Bone formation and osseointegration with titanium implant using granular- and block-type porous hydroxyapatite ceramics (IP-CHA).

The aim of the present study was to examine whether interconnected porous hydroxyapatite ceramics (IP-CHA) could be used as bone substitute for implant treatment in reconstructive surgery. We firstly assessed if surround of the titanium surface placed into granular or block-type IP-CHA can observe new bone formation in a rabbit bone defect model. Subsequently, osseointegration and stability of titanium implant inserted into block-type IP-CHA was investigated in a rabbit onlay graft model. Direct contact between new bone and the surface of the titanium in granular- or block-type IP-CHA was found in a rabbit bone defect. Further, new bone formation was found in direct contact with the implant surface in the block-type IP-CHA in an onlay graft model, and the implant stability quotient (ISQ) values were significantly increased after surgery. Therefore, IP-CHA may be a useful material for implant treatment in reconstructive surgery strategies.

Clinical evaluation of novel interconnected porous hydroxyapatite ceramics (IP-CHA) in a maxillary sinus floor augmentation procedure.

The aim of this study was to evaluate the clinical behavior and the histological aspects of interconnected porous hydroxyapatite ceramics (IP-CHA) in maxillary sinus floor augmentation procedures. A 59-year-old female patient received one-stage implant integration with right maxillary sinus floor augmentation with mixture grafts from the cortical bone and IP-CHA. Implant stability of each fixture increased at 9 months after fixture installation compared with the first operation and adequate fixation of each fixture could be obtained. Histological analysis revealed there was new bone formation in the majority of pores of IP-CHA. Moreover, on a panoramic radiograph taken at 33 months the mixture grafts were distinctly observed as a radiopacity in the right sinus cavity, and marked absorption of mixture grafts was not found. Our results suggest that IP-CHA has the potential to provide a major scaffold for osteoprogenitor cells and is a useful grafting material for maxillary sinus augmentation.

Transcriptional effect of a calmodulin inhibitor, W-7, on the ligninolytic enzyme genes in Phanerochaete chrysosporium.

We investigated the effects of a calmodulin (CaM) inhibitor, W-7, on the expression of lignin peroxidase (LiP) and manganese peroxidase (MnP) genes in Phanerochaete chrysosporium to consider the role of cam gene, which was upregulated in parallel with the total activities of LiP and MnP in our previous transcriptomic analysis. The addition of 100 µM W-7 to the fungal cultures repressed the total activities of LiP and MnP, whereas the addition of 100 µM W-5, which is a control drug of W-7, retained approximately half of them, indicating that the effect of W-7 was attributable to CaM inhibition. Real-time reverse transcription polymerase chain reaction analysis revealed that most of lip and mnp isozyme genes predicted from whole-genome data were significantly inhibited by W-7 at the transcription level (P ≤ 0.05). These results suggest that CaM has an important role for the expression of isozyme genes of LiP and MnP at the transcription level.

Changes in the gene expression of the white rot fungus Phanerochaete chrysosporium due to the addition of atropine.

We constructed a LongSAGE (Long Serial Analysis of Gene Expression) library from a 3-d culture of Phanerochaete chrysosporium supplemented with atropine, which inhibits the production of lignin-degrading enzymes. The library (the atropine library) contains 13,108 LongSAGE tags and 6,783 unique tags. The gene expression profile represented by the tags was compared with those of two previously constructed libraries, one of which was constructed using 2-d cultures in which the fungus had not yet produced ligninolytic enzymes (the 2-d library) and the other was constructed using 3-d cultures in which the fungus had just started to produce the enzymes (the 3-d library). We found a total of 595 genes that were at least twice more highly or at least twice less highly expressed in the 3-d library than in the 2-d library or the atropine library, and the fluctuations were statistically significant. The relationships among these 595 genes were considered using cluster analysis. Of the 595 genes, 164 showed expression patterns similar to those of four ligninolytic enzyme genes, which were more expressed on day 3 than under any other conditions. Many of these 164 genes comprised genes possibly involved in lignin degradation, lipid metabolism, xenobiotic degradation, stress response, or signal transduction pathways.

Expression of TPX2 in salivary gland carcinomas.

TPX2 is a microtubule-associated protein and is required for microtubule formation at kinetochores in mammalian cells. The purpose of this study was to clarify the expression of TPX2 mRNA and correlation between TPX2 and clinicopathological factors in salivary gland carcinomas. The expression of TPX2 mRNA was investigated in 20 human salivary gland carcinomas (8 mucoepidermoid carcinomas, 7 adenoid cystic carcinomas, 5 acinic cell carcinomas) and 6 normal submandibular glands using real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The mean expression level of TPX2 mRNA was higher in mucoepidermoid carcinomas (0.53+/-0.51) than in normal submandibular glands (0.047+/-0.029); a significant association was found (Mann-Whitney U test, P=0.0067). The mean expression levels of TPX2 were also higher in acinic cell carcinomas (0.45+/-0.49) and adenoid cystic carcinomas (0.28+/-0.22) than in normal submandibular glands. Statistical correlations were found (Mann-Whitney U test, P=0.028 and P=0.003, respectively). Correlation between expression of TPX2 and receptor for hyaluronan-mediated motility (RHAMM) was also investigated in this study. A significant association was found between the mRNA expression levels of TPX and RHAMM (Pearson's correlation coefficient by rank test, P=0.020). These results indicate that human TPX2 mRNA is closely linked to increased or abnormal cell proliferation in malignant salivary gland tumors.

Long serial analysis of gene expression for transcriptome profiling during the initiation of ligninolytic enzymes production in Phanerochaete chrysosporium.

To analyze the transcriptome profile during the initiation of manganese peroxidase (MnP) and lignin peroxidase (LiP) production in Phanerochaete chrysosporium, we constructed long serial analysis of gene expression (LongSAGE) libraries. A total of 13,666 tags (the number of cumulative counted tags) that included 6,945 unique tags (the number of distinct tags) were isolated from the day-3 culture, which just started the enzymes production and was 24 h after veratryl alcohol addition and oxygen-purge into the culture (day-2 culture). A total of 12,402 tags that included 6,396 unique tags were isolated from the day-2 culture, in which the activity of enzymes is not detected. The comparison of the two libraries suggested that 38 genes showed significant (p < or = 0.01) fourfold or greater upregulation; this included the MnP gene (mnp2, mnp3) and LiP H8 gene. On the other hand, 43 genes showed significant (p < or = 0.01) fourfold or greater downregulation. This LongSAGE analysis found many new candidate genes regulating the enzymes production.