Efficacy of Gene Modification in Placenta-Derived Mesenchymal Stem Cells Based on Nonviral Electroporation

Mesenchymal stem cell (MSC)-based therapy using gene delivery systems has been suggested for degenerative diseases. Although MSC-based clinical applications are effective and safe, the mode of action remains unclear. Researchers have commonly applied viral-based gene modification because this system has efficient vehicles. While viral transfection carries many risks, such as oncogenes and chromosomal integration, nonviral gene delivery techniques are less expensive, easier to handle, and safe, although they are less efficient. The electroporation method, which uses Nucleofection technology, provides critical opportunities for hard-to-transfect primary cell lines, including MSCs. Therefore, to improve the therapeutic efficacy using genetically modified MSCs, researchers must determine the optimal conditions for the introduction of the Nucleofection technique in MSCs. Here, we suggest optimal methods for gene modification in PD-MSCs using an electroporation gene delivery system for clinical application.

Upregulation of C-Reactive Protein by Placenta-Derived Mesenchymal Stem Cells Promotes Angiogenesis in A Rat Model with Cirrhotic Liver

Background and Objectives@#Liver cirrhosis is accompanied by abnormal vascular shunts. The Wnt pathway is essential for endothelial cell survival and proliferation. C-reactive protein (CRP), which is produced by hepatocyte, activates angiogenesis in cardiovascular diseases. @*Methods@#and Results: The expression of CRP in CCl 4 -injured rat livers was detected using qRT-PCR and Western blotting after transplantation of placenta-derived mesenchymal stem cells (PD-MSCs) into rats. To determine whether CRP functions in hepatic regeneration by promoting angiogenesis through the Wnt pathway, we detected VEGF and β-catenin in liver tissues and BrdU and β-catenin in hepatocytes by immunofluorescence. The expression levels of CRP, Wnt pathway-related and angiogenic factors were increased in CCl 4 -injured and PD-MSCs transplanted rat livers. In vitro, the expression levels of Wnt signaling and angiogenic factors were decreased in siRNA-CRP-transfected rat hepatocytes. @*Conclusions@#CRP upregulation by PD-MSCs participates in vascular remodeling to promote liver regeneration via the Wnt signaling pathway during hepatic failure.

Identification of microRNAs and their target genes in the placenta as biomarkers of inflammation / 대한생식의학회지

Objective@#Recently, microRNA (miRNA) has been identified both as a powerful regulator involved in various biological processes through the regulation of numerous genes and as an effective biomarker for the prediction and diagnosis of various disease states. The objective of this study was to identify and validate miRNAs and their target genes involved in inflammation in placental tissue. @*Methods@#Microarrays were utilized to obtain miRNA and gene expression profiles from placentas with or without inflammation obtained from nine normal pregnant women and 10 preterm labor patients. Quantitative real-time polymerase chain reaction and Western blots were performed to validate the miRNAs and differentially-expressed genes in the placentas with inflammation. Correlations between miRNA and target gene expression were confirmed by luciferase assays in HTR-8/SVneo cells. @*Results@#We identified and validated miRNAs and their target genes that were differentially expressed in placentas with inflammation. We also demonstrated that several miRNAs (miR-371a-5p, miR-3065-3p, miR-519b-3p, and miR-373-3p) directly targeted their target genes (LEF1, LOX, ITGB4, and CD44). However, some miRNAs and their direct target genes showed no correlation in tissue samples. Interestingly, miR-373-3p and miR-3065-3p were markedly regulated by lipopolysaccharide (LPS) treatment, although the expression of their direct targets CD44 and LOX was not altered by LPS treatment. @*Conclusion@#These results provide candidate miRNAs and their target genes that could be used as placental biomarkers of inflammation. These candidates may be useful for further miRNA-based biomarker development.

Immunomodulatory Effects of Placenta-derived Mesenchymal Stem Cells on T Cells by Regulation of FoxP3 Expression

The immunomodulatory effects of mesenchymal stem cells (MSCs) are an important mediator of their therapeutic effects in stem cell therapy and regenerative medicine. The regulation mechanism of MSCs is orchestrated by several factors in both intrinsic and extrinsic events. Recent studies have shown that the dynamic expression of cytokines secreted from MSCs control T cell function and maturation by regulating the expression of FoxP3, which figures prominently in T cell differentiation. However, there is no evidence that placenta-derived mesenchymal stem cells (PD-MSCs) have strong immunomodulatory effects on T cell function and maturation via FoxP3 expression. Therefore, we compared the expression of FoxP3 in activated T cells isolated from peripheral blood and co-cultured with PD-MSCs or bone marrow-derived mesenchymal stem cells (BM-MSCs) and analyzed their effect on T cell proliferation and cytokine profiles. Additionally, we verified the immunomodulatory function of PD-MSCs by siRNA-mediated silencing of FoxP3. MSCs, including PD-MSCs and BM-MSCs, promoted differentiation of naive peripheral blood T cells into CD4+CD25+FoxP3+ regulatory T (Treg) cells. Intriguingly, the population of CD4+CD25+FoxP3+ Treg cells co-cultured with PD-MSCs was significantly expanded in comparison to those co-cultured with BM-MSCs or WI38 cells (p < 0.05, p < 0.001). Dynamic expression patterns of several cytokines, including anti- and pro-inflammatory cytokines and members of the transforming growth factor-beta (TGF-β) family secreted from PD-MSCs according to FoxP3 expression were observed. The results suggest that PD-MSCs have an immunomodulatory effect on T cells by regulating FoxP3 expression.

Effects of selenium on the survival and invasion of trophoblasts / 대한생식의학회지

OBJECTIVE: Placental oxidative stress is known to be a factor that contributes to pregnancy failure. The aim of this study was to determine whether selenium could induce antioxidant gene expression and regulate invasive activity and mitochondrial activity in trophoblasts, which are a major cell type of the placenta. METHODS: To understand the effects of selenium on trophoblast cells exposed to hypoxia, the viability and invasive activity of trophoblasts were analyzed. The expression of antioxidant enzymes was assessed by reverse-transcription polymerase chain reaction. In addition, the effects of selenium treatment on mitochondrial activity were evaluated in terms of adenosine triphosphate production, mitochondrial membrane potential, and reactive oxygen species levels. RESULTS: Selenium showed positive effects on the viability and migration activity of trophoblast cells when exposed to hypoxia. Interestingly, the increased heme oxygenase 1 expression under hypoxic conditions was decreased by selenium treatment, whereas superoxide dismutase expression was increased in trophoblast cells by selenium treatment for 72 hours, regardless of hypoxia. Selenium-treated trophoblast cells showed increased mitochondrial membrane potential and decreased reactive oxygen species levels under hypoxic conditions for 72 hours. CONCLUSION: These results will be used as basic data for understanding the mechanism of how trophoblast cells respond to oxidative stress and how selenium promotes the upregulation of related genes and improves the survival rate and invasive ability of trophoblasts through regulating mitochondrial activity. These results suggest that selenium may be used in reproductive medicine for purposes including infertility treatment.

Decreased C-reactive protein induces abnormal vascular structure in a rat model of liver dysfunction induced by bile duct ligation

BACKGROUND/AIMS: Chronic liver disease leads to liver fibrosis, and although the liver does have a certain regenerative capacity, this disease is associated with dysfunction of the liver vessels. C-reactive protein (CRP) is produced in the liver and circulated from there for metabolism. CRP was recently shown to inhibit angiogenesis by inducing endothelial cell dysfunction. The objective of this study was to determine the effect of CRP levels on angiogenesis in a rat model of liver dysfunction induced by bile duct ligation (BDL). METHODS: The diameter of the hepatic vein was analyzed in rat liver tissues using hematoxylin and eosin (H&E) staining. The expression levels of angiogenic factors, albumin, and CRP were analyzed by real-time PCR and Western blotting. A tube formation assay was performed to confirm the effect of CRP on angiogenesis in human umbilical vein endothelial cells (HUVECs) treated with lithocholic acid (LCA) and siRNA-CRP. RESULTS: The diameter of the hepatic portal vein increased significantly with the progression of cirrhosis. The expression levels of angiogenic factors were increased in the cirrhotic liver. In contrast, the expression levels of albumin and CRP were significantly lower in the liver tissue obtained from the BDL rat model than in the normal liver. The CRP level was correlated with the expression of albumin in hepatocytes treated with LCA and siRNA-CRP. Tube formation was significantly decreased in HUVECs when they were treated with LCA or a combination of LCA and siRNA-CRP. CONCLUSION: CRP seems to be involved in the abnormal formation of vessels in hepatic disease, and so it could be a useful diagnostic marker for hepatic disease.

Epigenetic Alterations of IL-6/STAT3 Signaling by Placental Stem Cells Promote Hepatic Regeneration in a Rat Model with CCl4-induced Liver Injury

BACKGROUND: Human chorionic plate-derived mesenchymal stem cells (CP-MSCs) isolated from the placenta have been reported to demonstrate therapeutic effects in animal models of liver injury; however, the underlying epigenetic mechanism of this effect has not been elucidated. Thus, we investigated whether CP-MSCs influence epigenetic processes during regeneration of the injured liver. METHODS: CP-MSCs were engrafted into a carbon tetrachloride (CCl4)-injured rat model through direct transplantation into the liver (DTX), intrasplenic transplantation (STX), and intravenous transplantation via the tail vein (TTX). Non-transplanted (NTX) rats were maintained as sham controls. Liver tissues were analyzed after transplantation using immunohistochemistry, western blot analysis, and quantitative methylation-specific polymerase chain reaction. Proliferation and human interleukin-6 (hIL-6) enzyme-linked immunosorbent assays were performed using CCl4-treated hepatic cells that were co-cultured with CP-MSCs. RESULTS: The Ki67 labeling index, cell cyclins, albumin, IL-6, and gp130 levels were elevated in the CP-MSC transplantation groups. The concentration of hIL-6 in supernatants and the proliferation of CCl4-treated rat hepatic cells were enhanced by co-culturing with CP-MSCs (p<0.05), while the methylation of IL-6/IL-6R and STAT3 by CP-MSC transplantation decreased. CONCLUSION: These results suggest that administration of CP-MSCs promotes IL-6/STAT3 signaling by decreasing the methylation of the IL-6/SATA3 promoters and thus inducing the proliferation of hepatic cells in a CCl4-injured liver rat model. These data advance our understanding of the therapeutic mechanisms in injured livers, and can facilitate the development of cell-based therapies using placenta-derived stem cells.

Advanced Research on Stem Cell Therapy for Hepatic Diseases: Potential Implications of a Placenta-derived Mesenchymal Stem Cell-based Strategy / 한양의대학술지

The category of chronic liver diseases comprise one of the most common medical diagnoses worldwide. Currently, orthotopic liver transplantation is the only effective treatment for end-stage hepatic disease, but this procedure is associated with many problems, including donor scarcity, operative damage, high cost, risk of immune rejection and lifelong immunosuppressive treatments. Thus, the development of new therapies is highly desirable. Cell therapy with stem cells is increasingly being used to repair damaged tissue or to promote organ regeneration. Stem cells, which possess self-renewal activity as well as differentiation potential, can be categorized as embryonic stem cells (ESCs) or adult stem cells (ASCs), which include hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs). Recently, placenta-derived mesenchymal stem cells (PD-MSCs) have been reported, and they are attracting much interest in stem cell research for their multiple advantages: 1) no ethical concerns, 2) the ability to obtain abundant cell numbers, 3) multi-lineage differentiation potential, and 4) strong immunosuppressive properties. PD-MSCs differentiate into hepatocyte-like cells when exposed to hepatogenic differentiation-inducing conditions and PD-MSCs transplantation has been shown to enhance hepatic regeneration and/or survival in a rat hepatic failure model by suppressing the progression of fibrosis and apoptosis and activating autophagy. In this review, we will explain the characteristics of several kinds of PD-MSCs and discuss recent studies of the therapeutic potential of PD-MSCs in the repair of liver injury and their utility in regenerative medicine. Although many problems remain to be solved, many studies support the potential for human stem cell therapies, including PD-MSCs, as a promising new technology for the therapeutic regeneration of human liver intractably damaged due to chronic disease and/or toxic and environmental insult.

The role of autophagy in the placenta as a regulator of cell death / 대한생식의학회지

The placenta is a temporary fetomaternal organ capable of supporting fetal growth and development during pregnancy. In particular, abnormal development and dysfunction of the placenta due to cha nges in the proliferation, differentiation, cell death, and invasion of trophoblasts induce several gynecological diseases as well as abnormal fetal development. Autophagy is a catalytic process that maintains cellular structures by recycling building blocks derived from damaged microorganelles or proteins resulting from digestion in lysosomes. Additionally, autophagy is necessary to maintain homeostasis during cellular growth, development, and differentiation, and to protect cells from nutritional deficiencies or factors related to metabolism inhibition. Induced autophagy by various environmental factors has a dual role: it facilitates cellular survival in normal conditions, but the cascade of cellular death is accelerated by over-activated autophagy. Therefore, cellular death by autophagy has been known as programmed cell death type II. Autophagy causes or inhibits cellular death via the other mechanism, apoptosis, which is programmed cell death type I. Recently, it has been reported that autophagy increases in placenta-related obstetrical diseases such as preeclampsia and intrauterine growth retardation, although the mechanisms are still unclear. In particular, abnormal autophagic mechanisms prevent trophoblast invasion and inhibit trophoblast functions. Therefore, the objectives of this review are to examine the characteristics and functions of autophagy and to investigate the role of autophagy in the placenta and the trophoblast as a regulator of cell death.

Significance of Serum Antibody Test for Toxocariasis in Healthy Healthcare Examinees with Eosinophilia in Seoul and Gyeongsangnam-do, Korea

There have been numerous reports on the relationship between eosinophilia and toxocariasis. The present study investigated seropositive rates of toxocariasis among healthy people with or without eosinophilia in urban and rural areas, and assessed risk factors for positive antibody test. A total of 610 healthy people, who visited health check-up (Medicheck(R), Korea Association of Health Promotion), 310 from Seoul and 300 from Gyeongsangnam-do, were subjected for this study. Their serum samples were tested by ELISA with the crude antigen of Toxocara canis larvae. Cross-reactions with other tissue invading helminth antigens were also investigated. Total antibody positive rate of toxocariasis was 8.7% of the 610 subjects. When the subjects were grouped into 3 by their eosinophil counts, the antibody positive rates significantly differed by the groups; 5.9% (18/306) in the group500/microL (P=0.028). A total of 22 serum samples cross-reacted with other tissue-invading helminth antigens. A questionnaire analysis recognized drinking alcohol and smoking as significant risk factors of toxocariasis. In conclusion, toxocariasis antibody positive rate is correlated with eosinophil counts. It is recommended that healthy subjects with eosinophilia by routine health examination and risk factors undergo Toxocara serology by multiantigen ELISA to investigate etiology.

Intestinal Helminthic Infections in Striped Field Mice, Apodemus agrarius, from Two Southern Regions of Korea

The present study was performed to know the infection status of intestinal helminths in a most common species of field mice, Apodemus agrarius, from 2 southern regions of Korea. Total 133 and 103 mice were collected by the mouse trap in Hapcheon-gun, Gyeongsangnam-do and Gurye-gun, Jeollanam-do, respectively, from July 2005 to June 2006. The small intestine of each mouse was resected and longitudinally opened with a pair of scissors. The intestinal contents were washed with 0.85% saline until the supernatant became clear. Helminths were collected with naked eyes or under a stereomicroscope from the sediment of the intestinal content. More than 11 species of helminths (4 nematode spp., 5 trematode spp., and 2 cestode spp.) were recovered. Among these, heligmosomoid nematodes (97.5%) was the most highly and heavily infected species. As the members of trematodes, Plagiorchis muris, Brachylaima sp., Echinostoma hortense, Echinostoma cinetorchis, and unidentified echinostome larvae were found in the small intestines of 35 (14.8%), 12 (5.1%), 6 (2.5%), 1 (0.4%), and 1 (0.4%) mice respectively. Two species of tapeworms, Hymenolepis nana and Hymenolepis diminuta were also detected in 79 (33.5%) and 21 (8.9%) mice, respectively. Conclusively, heligmosomoid nematodes were the most prevalent (dominant) species among more than 11 helminth species detected, and Brachylaima sp. fluke is newly added in the list of intestinal trematodes in Korea.

Altered expression of norepinephrine transporter and norepinephrine in human placenta cause pre-eclampsia through regulated trophoblast invasion / 대한생식의학회지

OBJECTIVE: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. METHODS: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. RESULTS: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. CONCLUSION: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

Effects of hypoxia inducible factors-1alpha on autophagy and invasion of trophoblasts / 대한생식의학회지

OBJECTIVE: This study was undertaken to determine the effect of hypoxia inducible factor (HIF)-1alpha on the cell death, autophagy, and invasion of trophoblasts. METHODS: To understand the effect of HIF-1alpha, we inhibited HIF-1alpha using siRNA under normoxia and hypoxia conditions. Invasion assay and zymography were performed to determine changes in the invasion ability of HIF-1alpha. Western blotting and immunofluorescence were performed to determine some of the signal events involved in apoptosis and autophagy. RESULTS: There was no difference in cell death through the inhibition of HIF-1alpha expression by siRNA; however, the expression of LC3 and autophagosome formation increased. On the other hand, autophagy was increased, and the invasive ability of trophoblast cells decreased according to the inhibition of HIF-1alpha expression by siRNA. These experimental results mean that HIF-1alpha genes regulate the invasive ability of trophoblasts by increasing autophagy. CONCLUSION: This study contributes important data for understanding the mechanism of early pregnancy implantation and the invasive ability of trophoblasts by defining the relationship between the roles of HIF-1alpha and autophagy.

Chin augmentation with bone transplantation from the mandible / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore a new method for mentoplasty.</p><p><b>METHODS</b>The bilateral prominent mandibular angle or outer lamina was resected through the intraoral approach. The resected bone fragments were shaped and rigid fixed to the chin with miniplates and screws.</p><p><b>RESULTS</b>A total of 30 patients (28 females, 2 males) accepted chin augmentation with this method. The mandibular angle bone was used in 20 cases and the mandibular outer lamina was used in 10 cases. The operative results were satisfactory, and the patient's facial contour was improved substantially.</p><p><b>CONCLUSION</b>No rejection reaction was found after this procedure. Chin augmentation with autogenous mandibular bone is an ideal method for genioplasty.</p>

Genomic Imbalances in Ependymoma by Degenerate Oligonucleotide Primed PCR-Comparative Genomic Hybridization

BACKGROUND: The most consistent chromosomal abnormality in ependymomas, is loss of 22q (17-75%) and gain of 1q (0-50%). However, significance of this abnormality is uncertain. METHODS: Genomic imbalances in 27 Korean ependymomas, including 21 low grade ependymomas, 4 anaplastic and 2 myxopapillary ependymomas, were analyzed by degenerate oligonucleotide primed-PCR-comparative genomic hybridization. RESULTS: Common gains were found in 17 (63%), 20q (59%), 9q34 (41%), 15q24-qter (33%), 11q13 (30%), 12q23 (26%), 7q23-qter (26%), 16q23-qter (30%), 19 (26%), and 1q32-qter (22%). DNA amplification was identified in 12 tumors (44%). Chromosomal loss was a less common occurrence in our study, but was found in 13q (26%), 6q (19%), and 3 (11%). CONCLUSION: The recurrent gains or losses of the chromosomal regions which were identified in this study provide candidate regions that may be involved in the development and progression of ependymomas.

SO2 Effects on Glycoconjugates of the Respiratory Mucosa in Rat Nasal Cavity / 체질인류학회지

The experiments of this study was performed to investigate the effects of sulfur dioxide on the changes of glycoconjugates of respiratory system of the rat. Sprague -Dawley male rats weighing about 200 ~250g were divided into a control group and SO2 exposed groups. Again SO2 exposed groups were divided into 10 ppm, 25 ppm, 50 ppm, 100 ppm and 200 ppm subgroups according to concentrations of SO2 and each SO2 exposed groups were divided into 1, 3 and 6 hours groups. For the histological changes, H -E(hematoxylin -eosin) and PAS(periodic acid Schiff) staining were used and to investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL -1, sWGA, UEA -1, LCA and Con A) were applied. Generally, the effects of SO2 on the rat nasal respiratory region were more serious at the high concentrations. Moreover, as the exposed time was longer even at the low concentrations, the effects of SO2 were similar to those of high concentration. Compared with all SO2 concentrations, the longer exposed time was, the more serious the effects of SO2 were. In the SO2 exposed groups the binding of PNA, RCA -1 and UEA -1 of cilia in the nasal septal respiratory epithelium tended to increase in the 10 ppm and 25 ppm SO2 exposed groups but it tended to decrease in the 100 ppm and 200 ppm SO2 exposed groups. In the cytoplasm of columnar cells of nasal septal respiratory epithelium, Con A binding increased in all the SO2 exposed groups. In the goblet cells DBA, SBA, PNA, RCA -1 and UEA -1 binding increased remarkably in the 50 ppm SO2 exposed groups but it decreased largely or disappeared in the 100 ppm and 200 ppm SO2 exposed groups. The binding of SBA, PNA, BSL -1, UEA -1 and Con A in the intraepithelial mucous cells which were not detected in the control group, increased in the 25 ppm and 50 ppm SO2 exposed groups while it tended to decrease in the 100 ppm and 200 ppm SO2 exposed groups. The binding of sWGA increased according to the concentrations of SO2 were higher and exposed times were longer. In the superior nasal septal gland, the binding of PNA increased in the 50 ppm and 100 ppm SO2 exposed groups and that of Con A increased in the 25 ppm and 50 ppm SO2 exposed groups. In the inferior nasal septal gland, except for LCA, the binding of the other lectins increased remarkably in the 25 ppm and 50 ppm SO2 exposed groups but it tended to decrease in the 100 ppm and 200 ppm SO2 groups. In the mucous duct cells, the reaction of PNA and RCA -1 increased compared with that of the control group. And the reaction of BSL -1 and UEA -1 increased in the lower concentrations of 50 ppm SO2 exposed group but it decreased in the 100 ppm and 200 ppm SO2 exposed groups. The binding of Con A increased in the 25 ppm and 50 ppm SO2 exposed groups. Consequently, from the results above mentioned that SO2 affected serious changes on glycoconjugates metabolism in the nasal cavity.

The Role of CD44 in the Adhesion of Human Osteosarcoma to Hyaluronic Acid / 체질인류학회지

Osteoblast expresses a sequence of extracellular matrix during differentiation, suggest that multiple adhesion mechanisms regulate osteoblast differentiation and bone development. Hyaluronic acid (HA) is a glycosaminoglycan (GAG) which mainly distribute in cartilage and extracellular matrix (ECM). HA has very high molecular weights and their structures occupy large solvent domains which give solution of high viscosity. During early development and before tissue differentiation, HA can constitute the major structural macromolecule in the ECM, where it can promote both cell proliferation and migration. CD44 is a cell surface receptor for HA. The polymorphic family of integral membrane glycoproteins CD44 is found on a wide variety of cells. CD44's function in the cell membrane is transmembrane signalling between extracellular matrix and acin filament. In present study, HOS was used as a model to examine whether HOS adhere to HA, CD44 and GAGs on cell surface participate in the adhesion to HA and there is difference in CD44 expression at that time. HOS adhered to HA in the manner of time -dependent. After incubation for 180 minutes, about 90% of cells adhered to HA. When HOS was pretreated with anti -CD44 antibody, hyaluronidase and genistein, the adhesion rate was significantly decreased. CD44 was more expressed in HOS plated on HA -coated wells than BSA -coated wells. Taken together, HOS has a adhesive affinity to HA. CD44, GAGs on cell surface and tyrosine kinases play an important role in the adhesion of HOS to HA.

Lectin Histochemistry for Studying Glycoconjugates in Rat Lingual Salivary Glands during the Postnatal Development / 체질인류학회지

The developmental changes of the lingual salivary glands in the postnatal rats were examined by lectin histochemical methods. For the morphological changes, H-E and PAS staining were used. The biotinylated lectins used in the study were DBA, SBA, PNA, BSL-1, sWGA, RCA-1, UEA-1, Con A and LCA. The promordia and undifferentiated acini of the lingual glands were found in the mucous glands at 0 day suckling rat and the von Ebner's glands at 3 day suckling rat, respectively. The differentiation and maturation of the lingual glands were faster than those of the von Ebner's gland. The differentiation and proliferation of both glands were occurred remarkably at suckling periods rather than weaning periods. The lectin binding pattern of glandular promordia and undifferentiated serous acini in von Ebner's gland was weak in BSL-1 and weak to moderate in RCA-1. DBA and sWGA showed tendency to increase in 1 week suckling rat, but The binding reactivity of other lectins was disappeared except BSL-1 that was reacted tracely in 2 and 3 weak suckling and 4 week weaning rat. RCA-1, PNA, sWGA, BSL-1 and SBA of the differentiated serous acini were appeared in the 2 week suckling rat and SBA and sWGA was more intense. Especially, the reactivity of these lectins of suckling periods was showed more tendency to increase than that of weaning periods. The increase of PNA, SBA and BSL-1 was prominent during suckling and weaning periods. RCA-1 and sWGA were decreased in 5 week rat, increased in 6 week rat, and then decreased in adult rat. UEA-1 which was not shown from 0 day to 2 week was showed trace to moderate reactivity in some serous acini. Con A and PNA of glandular promordia and undifferentiated mucous acini were appeared trace or weak, and absent at 0 day suckling rat, but PNA reactivity was showed tendency to incerase at 3 day suckling rat. Other lectins of these promordia and acini were not showed reactivity. In the differentiated mucous acini at 0 day suckling rat, all mucous acini were weak to moderate with DBA, and some of mucous acini also were weak to moderate with BSL-1. Most mucous acini showed weak reactivity with SBA, but some mucous acini showed trace or weak reactivity with RCA, PNA, sWGA and BSL-1. The reactivity of BSL-1 and sWGA was increased from birth to 2 week and then decreased, and absent at 5 week. But it increased at 6 week. RCA-1 and PNA also increased in the acini up to 1 week. However, PNA reactivity was absent at 5 and 6 week. With RCA-1, the intensity of reactivity was increased. Differentiated mucous acini was reacted to increase with SBA from birth, the intensity was strong in weaning periods rather than suckling period. UEA-1 reactivity was showed to decrease from 1 week to 2 week and moderately increased from 3 week to 5 week, and thereafter decreased. DBA binding pattern was somewhat changed throughout the observation periods but it was predominent.

Expression of Laminin Chains in the Neuronal Cells of Mouse Brain

Laminin-1 is biologically active and can effect cellular proliferation, differentiation, migration, and apoptosis. In the central nervous system, neuronal cells are rarely reported to give positive reaction by laminin antibody staining. However, the original cell type which can produce the laminin molecule has not been well established. Since the neuronal cells of brain are derived from neuroectoderm, we thought that the neuronal cells should be able to produce the laminin molecules as other epithelial cells. In this study we aimed to explore whether the neuronal cells express the laminin chain mRNAs, and further to identify which types of laminin isoform are expressed at the specific sites of the brain structure. We found that neuronal cell was the important cell type in mouse brain, which could produce laminin isoforms. Although immunostainings disclosed reactivity of laminins in the basement membrane of capillaries as well as neuronal cells, mRNA expressions of laminins were intense only in the neuronal cells. It was relatively weak in the endothelial cells. Among neuronal cells the cortical cells of cerebrum, pyramidal cells of hippocampus, and Purkinje cells of cerebellum showed pronounced expression of laminin chain mRNA. Glial cells, especially astrocytes, were negative for laminin subtypes both in immunohistochemistry and in situ hybridization. Taken together, our data indicate that the neuronal cells of mouse brain actively produce laminin isoforms, and the resultant polymerized laminins are accumulated mainly in the basement membrane of capillaries. In conclusion, the results indicate that neuronal cells produce and utilize the different laminin chains to maintain the neurovascular environment of brain.

Lectin Histochemistry for Effects of N - Nitrosodimethylamine on Glycoconjugates in the Rat Lingual Salivary Glands / 체질인류학회지

The effect of NDMA after oral administration (17 mg/ml) on the glycoconjugates of lingual von Ebner's gland and mucous gland were investigated with lectin histochemical methods. For lectin histochemical studies, the biotinylated lectins (DBA, PNA, SBA, BSL -1, sWGA, RCA -1, LCA, UEA -1, and ConA) were applied. Lectin binding patterns of glycoconjugates of lingual von Ebner's gland showed the decreased affinity for DBA, PNA, BSL -1 and sWGA in NDMA -treated group compared with control group. The remarkable decrease of binding affinity of NDMA -treated group was observed in PNA for 12 and 24 hours, DBA for 96 hours, BSL -1 for 72 hours, and sWGA for 3 hours, while the striking decrease of BSL -1 and sWGA binding was observed in NDMA -treated group for 12 hours. But these decreases of binding were tended to recover in PNA and sWGA after 72 hours of NDMA treatment, and in DBA after 120 hours. The binding affinity of SBA and RCA -1 was decreased in NDMA -treated group for 3 hours, while the other NDMA -treated group showed an increased affinity. Especially, the increase of SBA binding was remarkable. There was a little change in binding affinity of UEA -1, LCA and Con A in NDMA -treated group. Lectin binding patterns of glycoconjugates of lingual mucous gland showed decreased affinities for SBA, sWGA and UEA -1 in NDMA -treated group. The striking decreases of binding affinity for NDMA -treated group was observed in SBA and sWGA for 3 hours, and UEA -1 for 3 and 24 hours. And the remarkable decreases of binding affinity for NDMA -treated group was found in SBA for 24 and 48 hours, sWGA for 48, 72 and 96 hours, and UEA -1 for 48 hours. These decreases of binding affinity of NDMA -treated group were tended to recover in SBA and UEA -1 after 96 hours and in sWGA after 120 hours. The binding affinity for PNA and ConA showed a little but not remarkable increase in NDMA - treated group, and LCA binding showed a little decrease following a little increase in NDMA - treated group. The affinity of DBA binding was decreased in NDMA -treated group for 12 hours and 24 hours, while the other NDMA -treated group showed an increased affinity. Especially, there was a remarkable increase in NDMA -treated group for 96 hours. From these results, it is suggested that the toxicity of NDMA may be related with the carcinogen of the rat tongue, and glycoconjugates are concerned with the repaire of the destruction of the lingual mucous acini.