• Korean Journal of Animal Reproduction
• /
• v.21 no.1
• /
• pp.61-69
• /
• 1997

The present study was carried out to investigate the effects of EGF and anti-EGF on early embryonic development and hatching in mice. Developmental and hatching rates of mouse em-bryos from 2-cell to morular stage which were cultured in Ham's FlO medium supplemented with EGF (1-1,000 ng/ml) or anti-EGF (whole serum diluted from 1:10 to 1:1,000) were compared to those of control When mouse early 2-cell embryos were cultured in the EGF supplemented medium, blastulation was accelerated compared with control. Hatching rate was also significantly (p

• Journal of the korean academy of Pediatric Dentistry
• /
• v.34 no.3
• /
• pp.438-446
• /
• 2007

Viable cells of periodontal ligament would be an important factor for the successful replantation of an avulsed tooth. Therefore, it is critical to choose the storage medium for the preservation of traumatically avulsed teeth. Growth factors and hormones could be considered for the therapeutic application of the maintenance of viable periodontal ligament fibroblasts (PDLFs). Epidermal growth factor (EGF) has been suggested as an important player for the regeneration and wound healing process on other tissues. Therefore, EGF was evaluated for the therapeutic application on avulsed teeth. In addition, the synergic effect of EGF with tri-iodothyronine (T3) and heparin-binding epidermal growth factor-like growth factor (HB-EGF). The cell proliferation of PDLFs was determined by MTT assay and increased dose-dependently up to 10 ng/ml in the presence of EGF. Maximum cellular growth was shown at the concentration of 10 ng/ml EGF. Also, EGF promoted the wound healing of PDLFs examined by in vitro wound healing assay. Combined effects of EGF with T3 or HB-EGF on the proliferation of PDLFs were also studied. Interestingly, EGF showed the synergic effect on the proliferation of PDLFs with T3 and HB-EGF. To find out the mechanism of the synergic effect of EGF and T3, the effect of T3 on the expression of endogenous EGF receptor was determined by RT-PCR. The result was that T3 enhanced the expression of EGF receptor in PDLFs. It suggested that EGF might be a good choice for a therapeutic application, which can be used as combination with T3 and HB-EGF.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.1
• /
• pp.13-20
• /
• 1997

The objective of this study was to determine the effect of EGF on the preimplantation development of mouse IVF embryos and their ICM and TE cell number. And also, we examined the expression of EGF-R protein on embryonic development by indirect immunofluorescence. The results obtained in these experiments were summarized as follows; Group culture (5 embryos/ 25 ${\mu}l$) showed more improved development rate to blastocyst than singly culture. This inferior development of singly cultured 2-cell embryos improved by the addition of EGF. Especially, 2-cell embryos cultured singly in 10 ng/ml of EGF (62.4%) indicated significant difference in development to blastocyst compared with control group (47.9%). Also, cell number of ICM and TE by differential labelling showed the increased pattern in the EGF treatment group. The stimulating effect of EGF with the development level was significantly increased after 4-cell stage (p<0.05). ICM proportion also showed the increased pattern with the developmental level in the EGF treatment group. In addition, expression of EGF-R by indirect immunofluorescence detected after 4-cell stage. Therefore, EGF could stimulate preimplantation mouse embryo development by binding with expressed EGE-R after 4-cell stage and produce the more increased ICM and TE cell number of blastocyst.

• Korean Journal of Animal Reproduction
• /
• v.20 no.3
• /
• pp.279-288
• /
• 1996

The objective of this study was to determine the effect of EGF on the development of IVM/IVF bovine embryos and their ICM and TE cell number. In addition, we examined the combined effect of EGF and coculture to the bovine embryo development and the expression of EGF-R protein on bovine embryos by indirect immunofluorescence. The results obtained in these experiments were summarized as follows: When the IVM/IVF 4- to 8-cell embryos were treated at 0, 1, 10, 100 ng/ml of EGF, EGF treatment group showed improved development to blastocyst and increased pattern of ICM and TE cell number compared with control, although there is not significantly different. The stimulating effect of EGF (10 ng/ml) to the develop ment level of IVM/IVF bovine embryos significantly increased development rate to blastocyst after 8-cell stage (p<0.05), although there is no significant effect to the increase of ICM and TE cell numbers. Also, expression of EGF-R on the bovine embryonic stage by indirect immunofluor escence presents after 4-cell stage and the intensity of the EGF-R staining was variable with the development progression. On the other hand, embryos cultured in coculture group added either with or without EGF commonly indicated the significant difference in development rate to blastocyst and Total cell number compared with control. These results suggest that the a addition of EGF to the coculture may stimulate the coculture effect between IVM/IVF bovineembryos and cumulus cells. Therefore, EGF could promote preimplantation bovine embryo development by binding with expressed EGF~R after 4-cell stage, and stimulate the production of embryotrophic factors from the coculture environment. Also, the present study showed that there was no significant effect of EGF to the increase of ICM and TE cell number although the rate of blastocyst significantly increased when treated with EGF after 8-cell stage (p<0.05).

• Korean Journal of Animal Reproduction
• /
• v.19 no.3
• /
• pp.217-226
• /
• 1995

The objective of this experiment was to test the effect of EG F on nuclear maturation of pig immature oocytes in vitro. Basic medium used TCM-199 supplemented with 0.2 mM pyruvate, 1 ${\mu}\textrm{g}$/ml estradiol-I7$\beta$ and 25 ${\mu}\textrm{g}$/ml gentamycin, this medium treated with EGF, FSH and FBS. Experiment 1 examined to the effect according to the addition of FSH or EGF (0, 1. 10 and 100 ng EGF/ml) in oocytes maturation. Nuclear maturation rates (M ll %) of 1, 10 and 100 ng EGF/ml (83.0. 8fi.7 and H7.5%) treatments were significantly higher than those of non- and FSH-treated groups (27.3 and 60.3%, p < 0. 001). Experiment 2 examined to the interactive effects of EGF. FSH or FBS during oocytes maturation. Nuclear maturation rates (M ll %) of EGF alone, EGF plus FSH, EGF plus FBS, FSH plus FBS, and EGF plus FSH added FBS treatments (86.7, 90.2, 87.1. 89.6% and 92.6%) were significantly higher than those of non, FSH, and FBS alone treatments (22.3, 52.2 and 42.3%, p < 0.001). Also, cumulus cells expansion of oocytes maturation was examined to total treatments. Normal cumulus cells expansion was shown by FSH plus FBS, EGF or EGF with FBS combination treatments, but cumulus cells of oocyte complexes were still clumped together in EGF-treated groups although they had separated from oocytes. However, EGF showed a positive on nuclear maturation. These results conclude that EGF alone can stimulate nuclear maturation in pig immature oocytes.

• Journal of Embryo Transfer
• /
• v.13 no.3
• /
• pp.245-249
• /
• 1998

The purpose of this study was to evaluate the effects of growth factors such as epidermal growth factor (EGF) and insulin-like growth factor-1 (IGF-1) on maturation of bovine follicular oocytes in vitro. Oocytes were recovered from the ovaries of slaughtered Hanwoos. The oocytes were matured in TCM 199 at 39$^{\circ}C$, 5% $CO_2$ in air. Growth factors were added to maturation medium as follows: control (no serum), EGF (10ng/m1, 50ng/ml or 100ng/m1), IGF-1 (100ng/m1) and EGF (50ng/ml) + IGF-1 (100ng/m1). The oocytes were placed onto a slide and stained with aceto-orcein dye. Nuclear maturation was evaluated and classified as germinal vesicle breakdown (GVBD), metaphase-I (MI) or metaphase-ll(Mll). Maturation rates were 37.9% (control), 45.8% (EGF, 10ng/m1), 55.8% (EGF, 50ng/ml), 44.4% (EGF, 100ng/m1), 46.7% (IGF-1, 100ng/m1) and 67.0% (IGF-1+EGF). The highest group developed to Mll stage was IGF-1+EGF treatment group (p<0.05). Therefore, nuclear maturation of bovine oocytes were affected by both of growth factors, and it seems to have a mutual activity between them.

• Journal of Pharmaceutical Investigation
• /
• v.27 no.2
• /
• pp.139-143
• /
• 1997

Epidermal growth factor (EGF) is a mitogen which activate the proliferation of basal cells in skin, which implicate the wound healing in severe skin damage such as burn. To carry out the preclinical test for the pharmacological action of EGF, EGF in transdermal delivery system must be stable. Since EGF is a protein susceptible to proteolysis and unstable in aqueous solution, in vitro stabilization of EGF is prerequisite for the formulation. In this study, effect of additives on the stability of EGF is investigated in vitro. The stability of EGF in aqueous solution was enhanced with the various water-soluble polysaccharides such as HPMC, sorbitol, mannitol and dextrin. EGF was successfully extracted from the ointment with 5% HPMC solution, and EGF in aqueous solution and ointment was also successfully stabilized with 5% HPMC. The ointments prepared with different amount of EGF were applied on the damaged dorsal skin of rats for the determination of optimal concentration of EGF. The ointment with EGF $(10\;{\mu}g/g)$ showed good wound healing action on the damaged skin of rats.

• KSBB Journal
• /
• v.7 no.3
• /
• pp.201-208
• /
• 1992

Cell surface binding of epidermal growth factor(EGF) to EGF receptors was studied for a series of site-directed receptor mutants transfected into B82 mouse fibroblasts. Scatchard plots for truncation mutant receptors significantly lost nonlinearity for truncations below residue 1022. Transient plots of dissociation kinetics exhibited biphasic behavior for all receptor types, but the fraction of receptor in slow-dissociating form was reduced by an order of magnitude for the truncation mutants below residue 1022. Comparison of dissociation kinetics between control cells and cells treated with Triton X-100 revealed no significant variation for the slow-dissociating receptor form, but a noticeable variation was observed for the fast-dissociating receptor form when EGF receptors were truncated below residue 991. These results suggest that high affinity of EGF binding at cell surface depend on the EGF receptor cytoplasmic region.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.45 no.2
• /
• pp.175-184
• /
• 2019

The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.

• Korean Journal of Animal Reproduction
• /
• v.17 no.1
• /
• pp.49-56
• /
• 1993

Mouse mammary epithelial cells(NMuMG) were plated onto 24 well phates(100,000 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, EGF)0~100ng/ml) was added simultaneously with IGF-I(10ng/ml), 1$\mu$M photoreactive cAMP(4,5-dimethoxy-2-nitrobenzyl adenosine-3',5' cyclic monophosphate, DMNB) or IGF-I plus DMNB. After 2 hours, the cells were expposed to UV light(300nm, 3 second pulse0 in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml, 18~19 hours after UV exposure). Without IGF-I or DMNB, EGF(10 or 100ng/ml) increased DNA synthesis from 8,362 dpm/well in control to 16,345 or 18,684 dpm/well with EGF(pooled SE=1,239 dpm/well, P<0.05). IGF-I or IGF-I plus DMNB alone increased DNA synthesis from 8,362 dpm/well in control to 17,307 or 20,427 dpm/well, respectively(P<0.05). Addition of IGF-I, DMNB or IGF-I plus DMNB into 0~100ng/ml EGF did not significantly change the shape of dose response curve of EGF alone. In other experiment, EGF or IGF-I plus DMNB into 10ng/ml EGF group exhibited interaction effect in DNAsynthesis [EGF(10ng/ml)=18,497; IGF-I＋EGF=22,837; DMNB＋EGF=20,658 ; IGF-I＋DMNB＋EGF=29,658, pooled SE=1,055, P<0.05]. These results indicate that simultaneous activation of EGF, IGF-I and intracellular cAMP interact in DNA synthesis of mouse mammary epithelial cells.