A blood test is a laboratory analysis performed on a blood sample that is usually extracted from a vein in the arm using a needle, or via fingerprick. They are used to determine physiological and biochemical states, such as disease, mineral contents, drug effectiveness, and organ function. Although the term blood test is used, most routine tests (except for most haematology) are done on plasma or serum, instead of blood cells. Main advantage of using saliva in diagnostics is easy and non invasive sample taking compared to peripheral blood. According to the study published, saliva contains more than 20 percent of the proteins found in blood. The purpose of present study is to compare biochemical enzymes in saliva and in blood serum and to evaluate the usefulness of saliva specimens instead of blood in dental clinic. The saliva from 215 healthy over 50 years of aged people lived in Dong-gu district, Gwangu city was collected and the analysis was performed by six enzyme-linked immunosorbent assays (ELISA). ELISA results were compared with blood chemistry results. The values or patterns on Alanine Aminotransperase (ALT), Aspartate Aminotransperase (AST), Cholesterol and Triglyceride in saliva were not correlated with those in blood serum. However, Albumine and γ-glutamyl transpeptidase (γ-GTP) were followed the positive relationship with blood chemistry. These result showed that detection and identification of Albumine and γ-GTP level could be established by saliva ELISA analysis, so that ELISA assay on saliva could be useful alternative to serum testing.
Diabetic patients tend to exhibit delayed bone formation and osteoblast differentiation, which results in osteopenia. Recently, numerous clinical reports suggest that 635-nm light irradiation improves bone regeneration and wound healing, and reduces pain in patients suffering from diabetes. The purpose of the present study was to test the hypothesis that 635-nm irradiation can influence bone formation by MC3T3-E1 osteoblasts cultured on high concentrations of glucose(25mmol/L D-glucose) in the presence or absence of phorbol 12-myristate 13-acetate(PMA), and to establish an in vitro pathological model of bone formation. The effect of 635-nm irradiation on bone formation was investigated using Alizarin Red S staining, and alkaline phosphatase enzyme activ ity and calcium deposition assays. In addition, gene expression of the o steogenic markers BMP-2, osterix and osteocalcin were assayed by RT-PCR. Calcium deposition by MC3T3-E1 cells was reduced in the presence of high concentrations of glucose or by PMA supplementation. However, 635-nm irradiation led to an increase in calcium deposition by MC3T3 cells, followed by increased bone mineralization. mRNA expression of BMP-2 and osterix at an early stage and of osteocalcin at a late stage was significantly upregulated by 635-nm irradiation in MC3T3-E1 cells supplemented with high concentrations of glucose. Irradiation at 635 nm increases bone mineralization in MC3T3-E1 cells cultured in vitro on high concentrations of glucose and alters osteogenic gene expression, which accelerates bone formation in hyperglycemic conditions.
Human gingival fibroblasts (hGFs) were reported to play an important role in inflammatory reactions to lipopolysaccharide (LPS) from P.gingivalis in the periodontal connective tissue. Although the biostimulatory effects of hyperbaric oxygen therapy, such as anti-inflammatory activity, have been reported, the pathological mechanism is not completely understood. This study examined the changes in the inflammatory cytokine profiles, which are produced after exposure to hyperbaric oxygen in P.gingivalis LPS-treated human gingival fibroblasts, and subsequently to examine the mitogen activated protein kinase (MAPK) pathway involved in cytokine production. Gingival fibroblasts with or without P.gingivalis LPS were exposed to hyperbaric oxygen, and the cytokine profiles in the supernatant were observed using a human inflammation antibody array. The expression of cyclooxyginase-2 (COX-2) protein, phosphorylation of extracellular signal-regulated kinase (ERK1/2), p38, and c-Jun-N-terminal kinase (JNK) MAPK by western blot analysis, and the amount of prostaglandin E2 (PGE2) in the supernatant by an enzyme-linked immunoassay were determined. COX-2 protein expression and PGE2productionwereincreasedsignificantlyintheP. gingivalis LPS-treated group, and were decreased by treating P. gingivalis LPS with hyperbaric oxygen. Treatment of P. gingivalis LPS in the gingival fibroblasts led an increase in the amount of pro-inflammatory-related cytokines interleukin-6 (IL-6) and IL-8 released, whereas hyperbaric oxygen inhibits the irrelease. Ananalysis of the MAPK signal transduction showed that hyperbaric oxygen induced a significant decrease in the level of P38 phosphorylation regardless of the presence or absence of LPS. In addition, hyperbaric oxygen promoted JNK phosphorylation, significantly in the presence of LPS. Hyperbaric oxygen can inhibit pro-inflammatory cytokines and mediate the MAPK signal pathway, and appears to be useful as an anti-inflammatory tool.
The pattern of wound healing process differs markedly according to the cell types. Gingival wounds heal more rapidly without scar, however dermal wounds show collagen laid down in thick disorganized patterns and keloid formation. This h as b een s uggested t o be d ue t o the presence of d ifferent E C M components a nd c ytokines a s well a s growth factors. The purpose of this study was to examine the differential expression of genes in connection with keloid formation in gingival fibroblasts (hGFs) and dermal fibroblasts (hDFs) in response to inflammation. In this study, we investigated the differences between hGFs and hDFs in the expression and production of cyclooxygenase (COX-2), prostaglandins E2 (PGE2), transforming growth factor (TGF)-β, collagens, matrix metalloproteinases (MMPs), and tissue inhibitors of matrix metalloproteinases (TIMPs) which play important roles in collagen deposition in wound healing. The hGFs and hDFs were primary cultured and allocated to arachidonic acid (AA) treatment group and control group. Protein and mRNA were extracted right after (0 hr) and 24 hr after AA treatment. At a defined concentration of AA in hGFs and hDFs, MTT assay was performed. The mRNA and protein expression levels of COX-2, TGF-β, collagen 1 and 3, MMP 1 and TIMP 1 were examined by Real-time PCR and Western blots. The amounts of PGE2 were measured by enzyme-linked immunosorbent assay (ELISA).The expression of COX-2 and TGF-β exhibited reduced levels in hGFs , but were increased in hDFs at 24 hr after AA treatment. Production of PGE2 was increased in hGFs and hDFs at right after AA treatment but, not changed at 24 hr after AA treatment. The protein and mRNA expression of collagen 1 and 3 were decreased in hGFs , whereas increased in hDFs at 24 hr AA treatment. Expression of MMP-1 protein was increased in hGFs at 24 hr but, was decreased in hDFs at 24 hr compared with that of control. The protein expression of TIMP-1 was decreased in hGFs but, was increased in hDFs at 24 hr compared with that of control. These observations demonstrate differential expression between gingival and dermal fibroblasts in regulation of collagenolytic capacity by extracellular matrix-associated genes in keloid formation associated with wound repair.
Candida albicans and their associated Candida species are opportunistic pathogens which exists as normal flora in the oral cavities of healthy individuals. In response to physiological changes in the host, these yeasts can become pathogenic, resulting in oral candidiasis. The rapid detection and identification of Candida species in clinical laboratories are extremely important for the management of patients with hematogenous candidiasis. The presently available culture and biochemical methods for detection and species identification of Candida are time-consuming and lack the required sensitivity and specificity. In this study, we have established a seminested PCR (snPCR) using universal and species-specific primers for detection of Candida species in saliva. The universal outer primers amplified the 3end of 5.8S ribosomal DNA (rDNA) and the 5end of 28S rDNA, including the internally transcribed spacer 2 (ITS2), generating 350- to 410-bp fragments from the four commonly encountered Candida spp., viz., C. albicans, C. tropicalis, C. glabrata, and C. parapsilosis. The saliva from 331 healthy and, over 50 years of aged people lived in Dong-gu, Gwangu city, was collected. Total DNA were extracted by Hoffman-Winston yeast total DNA prep. method and performed t he s nP CR. R esults appeared to b e negative on 292 people ( 88.2%), however, 2 6 people ( 7.9%) were p ositive Candida albicans, 6 people (1.8%) were positive Candida glabrata, 5 people (1.5%) were positive Candida tropicalis, and only 2 person (0.6%) were positive Candida parapsilosis. These result showed that detection and identification of Candida species could be established by saliva analysis, so that snPCR on saliva is useful method of diagnosis of clinical fields
Intracellular reactive oxygen species(ROS) produced in a various pathologic state was known to intermediate many cellular response such as inflammation. Recently, low level light irradiation by HeNe laser used in many clinical field could improve inflammatory state by scavenging intracellular ROS through photo-detachment/dissociation process. The purpose of this study is to investigate the differential effects of blue and red light irradiation on ROS scavenging effects. Immortalized human oral keratinocyte HaCat cells were used. Phorbol 12-myristate 13-acetate(PMA) was treated for inflammation. Red(635nm) and blue(470nm) light irradiation was carried out. To asses the intracellular ROS by light irradiation, confocal microscopic and flow cytometric assay with DCF fluorescence for total ROS and ESR spectrometry of DMPO-O2 - for superoxide anion were caried out. And microarray was performed for mRNA expression level. Released intracellular total ROS in PMA treated HaCat cell lines was dissociated efficiently by red light irradiation, while blue light irradiation did not. Rather, blue light irradiation increased ROS formation. For superoxide anion generated the first synthetic form of ROS, red light irradiation reduced its amount but blue light irradiation did not. In the mRNA expression in line with cyclooxygenase(COX) pathway, prostagrandin endoperoxide synthase 1(PTGS 1), prostagrandin endoperoxide synthase 2(PTGS 2) and phospholipase A2(PLA2) were increased by both light irradiation and they were decreased as time flows. And genes associated with ROS releasing, mRNA expressions of tumor necrosis factor receptor (TNFR) and interleukin 1beta(IL1B) were increased by 1 hour red light irradiation but did not by blue light irradiation. As a result, red and blue light irradiation showed different response in affecting the level of ROS. These findings indicate that red light rather than blue light is more useful for anti-inflammation in clinical field
The phytochemicals of many plants suggest their potential use as dietary supplements in cancer chemoprevention and chemotherapy. In the present study, antitumor activity of Cudrania tricuspidata, a plant native to East Asia, was investigated. Cell growth inhibition of the extract on HT-29 colorectal adenocarcinoma using MTT colorimetric assay was determined. Apoptosis on HT-29 cells was performed by DNA fragmentation analysis. PGE2 release was measured by enzyme immunoassay, because PGE2 is a key protumorigenic prostanoid in many human cancers. For the ROS scavenging activity, ROS level was detected by laser scanning confocal microscope. It was found that methanol extract of leaves inhibits cell viability by inducing apoptosis as evidenced by DNA fragmentation. Stem bark decreases synthesis of PGE2, inflammatory mediator. Fruits exhibited pronounced ROS scavenging activity. Taken together, these results suggest that Cudrania tricuspidata exerts growth inhibition and anti-oxidation on HT-29 cells through apoptosis, ROS scavenging respectively that it may have anti-cancer properties.
The purpose of the present study is to investigate the optimal wavelength, frequency and energy density for set up the photobiologic treatment of periodontal disease. To establish the present study, λ scan of 500㎚ to 900㎚ was used to search the optimal wavelength for maximal proliferation of human gingival fibroblasts. Cell proliferation assay was carried out as MTT assay. Light intensity of 0.8 to 3.25mW, frequency of 0 to 584㎐ and 0 to 2hours was applied for investigation of optimal energy density, frequency and applied duration. Finally, 628㎚ with 1mW/cm2 for 1hour of LED irradiation resulted in maximal proliferation of gingival fibroblasts. These results suggest that LED irradiation on gingival fibroblast show different proliferation according to the condition of irradiation, and demonstrate that LED irradiation can control the quantity of cell proliferation.
To investigate the differential expression of genes by 635nm LEDs irradiation in arachidonic acid-treated human gingival fibroblasts, cDNA microarray was carried out. Human gingival fibroblasts were primary cultured and arachidonic acid was treated to induce inflammation. 635nm of wave length was used for LEDs irradiation. The experimental group was categorized into four group ; control, only LEDs irradiation group, only arachidonic acid-treated group and arachidonic acid-treated with LEDs irradiation group. The expression of 8,078 genes were increased and the expression of 7,103 genes were decreased in only LEDs irradiation group. For arachidonic acid-treated with LEDs irradiation group, the expression of 6,815 genes were increased, while the expression of 8,031 genes were decreased comparing with only arachidonic acid-treated group. IL-13alpha2 receptor was the most expressed gene in LEDs irradiation group comparing with control, followed by MMP3. Genes which the most down regulated was BIRC3 in LEDs irradiation group. PLAB genes was the most up-regulated in arachidonic acid treated with LEDs irradation group, followed by ranked RARRES1. Considering the classification by cell function, genes associated with signal transduction were the most affected by LEDs irradiation, followed by the genes associated with nucleoside, nucleotide and nucleic acid metabolism. In arachidonic acid treated with LEDs irradiation, genes associated with signal transduction and protein metabolism were affected. Taken together, LEDs irradiation could affect various biological process and could identify many genes related to LEDs irradiation, which could be used for clinical application.