Chlorhexidine is a cationic bisbiguanide with broad antibacterial activity, and diverse activity encompassing gram-positive and gram-negative bacteria, yeasts, dermatophytes, and some lipophilic viruses. Chlorhexidine alters the permeability of the bacterial membrane. They also neutralize periodontal pathogens such as Streptococcus aureus, Porphyromonas gingivalis, and Prevotella intermedia. Porphyromonas gingivalis were cultured to isolate the Porphyromonas gingivalis outer membranes (OPG). Also, OPG were used to prepare large unilamellar liposomes with total lipids (OPGTL) extracted and prepared with the phospholipids mixture (PL). The effect of chlorhexidine digluconate on cell membrane kinetic changes was investigated using fluorescence polarization of n-(9-anthroyloxy) stearic acid with different rotational mobility depending on the probe substitution position (n) in the membrane phospholipid aliphatic chain. Chlorhexidine digluconate increased the hydrocarbon rotational mobility interior of the OPG, OPGTL and PL, but native and model membranes interfacial mobility were decreased. The sensitivity to increasing chlorhexidine digluconate effect on rotational mobility was proportional to the depth of probe position in descending order of 16-AP, 12-AS, 6-AS and 9-AS. Chlorhexidine digluconate disordering or ordering the effects on the membrane lipids may cause its bacteriostatic and bacteriocidal actions.

Herbal medicine has been the basis for medical treatments through much of human history, and such traditional medicine is still widely practiced today. Modern medicine makes use of many plant-derived compounds as the basis for pharmaceutical drugs. In traditionally, Achyranthes aspera, Safflower (Carthamus tinctorius) seed and Acanthopanax senticosus have been used for the treatment and prevention of bone-related diseases. In this study, we investigated the pharmacological effect of mixture of Achyranthes aspera, Safflower (Carthamus tinctorius) seed and Acanthopanax senticosus and the other herbs. Two types of enzymes were used to enhance the extraction components of amino acid, mineral content, free sugar, and flavor recovery in extracting natural herbal mixtures(NME). We evaluated regulation of osteogenic differentiation in human bone marrow mesenchymal stem cells using alkaline phosphatase staining, alizarin red S staining and RT-PCR. The CCK-8 assay indicated that NME had no cytotoxicity but increased cell survival. In addition, NME promoted the mineralization and expression of osteogenic differention marker genes in human bone marrow mesenchymal stem cells. Therefore, NME has an effect of promoting proliferation and osteogenic differentiation of human mesenchymal stem cell.

Nerve injury induced protein 1 (Ninjurin1) was originally described in neuroscience in which the expression of Ninjurin1 was regulated by Schwann cells and dorsal root ganglion neuronal cells of damaged nerve tissues. After the first discovery of Ninjurin1, the widespread expression of Ninjurin1 in adult and embryonic tissues have been observed including bone marrow, peripheral blood lymphocytes, thymus, and heart. Currently, the Ninjurin1 mediated positive regulation of pre-osteoclasts fusion and osteoclast development was reported. The bone homeostasis is dynamically balanced by bone-resorbing activity of osteoclast and bone-forming activity of osteoblast. Until now, the role of Ninjurin1 was never been described in osteoblastogenesis. Therefore, in this study, we have evaluated the expression and function of Ninjurin1 in osteoblast. The ample expression of Ninjurin1 was observed in bone marrow of mouse tibia sections but it was barely expressed in osteocytes. And also the expression levels of Ninjurin1 were gradually increased during osteoblast differentiation of calvarial pre-osteoblast, C2C12, and MC3T3-E1 cells. Importantly, the expression of Ninjurin1 was increased in the absence of osteogenic stimulus, BMP2, which suggests the cell density-dependent regulation of Ninjurin1. The controlled expression of Ninjurin1 by cell-density was evidently shown in not only pre-osteogenic osteoblast lineage cells but also in non-osteogenic cancer cells such as HeLa and A549 cells. In addition, the isoform-specific knockdown of Ninjurin1 remarkably reduced the alkaline phosphatase (ALP)-positive osteoblast differentiation. Thus, our results suggest a previously unappreciated mechanism of Ninjurin1 expression and also suggest its role on osteoblastogenesis.

The Hippo pathway was originally discovered in Drosophila by genetic screening and it has been shown to be conserved in various organisms including human. Until now, the essential roles of Hippo pathway in regulating cell proliferation, apoptosis, tumorigenesis, and organ size control is extensively studied. Currently, Mats1/2 (Mob1a/1b), one of the important components in Hippo pathway, mutant mice were generated which has abnormal phenotype such as resistance to apoptosis and spontaneous tumorigenesis. Of note, Mats1/2 mutant mice also showed dental malocclusion. Therefore, in this study, we have evaluated the bone phenotype of Mats1/2 mutant mice. Although the mRNA expressions of Mats1 or Mats2 were observed in both osteoclastogenesis and osteoblastogenesis, the increase of Mats1 level was most prominent during osteoblastogenesis. The RANKL-induced osteoclast differentiation from bone marrow-derived macrophages (BMMs) was unaltered upon Mats1/2 mutation; however, the osteoblast differentiation using calvarial pre-osteoblasts was significantly reduced in Mats1/2 mutant mice compare to that of wild type mice. In accordance with in vitro results, Mats1/2 mutant mice showed decreased bone volume as well as increased trabecular separation in μCT analyses. These results may provide novel prospect of the probable linkage between Hippo pathway and bone homeostasis.

Connective tissue growth factor (CTGF, CCN2) is one of the multi-functional secreted proteins which belong to CCN family of cysteine-rich growth factors. CTGF is known to have pivotal roles in embryonic endochondral ossification but its role in relevance to periodontitis is never been determined. To identify new molecular mediators associated with periodontitis-induced bone resorption, we have analyzed publicly available GEO database and found the markedly augmented CTGF mRNA expression in periodontitis gingival tissues. The existence of CTGF significantly enhanced mature osteoclasts survival which accompanied by reduction in TUNEL-positive nuclei and PARP cleavage. These results may provide another line of evidence the CTGF mediated prolonged osteoclast survival and subsequent increased bone resorption in the periodontitis patients.

Visfatin is a pro-inflammatory cytokine, which is thought to play a central role in systemic inflammation and the pathogenesis of obesity related diseases. Only a few studies investigated the effect of visfatin on human cancers. Furthermore, there have been no studies on the association between the expression of visfatin in OSCC tissue and its effect on OSCC patients. Hence, the present study analyzed the expression of visfatin in OSCC from Korean patients. Immunohistochemistry for visfatin was performed using 12 normal oral mucosas (NOM), 16 oral leukoplakias (with/without dysplasia), and 58 OSCC patients samples. Immunoreactivity was semi-quantitatively scored and the correlation between the expression of visfatin and clinicopathological parameters of OSCC patients was analyzed. The immunohistochemical analysis demonstrated that the expression level of visfatin increased in OSCC alone (p<0.05). Moreover, the immunoexpression score of visfatin was significantly correlated with TNM stage of OSCC patients. Our findings suggested that visfatin can play a certain role in the pathogenesis of OSCC. In addition, visfatin was associated with the tumor progression of OSCC patients and may act as independent biomarker of OSCC.