Man-Il Huh - Academia.edu (original) (raw)

Papers by Man-Il Huh

Journal of Cellular Biochemistry, 2007

Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Wester... more Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Western blotting, and RT-PCR analysis to investigate the roles of matrix metalloproteinases (MMPs) and the related ''a disintegrin and metalloproteinase'' (ADAM) family proteinases in chick corneal development. While MMP-13 was expressed in developing chick corneas from embryonic day (ED) 5 to ED 10, its inhibitor, tissue inhibitors of metalloproteinase-1 (TIMP-1), was expressed from ED 18 to 2 days post-hatching (P2). Early MMP-13 activity may be associated with degradation of type IX collagen from the primary stroma, which loosens the collagen fibrils and facilitates neural crest (NC) cell migration. The membrane-bound and secreted forms of ADAM10 were both detected throughout corneal development, and active ADAM10 formed a cleavage complex with CD44v6, a CD44 splice variant that is a major cell surface adhesion molecule for hyaluronic acid (HA) and has been implicated in cell migration. Both CD44v6 and its ectodomain cleavage products were detected from ED 5 to ED 14, and a broad-spectrum MMP inhibitor blocked ectodomain cleavage in cultured stromal cells. These findings suggest that ADAM10 mediates CD44v6 cleavage in the developing cornea, facilitating NC cellderived mesenchymal cell migration. Finally, we identified high levels of active membrane-type 3-MMP (MT3-MMP) in developing corneas at ED 7, ED 14, and ED 18. MT3-MMP takes part in MMP-2 activation and possibly also CD44v6 shedding, suggesting that this pathway may be involved in cell migration. These findings collectively show for the first time that multiple MMPs, ADAMs, and TIMPs appear to functionally interact during corneal development.

Acta Histochemica, 2013

Although several matrix metalloproteinases (MMPs) have been implicated in testis development, the... more Although several matrix metalloproteinases (MMPs) have been implicated in testis development, the presence of MMP-13 protein has not been directly substantiated in the male avian gonads. In this study, we examined the expression patterns of MMP-13 and MMP inhibitors, TIMP-1 and TIMP-2, in immature (4weeks), pre-pubertal (16weeks), and mature (1year) chicken testes. Using RT-PCR analysis, we observed that MMP-13 mRNA was expressed in immature testis. In Western blot analysis, the expression level of MMP-13 protein peaked in the immature testes during marked tissue remodeling, whereas it gradually decreased during testis maturation. High expression levels of TIMP-1 (34-kDa) and TIMP-2 (55-kDa) were detected only in immature and pre-pubertal testes and not in adult testis. Four different forms of TIMP-2 protein were differentially detected in the testes of growing and adult chicken. Using immunohistochemistry we localized both secreted and intracellular forms of MMP-13, TIMP-1, and TIMP-2 proteins. These proteins were temporally and spatially distributed in growing and adult testes, and all their expression levels were similar to the expression profile of Western blot results. These findings suggest that age-related changes of MMP-13 with balance of TIMPs act in concert to effect the controlled testicular remodeling and maturation.

Evidence-based complementary and alternative medicine : eCAM, 2016

Inhibition of osteoclast differentiation and bone resorption is a therapeutic strategy for the ma... more Inhibition of osteoclast differentiation and bone resorption is a therapeutic strategy for the management of postmenopausal bone loss. This study investigated the effects of Rhus javanica (R. javanica) extracts on bone marrow cultures to develop agents from natural sources that may prevent osteoclastogenesis. Extracts of R. javanica (eGr) cocoons spun by Rhus javanica (Bell.) Baker inhibited the osteoclast differentiation and bone resorption. The effects of aqueous extract (aeGr) or 100% ethanolic extract (eeGr) on ovariectomy- (OVX-) induced bone loss were investigated by various biochemical assays. Furthermore, microcomputed tomography (µCT) was performed to study bone remodeling. Oral administration of eGr (30 mg or 100 mg/kg/day for 6 weeks) augmented the inhibition of femoral bone mineral density (BMD), bone mineral content (BMC), and other factors involved in bone remodeling when compared to OVX controls. Additionally, eGr slightly decreased bone turnover markers that were inc...

Tissue Engineering and Regenerative Medicine, 2016

Investigative Opthalmology & Visual Science, 2015

The current study was done to determine the role of lipocalin-2 (LCN2) in the pathogenesis of dem... more The current study was done to determine the role of lipocalin-2 (LCN2) in the pathogenesis of demyelinating optic neuritis using an experimental autoimmune optic neuritis (EAON) model. The EAON was induced by subcutaneous immunization with an emulsified mixture of myelin oligodendrocyte glycoprotein (MOG35-55) peptide in mice. The LCN2 expression was examined in the optic nerve after MOG peptide injection. Degree of demyelination, inflammatory infiltration, glial activation, and expression profile of inflammatory mediators in the optic nerve were compared between LCN2 knockout (KO) animals and wild-type littermates by histological analysis and real-time PCR following EAON induction. Plasma levels of LCN2 in patients with optic neuritis were measured by ELISA. The expression of LCN2 was notably increased in the optic nerve after EAON induction. Expression of LCN2 was colocalized with reactive astrocytes. A significant reduction of demyelination, inflammatory infiltration, and gliosis was demonstrated in the optic nerve of LCN2 KO mice. The LCN2 KO mice also showed markedly reduced gene expression associated with the M1-polarized glia phenotype and toll-like receptor signaling in the optic nerve. The LCN2 levels in plasma were significantly higher in optic neuritis patients (71.6 ± 10.6 ng/mL) compared to healthy controls (37.4 ± 9.1 ng/mL, P = 0.0284). In this study, we demonstrated a significant induction of LCN2 expression in astrocytes of the optic nerve following EAON induction. Our results imply that astrocyte-derived LCN2 may have a pivotal role in the development of demyelinating optic neuritis, and LCN2 can be a therapeutic target to alleviate immune and inflammatory damage in the optic nerve.

Tissue Engineering and Regenerative Medicine, 2014

ABSTRACT The purpose of this study is to evaluate the effect of conditioned media (CM) collected ... more ABSTRACT The purpose of this study is to evaluate the effect of conditioned media (CM) collected from human amniotic fluid-derived stem cells (hAFSCs) on in vitro wound healing and on expression change of MMP-1 and procollagen 1A by ultra-violet A (UVA) irradiation. The conditioned media (CM) were collected from hAFSCs cultured with DMEM/F-12 serum free media. AFSC-CM were tested using in vitro wound healing model, and the results demonstrated that AFSC-CM facilitated cell proliferation in the dermal fibroblast (132.72 ± 7.48%) and skin epidermal cell (125.95 ± 4.61%), and accelerated cell migration after wound was generated. In order to evaluate the effect of AFSC-CM treatment on gene expressions changed by UVA irradiation, AFSC-CM and CM collected from culture of epidermal cell after UVA irradiation were used in dermal fibroblast culture. qRT-PCR data indicated that upregulated MMP-1 expression by UVA was down-regulated and down-regulated procollagen 1A expression was recovered by treatment with AFSC-CM. Collectively, the results suggest that hAFSC-CM has a potential to improve skin regeneration which was damaged by photo-aging.

Proceedings of the National Academy of Sciences, 2014

The mechanism by which the 8q24 MYC enhancer region, including cancer-associated variant rs698326... more The mechanism by which the 8q24 MYC enhancer region, including cancer-associated variant rs6983267, increases cancer risk is unknown due to the lack of protein-coding genes at 8q24.21. Here we report the identification of long noncoding RNAs named cancer-associated region long noncoding RNAs (CARLos) in the 8q24 region. The expression of one of the long noncoding RNAs, CARLo-5, is significantly correlated with the rs6983267 allele associated with increased cancer susceptibility. We also found the MYC enhancer region physically interacts with the active regulatory region of the CARLo-5 promoter, suggesting long-range interaction of MYC enhancer with the CARLo-5 promoter regulates CARLo-5 expression. Finally, we demonstrate that CARLo-5 has a function in cell-cycle regulation and tumor development. Overall, our data provide a key of the mystery of the 8q24 gene desert.

NeuroImage, 2012

In the present study, we report a new method of manganese enhanced magnetic resonance imaging (ME... more In the present study, we report a new method of manganese enhanced magnetic resonance imaging (MEMRI) using intratympanic (IT) manganese administration. We explore Mn 2+ uptake from the middle ear cavity into the cochlea through mechanically gated ion channels of the hair cell and also functional auditory tract tracing without the use of excessive auditory stimuli for a long time period outside the scanner. After manganese administration in animals with normal hearing and unilateral deafness, T1-weighted MR images were obtained for up to 48 h with a 3.0 T MR imager. In normal rats, the mean signal-to-noise ratio (SNR) at each region of interest on the auditory pathway was significantly higher in the IT injection group than in the intraperitoneal (IP) injection group (P b 0.05). Furthermore, the cochlea showed Mn 2+ signal enhancement only in the IT injection group. In unilateral deafness rats, the IT injection of Mn 2+ into the deaf-side middle ear cavity demonstrated signal enhancement in the cochlea but not in other auditory structures without axonal transport of Mn 2+ along the auditory pathway. On the other hand, the IT injection of Mn 2+ into the normal-side middle ear cavity demonstrated that the mean SNRs at the cochlea, cochlear nucleus, superior olivary complex, lateral lemniscus and inferior colliculus were significantly higher in the ipsilateral auditory pathway than in the contralateral pathway (P b 0.05). For the IP injection group, the mean SNRs at each auditory structure, except the cochlea, increased bilaterally. In conclusion, the present work demonstrated the potential advantages of a new IT MEMRI over conventional systemic injection strategies in that (i) the functional auditory tract tracing initiated by the hair cell function is possible and (ii) the axonal transport of Mn 2+ ions by trans-synaptic activity is possible without auditory stimulation for a long time period outside MR scanner.

Journal of Materials Chemistry, 2010

The work is directed toward the synthesis and characterization of gold nanoparticles coated by Gd... more The work is directed toward the synthesis and characterization of gold nanoparticles coated by Gd-chelate (Au@ GdL), where L is a conjugate of diethylenetriamine-N, N, N′, N′′, N′′-pentaacetic acid (DTPA) and penicillamine. Au@ GdL is obtained by the ...

Journal of Cellular Biochemistry, 2007

Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Wester... more Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Western blotting, and RT-PCR analysis to investigate the roles of matrix metalloproteinases (MMPs) and the related ''a disintegrin and metalloproteinase'' (ADAM) family proteinases in chick corneal development. While MMP-13 was expressed in developing chick corneas from embryonic day (ED) 5 to ED 10, its inhibitor, tissue inhibitors of metalloproteinase-1 (TIMP-1), was expressed from ED 18 to 2 days post-hatching (P2). Early MMP-13 activity may be associated with degradation of type IX collagen from the primary stroma, which loosens the collagen fibrils and facilitates neural crest (NC) cell migration. The membrane-bound and secreted forms of ADAM10 were both detected throughout corneal development, and active ADAM10 formed a cleavage complex with CD44v6, a CD44 splice variant that is a major cell surface adhesion molecule for hyaluronic acid (HA) and has been implicated in cell migration. Both CD44v6 and its ectodomain cleavage products were detected from ED 5 to ED 14, and a broad-spectrum MMP inhibitor blocked ectodomain cleavage in cultured stromal cells. These findings suggest that ADAM10 mediates CD44v6 cleavage in the developing cornea, facilitating NC cellderived mesenchymal cell migration. Finally, we identified high levels of active membrane-type 3-MMP (MT3-MMP) in developing corneas at ED 7, ED 14, and ED 18. MT3-MMP takes part in MMP-2 activation and possibly also CD44v6 shedding, suggesting that this pathway may be involved in cell migration. These findings collectively show for the first time that multiple MMPs, ADAMs, and TIMPs appear to functionally interact during corneal development.

Journal of Cellular Biochemistry, 2009

In this study, temporal and spatial distribution of three TGF-b isoforms and their downstream sig... more In this study, temporal and spatial distribution of three TGF-b isoforms and their downstream signaling pathways including pSmad2 and p38MAPK were examined during fibrotic wound repair. In normal chick corneas, TGF-b1, -2, and -3 were weakly detected in Bowman's layer (BL). In healing corneas, TGF-b1 was primarily deposited in the fibrin clot and the unwounded BL. TGF-b2 was highly expressed in healing epithelial and endothelial cells, and numerous active fibroblasts/myofibroblasts. TGF-b3 was mainly detected in the unwound region of basal epithelial cells. a-Smooth muscle actin (a-SMA) was initially appeared in the posterior region of repairing stroma at day 3, and was detected in the entire healing stroma by day 7. Notably, a-SMA was absent in the central region of healing stroma by day 14, and its staining pattern was similar to those of TGF-b2 and p38MAPK. By contrast, pSmad2 was mainly detected in the fibroblasts. In normal cornea, laminin was mainly detected in both epithelial basement membrane (BM) and Descemet's membrane (DM). By contrast to reconstitution of the BM in the wound region, the DM was not repaired although endothelial layer was regenerated, indicating that high levels of TGF-b2 were released into the posterior region of healing stroma on day 14. High levels of a-SMA staining, shown in cultured repair stromal cells from healing corneas on day 14 and in TGF-b2 treated normal stromal cells, were significantly reduced by p38MAPK inhibition. Collectively, this study suggests that TGF-b2-mediated myofibroblast transformation is mediated, at least partly, by the p38MAPK pathway in vivo.

Journal of Cellular Biochemistry, 2013

Here, we examined the role of ADAM10 during retinal cell differentiation in retinal sections and ... more Here, we examined the role of ADAM10 during retinal cell differentiation in retinal sections and in vitro cultures of developing chick retinal cells from embryonic day 6 (ED6). Immunohistochemistry showed that ADAM10 is abundantly expressed in the inner zone of neuroblastic layer at ED5, and it becomes more highly expressed in the ganglion cell layer at ED7 and ED9. Western blotting confirmed that ADAM10 was expressed as an inactive pro-form that was processed to a shorter, active form in control cultured cells, but in cultures treated with an ADAM10 inhibitor (GI254023X) and ADAM10-specific siRNA, the level of mature ADAM10 decreased. Phase-contrast microscopy showed that long neurite extensions were present in untreated cultures 24 h after plating, whereas cultures treated with GI254023X showed significant decreases in neurite extension. Immunofluorescence staining revealed that there were far fewer differentiated ganglion cells in ADAM10 siRNA and GI254023X-treated cultures compared to controls, whereas the photoreceptor cells were unaltered. The Pax6 protein was more strongly detected in the differentiated ganglion cells of control cultures compared to ADAM10 siRNA and GI254023X-treated cultures. N-cadherin ectodomain shedding was apparent in control cultures after 24 h, when ganglion cell differentiation was observed, but ADAM10 siRNA and GI254023X treatment inhibited these processes. In contrast, N-cadherin staining was strongly detected in photoreceptor cells regardless of ADAM10 siRNA and GI254023X treatment. Taken together, these data indicate that the inhibition of ADAM10 can inhibit Pax6 expression and N-cadherin ectodomain shedding in retinal cells, possibly affecting neurite outgrowth and ganglion cell differentiation.

Biomaterials, 2011

Biomedical applications of magnetic nanoparticles (MNP), including superparamagnetic nanoparticle... more Biomedical applications of magnetic nanoparticles (MNP), including superparamagnetic nanoparticles, have expanded dramatically in recent years. Systematic and standardized cytotoxicity assessment to ensure the biosafety and biocompatibility of those applications is compulsory. We investigated whether exposure to static magnetic field (SMF) from e.g. magnetic resonance imaging (MRI) could affect the cytotoxicity of superparamagnetic iron oxide (SPIO) nanoparticles using mouse hepatocytes and ferucarbotran, a liver-selective MRI contrast agent as a model system. We show that while the SPIO satisfied the conventional cytotoxicity assessment, clinical doses combined with SMF exposure exerts synergistic adverse effects such as reduced cell viability, apoptosis, and cell cycle aberrations on hepatocytes in vitro and in vivo. Concomitant treatments with the SPIO and SMF generated SPIO aggregates, which demonstrated enhanced cellular uptake, was sufficient to induce the cytotoxicity without further SMF, emphasizing that the SPIO aggregates were the predominant source of the cytotoxicity. Interestingly, the apoptotic effect was dependent on levels of reactive oxygen species (ROS) and SPIO uptake while the reduced cell viability was independent of these factors. Moreover, long-term monitoring showed a significant increase in multinuclear giant cells in the cells concomitantly treated with the SPIO and SMF compared with the control. The results demonstrate that the SPIO produces unidentified cytotoxicity on liver in the presence of SMF and the SPIO aggregates predominantly exert the effect. Since aggregation of MNP in biological milieu in the presence of strong SMF is inevitable, a fundamentally different approach to surface fabrication is essential to increase the biocompatibility of MNP.

Journal of Cellular Biochemistry, 2007

Collagenase-1 is a protease expressed by active fibroblasts that is involved in remodeling of the... more Collagenase-1 is a protease expressed by active fibroblasts that is involved in remodeling of the extracellular matrix (ECM). In this study, we characterize the intracellular signaling mechanism of collagenase-1 production by IL-1a in subcultured normal fibroblasts (NF) from uninjured normal corneas, compared to that in repair wound fibroblasts (WF). In NF, collagenase-1 was induced specifically after the exogenous addition of IL-1a via activation of ERK and p38MAPK. Collagenase-1 expression was strongly suppressed upon treatment with either a MEK or p38MAPK inhibitor. In contrast, repair WF constitutively synthesized both IL-1a and collagenase-1. Combined treatment with both mitogen-activated protein kinase (MAPK) inhibitors dramatically reduced collagenase-1 synthesis, while individual MEK1 or p38 inhibitors weakly modulated the collagenase-1 level. The results indicate that both pathways are crucial in the regulation of collagenase-1 synthesis. Furthermore, an IL-1a receptor antagonist (IL-1ra) could not abolish constitutive collagenase-1 synthesis, even at high doses, suggesting that other cytokines/factors are additionally involved in this process. We propose that induction of collagenase-1 by IL-1a in both WF and NF depends on a unique combination of cell type-specific signaling pathways.

Journal of Cellular Biochemistry, 2007

Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Wester... more Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Western blotting, and RT-PCR analysis to investigate the roles of matrix metalloproteinases (MMPs) and the related ''a disintegrin and metalloproteinase'' (ADAM) family proteinases in chick corneal development. While MMP-13 was expressed in developing chick corneas from embryonic day (ED) 5 to ED 10, its inhibitor, tissue inhibitors of metalloproteinase-1 (TIMP-1), was expressed from ED 18 to 2 days post-hatching (P2). Early MMP-13 activity may be associated with degradation of type IX collagen from the primary stroma, which loosens the collagen fibrils and facilitates neural crest (NC) cell migration. The membrane-bound and secreted forms of ADAM10 were both detected throughout corneal development, and active ADAM10 formed a cleavage complex with CD44v6, a CD44 splice variant that is a major cell surface adhesion molecule for hyaluronic acid (HA) and has been implicated in cell migration. Both CD44v6 and its ectodomain cleavage products were detected from ED 5 to ED 14, and a broad-spectrum MMP inhibitor blocked ectodomain cleavage in cultured stromal cells. These findings suggest that ADAM10 mediates CD44v6 cleavage in the developing cornea, facilitating NC cellderived mesenchymal cell migration. Finally, we identified high levels of active membrane-type 3-MMP (MT3-MMP) in developing corneas at ED 7, ED 14, and ED 18. MT3-MMP takes part in MMP-2 activation and possibly also CD44v6 shedding, suggesting that this pathway may be involved in cell migration. These findings collectively show for the first time that multiple MMPs, ADAMs, and TIMPs appear to functionally interact during corneal development.

Acta Histochemica, 2013

Although several matrix metalloproteinases (MMPs) have been implicated in testis development, the... more Although several matrix metalloproteinases (MMPs) have been implicated in testis development, the presence of MMP-13 protein has not been directly substantiated in the male avian gonads. In this study, we examined the expression patterns of MMP-13 and MMP inhibitors, TIMP-1 and TIMP-2, in immature (4weeks), pre-pubertal (16weeks), and mature (1year) chicken testes. Using RT-PCR analysis, we observed that MMP-13 mRNA was expressed in immature testis. In Western blot analysis, the expression level of MMP-13 protein peaked in the immature testes during marked tissue remodeling, whereas it gradually decreased during testis maturation. High expression levels of TIMP-1 (34-kDa) and TIMP-2 (55-kDa) were detected only in immature and pre-pubertal testes and not in adult testis. Four different forms of TIMP-2 protein were differentially detected in the testes of growing and adult chicken. Using immunohistochemistry we localized both secreted and intracellular forms of MMP-13, TIMP-1, and TIMP-2 proteins. These proteins were temporally and spatially distributed in growing and adult testes, and all their expression levels were similar to the expression profile of Western blot results. These findings suggest that age-related changes of MMP-13 with balance of TIMPs act in concert to effect the controlled testicular remodeling and maturation.

Evidence-based complementary and alternative medicine : eCAM, 2016

Inhibition of osteoclast differentiation and bone resorption is a therapeutic strategy for the ma... more Inhibition of osteoclast differentiation and bone resorption is a therapeutic strategy for the management of postmenopausal bone loss. This study investigated the effects of Rhus javanica (R. javanica) extracts on bone marrow cultures to develop agents from natural sources that may prevent osteoclastogenesis. Extracts of R. javanica (eGr) cocoons spun by Rhus javanica (Bell.) Baker inhibited the osteoclast differentiation and bone resorption. The effects of aqueous extract (aeGr) or 100% ethanolic extract (eeGr) on ovariectomy- (OVX-) induced bone loss were investigated by various biochemical assays. Furthermore, microcomputed tomography (µCT) was performed to study bone remodeling. Oral administration of eGr (30 mg or 100 mg/kg/day for 6 weeks) augmented the inhibition of femoral bone mineral density (BMD), bone mineral content (BMC), and other factors involved in bone remodeling when compared to OVX controls. Additionally, eGr slightly decreased bone turnover markers that were inc...

Tissue Engineering and Regenerative Medicine, 2016

Investigative Opthalmology & Visual Science, 2015

The current study was done to determine the role of lipocalin-2 (LCN2) in the pathogenesis of dem... more The current study was done to determine the role of lipocalin-2 (LCN2) in the pathogenesis of demyelinating optic neuritis using an experimental autoimmune optic neuritis (EAON) model. The EAON was induced by subcutaneous immunization with an emulsified mixture of myelin oligodendrocyte glycoprotein (MOG35-55) peptide in mice. The LCN2 expression was examined in the optic nerve after MOG peptide injection. Degree of demyelination, inflammatory infiltration, glial activation, and expression profile of inflammatory mediators in the optic nerve were compared between LCN2 knockout (KO) animals and wild-type littermates by histological analysis and real-time PCR following EAON induction. Plasma levels of LCN2 in patients with optic neuritis were measured by ELISA. The expression of LCN2 was notably increased in the optic nerve after EAON induction. Expression of LCN2 was colocalized with reactive astrocytes. A significant reduction of demyelination, inflammatory infiltration, and gliosis was demonstrated in the optic nerve of LCN2 KO mice. The LCN2 KO mice also showed markedly reduced gene expression associated with the M1-polarized glia phenotype and toll-like receptor signaling in the optic nerve. The LCN2 levels in plasma were significantly higher in optic neuritis patients (71.6 ± 10.6 ng/mL) compared to healthy controls (37.4 ± 9.1 ng/mL, P = 0.0284). In this study, we demonstrated a significant induction of LCN2 expression in astrocytes of the optic nerve following EAON induction. Our results imply that astrocyte-derived LCN2 may have a pivotal role in the development of demyelinating optic neuritis, and LCN2 can be a therapeutic target to alleviate immune and inflammatory damage in the optic nerve.

Tissue Engineering and Regenerative Medicine, 2014

ABSTRACT The purpose of this study is to evaluate the effect of conditioned media (CM) collected ... more ABSTRACT The purpose of this study is to evaluate the effect of conditioned media (CM) collected from human amniotic fluid-derived stem cells (hAFSCs) on in vitro wound healing and on expression change of MMP-1 and procollagen 1A by ultra-violet A (UVA) irradiation. The conditioned media (CM) were collected from hAFSCs cultured with DMEM/F-12 serum free media. AFSC-CM were tested using in vitro wound healing model, and the results demonstrated that AFSC-CM facilitated cell proliferation in the dermal fibroblast (132.72 ± 7.48%) and skin epidermal cell (125.95 ± 4.61%), and accelerated cell migration after wound was generated. In order to evaluate the effect of AFSC-CM treatment on gene expressions changed by UVA irradiation, AFSC-CM and CM collected from culture of epidermal cell after UVA irradiation were used in dermal fibroblast culture. qRT-PCR data indicated that upregulated MMP-1 expression by UVA was down-regulated and down-regulated procollagen 1A expression was recovered by treatment with AFSC-CM. Collectively, the results suggest that hAFSC-CM has a potential to improve skin regeneration which was damaged by photo-aging.

Proceedings of the National Academy of Sciences, 2014

The mechanism by which the 8q24 MYC enhancer region, including cancer-associated variant rs698326... more The mechanism by which the 8q24 MYC enhancer region, including cancer-associated variant rs6983267, increases cancer risk is unknown due to the lack of protein-coding genes at 8q24.21. Here we report the identification of long noncoding RNAs named cancer-associated region long noncoding RNAs (CARLos) in the 8q24 region. The expression of one of the long noncoding RNAs, CARLo-5, is significantly correlated with the rs6983267 allele associated with increased cancer susceptibility. We also found the MYC enhancer region physically interacts with the active regulatory region of the CARLo-5 promoter, suggesting long-range interaction of MYC enhancer with the CARLo-5 promoter regulates CARLo-5 expression. Finally, we demonstrate that CARLo-5 has a function in cell-cycle regulation and tumor development. Overall, our data provide a key of the mystery of the 8q24 gene desert.

NeuroImage, 2012

In the present study, we report a new method of manganese enhanced magnetic resonance imaging (ME... more In the present study, we report a new method of manganese enhanced magnetic resonance imaging (MEMRI) using intratympanic (IT) manganese administration. We explore Mn 2+ uptake from the middle ear cavity into the cochlea through mechanically gated ion channels of the hair cell and also functional auditory tract tracing without the use of excessive auditory stimuli for a long time period outside the scanner. After manganese administration in animals with normal hearing and unilateral deafness, T1-weighted MR images were obtained for up to 48 h with a 3.0 T MR imager. In normal rats, the mean signal-to-noise ratio (SNR) at each region of interest on the auditory pathway was significantly higher in the IT injection group than in the intraperitoneal (IP) injection group (P b 0.05). Furthermore, the cochlea showed Mn 2+ signal enhancement only in the IT injection group. In unilateral deafness rats, the IT injection of Mn 2+ into the deaf-side middle ear cavity demonstrated signal enhancement in the cochlea but not in other auditory structures without axonal transport of Mn 2+ along the auditory pathway. On the other hand, the IT injection of Mn 2+ into the normal-side middle ear cavity demonstrated that the mean SNRs at the cochlea, cochlear nucleus, superior olivary complex, lateral lemniscus and inferior colliculus were significantly higher in the ipsilateral auditory pathway than in the contralateral pathway (P b 0.05). For the IP injection group, the mean SNRs at each auditory structure, except the cochlea, increased bilaterally. In conclusion, the present work demonstrated the potential advantages of a new IT MEMRI over conventional systemic injection strategies in that (i) the functional auditory tract tracing initiated by the hair cell function is possible and (ii) the axonal transport of Mn 2+ ions by trans-synaptic activity is possible without auditory stimulation for a long time period outside MR scanner.

Journal of Materials Chemistry, 2010

The work is directed toward the synthesis and characterization of gold nanoparticles coated by Gd... more The work is directed toward the synthesis and characterization of gold nanoparticles coated by Gd-chelate (Au@ GdL), where L is a conjugate of diethylenetriamine-N, N, N′, N′′, N′′-pentaacetic acid (DTPA) and penicillamine. Au@ GdL is obtained by the ...

Journal of Cellular Biochemistry, 2007

Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Wester... more Tissue remodeling is central to embryonic development. Here, we used immunohistochemistry, Western blotting, and RT-PCR analysis to investigate the roles of matrix metalloproteinases (MMPs) and the related ''a disintegrin and metalloproteinase'' (ADAM) family proteinases in chick corneal development. While MMP-13 was expressed in developing chick corneas from embryonic day (ED) 5 to ED 10, its inhibitor, tissue inhibitors of metalloproteinase-1 (TIMP-1), was expressed from ED 18 to 2 days post-hatching (P2). Early MMP-13 activity may be associated with degradation of type IX collagen from the primary stroma, which loosens the collagen fibrils and facilitates neural crest (NC) cell migration. The membrane-bound and secreted forms of ADAM10 were both detected throughout corneal development, and active ADAM10 formed a cleavage complex with CD44v6, a CD44 splice variant that is a major cell surface adhesion molecule for hyaluronic acid (HA) and has been implicated in cell migration. Both CD44v6 and its ectodomain cleavage products were detected from ED 5 to ED 14, and a broad-spectrum MMP inhibitor blocked ectodomain cleavage in cultured stromal cells. These findings suggest that ADAM10 mediates CD44v6 cleavage in the developing cornea, facilitating NC cellderived mesenchymal cell migration. Finally, we identified high levels of active membrane-type 3-MMP (MT3-MMP) in developing corneas at ED 7, ED 14, and ED 18. MT3-MMP takes part in MMP-2 activation and possibly also CD44v6 shedding, suggesting that this pathway may be involved in cell migration. These findings collectively show for the first time that multiple MMPs, ADAMs, and TIMPs appear to functionally interact during corneal development.

Journal of Cellular Biochemistry, 2009

In this study, temporal and spatial distribution of three TGF-b isoforms and their downstream sig... more In this study, temporal and spatial distribution of three TGF-b isoforms and their downstream signaling pathways including pSmad2 and p38MAPK were examined during fibrotic wound repair. In normal chick corneas, TGF-b1, -2, and -3 were weakly detected in Bowman's layer (BL). In healing corneas, TGF-b1 was primarily deposited in the fibrin clot and the unwounded BL. TGF-b2 was highly expressed in healing epithelial and endothelial cells, and numerous active fibroblasts/myofibroblasts. TGF-b3 was mainly detected in the unwound region of basal epithelial cells. a-Smooth muscle actin (a-SMA) was initially appeared in the posterior region of repairing stroma at day 3, and was detected in the entire healing stroma by day 7. Notably, a-SMA was absent in the central region of healing stroma by day 14, and its staining pattern was similar to those of TGF-b2 and p38MAPK. By contrast, pSmad2 was mainly detected in the fibroblasts. In normal cornea, laminin was mainly detected in both epithelial basement membrane (BM) and Descemet's membrane (DM). By contrast to reconstitution of the BM in the wound region, the DM was not repaired although endothelial layer was regenerated, indicating that high levels of TGF-b2 were released into the posterior region of healing stroma on day 14. High levels of a-SMA staining, shown in cultured repair stromal cells from healing corneas on day 14 and in TGF-b2 treated normal stromal cells, were significantly reduced by p38MAPK inhibition. Collectively, this study suggests that TGF-b2-mediated myofibroblast transformation is mediated, at least partly, by the p38MAPK pathway in vivo.

Journal of Cellular Biochemistry, 2013

Here, we examined the role of ADAM10 during retinal cell differentiation in retinal sections and ... more Here, we examined the role of ADAM10 during retinal cell differentiation in retinal sections and in vitro cultures of developing chick retinal cells from embryonic day 6 (ED6). Immunohistochemistry showed that ADAM10 is abundantly expressed in the inner zone of neuroblastic layer at ED5, and it becomes more highly expressed in the ganglion cell layer at ED7 and ED9. Western blotting confirmed that ADAM10 was expressed as an inactive pro-form that was processed to a shorter, active form in control cultured cells, but in cultures treated with an ADAM10 inhibitor (GI254023X) and ADAM10-specific siRNA, the level of mature ADAM10 decreased. Phase-contrast microscopy showed that long neurite extensions were present in untreated cultures 24 h after plating, whereas cultures treated with GI254023X showed significant decreases in neurite extension. Immunofluorescence staining revealed that there were far fewer differentiated ganglion cells in ADAM10 siRNA and GI254023X-treated cultures compared to controls, whereas the photoreceptor cells were unaltered. The Pax6 protein was more strongly detected in the differentiated ganglion cells of control cultures compared to ADAM10 siRNA and GI254023X-treated cultures. N-cadherin ectodomain shedding was apparent in control cultures after 24 h, when ganglion cell differentiation was observed, but ADAM10 siRNA and GI254023X treatment inhibited these processes. In contrast, N-cadherin staining was strongly detected in photoreceptor cells regardless of ADAM10 siRNA and GI254023X treatment. Taken together, these data indicate that the inhibition of ADAM10 can inhibit Pax6 expression and N-cadherin ectodomain shedding in retinal cells, possibly affecting neurite outgrowth and ganglion cell differentiation.

Biomaterials, 2011

Biomedical applications of magnetic nanoparticles (MNP), including superparamagnetic nanoparticle... more Biomedical applications of magnetic nanoparticles (MNP), including superparamagnetic nanoparticles, have expanded dramatically in recent years. Systematic and standardized cytotoxicity assessment to ensure the biosafety and biocompatibility of those applications is compulsory. We investigated whether exposure to static magnetic field (SMF) from e.g. magnetic resonance imaging (MRI) could affect the cytotoxicity of superparamagnetic iron oxide (SPIO) nanoparticles using mouse hepatocytes and ferucarbotran, a liver-selective MRI contrast agent as a model system. We show that while the SPIO satisfied the conventional cytotoxicity assessment, clinical doses combined with SMF exposure exerts synergistic adverse effects such as reduced cell viability, apoptosis, and cell cycle aberrations on hepatocytes in vitro and in vivo. Concomitant treatments with the SPIO and SMF generated SPIO aggregates, which demonstrated enhanced cellular uptake, was sufficient to induce the cytotoxicity without further SMF, emphasizing that the SPIO aggregates were the predominant source of the cytotoxicity. Interestingly, the apoptotic effect was dependent on levels of reactive oxygen species (ROS) and SPIO uptake while the reduced cell viability was independent of these factors. Moreover, long-term monitoring showed a significant increase in multinuclear giant cells in the cells concomitantly treated with the SPIO and SMF compared with the control. The results demonstrate that the SPIO produces unidentified cytotoxicity on liver in the presence of SMF and the SPIO aggregates predominantly exert the effect. Since aggregation of MNP in biological milieu in the presence of strong SMF is inevitable, a fundamentally different approach to surface fabrication is essential to increase the biocompatibility of MNP.

Journal of Cellular Biochemistry, 2007

Collagenase-1 is a protease expressed by active fibroblasts that is involved in remodeling of the... more Collagenase-1 is a protease expressed by active fibroblasts that is involved in remodeling of the extracellular matrix (ECM). In this study, we characterize the intracellular signaling mechanism of collagenase-1 production by IL-1a in subcultured normal fibroblasts (NF) from uninjured normal corneas, compared to that in repair wound fibroblasts (WF). In NF, collagenase-1 was induced specifically after the exogenous addition of IL-1a via activation of ERK and p38MAPK. Collagenase-1 expression was strongly suppressed upon treatment with either a MEK or p38MAPK inhibitor. In contrast, repair WF constitutively synthesized both IL-1a and collagenase-1. Combined treatment with both mitogen-activated protein kinase (MAPK) inhibitors dramatically reduced collagenase-1 synthesis, while individual MEK1 or p38 inhibitors weakly modulated the collagenase-1 level. The results indicate that both pathways are crucial in the regulation of collagenase-1 synthesis. Furthermore, an IL-1a receptor antagonist (IL-1ra) could not abolish constitutive collagenase-1 synthesis, even at high doses, suggesting that other cytokines/factors are additionally involved in this process. We propose that induction of collagenase-1 by IL-1a in both WF and NF depends on a unique combination of cell type-specific signaling pathways.