1.Biological Characteristics of Human Periodontal Ligament Cells.
Gwi woon PARK ; Hyung shik SHIN ; Hyung keun YOU
The Journal of the Korean Academy of Periodontology 1997;27(2):291-303
Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at 37degrees C, 5% CO2, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity, and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.
Alkaline Phosphatase
;
Bicuspid
;
Calcitonin
;
Collagen
;
Collagen Type I
;
Fibroblasts
;
Humans*
;
Humidity
;
Immunohistochemistry
;
Incubators
;
Osteocalcin
;
Osteonectin
;
Periodontal Ligament*
;
Population Characteristics*
;
Regeneration
2.Effect of Extract of Seeds of Carthamus tinctorius L. on Mineralization in Periodontal Ligament Cells and Osteoblastic Cells.
Jung Goo KANG ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1998;28(3):475-489
The main goal for the treatment of periodontal diseases is the regeneration of lost cementum, bone and connective tissue. Clinical and histological research suggests that it is possible to restore periodontal structures. Seeds of Carthamus tinctorius L. has been used for the treatment of bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine the effect of extract of seeds of Carthamus tinctorius L. on mineralization in periodontal ligament cells and osteoblastic cells. Periodontal ligament cells were primarily obtained from a extracted premolars with non-periodontal diseases. Osteoblastic cells were obtained from calvariae of a fetal rat. Cells were cultured with DMEM at 37degrees C with 5% CO2 in 100% humidity incubator. Alkaline phosphatase(ALP) level and the number of calcification nodules were examined and western blot analysis using osteonectin was performed. Measurements of ALP levels and calcification nodules showed that extract of seeds of Carthamus tinctorius L. had significantly higher activity than control in all of both cells. In western blot analysis, protein expression of osteonectin indicated that extract of seeds of Carthamus tinctorius L. showed an increased pattern than control in all of both cells. From the above results, it seems that extract of seeds of Carthamus tinctorius L. has excellent effect on mineralization in periodontal ligament cells and osteoblastic cells.
Animals
;
Bicuspid
;
Blotting, Western
;
Carthamus tinctorius*
;
Carthamus*
;
Connective Tissue
;
Dental Cementum
;
Fractures, Bone
;
Humidity
;
Incubators
;
Osteoblasts*
;
Osteonectin
;
Osteoporosis
;
Periodontal Diseases
;
Periodontal Ligament*
;
Rats
;
Regeneration
;
Skull
3.An Experimental Study on the Biological Specificity of Nitric Oxide and Nitric Oxide Synthetase in Periodontium-Related Cells.
Hyung Jin YOON ; Dong Whan YOON ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1997;27(4):883-908
Bone remodeling is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. The process is tightly regualted at the local level by an incompletely known netwotk of peptide and non-peptide fators. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bioregualtory molecule in a variety of tissue, but little is known about its possible role in periodontal tissue. The purpose of this study is to investigate the expression of nitric oxide synthetase(NOS) in inflamed gingiva and the effects of cytokine on the expression of NOS protein. The expression of NOS in gingival tissue was evaluated by immunohistochemical staining for NOS1, NOS2, NOS3. The effect of cytokine on the expression of NOS in human periodontal ligament cells and osteoblast-like HOS cells by western blot analysis. Further, we studied that NO functions in periodontal ligment cells as a regulatory molecule. PDL cells incubated with NOS inhibitor and donor. The protein expression, type I collagen & non-collagenous protein, nitrate production and cell proliferation were evaluated The results were as follows. 1. NOS1, NOS2, NOS3 was rarely distributed in healthy gingiva, but stronger stained in gingival epithelium, endothelial cells, and mononuclear cells of inflammed gingiva. 2. The cytokine stimulated NOS1, and NOS3 protein were not inducing or inhibitory effect to compared with control in PDL and HOS cells. 3.Incubation of cells with combination of TNF-alpha, IFN-gamma, LPS result in a time dependant increase in NOS2 expression, reaching a maximal level after 24 hours of stimulation. 4. The osteonectin protein inhibitory effect of NMA, inhibitor of NOS, was reversed by Larginine in dose dependant manner. 5. NMA decreased cell proliferation and nitrate production, but the inhibitory efffect of NMA was also prevented by the NO donor, sodium nitropruiside. These results suggest that exogenously synthesized NO was playing a stimulating effect on cell proliferation or on non-collagenous protein expression. Therefore NO have an important role in mediation of localized bone destruction associated inflammatory bone disease such as periodontitis
Arginine
;
Blotting, Western
;
Bone Diseases
;
Bone Remodeling
;
Bone Resorption
;
Cell Proliferation
;
Collagen Type I
;
Endothelial Cells
;
Epithelium
;
Gingiva
;
Humans
;
Negotiating
;
Nitric Oxide Synthase*
;
Nitric Oxide*
;
Osteogenesis
;
Osteonectin
;
Periodontal Ligament
;
Periodontitis
;
Sensitivity and Specificity*
;
Sodium
;
Tissue Donors
;
Tumor Necrosis Factor-alpha
4.Effects of Olibanum Extracts on the Activity and Differentiation of MC3T3-E1 Cells.
Sang Heon HAN ; Myoung Dong KIM ; Seung Han YOU ; Yong Ouk YOU ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 2001;31(2):287-298
Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use have been studied for their capacity of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum has the effects to hemostasis, analgesic and anti-inflammatory, and it also has been traditionally used as a drug for the treatment of bone disease in oriental medicine. The purpose of the present study was to investigate the effects of Olibanum extracts on the activity and differentiation of MC3T3-E1 cells, alkaline phosphatase(ALP) synthesis, formation of bone nodules and expression of type I collagen of MC3T3-E1 cells. To examine the cellular activity, MC3T3-E1 cells were cultured with alpha-MEM(control) and each concentration of Olibanum for 2 days and 4 days. To compare the ALP synthesis, MC3T3-E1 cells were cultured with alpha-MEM(negative control), dexamethasone(positive control), and each concentration of Olibanum for 2 days and 4 days. To compare the bone nodule formation, MC3T3-E1 cells were cultured for 21 days, and to compare the type I collagen expression, MC3T3-E1 cells were cultured for 4 days. The cellular activity of MC3T3-E1 cells treated with 1 microgram/ml of Olibanum extracts was significantly increased at 4-day(p<0.05) to control. The activity of ALP in MC3T3-E1 cells treated with 1 microgram/ml Olibanum extracts was significantly increased at 4-day(p<0.05). All the experimental groups showed much more bone nodule formation than control groups. The group treated with 1 microgram/ml of Olibanum extracts was the highest bone nodule formation, and showed much more type I collagen expression than negative control. These results indicate that Olibanum extracts may be considered effective in the activity and differentiation of MC3T3-E1 cells.
Bone Diseases
;
Boswellia*
;
Collagen Type I
;
Hemostasis
;
Medicine, East Asian Traditional
5.Effects of Several Biodegradable Controlled-Release Local Delivery Drugs on the Treatment of Periodontitis.
Dae Seung YANG ; Kyung Tae YOU ; Sung Hee PI ; Myung Yeon LEE ; Yong Ouk YOU ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1999;29(4):725-735
The present study was performed to evaluate the clinical effects following local application of 30% minocycline strip(polycaprolactone), 2% minocycline gel(hydro-carbon gel) and 12% minocycline strip(polylactide, Minodent) to augment scaling and root planing in patients with chronic adult periodontitis. Forty teeth with periodontitis were enrolled in the study anddistributed into 4 groups including control group. All patients performed standardized oral hygiene instructions and mechanical debridement at the beginning of the study and then each local delivery drugs were inserted into periodontal pocket in each groups. Examinations regarding plaque index(PI), papillary bleeding index (PBI), probing pocket depth (PPD) were carried out at 0, 2, 4 weeks. All experimental groups showed statistically significant differences between baseline and 2 and 4 weeks in every clinical indices. Especially, 30%minocycline strip and Minodent group showed a significant improvement in PBI at 2 weeks and in PPD at 2 and 4 weeks. In conclusion, highly bio-resorbable Minodent delivered subgingivally as an adjunct to scaling and root planing induces better clinical effects for periodontal health than 2% minocycline gel and control group.
Adult
;
Male
;
Female
;
Humans
6.Effect of Mechanical Stress on the Proliferation and Expression of Cell Cycle Regulators in Human Periodontal Ligament Cells.
Hyung Keun YOU ; Hyung Shik SHIN ; Gene LEE ; Byung Moo MIN
The Journal of the Korean Academy of Periodontology 1999;29(3):593-605
No abstract available.
Cell Cycle*
;
Humans*
;
Periodontal Ligament*
;
Stress, Mechanical*
7.Effects of Several Herbal Medicines on Alkaline Phosphatase Activity in Human Fetal Osteoblasts.
Myoung Ku LEE ; Hee In CHOI ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 2003;33(1):49-60
Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.
Albizzia
;
Alkaline Phosphatase*
;
Bone and Bones
;
Bone Diseases
;
Dexamethasone
;
Genetic Engineering
;
Humans*
;
Intercellular Signaling Peptides and Proteins
;
Morinda
;
Osteoblasts*
;
Peptides
8.Effects of some herbal drugs on gingival fibroblast and periodontal ligament cellular activity.
Jin Soo DOO ; Jung Ku KANG ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1997;27(3):459-468
Healing of periodontal tissues require the migration and proliferation of gingival fibroblasts and periodontal ligament cells. There is many evidences that the some agents like cytokines and polypeptide growth factors are mediate these cellular events in wound healing. Recently someone is interested in herbal drugs on periodontal tissue healing processes. The purpose of this study was to examine the effects of 4 herbal drugs, Carthami Flis, Moutan Radicis Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. on human gingival fibroblasts and periodontal ligament cells. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. The powder from extracted herbal drugs were prepared with distilled water. Cells were cultured with DMEM at 37degrees C, 5% CO2, 100% humidity incubator, and treated with each herbal drugs with proper concentration for 1, 2, and 3 days. The cell activity was determined by ELISA reader using MTT assay. There was the most significant elevation in 10(-3)g/ml of almost herbal drugs on cellular activities. The result of this study demonstrated that Carthami Flis, Moutan Radicis Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. appears to have beneficial effect on healing process after periodontal treatment.
Bicuspid
;
Carthamus tinctorius
;
Cytokines
;
Enzyme-Linked Immunosorbent Assay
;
Fibroblasts*
;
Humans
;
Humidity
;
Incubators
;
Intercellular Signaling Peptides and Proteins
;
Periodontal Ligament*
;
Water
;
Wound Healing
9.Effect of Smoking on Adult Periodontitis after Non-surgical Periodontal Therapy.
Young Chae PARK ; Jung Ku KANG ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1997;27(2):305-315
The purpose of this study was to investigate the effects of smoking on adult periodontitis after non-surgical periodontal therapy. The study population consisted of 40 patients with moderate to advanced periodontitis. Smokers(n=20) were defined as individuals smoking at least twenty cigarettes per day at the time of the initial examination. The non-smoking group(n=20) consisted of individuals who were not smoking at the initial examination. The average age was 42.4 years for the smoking and non-smoking group. Examination regarding plaque index, gingival index, pocket depth and contrast phase microscope were performed. Evaluation were made at the first, the second and the fourth weeks after periodontal non-surgical therapy. The results were as follows: 1. Clinical indices including plaque index, gingival index, and pocket depth were decreased in both smoking and non-smoking group at the first, the second, and the fourth weeks. Especially, clinical indices of non-smokers were more significantly decreased than those of smokers. 2. Non-motile rods were increased and motile rods were reduced at the fourth week. spirochetes were reduced significantly in the non-smoking group at the fourth week. These results suggest that smoking play a minor role in adult periodontitis after non-surgical periodontal therapy.
Adult*
;
Chronic Periodontitis*
;
Humans
;
Periodontal Index
;
Periodontitis
;
Smoke*
;
Smoking*
;
Spirochaetales
;
Tobacco Products
10.Effects of a xenographic bovine bone on the bone mineralization in human fetal osteoblasts.
Ki Jong SUN ; Ha Na HYUN ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 2002;32(4):801-809
The ultimate goal of periodontal therapy is to promote the regeneration of lost periodontal tissue, there have been many attempts to develop a method to achieve this goal, but none of them was completely successful. The purpose of this study is to evaluate the effects of Bio-Oss(R) on alkaline Phosphatase (ALP) activity in human fetal osteoblasts (hFOB1). The results of this study were as follows, in ALP Activity, 100 microgram/ml Bio-Oss(R) treated group showed significantly increased value than negative control group, but positive group(10(-7) M dexamethasone treated group) showed the highest ALP activity at 3 day. In mineralization assay, numerous mineralized nodules were identified as darkly stained spots in 100 microgram/ml Bio-Oss(R) treated group than two control groups, whereas a small number of mineralized nodules were showed in the positive control. ALP may relate to the initial phase of bone nodule formation. On the basis of these results, this study showed Bio-Oss(R) is capable of accelerating new bone formation through hFOB1 differentiation in vitro.
Alkaline Phosphatase
;
Calcification, Physiologic*
;
Dexamethasone
;
Humans*
;
Osteoblasts*
;
Osteogenesis
;
Regeneration