations in a randomized controlled clinical study. METHODS: 63 approximal carious lesions were restored in 38 patients by two experienced operators. The carious dentin and approximal demineralized enamel were completely removed for the tunnel restoration leaving the marginal ridge intact. The occlusal opening was enlarged to the center of the tooth to make access easy. Conservative cavity preparation was performed for the Class II restorations. All cavities were restored with Scotchbond Multipurpose Plus and Z250. At baseline, 1- and 2-year recalls, the restorations were evaluated clinically by two examiners using a modified USPHS rating system and radiographically for recurrent, caries. RESULTS: Both types of restorations performed well clinically, and there was no significant difference (P> 0.05) by the chi square test in any category. Recurrent caries was not detected. Only one tunnel restoration had to be replaced with a Class II restoration because of falling of the restoration with the marginal ridge after 2 years. However, no significant difference in the survival rates (P> 0.05) could be demonstrated between the restorations by the log-rank test.     

Effects of plaque control on the patency of dentinal tubules: an in vivo study in beagle dogs

BACKGROUND: The aim of this in vivo study was to evaluate the effects of plaque control on the patency of dentinal tubules using vital teeth of beagle dogs. METHODS: Class V cavities were prepared on the cervical areas of the mandibular and maxillary molars in each dog with a diamond point. To simulate the state of dentinal hypersensitivity, the teeth were etched with 50% citric acid for 2 minutes to obtain patent dentinal tubules. Plaque control was achieved by brushing the left-side teeth every day, whereas no plaque control was performed for the right-side teeth. A dentin biopsy was performed after 1, 2, and 3 weeks using the cylindrical diamond point to obtain dentin specimens. RESULTS: In the plaque control group, some of the dentinal tubules were occluded with precipitate (Ca/P=1.49), and the diameter of the dentinal tubules decreased from 2.42+/-0.33 microm (mean+/-SD) to 1.11+/-0.51 microm after 7 days, although most of the dentinal tubules remained open. In contrast, no precipitate was observed in the dentinal tubules of the non-plaque control group. Also, the diameter of the dentinal tubules increased from 2.42+/-0.33 to 2.9+/-0.49 microm, due to the demineralization of the peritubular and intertubular dentin. CONCLUSION: Plaque control plays a key role in reducing the patency of dentinal tubules and, therefore, might promote the natural repair of dentinal hypersensitivity.     

Inhibitory effects of tumor necrosis factor-alpha on migration of human periodontal ligament cells

BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is associated with chronic gingival inflammation and is suspected to influence periodontal destruction. However, the exact roles of TNF-alpha in wound healing and periodontal tissue regeneration are largely unknown. In the present study, we examined the effects of TNF-alpha on migration and proliferation of human periodontal ligament (PDL) cells. METHODS: PDL cells were cultured in the presence of TNF-alpha to determine its effects on cellular migration and proliferation. The protein expression profiles of alpha5 and beta1 integrin subunits and their related molecules, paxillin and focal adhesion kinases (FAK), were investigated. Gene expression of fibronectin also was assayed. Further, the activation of Rho-family small guanosine triphosphate (GTP)-binding protein (RhoA) was evaluated using a GTP-loading pull-down assay, and focal adhesion formation by PDL cells after transfection with the expression vector of paxillin-fused green fluorescent protein (GFP) also was observed with confocal microscopy. RESULTS: Cellular migration was impaired by TNF-alpha and recovered following the addition of anti-TNF-alpha antibodies. In contrast, PDL cell proliferation was not affected by TNF-alpha. TNF-alpha upregulated the expression of the alpha5 and beta1 integrin subunits, whereas fibronectin was not overexpressed. Phosphorylation of paxillin and FAK by PDL cells was induced, and RhoA activation also was induced. Confocal microscopic analysis revealed that TNF-alpha induced focal adhesion and stress fiber formation in all parts of the cells. CONCLUSION: Our results suggested that TNF-alpha impairs cellular migration by enhancing cellular adhesive ability following significant focal adhesion and stress fiber formation.