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>

Effect of Single Growth Factor and Growth Factor Combinations on Differentiation of Neural Stem Cells

Choi, Kyung-Chul,Yoo, Do-Sung,Cho, Kyung-Sock,Huh, Pil-Woo,Kim, Dal-Soo,Park, Chun-Kun The Korean Neurosurgical Society 2008 Journal of Korean neurosurgical society Vol.44 No.6

Objective : The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF). insulin growth factor-I (IGF-I). brain-derived neurotrophic factor (BDNF). and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Methods : Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using $\beta$-tubulin III and NeuN as neural markers. Results : Neural differentiations in the presence of individual growth factors for $\beta$-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-I, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for $\beta$-tubulin III and 3.1% for NeuN. For $\beta$-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1 %; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Conclusion : Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and post-mitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.

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>

일반 논문 : 종단적 자료를 활용한 벤처기업의 생존 및 성장 영향요인에 관한 실증연구

설원식 ( Won Sik Sul ),홍길표 ( Kil Pyo Hong ) 한국중소기업학회 2011 中小企業硏究 Vol.33 No.1

본 연구에서는 벤처기업의 생존에 영향을 주는 요인이 성장에도 동일한 영향을 주거나 혹은 그렇지 않은 관계가 존재하는지를 파악하기 위해 벤처기업의 생존 영향요인과 성장 영향요인을 연계시켜 분석하였다. 이를 위해 2003년과 2004년 벤처기업 경영실태 설문조사에 응했던 기업을 대상으로 설문조사 이후 4∼5년이 경과한 2008년 기준 생존 여부를 조사해 생존 영향요인을 분석했다. 또한 2003년 설문에 응답한 기업 중 2008년까지 생존한 기업을 대상으로 4년이 경과한 2007년까지 실제로 벤처기업이 얼마나 성장했는가를 측정하여 분석함으로써 성장 영향요인을 검증하였다. 연구의 주요 결과는 다음과 같다. 경영자 특성요인은 전반적으로 벤처기업의 생존요인이면서 성장요인으로 작용하는 것으로 나타났다. 한편 대표이사의 지식수준이나 대기업과의 네트워킹 능력은 벤처기업의 생존요인으로는 중요한 의미를 가지지만, 성장요인으로는 큰 영향을 미치지 않음을 발견하였다. 반면, 연구개발투자는 벤처기업의 생존에는 별 영향을 미치지 않지만, 성장과는 유의한 양(+)의 상관관계를 갖는 것으로 나타났다. 벤처기업의 업력 및 기업규모는 기대했던 것처럼 벤처기업의 생존과 성장에 서로 다른 방향의 영향을 미침을 확인하였다. 본 연구는 그간 상호 연계성을 갖지 않은 채 각각 분리되어 연구되어 왔던 벤처기업 생존 영향요인과 성장 영향요인을 비교·분석했다는 점에서 의의를 가지며, 연구결과는 벤처기업의 경영자나 벤처기업 정책결정자들에게 많은 실무적 시사점을 제시해 줄 수 있을 것으로 기대한다. Few attempts have been made to investigate the influences of and differences between survival and growth factors from an integrated perspective, whereas prior research has focused on the determinants of survival or growth respectively for new SMEs and venture firms. Do determinant factors of a firm`s survival also play an identical role in the growth of the firm? Conversely, are there factors which influence positively on a firm`s survival or deter a firm from growth? Using a group of more than 8,000 samples, we examined empirically whether determinant factors of survival and growth are identical in ventures. First of all, we analyzed survival factors by investigating whether firms in surveys conducted by the Small and Medium Business Administration in 2003 and 2004 survived until 2008. We investigated growth factors by analyzing the growth of those surviving venture firms during the four years after the previous surveys. This distinctive approach provided us with a new perspective to understand the determinant factors of growth. The factors we considered influencing survival and growth of venture firms were categorized based on firms` characteristics, such as organization, manager, technology, marketing and finance. Size and age of firms were the measures of organizational characteristics while ownership of founder and knowledge level of CEOs were those of managers characteristics. The capacity for technological innovation and investment in R&D were the proxy for assessing technological capabilities. Marketing characteristics were measured in terms of networking capacity with large firms and the level of globalization. Finally we used profitability and debt to equity ratio to assess financial characteristics. We analyzed the determinants of survival using binary logit model, where we divided sample firms into two categories, survival group and extinct group. Survival factors were sifted out through characteristics that were common to currently existing firms at the time of the 2003/2004 surveys. We investigated the determinant factors of growth using multiple regression model, where we computed growth rate of sales for each venture firm between 2003 and 2007, and used the growth rates as a dependent variable in regression analysis. Determinant factors influencing growth of venture firms were analyzed based on the firm characteristics mentioned above. The empirical results are summarized as follows. First of all, the hypothesis with regard to organizational characteristics, was partly supported, in that their effects on survival and growth were contrary to each other. We measured organizational characteristics by business history as well as size of firms using total assets and the number of employees. Business history measured by the number of years since incorporation, in particular, has a positive relationship on survival while it has a negative effect on growth rate in line with the hypothesis. Since start-up companies have the most considerable disadvantages in their early stages, chances of survival are substantially increased as business years roll on. The growth rate, meanwhile, slows down as years roll on, since start-up companies diverge from enterprising management in their early stage and intend to pursue stability. The size of ventures turns out to have a positive effect on survival, offsetting the disadvantages of startup by benefits from economies of scale, although size does not have a significant relationship with growth rate. Manager characteristics mostly work as a survival and growth factor. Founder ownership which is closely related to agency cost also turns out to be a survival and growth factor with statistical significance. It is interpreted as that founder ownership has a positive impact since potential agency cost between owner and manager would decrease when a founder owns and runs a venture. Academic background as a proxy for knowledge level of CEO is a survival factor, albeit statistically insignificant as a growth factor. We understand that the higher knowledge level a manager has, the more embedded technology and knowledge he or she is likely to have. High knowledge level would enhance a firm`s viability in the face of shocks in the external environment as shown in prior research. The empirical result also indicates that the capacity for technology innovation measured by the number of registered patents and patent applications turns out to have a positive effect on survival and a negative effect on growth. Although this study has the limitation that we narrowed the capacity for technology innovation down to the number of patents, the findings on growth rate is surprisingly different from our expectation beforehand. Meanwhile, the growth of ventures is positively related to R&D investment while R&D investment is not related to survival. It occasionally happens in the field that R&D investment adds to the burden on cash flow and rather threatens a firm`s survival when the investment does not lead to sales. Therefore, further research may need to distinguish R&D investment by sales. The impact of marketing on the survival and growth of venture firms was verified based on the networking capacity with large firms and the level of globalization. Networking capacity with large firms measured by weighted supply contracts did not work as a growth factor but worked as a survival factor. It seemed that supply contracts to big enterprises provided only stable sales for survival but they guaranteed neither high return for growth nor rapid sales growth. Globalization was measured by the experience of foreign market entry, where we expected that the level of globalization would have positive impact on growth since venture firms which entered overseas market would have new market opportunities. The empirical result, however, did not present a statistically significant relationship between growth and the level of globalization of venture firms. Finally, it is hypothesized that financial characteristics such as profitability and debt to equity ratio would influence the survival of venture firms. The deficiency of financial resources was expected to become a risk factor for the survival of venture firms, when firms did not have sizable internal capital and could not utilize external capital market. Debt to equity ratio is negatively related to survival while profitability has no significant effect on it. High debt to equity ratio might lead to financial distress threatening survival since the increased burden of interest payments might aggravate cashflows and high debt to equity ratio made it difficult to find new sources of finance. Prior research has separated survival factors and growth factors without considering any linkage between them. This study is meaningful in that we empirically verified survival factors and growth factors intended for a group of ventures with an integrated approach. We started the research from the question of whether factors contributing to survival of firms would also have identical impact on growth as well. Thus we examined deterministic factors for survival and growth and hypothesized the relationship between the factors and their impacts on survival and growth of venture firms respectively. Using time series data of 8,000 sample firms, we sifted out factors such as founder ownership, which have a positive impact on survival and growth simultaneously. Unique factors such as firm age and capacity for technological innovation by proxy of the number of patents, are presented as determinants which have a positive impact on survival and a negative impact on growth. In addition, factors which have influence not on growth but on survival and vice versa are verified based on the empirical findings. The results provide managers in venture firms and policy makers with managerial implications. Furthermore, this research lays the groundwork for further study by investigating the survival and growth of ventures from a comprehensive perspective and contributes to develop new research models leading to extending related theories. There are, however, a few problems that remain to be explored since we limited survival factors and growth factors to internal characteristics of venture firms out of a variety of organizational characteristics. This research concludes that the characteristics of both external environments of an organization as well as internal management characteristics need to be taken into account to fully explain the determinants of survival factors and growth factors of venture firms for theoretical expansion as Romanelli (1989) suggested earlier.

Priming of mononuclear cells with a combination of growth factors enhances wound healing <i>via</i> high angiogenic and engraftment capabilities

Jin, Enze,Kim, Jong-Min,Kim, Sung-Whan John WileySons Ltd 2013 Journal of cellular and molecular medicine Vol.17 No.12

<P>Recently, we demonstrated that a specific combination of growth factors enhances the survival, adhesion and angiogenic potential of mononuclear cells (MNCs). In this study, we sought to investigate the changes of the angiogenic potential of MNCs after short-time priming with a specific combination of growth factors. MNCs were isolated using density gradient centrifugation and incubated with a priming cocktail containing epidermal growth factor (EGF), insulin-like growth factor (IGF)-1, fibroblast growth factor (FGF)-2, FMS-like tyrosine kinase (Flt)-3L, Angiopoietin (Ang)-1, granulocyte chemotactic protein (GCP)-2 and thrombopoietin (TPO) (all 400 ng/ml) for 15, 30 and 60 min. Wounds in nonobese diabetic-severe combined immune deficiency (NOD-SCID) mice were created by skin excision followed by cell transplantation. We performed a qRT-PCR analysis on the growth factor–primed cells. The angiogenic factors vascular endothelial growth factor (VEGF)-A, FGF-2, hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF) and interleukin (IL)-8 and the anti-apoptotic factors IGF-1 and transforming growth factor-β1 were significantly elevated in the MNCs primed for 30 min. (T30) compared with the non-primed MNCs (T0). The scratch wound assay revealed that T30- conditioned media (CM) significantly increased the rate of fibroblast-mediated wound closure compared with the rates from T0-CM and human umbilical vein endothelial cells (HUVEC)-CM at 20 hrs. <I>In vivo</I> wound healing results revealed that the T30-treated wounds demonstrated accelerated wound healing at days 7 and 14 compared with those treated with T0. The histological analyses demonstrated that the number of engrafted cells and transdifferentiated keratinocytes in the wounds were significantly higher in the T30-transplanted group than in the T0-transplanted group. In conclusion, this study suggests that short-term priming of MNCs with growth factors might be alternative therapeutic option for cell-based therapies.</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.

Hormonal Regulation of Insulin-Like Growth Factor Binding Protein Secretion by a Bovine Mammary Epithelial Cell Line

Kim, W.Y.,Chow, J.C.,Hanigan, M.D.,Calvert, C.C.,Ha, J.K.,Baldwin, R.L. Asian Australasian Association of Animal Productio 1997 Animal Bioscience Vol.10 No.2

A mammary epithelial cell line (MAC-T) established as a model for lactation was utilized to identify and characterize effects of various hormones upon insulin-like growth factor binding protein secretion. Ligand and immunoblot analyses of conditioned media indicated that insulin-like growth factor binding protein-2 was secreted by MAC-T cells. Insulin-like growth factor-I stimulated insulin-like growth factor binding protein-2 secretion in a dose-dependent manner, but prolactin and bovine somatotropin did not alter insulin-like growth factor binding protein-2 secretion. Insulin increased and cortisol decreased insulin-like growth factor binding protein-2 secretion. Effects of insulin-like growth factor-I on insulin-like growth factor binding protein-2 secretion support previous studies using primary cultures of bovine mammary cells and bovine fibroblasts. Effects of cortisol and insulin on insulin-like growth factor binding protein-2 secretion may be explained by changes in protein synthesis. In addition, supraphysiological doses of insulin can cross-react with the insulin-like growth factor-I receptor and stimulate insulin-like growth factor binding protein-2 secretion. MAC-T cells provide a model system to study mechanisms that regulate local insulin-like growth factor-I bioactivity.

성장호르몬 결핍증 진단에 있어서 혈중 insulin-like growth factor-I 및 insulin-like growth factor binding protein-3 농도의 진단적 유용성에 대한 연구

지근하,이정녀,정우영 대한소아청소년과학회 2008 Clinical and Experimental Pediatrics (CEP) Vol.51 No.12

Purpose:This study aimed to determine the best cutoff line for insulin-like growth factor (IGF)-I and insulin-like growth factor binding protein (IGFBP)-3 to discriminate between growth hormone deficiency (GHD) patients and the control group. Methods:Two hundred thirty subjects with normal controls (129 boys and 101 girls, aged 7-15 years), 14 patients with complete GHD (12 boys and 2 girls), and 17 patients with partial GHD (9 boys and 8 girls) were studied. IGF-I serum concentrations were measured by radioimmunoassay (RI), and IGFBP-3 concentrations were measured by immunoradiometric assay (IRMA). Results:The receiver operating characteristic (ROC) plot analysis showed that the best IGF-I and IGFBP-3 cutoff line was at -1 standard deviation (SD). By comparing IGF-I serum levels of GHD children within 1 SD of normal control, we determined the sensitivity (S) (87.5-100%) and specificity (Sp) (80-84.6%) according to the age group. For IGFBP-3, we determined the following values: S (58.7-85.7%) and Sp (79.2-85.5%). Eleven of 14 patients with complete GHD (78.5%) and 16 of 17 patients with partial GHD (94.1%) had IGF-I concentrations equal to or below -1 SD of the control group mean. Ten of 12 complete GHD children (83.3%) and 13 of 17 partial GHD children (76.5%) had IGFBP-3 concentrations equal or below -1 SD of the control group mean. Conclusion:We conclude that the measurement of IGF-I and IGFBP-3 concentrations might provide essential supplementary data in the diagnostic evaluation of patients with GHD. Our results support the need to use cutoff lines based on below -1 SD of the control. (Korean J Pediatr 2008;51:1329-1335) Purpose:This study aimed to determine the best cutoff line for insulin-like growth factor (IGF)-I and insulin-like growth factor binding protein (IGFBP)-3 to discriminate between growth hormone deficiency (GHD) patients and the control group. Methods:Two hundred thirty subjects with normal controls (129 boys and 101 girls, aged 7-15 years), 14 patients with complete GHD (12 boys and 2 girls), and 17 patients with partial GHD (9 boys and 8 girls) were studied. IGF-I serum concentrations were measured by radioimmunoassay (RI), and IGFBP-3 concentrations were measured by immunoradiometric assay (IRMA). Results:The receiver operating characteristic (ROC) plot analysis showed that the best IGF-I and IGFBP-3 cutoff line was at -1 standard deviation (SD). By comparing IGF-I serum levels of GHD children within 1 SD of normal control, we determined the sensitivity (S) (87.5-100%) and specificity (Sp) (80-84.6%) according to the age group. For IGFBP-3, we determined the following values: S (58.7-85.7%) and Sp (79.2-85.5%). Eleven of 14 patients with complete GHD (78.5%) and 16 of 17 patients with partial GHD (94.1%) had IGF-I concentrations equal to or below -1 SD of the control group mean. Ten of 12 complete GHD children (83.3%) and 13 of 17 partial GHD children (76.5%) had IGFBP-3 concentrations equal or below -1 SD of the control group mean. Conclusion:We conclude that the measurement of IGF-I and IGFBP-3 concentrations might provide essential supplementary data in the diagnostic evaluation of patients with GHD. Our results support the need to use cutoff lines based on below -1 SD of the control. (Korean J Pediatr 2008;51:1329-1335)

폐암 환자와 폐암 세포주를 주입한 Nude Mice에서 종양성장인자의 변화

김상범,박성달,김송명,허방 고신대학교 의과대학 2010 고신대학교 의과대학 학술지 Vol.25 No.1

Background and purpose : The incidence of non-small cell lung cancer has decreased than past decades but the prevalence rate of squamous cell carcinoma is highest among the various types of lung cancer. There have been one of the most difficult problems of the medical areas because of the law rate of operative treatment by micrometastasis still. The released growth regulating factors from the tumor cells have heen researching targets recently. This study is to know the changes of the tumor growth factors by examining the change of transforming growth factor-β1 (TGF-β1) within plasma of nude mouse was measured after each of different quantity of cultured cell of squamous cell carcinoma was injected into plerual caviry of nude mouse in animal experiment, while comparing the values of TGF, Insulin-like growth factor and epidermal growth factor between experimental trial and clinical trial. Method : Between June of 2006 and June of 2007 at Kosin University Hospital, 17 patients were chosen for this study where 13 of them were pathologically diagnosed of squamous cell carcinoma after initial diagnosis of lung cancer followed by radical pneumonectomy and pathologic biopsy, while 4 patients were pathologically diagnosed of benign tumor. Blood sample was collected prior to a surgery from the lung cancer patients scheduled for surgery, and the sample was cryopreserved. During surgery, dense tumor tissue without necrosis was excised in a size of 5mm3 and quickly frozen to be used for experimental tissue, whereas the tissue, which was located farthest from the lesion and deemed free of cancer cell, was to be used for control tissue. And the tissue obtained from benign tumor or normal tissue of granuloma patient was classified as a control tissue B, while the sample from malignant tissue of lung cancer was categorized as experimental tissue M. In animal experiment with 15 male nude mice without immunity, 5 subjects without cancer cell were used for control group, another 5 subjects with of 2×106 of cancer cell line (SW-900 G IV) injected into intrathoracic were used for experimental group I, and the other 5 subjects injected with twice quantity of cancer cell line of group I were classified as experimental group II. After breeding the subjects in the clean room for 8 weeks, blood sample was collected from their heart, and quantitative test of plasma TGF-β was performed. For the test of tumor growth factor, human TGF-β ELISA kit was used for quantitation of TGF-β1&2, and active TM non-eztraction IGF kit was for quantitation of IGF-I&II, while GE healthcare kit was used for EGF. Results : In animal experiment, TGF-β1 was expressed within the serum of all nude mice from control group as well experimental group I and II, where control group was at 28.49 fmol/ml while experimental group I and II were at 32.19 fmol/ml and 42.36 fmol/ml respectively. In clinical trial, TGF-β1 was measured higher from experimental group with 40.9 fmol/ml than from control group with 28.5 fmol/ml, and TGF-β2 was lower from experimental group with 12.3 fmol/ml than from control group with 30.3 fmol/ml, while the difference was insignificant with EGF (p<0.05) where control group and experimental group were at 0.11 ng.ml and 0.12 ng/ml respectively. Moreover, TGF-β1 was at 40.88 fmol/ml from control tissue B and at 15.55 fmol/ml from experimental tissue M, where as TGF-β2 was at 12.31 fmol/ml from control tissue B and 23.95 fmol/ml from experimental tissue M (p<0.05). TGF-β1 within the serum of control group was at 812.4 fmol/ml before surgery and at 989.1 fmol/ml after surgery, which showed a slight amplifiction, while TGF-β2 was at 16.8 fmol/ml before surgery and at 20.2 fmol/ml after surgery, exhibiting a slight increase (p<0.05). Serum IGF-I was at 117.9 ng/ml before surgery and at 214.4 ng/ml after surgery, and IGF-II was at 552.8 ng/ml and 552.8 ng/ml after surgery, while EGF showed insignificant changes between before surgery at 0.67 ng/ml and after surgery at 0.74 ng/ml (p<0.05). In lung cancer stage Ia,b, TGF-β1 was decreased through surgery as it was at 972.94 mol/ml before and 866.92 mol/ml after, and TGF-β2 was also decreased as measured at 42.40 mol/ml before surgery and 38.24 mol/ml after surgery. In stage IIIa,b, TGF-β1 was decreased as measured at 980.31 mol/ml before surgery and at 928.18 mol/ml, while TGF-β2 showed not much changes (p<0.05). In lung cancer stage Ia,b, serum IGF-I was decreased as measured at 112.7 mol/ml before surgery and at 87.92 mol/ml after surgery. and IGF-II was also decreased as measured 468.78 ng/ml before surgery and at 358.59 ng/ml after surgery (p<0.05). In stage III,b, IGF-I showed not much changes, but IGF-II was at 438.31 ng/ml before surgery and 404.2 ng/ml after surgery, where EGF was not all influenced by the phase. Conclusion : In the experiment with nude mice, TGF-β1 was increased as the number of cancer cell was increased after injection of cancer cell. The number of cancer cell from early tumor, such as Ia,b from post-surgery clinical lung cancer patients, was reduced after the surgery, and TGF-β1 was also reduced as the cancer cell was reduced as well. Therefore, it was believed that TGF-β1 can be a feasible prognostic factor for early diagnosis of lung cancer at Ia,b of early phase

Leukocyte-Poor Platelet-Rich Plasma-Derived Growth Factors Enhance Human Fibroblast Proliferation In Vitro

노규철,Xiao Ning Liu,Zhong Zhuan,양철중,김용태,이근우,최경호,김경옥 대한정형외과학회 2018 Clinics in Orthopedic Surgery Vol.10 No.2

Background: Leukocyte-poor platelet-rich plasma (LP-PRP) from peripheral blood is currently used as a concentrated source of growth factors to stimulate repair at sites of soft tissue injury. Fibroblasts are primary mediators of wound healing. Thus, we aimed to assess the positive effect of LP-PRP on human fibroblast proliferation in vitro . Methods: LP-PRP was prepared from 49 donors. The fibroblasts were seeded, and at 24 hours after seeding, 1 × 107/10 μL LP-PRP was added once to each well. The cells were harvested 10 times during study period at our planned points, and we examined cell proliferation using the water-soluble tetrazolium salt-1 assay. We collected the supernatants and measured the amount of growth factors such as platelet-derived growth factor (PDGF)-AB/BB, insulin-like growth factor-1 (IGF-1), transforming growth factor-β1 (TGF-β1), and vascular endothelial growth factor (VEGF), which are known to be involved in wound healing processes, by multiplex assay. Results: Human fibroblasts treated with LP-PRP showed a significant increase in proliferation when compared to untreated controls (p < 0.001 at days 4, 6, and 8). Multiplex cytokine assays revealed various secretion patterns. PDGF-AB/BB appeared at early time points and peaked before fibroblast proliferation. IGF-1 and TGF-β1 secretion gradually increased and peaked on days 4 and 6 post-treatment. The early VEGF concentration was lower than the concentration of other growth factors but increased along with cell proliferation. Conclusions: Platelets in LP-PRP release growth factors such as PDGF, IGF-1, TGF-β1 and VEGF, and these growth factors have a promoting effect for human fibroblast proliferation, one of the important mediators of wound healing. These results suggest that growth factors derived from LP-PRP enhance the proliferation of human fibroblast.