Osseointegration of the titanium implant coated with rhTGF-beta2/PLGA particles by electrospray: a preliminary microCT analyzing rabbit study / 대한치과보철학회지

PURPOSE: This preliminary rabbit study was conducted to evaluate the effect of recombinant human transforming growth factor-beta2 (rhTGF-beta2)/poly lactic-co-glycolic acid (PLGA) coating on osseointegration of the titanium (Ti) implant. MATERIALS AND METHODS: Eight Ti implants were anodized with 300 voltages for three minutes. Four of those were coated with rhTGF-beta2/PLGA by an electrospray method as the experimental group. The implants were placed into tibiae of four New Zealand rabbits, two implants per a tibia, one implant per each group. After 3 and 6 weeks, every two rabbits were sacrificed and micro-computed tomography (microCT) was taken for histomorphometric analysis. RESULTS: In scanning electron microscope (SEM) image, the surface of rhTGF-beta2/PLGA coated Ti implant showed well distributed particles. Although statistically insignificant, microCT analysis showed that experimental group has higher bone volume / total volume (BV/TV) and trabecular thickness (Tb.Th) values relatively. Cross sectional view also showed more newly formed bone in the experimental group. CONCLUSION: In the limitation of this study, rhTGF-beta2/PLGA particles coating on the Ti implant show the possibility of more favorable quantity of newly formed bone after implant installation.

Effects of Maternal Hyperthermia on the Palatal Mesenchyme Development of Hsp70 Knock-out Mice Fetuses / 체질인류학회지

To investigate the effects of maternal hyperthermia on the development of the palate, pregnant Hsp70 knock-out mice at gestational day (GD) 8.5 were immersed in 43degrees C water bath until their body core temperature reached at 43degrees C. Thereafter, pregnant mice were given more 5 minutes hyperthermic exposure. Heat-untreated Hsp70 WT mice fetuses were used as the control group. Fetuses were collected at embryonic day 13.5, 14.5 and 15.5 (E13.5, E14, 5 and E15.5). Heads followed by removal of the mandible and the tongue were obtained and photographed for palatal development. Developing palates were processed for histological and immunohistochemical studies. Tissue sections were immunostained for TGF-beta2, FGF-8 and fibronectin, and observed with light microscope. The obtained results were as follows: Cleft palate was formed in heat-treated Hsp70 KO fetuses at E14.5 and E15.5. Immunohistochemical findings indicated that TGF-beta2 expression of the experimental fetuses were more delayed than that of the control fetuses. Mesenchyme under the medial edge epithelium (MEE) and cells of MEE showed continuously strong positive TGF-beta2 reactivity at E15.5. FGF-8 was revealed in both of the mesenchyme and the epithelium at the same time. FGF-8 immunoreactivity in the mesenchyme and the epithelium of the heat-treated fetuses showed strong reactivity at E15.5. In the experimental fetuses fibronectin was revealed the mesenchyma and basal lamina at E15.5. Taken together, it is suggested that maternal hyperthermia induces continuous expression of TGF-beta2 and FGF-8 in the mesenchyme and delayed expression of fibronectin. These should affect the normal palatogenesis and result in cleft palate.

Inhibitory Effect of Gefitinib on the Proliferation of Lens Epithelial Cells

PURPOSE: To evaluate the effect of Gefitinib used for the treatment of non-small cell lung cancer on the proliferation of the lens epithelial cells and the activities of growth factors. METHODS: The cell samples were divided into a control group cultured in DMEM and three experimental groups. Group I was exposed to Gefitinib for 3 minutes; group II was exposed to growth factors; and for group III growth factors were added to each concentration of Gefitinib. MMT assays, BrdU staining and morphologically observations with a phase-contrast microscope were used to verify the degrees of cell proliferation. Western blot analysis was conducted to confirm the effects of Gefitinib on extracellular signal-regulated kinase phosphorylation and on type I collagen production. RESULTS: Lens epithelial cell proliferation in experimental group I was reduced to 76% and 56% for 1.00 micro M and 10.00 micro M of Gefitinib, respectively. Experimental group II showed a 140% increase in cell proliferation with EGF treatment. Experimental group III exhibited decreased lens epithelial cell proliferation with both EGF (86% and 66%), and TGF-beta2 (78% and 59%). BrdU staining demonstrated a significant decrease in cell proliferation in groups I and III exposed to more than 1.0 micro M Gefitinib, while group II showed an increase compared to the control group. Moreover, Western blot analysis revealed inhibiting effects on ERK phosphorylation and type I collagen production. CONCLUSIONS: In the culture of human lens epithelial cells, Gefitinib inhibited cell proliferation when used at a concentration above 1.00 (M for 3 minutes. Furthermore, the effects of EGF and TGF-beta2 were also inhibited.

Modulatory Effect of TGF-beta 2 to Proliferative Kinetics of Fibroblast in Keloid and Hypertrophic Scar

Keloid and hypertrophic scar are dermal fibroproliferative disorders characterized by an overabundant deposition of collagen. Recently these dermal proliferative disorders have been linked clinically to the cytokine transforming growth factor beta(TGF-beta), and in vitro tests have shown it to be responsible for the activation of fibroblasts and their production and deposition of collagen. By using an established in vivo animal model of proliferative scarring, these scars were examined. Proliferative scar specimens were implanted into athymic, asplenic nude rats and isolated in sandwich island flap based on the superficial inferior epigastric pedicle. After establishment of the transferred flap, the scars were injected with varying doses of TGF-beta 2 or vehicle for 5 consecutive days and then again on days 10, 15, and 20. The specimens were measured weekly during the period of dosing, and a biopsy was acquired on days 30 and 60. Fibroblasts from the explanted biopsies and the original scars were grown in cell culture, and cell proliferation studies were performed and the results compared. There was a dose response to TGF-beta 2, with 200ng showing the greatest effect. From the original scar specimens, keloid scars demonstrated the greatest cell proliferation kinetics-significantly faster than hypertrophic scars. After treatment with TGF-beta 2, keloids showed an increase in their cell proliferation kinetics compared with vehicle alone. This was not demonstrated with the hypertrophic scars. Elevated levels of TGF-beta 2 are a major contributing factor to the process of proliferative scars, but because hypertrophic scars do not result in an equally increased response to this cytokine, a truly causative role for this cytokine cannot be promulgated. Rather, it is rather combination of the proliferative scar fibroblasts' abnormal response to TGF-beta 2 stimulation and elevated levels of this cytokine that controls more accurately the process of keloid formation.

The Effect of Imatinib (Gleevec(R)) on The Proliferation of Lens Epithelial Cells and The Activities of Growth Factors

PURPOSE: To evaluate the effect of Imatinib which is tyrosine kinase inhibitor used for the treatment of chronic myelogenous leukemia on the proliferation of the lens epithelial cells and the activities of growth factors. METHODS: In the experimental group I, the cells were exposed to Imatinib for 3, 5 min with various concentration. In the experimental group II, the cells were cultured with PDGF, bFGF, TGF-beta2. In the experimental group III, the cells were exposed to Imatinib for 3, 5 min with the various concentration in the presence of each growth factor. The cell viability was assessed by MTT assay. And the cell proliferation were evaluated by BrdU staining. The phosphorylation of ERK (Extracellular-signal regulated kinase) and the amount of collagen type I produced by TGF-beta2 were analyzed with western blot. RESULTS: MTT assay showed that the viability of lens epithelial cells was decreased about 50% at the concentration above Imatinib 30 micro M for 5 mins exposure in group I. Both PDGF and bFFG induced significantly increased cell viability in group II. The group III that was treated with both PDGF and bFGF showed the decrease of cell viability after being exposed to Imatinib. As the concentration of Imatinib increased, BrdU incorporation of experimental group I was decreased compared with control group. It also found that The BrdU incorporation of experimental group III was also decreased compared with experimental group II. The phosphorylation of ERK 1/2 and the amount of collagen type I production was significantly decreased in addition of Imatinib 20 to 30 micro M for 3 mins exposure. CONCLUSIONS: The proliferation of lens epithelial cells could be inhibited by Imatinib. And the activities of PDGF, bFGF, TGF-beta2 were also inhibited by Imatinib.

Effects of Corneal Neovascularization on TNF-alpha and TGF-beta 2 mRNA Expression in Endotoxin Induced Uveitis

PURPOSE: We tried to evaluate the expression of TNF-alpha and TGF-beta 2 mRNA during the course of the endotoxin induced uveitis (EIU) in rat after inducing neovascularization on cornea. METHODS: Reverse transcription followed by polymerase chain reaction amplification was used to determine relative mRNA level in two ocular tissues (iris-ciliary body, cornea) at 0, 1, 3, 6, 12, 48 hours after subcutaneous injection of 200 ug of Escherichia coli endotoxin. RESULT: Rat with corneal neovascularization showed somewhat different pattern of expression. TGF-beta 2 mRNA expression was showed some fluctuations in cornea. TGF-beta 2 mRNA expression in cornea was decreased within 3 hours after endotoxin treatment and increased gradually after 6 hours until 48 hours. CONCLUSION: TGF-beta2 might be one of the important cytokines in EIU, especially animals with neovascularization on cornea.

Expression of Transforming Growth Factor -beta1, -beta2 in Human Endometrium of The Uterine Adenocarcinoma / 대한부인종양콜포스코피학회잡지

OBJECTIVE: To determine the differences of expression of Transforming Growth Factor (TGF)-beta1 and TGF-beta2 in the human proliferative, secretory, menopausal endometrium, and in hyperplasia and adenocarcinoma of the endometrium. MATERIALS AND METHODS: Fifty patients were divided into 5 groups. Twenty samples were collected from patients with endometrial hyperplasia (n=l0) and adenocarcinoma(n=10) after hysterectomy. Thirty samples were collected from the normal menstrual cycle and the menopausal women as a control group. The histological types of endometrium were proliferative(n=10), secretory(n=10), menopausal(n=10). Immunohistochemical staining was performed through the use of monoclonal antibodies against anti-human TGF-beta1/ beta2 polycolonal IgG rabbit antibody. Expression of TGF-beta1 and TGF-beta2 were judged positive when the staining revealed color development in 5% or more. Specimens were rated absent, trace, weak, moderate and intense. Then, they were scored 4 in case of intense positive for TGF-beta1 and TGF-beta2, and '0' in case of absent for TGF-beta1 and TGF-beta2 . RESULTS: Mean scores of TGF-beta1 in glandular cell were proliferative(1.0), secretory (2.3), menopausal endometrium(1.0), endometrial hyperplasia(0) and adenocarcinoma(0). Expression of TGF-beta2 in glandular cell were proliferative(3.8), secretory(3.3), menopausal endometrium(2.8), endometrial hyperplasia(0.3) and adenocarcinoma(0.3). No specific different expression of TGF-beta1 and TGF-beta2 was found between stromal, vascular and myometrium. However, there was similar expression of TGF-beta1 and TGF-beta2 in endometrial hyperplasia and adenocarcinoma gmup. CONCLUSIONS: TGF-beta1 and TGF-beta2 may have important roles to suppress the development of the precancerous or cancerous lesions in endometrial glandular cells.

The Effects of TGF-beta2 and bFGF on the Proliferation of Retinal Pigment Epithelial Cells

This study was undertaken to document the effect of transforming growth factor-beta2 (TGF-beta2(TGF-beta2) and basic fibroblast growth factor (bFGF) on the proliferation of pig retinal pigment epithelial cells (RPE). Whereas bFGF increased the proliferation, TGF-beta2 showed the inhibitory effect on the proliferation The inhibitory effect of TGF-beta2 disappeared in RPE subcultured with 10ng/ml of bFGF. Both TGF-beta2- and bFGF-specific antisense oligonucleotides blocked the autocrine effect of the growth factors. PLC-71 -specific antisense oligonucleotide inhibited the effect of TGF-beta2 and bFGF. Genistein inhibited the effect of TGF-beta2 and bFGF in dose-dependent man, ner. The data suggest the involvement. of in PLC-/1 and tyrosine kinase in signalling.

The Expression of TGF-beta1 and TGF-beta2 in Prostatic Carcinoma / 대한암학회지

Transfonning growth factor-beta (TGP-beta) is a multipotent growth factor affecting development, homeostasis, and tissue repair. We evaluate the significance of the expression of TGF-beta1 and TGF-beta2 and correlation with prognostic factors in prostate cancer. MATERIALS AND METHODS: In order to investigate the expression of TGF-beta1 and TGF-beta2, we analyzed Immunohistochemical staining from paraffin blocks of 22 cases of the prostate carcinoma and adjacent normal prostate. RESULTS: The expressions of TGF-beta1 and TGF-beta2 were noted in the cytoplasm of tumor cells, normal epithelial cells and stroma. The staining intensity and areas were examined and scored from 0 to 5. The TGF-beta1 staining scores of the tumor cells were higher than that of the adjacent normal epithelial cells (p=0.001). The TGF-beta2 staining scores of the tumor cells were also higher than that of the adjacent normal epithelial cells (p=0.003). However, there were no correlation between tumor surrounding stroma and normal stroma in TGF-beta1 and TGF-beta2 staining scores. The serum PSA level, the clinical stage, the Gleason score and the lymph node metastasis of the tumor did not correlated with the staining score of TGF-beta1 and TGF-beta2. CONCLUSION: These results indicate that the prostatic cancer was associated with alteration of TGF-beta1 and TGF-beta2 expression by prostatic epithelial cells which play a role in prostatic carcinogenesis.

Immunohistochemical Localization of Transforming Growth Factor-beta1, beta2 in the Endometrium of Fertile and Infertile Patients / 대한산부인과학회잡지

No abstract available.

Immunohistochemical Study of Transforming Growth Factor-beta2(TGF-beta2) and Vascular Endothelial Growth Factor(VEGF) after Laser Photocoagulation in the Ocular Tissues of White Rats

To explain a part of the machanism of regression of new vessels after laser photocoagulation in retinal vascular occlusive diseases causing neovascularization, particularly for diabetic retinopathy, retinal vein occlusion, retinopathy of prematurity, and age-related macular degene- ration, we performed this study. Photocoagulation was done with argon green laser on the right superofrontal retina of the 15 eyes of white rats. On the first, third, and sixth day after laser photocoagulation, both eyes of 5 rats were enucleated and stained with polyclonal anti-transforming growth factor-beta2(Anti-TGF-beta 2) and anti-vascular endothelial growth factor(Anti-VEGF) antibody by immuno peroxidase technique. TGF-beta2 and VEGF were not expressed in retina of normal control eyes. After laser photocoagulation, degree of expression of TGF-beta2 and VEGF increased in 24 hours, only in and adjacent to the photocoagulated area. Thereafter, degree of expression of both factor decreased, especially that of VEGF decreased much more than that of TGF-beta2. To elucidate the exact mechanism, more qualitative and quantitative analysis will be necessary.