Differential Expression Profiling of Salivary Exosomal microRNAs in a Single Case of Periodontitis - A Pilot Study

Exosomes are Nano-sized lipid vesicles secreted from mammalian cells containing diverse cellular materials such as proteins, lipids, and nucleotides. Multiple lines of evidence indicate that in saliva, exosomes and their contents such as microRNAs (miRNAs) mediate numerous cellular responses upon delivery to recipient cells. The objective of this study was to characterize the different expression profile of exosomal miRNAs in saliva samples, periodically isolated from a single periodontitis patient. Unstimulated saliva was collected from a single patient over time periods for managing periodontitis. MicroRNAs extracted from each phase were investigated for the expression of exosomal miRNAs. Salivary exosomal miRNAs were analyzed using Affymetrix miRNA arrays and prediction of target genes and pathways for its different expression performed using DIANA-mirPath, a web-based, computational tool. Following the delivery of miRNA mimics (hsa-miR-4487, -4532, and -7108-5p) into human gingival fibroblasts, the expression of pro-inflammatory cytokines and activation of the MAPK pathway were evaluated through RT-PCR and western blotting. In each phase, 13 and 43 miRNAs were found to be differently expressed (|FC| ≥ 2). Among these, hsa-miR-4487 (|FC|=9.292005) and hasmiR-4532 (|FC|=18.322697) were highly up-regulated in the clinically severe phase, whereas hsa-miR-7108-5p (|FC|= 12.20601) was strongly up-regulated in the clinically mild phase. In addition, the overexpression of miRNA mimics in human gingival fibroblasts resulted in a significant induction of IL-6 mRNA expression and p38 phosphorylation. The findings of this study established alterations in salivary exosomal miRNAs which are dependent on the severity of periodontitis and may act as potential candidates for the treatment of oral inflammatory diseases.

Association of gingival biotype with the results of scaling and root planing

PURPOSE: The concept of gingival biotype has been used as a predictor of periodontal therapy outcomes since the 1980s. In the present study, prospective and controlled experiments were performed to compare periodontal pocket depth (PPD) reduction and gingival shrinkage (GSH) after scaling and root planing (SRP) according to gingival biotype. METHODS: Twenty-five patients diagnosed with chronic periodontitis participated in the present study. The PPD and GSH of the labial side of the maxillary anterior teeth (from the right canine to the left canine) were evaluated at baseline and 3 months after SRP. Changes in the PPD following SRP were classified into 4 groups according to the gingival thickness and initial PPD. Two more groups representing normal gingival crevices were added in evaluation of the GSH. The results were statistically analyzed using the independent t-test. RESULTS: In the end, 16 patients participated in the present study. With regard to PPD reduction, there were no significant differences according to gingival biotype (P>0.05). Likewise, sites with a PPD of over 3 mm failed to show any significant differences in the GSH (P>0.05). However, among the sites with a PPD of under 3 mm, those with the thin gingival biotype showed more GSH (P<0.05). CONCLUSIONS: PPD changes after SRP were not affected by gingival biotype with either shallow or deep periodontal pockets. GSH also showed equal outcomes in all the groups without normal gingival crevices. The results of SRP seem not to differ according to gingival biotype.

Retinoic Acid Increases the Cell Cycle Progression of Human Gingival Fibroblasts by Increasing Cyclin E and CDK 2 Expression and Decreasing p21WAF1/CIP1 and p16INK4A Expression

Retinoic acid plays an important role in the regulation of cell growth and differentiation. In our present study, we evaluated the effects of all-trans retinoic acid (RA) on cell proliferation and on the cell cycle regulation of human gingival fibroblasts (HGFs). Cell proliferation was assessed using the MTT assay. Cell cycle analysis was performed by flow cytometry, and cell cycle regulatory proteins were determined by western blot. Cell proliferation was increased in the presence of a 0.1 nM to 1 microM RA dose range, and maximal growth stimulation was observed in cells exposed to 1 nM of RA. Exposure of HGFs to 1 nM of RA resulted in an augmented cell cycle progression. To elucidate the molecular mechanisms underlying cell cycle regulation by RA, we measured the intracellular levels of major cell cycle regulatory proteins. The levels of cyclin E and cyclin-dependent kinase (CDK) 2 were found to be increased in HGFs following 1 nM of RA treatment. However, the levels of cyclin D, CDK 4, and CDK 6 were unchanged under these conditions. Also after exposure to 1 nM of RA, the protein levels of p21WAF1/CIP1 and p16INK4A were decreased in HGFs compared with the control group, but the levels of p53 and pRb were similar between treated and untreated cells. These results suggest that RA increases cell proliferation and cell cycle progression in HGFs via increased cellular levels of cyclin E and CDK 2, and decreased cellular levels of p21WAF1/CIP1 and p16INK4A.

The clinical assessment of aggressive periodontitis patients

PURPOSE: Few epidemiologic studies have investigated aggressive periodontitis in Koreans, but such studies of disease prevalence and other clinical characteristics would be invaluable in providing proper treatment. The aim of this study was to assess the prevalence of aggressive periodontitis and to measure the extent of associated periodontal breakdown. METHODS: The study population consisted of 1,692 patients who visited the Department of Periodontology, Wonkwang Daejeon Dental Hospital from January to December, 2010. Clinical parameters (probing depth, gingival recession, periodontal attachment loss) were measured by a single examiner, and radiographic examination was performed at the baseline. RESULTS: Twenty-eight (1.65%) patients showed clinical features of aggressive periodontitis, of which 27 patients exhibited the generalized form, and 1 exhibited the localized form. There was no significant difference between the percentage of male and female patients. The probing pocket depth of the maxillary first molar was deeper than that of the other teeth and gingival recession was also the most serious at the maxillary first molar. The periodontal attachment loss was the highest at the maxillary first molar. The average number of missing teeth was 1.29 per subject. Loss of the second molar was prominent. CONCLUSIONS: Within the limitations of this study, the periodontal breakdown evaluated by attachment loss was found to be most severe at the first molars of aggressive periodontitis patients. However, further large scale multicenter studies are necessary to access more precise data, including prevalence.

Healing pattern of the mucous membrane after tooth extraction in the maxillary sinus

PURPOSE: To investigate the healing pattern of the mucous membrane after tooth extraction necessitated by periodontal disease in the maxillary sinus. METHODS: One hundred and three patients with 119 maxillary sinuses were investigated. Before implant placement, cone-beam computed tomography (CT) scanning was performed. The causes of extraction, the time elapsed since extraction, smoking, periodontal disease in adjacent teeth, and gender were recorded. In addition, the thickness of the mucous membrane of the maxillary sinus and the height of residual alveolar bone at the extracted area were calculated from CT images. RESULTS: The thickness of the mucous membrane in the periodontal disease group (3.05+/-2.71 mm) was greater than that of the pulp disease group (1.92+/-1.78 mm) and the tooth fracture group (1.35+/-0.55 mm; P<0.05). The causes of extraction, the time elapsed since extraction, and gender had relationships with a thickening of the mucous membrane of the maxillary sinus (P<0.05). In contrast, the height of the residual alveolar bone at the extracted area, periodontal disease in adjacent teeth, and smoking did not show any relation to the thickening of the mucous membrane of the maxillary sinus. CONCLUSIONS: The present study revealed distinct differences in healing patterns according to the causes of extraction in the maxillary sinus, especially periodontal disease, which resulted in more severe thickening of the mucous membrane.

Healing pattern of the mucous membrane after tooth extraction in the maxillary sinus

PURPOSE: To investigate the healing pattern of the mucous membrane after tooth extraction necessitated by periodontal disease in the maxillary sinus. METHODS: One hundred and three patients with 119 maxillary sinuses were investigated. Before implant placement, cone-beam computed tomography (CT) scanning was performed. The causes of extraction, the time elapsed since extraction, smoking, periodontal disease in adjacent teeth, and gender were recorded. In addition, the thickness of the mucous membrane of the maxillary sinus and the height of residual alveolar bone at the extracted area were calculated from CT images. RESULTS: The thickness of the mucous membrane in the periodontal disease group (3.05+/-2.71 mm) was greater than that of the pulp disease group (1.92+/-1.78 mm) and the tooth fracture group (1.35+/-0.55 mm; P<0.05). The causes of extraction, the time elapsed since extraction, and gender had relationships with a thickening of the mucous membrane of the maxillary sinus (P<0.05). In contrast, the height of the residual alveolar bone at the extracted area, periodontal disease in adjacent teeth, and smoking did not show any relation to the thickening of the mucous membrane of the maxillary sinus. CONCLUSIONS: The present study revealed distinct differences in healing patterns according to the causes of extraction in the maxillary sinus, especially periodontal disease, which resulted in more severe thickening of the mucous membrane.

Clinical study of guided bone regeneration of extracted socket with PLA/PGA membrane and silk fibroin membrane / 대한치주과학회지

PURPOSE: This study was designed to compare the bond regeneratiom effects of treatment using silk fibroin membrane ( Nanogide-S (R)) resorbable barrier with control group treated by polyactic acid / polylacticglycolic acid membrane(Biomesh (R) ) METHODS:44 severe bone loss on extraction socket from 44 patients were used in this study. In experimental group 22 sites of them were treated by silk fibrin membrane as and the other 22 sites were treated by polyactic acid/ polylacticglycolic acid membrane as a control group. Clinical parameters including recovered bone width, length and radiographic parameter of vertical length were evlauated at base line and 3 months after surgery. RESULTS: 1) Severe bone width, length was significantlly decreased in two group. 2) Bone width, length was significantlly decreased in two group. 3) Decreased bone width, length and radiographic examination differences between group. CONCLUSIONS: On the basis of these results, silk fibrin resorbable membrane has similar bone regeneration ability to polyactic acid / polylacticglycolic acid membrane in guided bone regeneration for severe bone loss defect on extraction socket.

Effect of poly(lactide-co-glycolide) (PLGA) on bone regeneration in rabbit calvaria / 대한치주과학회지

PURPOSE: The purpose of this study is to histologically and histomorphometrically evaluate the effect of PLGA on bone regeneration compared with bone graft material. METHODS:The experimental study was conducted in 10 rabbits with 2 different healing periods of 2 and 4 weeks. Following surgical exposure of the calvarium, 4 circular bone defects with a diameter of 4.6mm were formed. Rabbits were divided into control group, test groups I, and II. 10 defects assigned to the test group I were grafted with Nu-oss and other 10 defects assigned to the test group II were grafted with PLGA. The rest of the defects were in the negative control group. At 2nd and 4th week after surgery, 10 rabbits were sacrificed through intracardiac perfusion and then specimens were obtained. Histological analysis was performed following staining with trichorme and transversal sectioning of the calvarial bone. RESULTS: A group which used PLGA showed tissue reactions characterized by severe inflammation, rather than distinctive new bone formation. CONCLUSIONS: The present experimental investigations have failed to prove any beneficial effects of PLGA. PLGA used in this study exhibited foreign body reactions and a less favorable pattern of new bone formation in comparison to control group. CONCLUSION: PLGA did not function as scaffold. Further investigations of many types of micro PLGA that could improve its potential in GBR procedures are needed.

Inhibitory effects of Enterococcus faecium isolated from Korean infants on oral pathogens / 대한치주과학회지

PURPOSE: The probiotic effects of lactic acid bacteria have widely been researched in diverse human pathogens, but only a few effects are reported against oral pathogens. The antimicrobial effects of the Enterococcus faecium 7413 isolated from Korean infants on the 9 pathogen including 6 oral streptococci were investigated the clinical use of the antimicrobial peptide for oral microflora control. MATERIALS AND METHODS: E. faecium 7413 was identified by morphological, biochemical tests and 16S rDNA sequence analysis. Inhibitory effects of culture supernatants were determined for their ability to grow on agar plate containing pathogenic bacteria. RESULT: The culture supernatant of Enterococcus faecium 7413 showed inhibitory effects on oral pathogens, namely Streptococcus pyogenes KCTC 3556, S. pneumoniae KCTC 5080, S. mutans ATCC 25175, S. anginosus ATCC 33397, S. constellatus KCTC 3268, S. intermedius ATCC 27823 and Shigella flexneri KCTC 2008. Whereas it did not affect the multiplication of E. coli strains, KCTC 1041 and ATCC 43894. CONCLUSION: The data obtained in this study could be useful for future development of effective probiotics allowing prevention for oral pathogens.

Connective tissue graft for root coverage / 대한치주과학회지

PURPOSE: The treatment of gingival recessions is needed to reduce root sensitivity and improve esthetical satisfaction. Several surgical techniques have been proposed to achieve these goals. The use of connective tissue grafts has made esthetic root coverage a predictable procedure. Numerous clinical studies have represented that using connective tissue grafts to cover exposed root surface showed high success rates. This is a case report which demonstrates the technique to obtain root coverage of a buccal recession defect. MATERIALS AND METHODS: A 35-year-old patient with a high level of oral hygiene was selected for the study. This patient had one Class I Miller recession defect in the mandible. A coronally advanced flap in combination with the connective tissue graft was chosen for the treatment. After surgery, the patient was told to visit the hospital once a week for his oral management and professional prophylaxis. The depth of initial recession was 4.0 mm. RESULT: After three months, it reduced to 0.0 mm, and the average recession reduction was 4.0 mm. The average root coverage was 100%. CONCLUSION: The connective tissue graft is both effective and predictable way to produce root coverage in increasing the width of CAL and KT of various adjacent gingival recessions.

Comparative S tudy Of Osseointergration On Different Immediate Implants In Extraction Sockets Of Beagle Dogs / 대한치주과학회지

Recently, immediately after losing teeth, implant placement has been greatly attempted. Implant can help restoration of tooth functions within short time. This study was an attempt to examine the extent of osseointergation when the implants will be placed immediately after teeth extraction using domestic implant systems. Implants were inserted in beagle dogs and evaluated the clinical, radiological, histological and histomorphometric assay at 6 weeks and 12 weeks. For experimental materials, STAGE-1(R)(4.1x8mm, Lifecore, USA), SS-III(R)(4.0x8mm, OSSTEM, Korea) and IFI(R)(4.0x8 mm, DIO, Korea) implants treated with RBM were placed. All the placed site showed normal results without fail and inflammation clinically and radiologically. As a result of measurement by periotest, it showed -2 ~ -5 and stable status comprehensively. There was no statistically significant difference among implants(p<0.05). Bone tissue adjacent to implant showed increased marrow tissue at 6 weeks. Nevertheless, osteogenic structure was not observed remarkably. In a 12 weeks opinion, bone tissue composed of osseointegration along implant interface showed significantly decreased marrow tissue containing central vessels unlike a 6 weeks opinion and matured compact bone whose osteogenic structure is well formed. BIC were 42.4%, 32.0% and 34.9%, respectively in 6 weeks and there was no statistically significant difference among groups(p<0.05). In 12 weeks, BIC were 58.8%, 61.9% and 57.5%, respectively and there was no statistically significant difference among groups(p<0.05). It is considered that all 3 implant systems are suitable for immediate implant placement.

Study on Osseointegration of the Immediate Placement Using Korean Implant Systems in Beagle Dog / 대한치주과학회지

This study was attempted to evaluate home-manufactured implants by placing Stage-1(R) Implant (Lifecore. Co., USA) whose surface is treated with RBM that has already been varified clinically, Chaorum(R) Implant(Chaorum Co., Korea) whose surface treatment is same as that of Stage-1 Implant and Atlas(R) Implant(Cewellmedi Co., Korea) whose surface is treated with anodic oxidation immediately after the teeth of experimental animals were extracted to compare histological findings among them. Stage-1 Implant(diameter: 3.5mm, length: 10mm), Chaorum Implant(diameter: 3.3mm, length: 8.5mm) and Cowell medi Implant(diameter: 4.0mm, length: 8.0mm) were placed into the mandible premolars of 2 adult beagle dogs immediately after their teeth were extracted, and then histological findings were analyzed at 6 weeks. After those implants were inserted directly after their teeth were extracted, the results of periotest were recorded, radiography was done, the subjects went through thorough control for 6 weeks, and then comparison among periotest, radiography and histological finding was made. After comparison of those findings, the values of periotest were satisfactory and bone healing was relatively satisfactory on radiography at 6 weeks. For osseointegration with the bone tissue, Stage-1 was 45.3%, Chaorum 55.3%, and Cowellmedi 52.5%, which was a satisfactory result. Although implant surgery immediately after teeth were extracted involves difficulties among recent implant surgeries, it is being frequently used in that it may reduce surgery hours, the frequency of surgery, and bone loss for patients. This experiment was conducted to evaluate the technological levels of home-manufactured implants that have been remarkably developed in recent years and in conclusion, those implants showed nearly similar result.

Study on osseointegration of a Korean Transitional implant system in beagle dog / 대한치주과학회지

PURPOSE: Recently, dental implant systems have been widely used for the treatment of the extraction site, but we have been confronted with many limitations in esthetics, phonetics and function. Transitional implants(TI) were developed as a method of providing fixed provisional restorations during conventional implant healing. Until now, little data have been provided on korean transitional implants. The purpose of this study is to evaluate the implant placement site histologically after 4 weeks and 8 weeks. MATERIALS AND METHODS: Test group( IntermetzzoTM MEGAGEN, KOREA) and control group(Mini Drive Lock, Intra Rock, U.S.A.) were immediately placed in interseptal or interproximal bone of beagle dog after mandibular premolars extraction, and had a healing period with non-submerged state but without loading, Both TI surfaces were composed of rough surfaces. RESULTS: In the test group, the average percentage of BIC were respectively 39.40%(SD7.35) after 4 weeks and 44.05%(16.76) after 8 weeks, and In the control group were 50.75%(1.48) and 59.40%(0.00). DISCUSSION: We evaluated the initial ability of the osseointegration of TI through this study. Because TI is placed with a conventional implant simultaneously and loaded immediately, the ability of osseointegration is a very important factor for the success of TI during the initial healing phase. CONCLUSION: The results of the histological evaluation of these two groups were similar to those mentioned in other studies for osseointegration of implant.

Effects of biphasic calcium phosphate on bone formation in human fetal osteoblasts / 대한치주과학회지

No abstract available.

Effect of the Physiologically Active Compounds in Phlomidis Radix on Cell Cycle Regulation in Human Gingival Fibroblasts / 대한치주과학회지

The purpose of present study was to investigate the effects of physiologically active compound (SD62-122) from Phlomidis Radix on the cell cycle progression and its molecular mechanism in human gingival fibroblasts(HGFs). For this purpose, fibroblasts were isolated and cultured from excisioned gingiva during crown lengthening procedure in healthy adult. The following parameter were evaluated that there are cell number counting, MTT assay, cell cycle progression, western blot analysis. The cell number and MTT assay of primary cultured fibroblast was not increased at 2 days but significant increased compare to negative control at 3days(p<0.05). S phase was increased and G1 phase decreased in both 10(-8)M and 10(-9)M of SD62-122 in cell cycle analysis. The cell cycle regulation protein levels of Cyclin D1, Cyclin E, cdk 2, cdk 4 and cdk 6 were increased compare to control in both 10(-8)M and 10(-9)M of SD62-122. The protein levels of p21 and p53 were decreased compare to control, but the level of pRb was not changed compare to control in 10(-9)M of SD2-122. These results suggested that physiologically active compound (SD62-122) isolated from Phlomidis Radix increases the cell proliferation and cell cycle progression in HGFs, which is linked to increased cell cycle regulation protein levels of Cyclin D1, Cyclin E, cdk 2, cdk 4 and cdk 6, and decreased the levels of p21, p53.

A Study of Safflower Seed Extracts on Bone Formation in Vitro / 대한치주과학회지

The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and the repair of function. For more than a decade there have been many efforts to develop ma- terials and methods of treatment to promote periodontal tissue regeneration. Recently many efforts are concentrated on the regeneration potential of material used in traditional medicine. Safflower(Carthamus tinctorius L.) seed extract(SSE) have long clinically used in Korea to promote bone formation and prevent osteoporosis. The purpose of this study was to examine the effects of SSE on bone formation in human osteoblastic cell line. Human fetal osteoblastic cell line(hFOB 1.19) was cultured with DMEM and SSE(1microgram/ml, 10microgram/ml, 100microgram/ml, 1mg/ml) at 34degrees C with 5% CO2 in 100% humidity. The proliferation, differentiation of the cell was evaluated by several experiments. Cell proliferation was significantly increased at 10microgram/ml, 100microgram/ml, 1mg/ml of SSE after 3 and 7 days incubation(p<0.05). Cell spreading assay was significantly increased at 100microgram/ml of SSE after 3 days and 1microgram/ml, 10microgram/ml, 100microgram/ml, 1mg/ml of SSE after 7 days(p<0.05). Alkaline Phosphatase(ALP) level was significantly increased in 10microgram/ml, 100microgram/ml of SSE(p<0.05). Collagen synthesis was significantly increased at 10microgram/ml, 100microgram/ml, 1mg/ml of SSE(p<0.05). A quantified calcium accumulation was significantly increased at 10microgram/ml, 100microgram/ml of SSE(p<0.05). ALP and osteocalcin mRNA was expressed in 100microgram/ml of SSE by RT-PCR. These results indicate that SSE are capable of increasing osteoblasts mineralization and may play an important role in bone formation.

Study on the Biological Characteristics of Cultured Osteoblasts Derived from Alveolar Bone / 대한치주과학회지

Osteoblasts from alveolar bone may have an important role in the bone regeneration for periodontium, but their culture and characterization are not determined yet. The purpose of this study was to investigate the biological characteristics of primary explant cultured osteoblasts(PECO) from alveolar bone. Osteoblasts were isolated and cultured from alveolar socket of extracted tooth in children. To compare the characteristics, osteoblasts and gingival fibroblasts were cultured with DMEM at 37degrees C, 5% CO2, 100% humidity incubator, and human fetal osteoblasts cell line(hFOB1) were cultured with DMEM at 34degrees C, 5%, CO2, 100% humidity incubator. To characterize the isolated bone cells, morphologic change, cell proliferation and differentiation were measured. Morphology of PECO was small round body or cuboidal shape on inverted microscope and was similar with hFOB1. PECO became polygonal shape with stellate and had an amorphous shape at 9th passage in culture. PECO had significantly higher activity than that of gingival fibroblasts and hFOB1 in alkaline phosphatase activity. The expression of osteocalcin and bone sialoprotein in PECO was notably increased when compared with hFOB1 and gingival fibroblasts. These result indicated that PECO from alveolar bone in children has an obvious characteristics of osteoblast, may be applied for the regeneration of bone.

The Effects of Dichloromethane fraction of Phlomodis Radix(DFPR) on differentiation of Mouse Calvarial Cell / 대한치주과학회지

The purpose of this study was to evaluate the effects of DFPR on differentiation of mouse calvarial cell in vitro, to examine the possibility for periodontal regeneration. 10microgram/ml of DFPR was used as experimental concentration. osteogenic medium only was assigned as control, Experimental 1 was supplemented with 10nM dexamethasone, Experimental 2 with 10microgram/ml DFPR and Experimental 3 with 10nM dexamethasone + 10microgram/ml DFPR. cellular activity was evaluated by MTT method at 8, 12, 16 days, expression of mRNA of ALP, osteopontin, osteocalcin, collagen type-1 was detected by RT-PCR method at 4, 8, 12, 16 days of culture . extent of mineralization was observed by Von Kossa staining at 16 day of culture. The results are as follows 1)Any acceleration of differentiation was not observed at expression of differentiation marker, 2) Decrease in expression of extracelluar matrix and in bone nodule formation was observed The results suggested that DFPR have negative effect on the rate of differentiation on rat calvarial cell, decrease extracelluar matrix formation ,decrease bone nodule formation. Ongoing studies are necessary in order to determine effect of DFPR on periodontal regeneration.

Effects of Dichloromethane Fraction of Phlomidis Radix on Bone Formation in Human Fetal Osteoblasts / 대한치주과학회지

The ideal goal of periodontal therapy is the regeneration of periodontal tissue and repair of function. Although it is very difficult to attain this goal, recent advances in periodontal wound healing concepts encourage hope reaching it. Recently many efforts are concentrated on the regeneration potential of material used in traditional Korean medicine. Phlomidis Radix has been used for the treatment of blood stasis, bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine effects of dichloromethane fraction Phlomidis Radix on Bone Formation in Human Fetal Osteoblasts. Human fetal osteoblastic cell line(hFOB1 1.19 ; American Type Culture Collection, Manassas, VA) were used and cells were cultured containing DMEM and dichloromethane fraction Phlomidis Radix(100 ng/ml, 1 microgram/ml, 10 microgram/ml) at 34degrees C with 5% CO2 in 100% humidity. MTT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examnine the mineralization. Also bone calcification nodules were evaluated. The cellular activity of hFOB1 was increased in 100 ng/ml, 1 microgram/ml, 10 microgram/ml of dichloromethane fraction of Phlomidis Radix and especially significant increation was showed in 100 ng/ml of dichloromethane fraction of Phlomidis Radix at 6days (p<0.05). ALP level of hFOB1 was significantly increased in 100 ng/ml, 1 microgram/ml, 10 microgram/ml of dichloromethane fraction of Phlomidis Radix and especially more increation was showed in 10 microgram/ml of dichloromethane fraction of Phlomidis Radix (p<0.05). Calcification nodules of hFOB1 significantly increased in 10 microgram/ml of dichloromethane fraction of Phlomidis Radix at 21days of incubation (p<0.05). These results indicate that dichloromethane fraction of Phlomidis Radix has excellent effects on mineralization of hFOB1.

Effects of Ginseng Radix on the Cell Cycle Regulation in Human Fetal Osteoblast / 대한치주과학회지

Ginseng Radix(GR) had been used widely from oriental medicine and the effects of it have been investigated by many researchers. The purpose of present study was to investigate the effects of GR on the cell cycle progression and its molecular mechanism in human fetal osteoblast. The results were as follows. Increased cell proliferation was observed in cells exposed to 100 ng/ml, 10 ng/ml of GR-1 at 12 hours and 24 hours, 1 microgram/ml of GR-1 at 48 hours, and 100 microgram/ml, 10 microgram/ml of GR-2 at 12 hours, all treatment groups of GR-2 at 24 hours(p<0.05). S phase and G1 phase was increased in the group of treated with 100 ng/ml of GR-1, with 10 microgram/ml and 1 microgram/ml of GR-2, with 100 microgram/ml and 10 microgram/ml of GR-3 in the cell cycle analysis. The cell cycle regulation protein levels of Cyclin D1, Cyclin E, CDK 2, CDK 4 and CDK 6 were increased in the group of treated with 1 microgram/ml and 100 ng/ml of GR-1, with 10 microgram/ml and 1 microgram/ml of GR-2, with 100 microgram/ml and 10 microgram/ml of GR-3. On the other hand, p21 was decreased in the treatment group with 1 microgram/ml and 100 ng/ml of GR-1, with 10 microgram/ml and 1 microgram/ml of GR-2, 10 microgram/ml of GR-3, and p53 and p16 was decreased in the treatment group with 100 ng/ml of GR-1, 100 microgram/ml GR-3 and pRb was decreased in the all treatment groups except 1 microgram/ml of GR-1. These results suggested that GR increases the cell proliferation and the cell cycle progression in human fetal osteoblast, which is linked to increased cell cycle regulation protein levels of Cyclin D1, Cyclin E, CDK 2, CDK 4, CDK 6 and decreased cell cycle regulation protein levels of p21, pRb.