CD81 belongs to thetetraspanin family of cell-surface proteins, which contain four transmembrane domains and two outer-membrane loops. CD81 has been shown to be up-regulated in synoviocytes in rheumatoid arthritis (RA) and to promote synoviolin expression in the progression of RA. Recently, we showed that a small interfering RNA (CD81siRNA) targeting the gene encoding CD81 ameliorates arthritis in collagen-induced arthritic (CIA) rats. CD81 siRNA also decreased the expression of TNF-α in SW982 cells. Additionally, we established hybridoma cell lines producing monoclonal antibodies (MAbs) against CD81 and demonstrated that some of these MAbs impaired the proliferation of C6 glioma cells. Here, we report cloning of the cDNA encoding one such MAb (H chain and L chain) from hybridoma cells and construction of mouse-rat chimeric genes for expression of the chimeric antibody. This chimeric antibody, as expressed by CHO cells, bound to CD81 at the target’s outer-membrane loop (LEL).

Cartilage degradation frequently occurs during the postmenopausal period due to a decrease in type 2 collagen and constitutes a condition affected by the level of estrogen in the body. The purpose of this study is to prove that such postmenopausal reductions in estrogen levels influence the type 2 collagen expression in cartilage. Oophorectomies were performed on 54 Rattus norvegicus subjects and each of which was subsequently assigned to one of six different groups. 17β-estradiol present in the blood of the subjects was examined by means of ELISA technique, while the expression of MMP-13 and type 2 collagen in their knee joint cartilage was assessed immunohistochemically. A significant post-oophorectomy decrease of 17β-estradiol was detected in subjects (p<0.05), while the expression of MMP-13 increased appreciably (p<0.05) and a marked decrease in type 2 collagen expression occurred (p<0.05). The reduction in 17β-estradiol promoted an increase in the expression of MMP-13 and a decrease in type 2 collagen expression.

We have found newly synthesized peptide derived from commercially available Emdogain^® for periodontal tissue regeneration therapies in our previous study. That synthesized peptide consisted of seven amino acid sequence, WYQNMIR, which is exon 5 of porcine amelogenin and had the same effects as Emdogain^®. The amino acid sequence of human amelogenin exon 5 is WYQSIR which both N (asparagine) and M (methionine) in porcine amelogenin are replaced by S (serine). In the present study, we investigated the effect of a synthetic human amelogenin-derived peptide (HAP) consisting of WYQSIR on human periodontal ligament stem cells (PDLSCs) to be useful or not for the tissue regeneration. PDLSCs were isolated from third molars of adult donors. The effect of HAP on PDLSCs was investigated by culturing them in normal or osteogenic medium with and without HAP. Proliferation of the PDLSCs was evaluated using a cell proliferation assay after they had been treated with HAP (ranging from 1 ng/ml to 10,000 ng/ml) for 1, 3, 5, and 7 days. Osteogenic differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, calcium deposition, osteocalcin production, and mRNA expression of Runx2 and osteonectin after the PDLSCs had been treated with HAP (1,000 ng/ml) for 1, 7, 14, and 21 days. The number of viable cells was significantly increased in the presence of HAP in normal medium. Compared to cells cultured without HAP in osteogenic medium, cells cultured with HAP showed significantly increased ALP activity, calcium deposition, osteocalcin production, and Runx2 mRNA expression. The results show that HAP enhances proliferation and osteogenic differentiation of PDLSCs. The present study suggests that HAP may be a useful material for periodontal tissue regeneration.

The purpose of this study was to evaluate sella turcica morphology, calculate its size and area in subjects with Down syndrome (DS), and compare them with normal subjects. A total of 100 (50 normal and 50 DS) Malay subjects who had their computed tomography (CT) scan at the Radiology Department, Hospital Universiti Sains Malaysia (HUSM) for ordinary diagnosis. The selected age groups were divided into four groups as children (0-6 years), pre-adolescents (7-12 years), adolescents (13-20 years) and adults (21-35 years). The images were processed using Mimics V17.0 software. Conventional measurements included three different heights of the sella turcica (anterior, posterior, median), its length, diameter, area and width, measured in relation to the Frankfort reference line (FH). Morphometric methods were used to assess sella shape. Significant differences were found between control and DS groups in most parameters except for the sella height anterior and median, and sella area. No significant differences in size of the sella were found for control and DS groups in most parameters between genders. When age was evaluated, there were significant differences among some age groups and non-significant in other. The study found that sella turcica in DS presented with a three different shapes: U shape (50%), J shape (42%) and shallow (8%). The size and shape of the sella turcica is different between control and DS subjects. It was found that males had more significant differences in measured parameters compared to female. Among the age groups, the children age between 0-12 years old showed highly significant difference between DS and normal individuals, compared to adolescent and adults groups.

The aim of this study was to determine the potential for an additive effect from combining treatments with tolerogenic dendritic cells (tDCs) and etanercept (ETN) in a case of established type II collagen-induced arthritis (CIA). CIA mice were treated with type II collagen-pulsed tDCs either alone or in combination with ETN. We compared the effects of monotherapy with tDCs or ETN with the effect of combination treatment on the resultant rheumatoid arthritis using histopathological analyses and measurements of paw volumes, regulatory T cell populations, and cytokine levels. The clinical and histologic analysis revealed an additive effect from the combination therapy. Monotherapy with tDCs showed a tendency to ameliorate arthritis symptoms, while monotherapy with ETN reduced the paw swelling at 3 days after administration, but was not effective toward the end of the experimental period. The combined treatment group also showed a marked inducement of the regulatory T cells and Th2 cytokines that are affected by tDCs compared with those of either untreated or ETN-treated CIA mice. ETN administered in combination with tDCs provided a substantially higher level of clinical benefit at the cellular level to CIA mice than the administration of either of these agents alone.

A novel chemically curable, resin-modified-type mineral trioxide aggregate (MTA) material polymerized by tri-n-butylborane (PCX-TBB), and light-curable, resin-modified-type MTA material (TheraCal LC^®) clinically useful were evaluated in terms of solubility, calcium-release ability, compressive strength and apatite-forming ability. The solubility of PCX-TBB conformed to ISO 6876, whereas that of TheraCal LC^® did not conform to the abovementioned standard. The calcium-release ability of PCX-TBB and TheraCal LC^® showed no difference after 1 day of immersion in purified water. However, after 30 days of immersion, the calcium concentration of PCX-TBB increased and became higher than that of TheraCal LC^®. The compressive strength of PCX-TBB was stable even if immersed in simulated body fluid (SBF) for 30 days. In contrast, the compressive strength of TheraCal LC^® showed a tendency to decrease slightly by immersion in SBF. Furthermore, the apatite-forming ability of PCX-TBB was determined to be superior to that of TheraCal LC^®. The results presented herein suggest that PCX-TBB has potential as a superior MTA material and may have improved hard tissue-induction ability compared to that of TheraCal LC^®.

In recent years, the application of conditioned medium (CM) in regenerative medicine has received much attention. However, the incidence of cancer in Japan is continuing to rise, with one in two people having cancer, among those who may be candidates for CM treatment and may be affected by or have a risk of cancer. We investigated the effects of human dental pulp stem cell (DPSC)-derived CM on human oral squamous cell carcinoma (OSCC) cell lines. CM was extracted from DPSC isolated from human dental pulp, and cell proliferation rates upon contact with OSCC cell lines were compared. Furthermore, chemosensitivity was evaluated with the collagen gel droplet embedded culture drug sensitivity test and was compared using Dulbecco’s modified Eagle’s medium (DMEM). To test the effect of DPSC-CM on xenograft tumors, an in vivo comparative study was conducted to investigate the tumor proliferation rate upon DPSC-CM and DMEM administration in tumor-bearing nude mice. The tumor growth factor production in culture medium over time was measured with enzyme-linked immunosorbent assay (ELISA) and was compared using DMEM. Vascular endothelial growth factor (VEGF) level significantly increased in the DPSC-CM contact group with ELISA. Therefore, VEGF-A-mRNA expression in the OSCC cell line was studied with real-time polymerase chain reaction for comparison. No significant differences were observed in the cell proliferation rate or drug sensitivity between different culture media in each cell line or in in vivo tumor proliferation rate. However, VEGF level contained in the cultured medium was significantly higher than that in the DPSC-CM group. VEGF-A-mRNA level in OSCC cell lines was significantly higher in the DPSC-CM group, which increased over time. These results suggested that exposure to DPSC-CM does not immediately affect tumor growth or drug resistance, but induces VEGF overexpression in tumor cells.

In the field of forensic odontology, not only personal identification using oral conditions, including dental treatment marks and DNA typing, but also species identification, age or sex estimation using cranial or partial bones, and time after death may also be applicable as estimation methods. Among these many tasks, one of the most difficult is species identification of fragmented calvarium. This is because the calvarium has poor morphological features, except that it is a flat bone, and few reports have described how to differentiate it. In this study, a simple identification method for turtle shell (carapace and plastron) whose morphological characteristics closely resembled those of the calvarium was applied. As a result, in an enlarged image obtained using a stereoscopic microscope, the characteristics of each suture pattern could be confirmed. In the decalcified and non-decalcified bone specimens, the difference in the inner/outer laminar structure and the specific structural difference of the cancellous bone-equivalent part were confirmed. Furthermore, most of the features could be obtained in the destructive inspection when it was examined using micro CT imaging to determine whether discrimination by nondestructive inspection was possible or not. No significant difference in ingredients was found using the Electron Probe X-ray Micro Analyzer. The calvarium was more calcified than the carapace but less calcified than the plastron. From these results, we suggest that micro CT imaging is effective for discriminating between the calvarium and turtle shell in a short time.

The aim of this study was to synthesize a polymeric nanocarrier system loaded with superparamagnetic iron oxide nanoparticles (SPION) and the sulforaphane(SFN) through a cross-linking method in order to protect the SFN. According to this experiment, the drug-loading content and encapsulation efficiency were 29.97±0.08% and 0.34±0.41%, respectively. The total cumulative amount of SFN, released from the nanocarrier in vitro was 87.90% over a 72 h period. MTT assays indicated that the SPION-SFN-loaded nanocarrier exhibited a much higher in vitro antitumor efficacy than free SFN. Together, these results indicate that SPION-loaded nanocarrier exhibits good drug-loading ability and sustained-release function.

In this paper, the micro-structured hydroxyapatite (HA)/TiO₂ coating was firstly produced on titanium surface by one-step micro-arc oxidation (MAO), and then bone morphogenic protein-2 (BMP-2)-encapsulated chitosan (CS) coating was prepared on the HA/TiO₂ surface by dip-coating method to endow the bioactive HA/TiO₂ coating with good antibacterial property and improved biological properties. The bonding strength between coatings was studied by scratch method. The degradability of CS, BMP-2 release behavior, bioactivity, biocompatibility and antibacterial property of the BMP-2/CS/HA/TiO₂ composite coating were examined by in vitro tests. The results showed that, after loading BMP-2/CS on HA/TiO₂ surface, the surface roughness and wettability decreased but the bonding strength between the HA coating and titanium substrate was improved. CS, as a suitable carrier for sustained release of BMP-2, can control the release of BMP-2 continuously for 4 weeks long in a simulated human serum, and in PBS solution BMP-2 can be released more slowly. In addition, CS endowed the BMP-2/CS/HA/TiO₂ composite coating with good antibacterial activity. The BMP-2/CS/HA/TiO₂ showed good bioactivity, and it was also beneficial for cell adhesion, spreading and proliferation. Based on the above results, BMP-2/CS/HA/TiO₂ has great potential to be applied in clinic.

This study aimed to test stress relaxation and creep of the femur in osteoporotic rats after bone mesenchymal stem cell (BMSC) intervention and to provide a viscoelastic basis for the prevention and treatment of osteoporosis. The osteoporotic animal models prepared by ovariectomy were intervened with BMSCs and alendronate sodium tablets (ASTs), respectively, for 30 days, and then compared for serum calcium (Ca), phosphorus (P), alkaline phosphatase (ALP), estradiol (E2), estrogen receptor (ER), superoxide dismutase (SOD), and malondialdehyde (MDA) levels, as well as for stress relaxation, creep, and bone mineral density (BMD). BMD and Ca, ALP, E2, ER, and SOD levels, as well as 7200 s stress reduction and 7200 s strain increase, were statistically higher in BMSC group than in MOD and AST groups (p < 0.05). However, P and MDA levels in BMSC group were lower than those in MOD and AST groups. BMSC intervention can improve bone quality and restore bone viscoelasticity in osteoporotic rat models.