Silica is known as a promising osteoconductive material, and polycaprolactone is a bioactive and degradable material. The purpose of this study was to monitor the differentiation of MC3T3-E1 cells cultured on the layer-built silica/poly caprolactone non-woven fabric produced by electrospinning. Non-woven fabric (silica, polycaprolactone, PSP, SPS) was made by electrospinning and they were inserted in the 48 well cell culture plate. MC3T3-E1 cells were prepared by subculture. Cells were seeded to each well 1x10(5) concentration per well. Dulbecco's modified eagle medium with 10% FBS and 1% antibiotic-antimycotic solution was used. Confocal laser scanning microscope was taken 4 hours after incubation (95% air, 5% CO2, 37degrees C). Cell proliferation was monitored by spectrophotometer on 1, 7, 14 days, and the morphology of the growing cells was observed by field emission scanning electron microscope. To monitor the differentiation of osteoblasts on the materials, MC3T3-E1 cells were incubated in 48 well culture plate after seeding with the density of 1x10(5) concentration. Then ELISA kit & EIA kit were used on to assess osteocalcin and osteopontin expression respectively. The other conditions were the same as above. MC3T3-E1 cells were proliferated well on all of the materials. There were no statistical differences among them. The osteopontin expression of silica, PSP, SPS was significantly higher than other groups on day 3 (p<0.05), but after that time, there were no statistically signigicant differences. The osteocalcin expression was significantly higher in silica and PSP than other groups on day 14. These findings show that PSP was as good as silica on the effect of osteoblast differentiation. The PSP non-woven fabric may have the possibility as bone graft materials.
Cell Culture Techniques ; Cell Proliferation ; Eagles ; Enzyme-Linked Immunosorbent Assay ; Osteoblasts ; Osteocalcin ; Osteopontin ; Silicon Dioxide ; Transplants

This study evaluated the possibility of the 3-dimensional attachment of human periodontal ligament fibroblasts to a periodntally involved root surface after an EDTA treatment in vitro. The human PDL fibroblasts were isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37degrees C in humidified air containing 5% CO2. Eight single-rooted teeth were obtained from patients diagnosed with periodotitis. After scaling and root planing, four teeth were etched with 24% ethylenediaminetetracetic acid (EDTA) for two minutes (Experimental group). The other four teeth were not treated with EDTA and were used as the control group. The human PDL fibroblasts were placed in the total root surface and cultured for 4 weeks. The teeth were fixed in 2.5% glutaraldehyde in PBS before preparation for the scanning electron microscopy (SEM) examination. The human PDL fibroblasts showed a healthy morphology on the root surfaces treated with EDTA (Experimental group) and a relatively unhealthy appearance on the treated root surfaces (Control group). This suggests that EDTA favorably affects the 3-dimensional attachment of human PDL fibroblasts cultured on the root surfaces, which may play an important role in periodontal healing and regeneration.
Eagles ; Edetic Acid* ; Fibroblasts* ; Glutaral ; Humans* ; Microscopy, Electron, Scanning ; Periodontal Ligament* ; Regeneration ; Root Planing ; Tooth

Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontal ligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37degrees C in humidified air with 5% CO2. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix. The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.
Acellular Dermis ; Eagles ; Fibroblasts ; Humans ; Microscopy, Electron, Transmission ; Periodontal Ligament ; Regeneration ; Tissue Engineering* ; Tooth

OBJECTIVE: To evaluat the effects of a culture medium with glucose in the presence of glutamine on the development of mouse embryos. METHODS: Two-cell embryos recovered from ICR mice at 48 hrs after hCG injection (mated just after hCG injection) were cultured in DMEM (with 20% hFF) supplemented with or without glucose on the presence of glutamine. Embryos were cultured under three different glucose regimens: (1) 0 mM (control); (2) 0.5 mM (group I); or (3) 3.15 mM (group II), and were analyzed at 24, 48, 72 and 96 hours intervals. Chi-square test (x2-test) was used to compare values of groups. RESULTS: No differences were found in the number of embryos showing morula (control: 37.5%; group I: 51.0%; group II: 48.4%), blastocyst (control: 21.5%; group I: 33.3%; group II: 34.4%) and blastocyst and hatching or hatched blastocyst (control: 81.9%; group I: 83.3%; group II: 82.8%) between groups at 24 hrs, 48 hrs or 72 hrs respectively. However at 96 hrs, the number of hatched and attached blastocyst was significantly higher in group I (82.3%) and II (78.5%) than control (63.2%; P<0.05). CONCLUSION: The addition of glucose (0.5 mM) to the DMEM, as energy source, improved the rate of development of late stage embryos in mice.
Animals ; Blastocyst ; Eagles* ; Embryonic Development* ; Embryonic Structures* ; Female ; Glucose* ; Glutamine* ; Mice* ; Mice, Inbred ICR ; Morula ; Pregnancy

This study was conducted to investigate the effects of Tissue Culture Medium 199 (TCM) and Dulecco's Modified Eagle Medium (DMEM) on the blastulation and grade of human oocytes on Vero cells in vitro. A cohort of 79 and 93 oocytes in metaphase II stage were used in TCM 199 and DMEM respectively. No differences were found in the nurser of oocytes showing two-pronuclei between TCM (82.3%) and DMEM (86.0%). The number of fertilized oocytes reaching the blastocyst was not significant in TCM (60.0%) and DMEM (63.1%).4 total of 89 blastocysts were categorized into the four grades (BG1, BG2, BG3 and early) depending on their morphology. The number of embryos achieving the blastocyst grade 1 (BG1) was significantly higher (p<0.05) in DMEM (50.8%) than TCM (15.0%). It is concluded that cultured oocytes in DMEM with glutamine on Vero cells should be significantly increased BG1.
Blastocyst ; Cohort Studies ; Eagles ; Embryonic Structures* ; Glutamine* ; Humans* ; Metaphase ; Oocytes ; Vero Cells*

BACKGROUND: The influencion the environment on a culture is expressed via four routes. (1) the nature of the substrate or phase on or in which the cells grow (2) the physicochemical and physiological constitution of the medium, (3) the constitution of the gas phase, and (4) the incubation temperature. Melanization is closely related to the constitution and amounts of amino acids in the medium. OBJECTIVE: The aim of this study is to investigate whether there are some differences of proliferation and melanization in cultured B,F, mouse melanoma cells according to different culture media. METHODS: We examined the color of cell pellet, cell morphology, electron microscopic findings, cell counts and melanin conlensin BgF mouse melanoma cells cultured in Dulbeccos modified Eagles medium(DMEM), F-10, MCDB 153, Minimal essential medium(MEM), and RPMI 1640, respectively. RESULTS: 1. The color of cell pellet., ringed from dark gray to light brown. The order of the darkness was DMEM, MEM, RPMI 1640, MCDB 153, and F-10 medium. 2. Most Bg, mouse melanoma cells had an epithelioid morphology, but a few cells in MCDB 153 medium showed dendrites. On the 4th day after culture, the cells in F-10 medium were larger than those in the other media. 3. In the electron microscopic. findings, BF, mouse melanoma cells in DMEM and MEM con tained numerous stage IV nelanosomes, however, those in RPMI 1640 and MCDB 153 medium contained a few, and those in F-10 medium did few. 4. The number of BF, mouse melanoma cells were 1.42 + 0.06 x 10", 1.42 + 0.12 x 10", l. 17 + 0.08 x 10, 0.73 0.06 x 10, 0.32 0.01 x 10, in RPMI 1640, DMEM, MEM, F 10, and MCDB 153 medium, respectively. 5. In the MTT assay, the order of the optical density of B,F, mouse melanoma cells in various media was as followings, DMEM, RPMI 1640, MEM, F-10, and MCDB 153. 6. Compared with the melanin contents of B;F, mouse melanoma cells in DMEM, they were 77.97% in MEM, 67.91% in RPMI 1640 and MCDB 153 medium, and 55.94% in F-10 medium. CONCLUSION: The phenotypic changes of BF, mouse melanoma cells were induced by various culture rnedia and were reversilvle. Since the phenotypes of cells can be changed by the culture media, researchers should choose the appropriate culture medium for the cells.
Amino Acids ; Animals ; Cell Count ; Constitution and Bylaws ; Culture Media ; Darkness ; Dendrites ; Eagles ; Melanins ; Melanoma* ; Mice* ; Phenotype

The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. To evaluate the effects of Dex growth and differentiation of MC3T3-E1 cells, cells were seeded in alpha-modified eagle medium containing 10% fetal bovine serum, 10mM beta-glycerophosphate , 50microgram/ml of ascorbic acid, with or without 10(-7)M Dex and examined cell proliferation activities, alkaline phosphatase activities, and bone nodule formation until 25days. The results were as follows: 1. In Dex group, cell proliferation activities were lower until 15 days compared to control group. Bone nodules formation were showed at 10 days. 2. In the time-response effect, ALP activities were increased until the 10 days in control groups thereafter decreased and ALP activities of Dex group were lower aspect than control group until the 10 days In this study, bone nodule formation of osteoblast-like cells were accelerated by Dex and cell proliferation activities, ALP activity of Dex group showed lower than control group. Dex was considered that it did suppress initial growth, but accerelate mineralization of osteoblast-like cells.
Alkaline Phosphatase ; Ascorbic Acid ; Bone and Bones ; Cell Proliferation ; Dexamethasone* ; Eagles ; Regeneration

PURPOSE: This study was performed to investigate the prevalence, morphology, and calcification pattern of the elongated styloid process in the Mathura population and its relation to gender, age, and mandibular movements. MATERIALS AND METHODS: The study analyzed digital panoramic radiographs of 2,706 adults. The elongated styloid process was classified with the radiographic appearance based on the morphology and calcification pattern. The limits of mandibular protrusion were evaluated for each subject. The data were analyzed by using a Student's t-test and chi-squared test with significance set at p=0.05. RESULTS: Bilateral elongation having an "elongated" type styloid process with a "partially mineralized" pattern was the most frequent type of styloid process. No correlation was found between styloid process type and calcification pattern on the one hand and gender on the other, although elongated styloid was more prevalent in older and male populations (p<0.05). Further styloid process elongation showed no effect on mandibular protrusive movement (p>0.05). CONCLUSION: Dentists should recognize the existence of morphological variation in elongated styloid process or Eagle syndrome apparent on panoramic radiographs. We found higher prevalence of elongated styloid process in the population of the Mathura region when compared with other Indian populations. The calcification of the styloid process was more common in the older age group with no correlation to gender, mandibular movement and site. "Type I" with a "partially calcified" styloid process was observed more frequently in the population studied.
Adult ; Dentists ; Eagles ; Hand ; Humans ; Male ; Ossification, Heterotopic ; Prevalence ; Radiography, Panoramic ; Temporal Bone

The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. This paper investigated the effect of growth factor on bone cells. Platelet-derived growth factor(PDGF) is the one of the polypeptide growth factor that has been reported as a biological mediator which regulates activities of the cell proliferation, migration and metabolism of undifferentiated mesenchymal cells. The purpose of this study is to evaluate the effects of PDGF on bone nodule formation and ALP activity of MC3T3-E1 cells. Cells were seeded at 1x10(5)cells/well in alpha-modified eagle medium containing 10% fetal bovine serum, 10ml beta-glycerophosphate and 50microgram/ml of ascorbic acid. PDGF 0, 0.1, 1, 10ng/ml were added to the cells at a confluent state and cultured for 3, 7, 14, 21, 28 days. We examined bone nodule formation and alkaline phosphatase activity. The results were as follows: There were bone nodule formation at day 21 both in control and all the experimental groups, and at day 28, all the experimental groups showed much more bone nodules than control groups. Compared to control-1 group, ALP activity was increased in PDGF 0.1ng/ml group and was decreased in 1,10ng/ml PDGF treated groups.(P< 0.05, P< 0.01) Compared to control-2, ALP activity was decreased in all the experimental groups except PDGF 0.1ng/ml in 21 day group. In the time-response effect, ALP activity was increased by the day 14 in all the experimental groups and thereafter ALP activity was decreased.(P<0.05, P< 0.01) In the dose-response effect, ALP activity was decreased as the dose of PDGF was increased, and after 21 day ALP activity was lowest in 1ng/ml group, ALP activity was highest in the day 7 in control group and 0.1ng/ml, 14 day experimental group. In conclusion, PDGF is considered more effective in the proliferation than differentiation of osteoblast-like cells, and it may be useful to study the combined effect of PDGF and other growth factors on osteoblast-like cells.
Alkaline Phosphatase ; Ascorbic Acid ; Bone and Bones ; Cell Proliferation ; Eagles ; Intercellular Signaling Peptides and Proteins ; Metabolism ; Regeneration

BACKGROUND: Engineered cell sheet transplantation has been considered an alternative physiological therapy for endocrine disorders. In this study, we attempted to fabricate functional human thyroid cell sheets using the engineering technology by culturing primary thyrocytes in free-feeder monolayers and assessed their proliferation and function in two different media. METHODS: The non-tumorous tissues (approximately 2 g) were dissected during surgery. Primary human thyroid cells were isolated by mechanical dispersion and treatment with isolation solution. The cells were cultured on tissue culture dishes or temperature-responsive culture dishes to induce the formation of detached cell sheets. RESULTS: Primary thyroid cells isolated from nine patients were positive for thyroid transcription factor 1, thyroglobulin (TG) and cytokeratin 7. Cell sheets with follicles were fabricated by cells incubated in both Dulbecco's Modified Eagle Medium (DMEM) and hepatocyte-defined medium (HDM) culture medium. The diameter and thickness of sheets fabricated in HDM were larger and thicker than those fabricated from DMEM. Furthermore, the cells incubated in HDM secreted higher levels of fT3 and fT4 than those incubated in DMEM. The thyroid peroxidase and TG mRNA of cells maintained in HDM were higher than those in cells maintained in DMEM. CONCLUSION: HDM appears suitable as a culture medium for maintaining primary thyrocytes and fabricating functional cell sheets. These in vitro findings may contribute to the development of appropriate culture conditions for human thyrocytes as well as engineered functional cell sheets.
Eagles ; Humans ; In Vitro Techniques ; Iodide Peroxidase ; Keratin-7 ; RNA, Messenger ; Thyroglobulin ; Thyroid Gland ; Transcription Factors