Effects of heparin-binding epidermal growth factor-like growth factor on cell repopulation and signal transduction in periodontal ligament cells after scratch wounding <i>in vitro</i>

Lee, J. S.,Kim, J. M.,Hong, E. K.,Kim, S.-O.,Yoo, Y.-J.,Cha, J.-H. Blackwell Publishing Ltd 2009 Journal of periodontal research Vol.44 No.1

<P>Background and Objective: </P><P>A growing amount of attention has been placed on periodontal regeneration and wound healing for periodontal therapy. This study was conducted in an effort to determine the effects of heparin-binding epidermal growth factor-like growth factor on cell repopulation and signal transduction in periodontal ligament cells after scratch wounding <I>in vitro</I>.</P><P>Material and Methods: </P><P>Human periodontal ligament cells were acquired from explant tissue of human healthy periodontal ligament. After the wounding of periodontal ligament cells, the change in expression of heparin-binding epidermal growth factor-like growth factor and epidermal growth factor receptors 1–4 mRNA was assessed. The effects of heparin-binding epidermal growth factor-like growth factor on periodontal ligament cell proliferation and repopulation were assessed <I>in vitro</I> via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and by photographing the injuries, respectively. Extracellular signal-regulated kinase (Erk)1/2, p38 and Akt phosphorylation was characterized via western blotting.</P><P>Results: </P><P>Scratch wounding resulted in a significant up-regulation of heparin-binding epidermal growth factor-like growth factor mRNA expression, whereas wounding had no effect on the expression levels of epidermal growth factor receptors 1–4. Interestingly, no expression of epidermal growth factor receptors 2 and 4 was detectable prior to or after wounding. Heparin-binding epidermal growth factor-like growth factor treatment promoted the proliferation and repopulation of periodontal ligament cells. The scratch wounding also stimulated the phosphorylation of Erk1/2 and p38, but not of Akt, in periodontal ligament cells, and heparin-binding epidermal growth factor-like growth factor treatment applied after wounding amplified and extended the activations of Erk1/2 and p38, but not of Akt. Furthermore, Erk1/2 inhibition blocked the process of cell repopulation induced by heparin-binding epidermal growth factor-like growth factor, whereas the inhibition of p38 delayed the process.</P><P>Conclusion: </P><P>These results indicate that heparin-binding epidermal growth factor-like growth factor may constitute a critical factor in the wound healing of human periodontal ligament cells by a mechanism that requires the activation of Erk1/2 via specific interaction with epidermal growth factor receptor 1.</P>

구강악안면영역에서 발생한 편평상피세포암종세포주에서 상피성장인자수용체의 발현 및 돌연변이에 관한 연구

김철환,김경욱 大韓顎顔面成形再建外科學會 2003 Maxillofacial Plastic Reconstructive Surgery Vol.25 No.2

Carcinogenesis has been considered as multiple stage process of denaturation of gene which control regulation of cell growth. The growth and differentiation of oral mucosal cell is controlled by growth factors which regulate development and differentiation of cell and apoptosis. It is well known that cell growth is not prevented, and secretion or reaction of growth factors is disturded in head and neck carcinoma including oral carcinoma. The epidermal growth factor is a polypeptide with potent mitogenic activitly of 6,045 daltons which has been shown to involve nuclear differentiation by trigger a cascade of intracellular ionic change and morphological transformation that it bind with epidermal growth factor receptor in surface of cells and delivering signal to inside of cells. The epidermal growth factor receptor is 170-kDa transmembrance phosphoglycoprotein. The extracelloular domain is binding site of epidermal growth factor, and intracellular domain possesses intrinsic tyrosine kinase astivity,which the stimulation can lead to autophosphylation of epidermal growth factor receptor and sequences lead to phosphylation of target protein in case of epidermal growth factor bind with target cell surface recrptor. Inrecently, eapression of epidermal growth factor receptor has been detected in oral squamous cell carcinoma, and overexpression of it is proved, and possibility of early markers of carcinogenesis in head and neck is presented. The many studies for the expression of epidermal growth factor receptor and relationship between it and prognosis of malignancy in carcinoma of breast or stomach have documented. But, in oral squamous cell carcinoma, the study for difference of amplication and mutation of epidermal growth factor receptor gene between primary carcinoma and merastastic carcinoma is rare. For study on mRNA expression of epidermal growth factor receptor,normal human oral keratinocyte, and cell lines of primary and metastastic oral squamous cell carcinomas were cultured, and then, electrophresis and RT-PCR(Reverse Transcriotion-polymerase Chain Reaction)were oerformed. The results were obstained as follows: The mutation of mRNA of EGF receptor were not detected in all oral squamous carcinoma cell lines. The expression of EGF receptor in oral squamous cell lines expressed 3-7 times higher than that of normal human oral keratinocyte of cytoplasmic domain. and 2-3times higher in extracellular domain. the expression of EGF receptor of cytoplasmic domain more expressed than that of extracellular domain in primary squamous cell line. The expression of EGF receptor in metastastic carcinoma cell lines is similis\ar or underexpressed than that of normal human oral keratinocyte cell lines. the expression of EGF receptor in the primary carcinoma cell lines expressed 2-5 times higher than that of metastastic carcinoma cell lines. From the results obtained in this study, the mutation have not mRNA of EGF receptor in carcinoma cell lines, and EGF receptor was overexpresses in early stage of carcinogemesis, and cytoplasmic domain of EGF receptor have relation with carcinogenesis, and EGF receptor have no relation with metastasis.

Enhanced regenerative healing efficacy of a highly skin-permeable growth factor nanocomplex in a full-thickness excisional mouse wound model

Bae, Il-Hong,Park, Jin Woo,Kim, Dae-Yong Dove Medical Press 2014 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.9 No.-

<P>Exogenous administration of growth factors has potential benefits in wound healing; however, limited percutaneous absorption, inconsistent efficacy, and the need for high doses have hampered successful clinical use. To overcome these restrictions, we focused on the development of a topical formulation composed of highly skin-permeable multimeric nanocomplex of growth factors. In the present study, we fused low-molecular-weight protamine (LMWP) with epidermal growth factor (EGF), insulin-like growth factor 1 (IGF-I), and platelet-derived growth factor A ligand (PDGF-A) (producing recombinant [r]LMWP-EGF, rLMWP-IGF-I, and rLMWP-PDGF-A, respectively) via genetic modification. Then, we used in vitro cell proliferation studies to assess the biological activity and the benefits of the combination. The LMWP-conjugated growth factors were complexed with low-molecular-weight heparin (LMWH) and formulated with Poloxamer 188 as a delivery vehicle. After confirming the enhanced skin permeability, in vivo studies were performed to assess whether the LMWP-conjugated growth factor nanocomplex formulations accelerated the healing of full-thickness wounds in mice. The LMWP-conjugated growth factors were biologically equivalent to their native forms, and their combination induced greater fibroblast proliferation. rLMWP-EGF showed significantly enhanced permeability and cumulative permeation, and the rates for rLMWP-IGF-I and rLMWP-PDGF-A, across excised mouse skin, were 124% and 164% higher, respectively, than for the native forms. The LMWP-fused growth factors resulted in formation of nanocomplexes (23.51±1.12 nm in diameter) in combination with LMWH. Topical delivery of growth factors fused with LMWP accelerated wound re-epithelialization significantly, accompanied by the formation of healthy granulation tissue within 9 days compared with a free–growth factor complex or vehicle. Thus, the LMWP-conjugated growth factor nanocomplex can induce rapid, comprehensive healing and may be a candidate wound-healing therapeutic.</P>

인간 황체화과립막세포에서 변형성장인자-α가 인슐린유사성장인자-2와 인슐린유사성장인자 결합단백질-1, 3의 분비 및 표피성장인자 수용체의 발현에 미치는 영향

강은희 ( Eun Hee Kang ),홍석호 ( Seok Ho Hong ),조준식 ( Jun Sik Cho ),김성훈 ( Sung Hoon Kim ),채희동 ( Hee Dong Chae ),김정훈 ( Chung Hoon Kim ),강병문 ( Byung Moon Kang ),남주현 ( Joo Hyun Nam ) 대한산부인과학회 2002 Obstetrics & Gynecology Science Vol.45 No.12

목적 : 변형성장인자-α (transforming growth factor-α, TGF-α)를 인간 난소 황체화과립막세포 (luteinized granulosa cell)에 처리하고 배양하여 TGF-α가 인간 난소 황체화과립막세포의 인슐린유사성장인자-Ⅱ (insulin- like growth factor-Ⅱ, IGF-Ⅱ), 인슐린유사성장인자 결합단백질-1, 3 (insulin-like growth factor binding protein-1, 3, IG Objective : To investigate the effects of transforming growth factor (TGF)-α on insulin-like growth factor (IGF)-α, insulin-like growth factor binding protein (IGFBP)-1, and 3 secretion and epidermal growth factor (EGF) receptor expression in cultured hum

Epidermal Growth Factor 와 Transforming Growth Factor-α가 인체 구강편평상피세포암 세포의 성장에 미치는 영향에 관한 실험적 연구

박영욱,Park, Young-Wook 대한악안면성형재건외과학회 1998 Maxillofacial Plastic Reconstructive Surgery Vol.20 No.4

Stimulatory effects of epidermal growth factor (EGF) and transforming growth $factor-{\alpha}$($TGF-{\alpha}$) on the growth of squamous cancer cell lines established from human oral cancer tissue with moderate differentiation were studied in vitro. After culturing in serum-free media for 24 hours, growth factors-EGF only, $TGF-{\alpha}$ only and EGF, $TGF-{\alpha}$ together-were added to the media and numbers of cells were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and compared with the control at 96, 144 hours. Each of EGF and $TGF-{\alpha}$ showed statistically significant stimulatory effects on the growth of cells respectively. Dose-dependent relationship of the stimulatory effects were not clearly demonstrated. The effects of EGF were higher than those of $TGF-{\alpha}$ and combinative administration showed higher effects than those of single uses. In conclusion, EGF may play an important and major role in differentiation and growth of human oral squamous cancer cells. $TGF-{\alpha}$, produced from cells activated by EGF, also can stimulate the cell growth and could be an alternative ligand for EGF receptor.

Epidermal Growth Factor와 Transforming Growth Factor-α가 인체 이행상피암 세포주의 성장에 미치는 영향에 관한 연구

신기용,김선진,최원호,최도연,박해영,이춘용,공구,우영남,김동한 大韓泌尿器科學會 1998 Korean Journal of Urology Vol.39 No.1

Human Epidermal Growth Factor Improves Atopic Disease-like Skin Lesions in DFE/DNCB Induced BALB/c Mice and Human Keratinocytes

김낭규,김성훈,박보현,최은주 한국운동생리학회 2022 운동과학 Vol.31 No.4

PURPOSE: Atopic skin disease is an inflammatory disease with a multifactorial pathogenesis that presents as itchy, dry skin. Human epidermal growth factor (EGF) plays an essential role in regulating cell growth, proliferation, and differentiation. Accordingly, this study aimed to investigate the effects of EGF on atopic diseases. METHODS: Atopic disease was induced by the Dermatophagoides farinae house dust mite extract and 2,4-dinitrochlorobenzene in BALB/c mice. The mice were divided into the following five atopic disease induction and treatment groups: control, atopic disease only, atopic disease+epidermal growth factor-1, atopic disease+epidermal growth factor-10, and positive control atopic disease+ceramide. We examined dermal and epidermal ear thickness, mast cell and T cell infiltration, serum immunoglobulin E and immunoglobulin G2a levels, and mRNA expression of pathogenic cytokines in murine ear tissue and human keratinocyte cells. RESULTS: Epidermal growth factor treatment increased epidermal and dermal ear thickness and inhibited mast and T cell infiltration in the ear in a dose-dependent manner. Treatment also suppressed immunoglobulin E and G2a levels and the mRNA expression of pathogenic cytokines in murine ear tissue and human keratinocytes. The therapeutic outcome of epidermal growth factor was stronger than that of the positive control ceramide. CONCLUSIONS: EGF may be beneficial in the treatment of atopic disease.

Epidermal Growth Factor가 흰 생쥐 표피 및 모발 성장에 미치는 영향에 관한 연구

이유신(Yoo Shin Lee),최지호(Jee Ho Choi) 대한피부과학회 1985 대한피부과학회지 Vol.23 No.6

Epiclerrnal growth factor(EGF), a low-molecular-weight polypeptide chain consisting of 53 amino acids, stimulates the growth and keratinization of epidermal cells. Recently, some authors reported that hair growth rate and follicular bulb growth were inhibited by EGF in neonatal mice. Since the hair and follicular cells are of epidermal origin, the effect of EGF would appear to be inconsistent with its role promoting the proliferation and differentiation of epidermal cells in vivo and in vitro. We have investigated the effects of EGF on the hair and epidermis in adult male mouse. The results are summarized as follows: l. 1'he hair length and maximum diameter were significantly reduced in EGF- treated regions compared with control group, and inhibition of hair growth was correlated with EGF dose. Thickness of the epidermis was significantly increased in EGF-treated regions compared with control group, and proliferation of the epidermis was correlated with EGF dose. 3 There were no significant differences in the hair length, maximum diameter, and egidermal thickness between untreated contralateral right mid-side regions of EGF-treated groups and control group.

창상조절인자들이 창상수축에 미치는 영향:FPCL model을 이용한 실험적 방법

조을제,남성한,박준석,김점용,이호남,김동현 大韓成形外科學會 1999 Archives of Plastic Surgery Vol.26 No.5

Many investigators have reported that collagen gel contraction reflects the mechanism of wound contraction. In 1995, Tsai et al. reported that hypertrophic scar-derived fibroblasts in a connective tissue model possessed the greatest contraction potency when compared with those of normal skin and normal oral mucosa-derived CTMs. In this study, we studied the effect of collagen gel contraction by growth factors such as epidermal growth factor, platelet-derived growth factor, transforming growth factor-β1, and transforming growth factor-β3, Skin fibroblasts used in this study were obtained from the explant of rat skin culture. Fibroblasts were cultured in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Growth factors were added per FPCL in the desired concentrations and we measured the collagen gel diameters in growth factor-treated FPCL on day 1,2,3, and 4 respectively after starting incubation. We examined the effects of EGF, PDGF, TGF-β₁, TGF-β₃ and the effects of combinations of TGF-β₁+ EGF, TGF-β₁+ PDGF, and TGF-β₁+ TGF-β₃ to contract a collagen gel. EGF has little influence on collagen gel contraction. TGF-β₁ and TGF-β₃ increase the collagen contraction. TGF-β₁ enhanced the contractility of collagen gel according to the concentrations. While TGF-β₃ alone had stimulatory contraction effects at low dose, high doses of TGF-β₃ decreased the potency of collagen gel contraction. A combination of TGF-β₁ and EGF minimally decrease TGF-β₁ activity. A combination of TGF-β₁and PDGF had an effect similar to TGF-β₁ activity. A combination of TGF-β₁ and TGF-β₃ decreased TGF-β₁ activity. According to reports that FPCL contraction is equivalent to the process of wound contraction, growth factors which enhance gel contraction may be related to wound contraction and wound healing. TGF-β₁ is reported to enhance scar formation in fetal wound. EGF accelerates wound healing and inhibits the promotion of hypertrophic scar formation. Compared to the effect of collagen gel contraction in this study, the combination of TGF-β₁ and TGF-β₃ that inhibited the promotion of collagen gel contraction are thought to diminish the formation of scar tissue. As well, EGF that has not enhanced collagen gel contraction is thought to diminish the production of scar tissue. We will study the interactive effects of TGF-β₃, EGF and TGF-β₁ on the contraction of collagen gels in the future.

Ligand-Independent Epidermal Growth Factor Receptor Overexpression Correlates with Poor Prognosis in Colorectal Cancer

Yun, Sumi,Kwak, Yoonjin,Nam, Soo Kyung,Seo, An Na,Oh, Heung-Kwon,Kim, Duck-Woo,Kang, Sung-Bum,Lee, Hye Seung Korean Cancer Association 2018 Cancer Research and Treatment Vol.50 No.4

<P><B>Purpose</B></P><P>Molecular treatments targeting epidermal growth factor receptors (EGFRs) are important strategies for advanced colorectal cancer (CRC). However, clinicopathologic implications of EGFRs and EGFR ligand signaling have not been fully evaluated. We evaluated the expression of EGFR ligands and correlation with their receptors, clinicopathologic factors, and patients’ survival with CRC.</P><P><B>Materials and Methods</B></P><P>The expression of EGFR ligands, including heparin binding epidermal growth factor-like growth factor (HBEGF), transforming growth factor (TGF), betacellulin, and epidermal growth factor (EGF), were evaluated in 331 consecutive CRC samples using mRNA <I>in situ</I> hybridization (ISH). We also evaluated the expression status of EGFR, human epidermal growth factor receptor 2 (HER2), HER3, and HER4 using immunohistochemistry and/or silver ISH.</P><P><B>Results</B></P><P>Unlike low incidences of <I>TGF</I> (38.1%), <I>betacellulin</I> (7.9%), and <I>EGF</I> (2.1%), <I>HBEGF</I> expression was noted in 62.2% of CRC samples. However, the expression of each EGFR ligand did not reveal significant correlations with survival. The combined analyses of EGFR ligands and EGFR expression indicated that the ligands‒/EGFR+ group showed a significant association with the worst disease-free survival (DFS; p=0.018) and overall survival (OS; p=0.005). It was also an independent, unfavorable prognostic factor for DFS (p=0.026) and OS (p=0.007). Additionally, HER4 nuclear expression, regardless of ligand expression, was an independent, favorable prognostic factor for DFS (p=0.034) and OS (p=0.049), by multivariate analysis.</P><P><B>Conclusion</B></P><P>Ligand-independent EGFR overexpression was suggested to have a significant prognostic impact; thus, the expression status of EGFR ligands, in addition to EGFR, might be necessary for predicting patients' outcome in CRC. </P>