To understand the corneal regeneration induced by bevacizumab, we investigated the structure changes of stroma and basement membrane regeneration. A Stick soaked in 0.5 N NaOH onto the mouse cornea and 2.5 mg/ml of bevacizumab was delivered into an alkali-burned cornea (2 ${\mu}l$) by subconjunctival injections at 1 hour and 4 days after injury. At 7 days after injury, basement membrane regeneration was observed by transmission electron microscope. Uneven and thin epithelial basement membrane, light density of hemidesmosomes, and edematous collagen fibril bundles are shown in the alkali-burned cornea. Injured epithelial basement membrane and hemidesmosomes and edematous collagen fibril bundles resulting from alkali-burned mouse cornea was repaired by bevacizumab treatment. This study demonstrates that bevacizumab can play an important role in wound healing in the cornea by accelerating the reestablishment of basement membrane integrity that leads to barriers for scar formation.
In this study, cement/PVDF hollow-fiber hybrid membranes were prepared via a mixed process of diffusion-induced phase separation and hydration. The presence of X-ray diffraction peaks of Ca(OH)2, an AFt phase, an AFm phase, and C-S-H phase confirmed the hydration reaction. Good hydrophilicity was obtained. The cross-sectional and surface morphologies of the hybrid membranes showed that an asymmetric pore structure was formed. Hydration products comprising parallel plates of Ca(OH)2, fibrous ettringite AFt, and granulated particles AFm were obtained gradually. For the hybrid membranes cured for different time, the pore-size distribution was similar but the porosity decreased because of blocking of the hydration products. In addition, the water flux decreased with hydration time, and carbon retention was 90% after 5 h of rejection treatment. Almost all the Zn2+ ions were adsorbed by the hybrid membrane. The above results proved that the obtained membrane could be alternative as basement membrane for separation application.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.30
no.4
/
pp.271-281
/
2004
Adenoid cystic carcinoma is malignant tumor in salivary gland, and its behavior is very invasive. Of all malignant tumor adenoid cystic carcinoma is occured in frequency of 4.4% in major salivary gland, and 1.29% in minor salivary gland. Histopathologically, adenoid cystic carcinoma is characterized by a cribriform appearance, and tubular form and solid nest type tumor can be seen. The tumor cell structure composed of modified myoepithelial cell, and basaloid cell. Extracellular matrix of this tumor cell contains variable ground substance with basement membrane component. Basement membrane matrix composed of collagen fibers, glycoproteins, proteoglycans, and its function is well known that it participate in differentiation, proliferation, and growth of tumor cell. Basement membrane molecule is essential for invasion of peripheral nerve, blood vessel, skeletal muscle in tumor cell of adenoid cystic carcinoma. In many studies, the tumor cell of adenoid cystic carcinoma containing modified myoepithelial cell participate in synthesis of proteoglycan. In this study, tissue sample of adenoid cystic carcinoma of human salivary gland were obtained from 15 surgical specimen, and all specimen were routinely fixed in 10% formalin and embedded. Serial $4-{\mu}m$ thick sections were cut from paraffin blocks. the histopathologic evaluation was done with light microscopy. And, the immunohistochemical staining, characteristics of glycosaminoglycan were observed. For biochemical analysis of glycosaminoglycan, isolation of crude glycosaminoglycan from tumor tissue and Western bolt analysis were carried out. With transmission electomicroscopy, tumor cell were observed. Biologic behavior of adenoid cystic carcinoma was observed with distribution and expression of basement membrane of glycosaminoglycan in tumor cells, The results obtained were as follows: 1. In immunohistochemical study, chondroitin sulfate is postively stained in tumor cell and interstitial space, dermatan sulfate is weakly stained in ductal cell. But keratan sulfate is negatively stained. 2. In immunohistochemical study, heparan sulfate is strong positive stained in tumor cell and basement membrane, especially in invasion area to peripheral nerve tissue. 3. In transmission electromicroscpic view, the tumor cells are composed modifed myoepithelial cells, and contains many microvilli and rough endoplasmic reticulum. 4. In Western blot analysis, the expression of glycosaminoglycan is expressed mostly in heparan sulfate. From the results obtained in this study, tumor cell of adenoid cystic carcinoma is composed modified myoepithelial cell, and glycosaminoglycan of basement membrane molecule of heparan sulfate and chondroitin sulfate mostly participate in the development and invasiveness of adenoid cystic carcinoma by immunohistochemical study and western blot analysis.
Journal of Physiology & Pathology in Korean Medicine
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v.17
no.5
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pp.1315-1320
/
2003
The purpose of this research was to investigate effects of Bambusae Caulis in Liquamen (BCL) on the synthesis of basement membrane proteins during proliferation and differentiation of 3T3-L1 cells. BCL has been used to relieve the cough and asthma, and remove phlegm in traditional oriental medicines. In recent years. it was studied for its antiinflammatory, antiallergenic. immune-modulating and anticarcinogenic capabilities. We have previously observed that glycyrrhizin stimulates the adipose conversion of 3T3-L1 cells. To investigate effects of BCL on the basement membrane proteins during proliferation and differentiation of 3T3-L1 cells, we have analyzed synthetic amounts of basement membrane components such as type IV collagen and BM40. BCL stimulated the synthesis and secretion of type IV collagen from both 3T3-L1 preadipocytes and adipocytes. The synthesis and secretion of BM40 was not affected by BCL. The continuous addition of BCL markedly stimulated cell growth and increased cell density. These results suggest an important role for type IV collagen in adipocyte differentiation.
Kim, Young-Ho;Park, Kwan-Kyu;Kim, Young-Man;Cho, Soo-Yeul
Applied Microscopy
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v.28
no.4
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pp.577-590
/
1998
Puromycin aminonucleoside (PAN) nephropathy was induced in a group of Sprague-Dawley rat by a single dose of intraperitoneal Injection to study an ultrastructural change of glomerulus. The experimental rats developed proteinuria three days after PAN injection. Electron microscopic studies of glomeruli showed the loss of epithelial foot processes, formation of cytoplasmic vacuoles, microvillous formation and increased numbers of lysosomes in the cytoplasm of podocytes. It is strongly suggested that proteinuria in PAN nephrosis may be primarily due to a glomerular epithelial lesion, leading to focal disarray of anionic sites or focal defects in the epithelial covering of the basement membrane. The loss of anionic sites in the basement membrane nay be caused by the foot process fusion and the epithelial detachment from the basement membrane.
Background and Objectives : Sulfur dioxide gas is one of the major airborne Pollutants noxious to human in industrialized countries. The most vulnerable areas in the human respiratory system were the trachea and main bronchi and a gradient of decreasing damage was observed in the peripheral tracheobronchial tree. Induced functional alteration was increased mucosal permeability, and morphological changes were epithelial sloughing, intracellular edema, mitochondrial swelling, widened intercellular spaces, and ciliary cytoplamic extrusions. The laminins are a family of extracellular matrix glycoproteins localized in the basement membrane. Their primary role is cell-matrix attachment, but many additional biologic activities, including Promoting cell growth and migration, tumor growth and metastasis, wound repair, and graft survival, have been demonstrated. Materials and Methods : Histologic changes and expression of laminin in tracheal mucosa sacrificed at 1 day, 2 day, 3 day, 1 week, 2 weeks, and 3 weeks after continued SO2 exposure of 250 ppm for 30 minutes a day(to 7week) were studied in rats. In this study, mild immune reaction for laminin was noted at the apical cytoplasm of epithelial cells and basement membrane one day after a 7 week $SO_2$ exposure. The cilia and nucleoi of epithelial cells were normal and no immune reaction was noted in Goblet cells. The lamina propria of the tracheal tissue was infiltrated by monocytes and lymphocytes. Results : At 24 hours after exposure, all tracheal cells except Goblet cells revealed a mild immune reaction for laminin. No immune reactions were noted in the basement membrane. At 72 hours after exposure, mild or moderate immune reactions for laminin was seen in the tracheal cell cytoplasm. Irregular faint immune reaction for laminin was noted in the basement membrane. At 1 week after exposure, strong immune reaction for laminin was detected over all tracheal cells, and the basement membrane was seen clearly. At 2~3 weeks after exposure, strong immune reaction for laminin was seen in all tracheal epithelial cells except Goblet cells and a mild immune reaction was partly revealed in the basement membrane. Conclusion : Our study suggests that 502 produces histologic damage on the tracheal mucosa. Longer duration after exposure of $SO_2$ makes more progressive healing on the tracheal mucosa and increased immunoreactivity for laminin.
The filtration barrier of kidney consists of endothelial cell, glomerular capillary, glomerular basement membrane, mesangial matrix, and podocyte. In aged rats, the morphological changes were shown in various parts, including the glomerulus. These changes were thickening of basement membrane and mesangial matrix, crescent formation of glomerular capillary, deformity of foot processes, glomerular sclerosis and obsolescence. But these glomerular morphologies are partial images or few serial images analysis. In this study, we examined the morphological alteration of glomerulus in the young and aged rats by light microscopy, transmission electron microscopy and three dimensional reconstruction. We were found in aged rat glomerulus, expansion of urinary space and mesangial matrix, thickening and degrading of glomerular basement membrane, decreasing in podocyte foot processes, fragmentation of podocytic nucleus membrane. These observations indicate that may provide useful data for investigating the pathogenesis of age-related dysfunction of kidney.
Purpose: The retention of the basement membrane complex, which was the unique feature of the acellular dermal matrix ($AlloDerm^{(R)}$), plays an important role in the normal process of wound healing. The present study was aimed to compare the healing of the acellular dermal matrix according to the graft method in the rabbit ear. Materials and methods: Six mature rabbits weighing about 3.0 kg were used, $10\;{\times}\;5\;mm$ sized subcutaneous pockets were created between the ear skin and the underlying perichondrium. In the control group, the acellular dermal matrix was grafted with the basement membrane facing toward the perichondrium. On the contrary, the acellular dermal matrix was grafted with the basement membrane facing toward the skin side in the experimental group I. In the experimental group II, the acellular dermal matrix was grafted like rolled configuration with basement membrane side in. The grafted site was picked at 3, 7, and 21 days after the graft. Serial sections were processed by H-E stain and examined under light microscopy to assess the healing patterns. Results: There was no distinct volume loss in the gross examination, but resorption was observed from the edge of the acellular dermal matrix in the histological examination. The space of resorption was replaced by the newly formed fibrous tissues and vessels. The inflammatory cells were more increased at 7 days after the graft than the early days. However, inflammation was decreased at 21 days after the graft. Regardless of the graft direction, no differences were observed between the control and the experimental group I in the healing patterns. Conclusion: These results suggest that the acellular dermal matrix can be used simply and effectively without regard to the graft direction as a substitute of autogenous material for repairing soft tissue defect.
Purpose: Large skin defect by various causes, should be covered by autologous skin graft. But, the donor site of autologous skin graft is limited and leaves permanent donor scar and contracture. There have been our trial to engineer artificial skin using allogenic dermis (AlloDerm) with basement membrane. Methods: Dermal and epidermal layer were separated by immersing in dipase solution for 30 minutes, and the separated layers were treated with 0.05% trypsin for 10 minutes. And then each layer was cultivated to fibroblasts and keratinocytes on a culture medium. Fibroblasts were first penetrated into basement membrane of allogenic dermis facing down, then allogenic dermis was flipped over to face up and keratinocytes were transplanted to allogenic dermis. Results: Observing artificial skin fabricated in vitro, we found following: 1) The artificial skin opened in air for 5 days formed epidermal layer. In dermal layer, fibroblast was distributed evenly among all. 2) The artificial skin opened in air for 30 days formed thicker and thicker, and it formed basement membrane, spinous and granular layers. PAS stain to confirm existence of basement membrane showed positive reaction. 3) Cytokeratin 10 stain to confirm the formation of epidermal layer showed positive reaction. 4) The formation of thick keratin, lamellar body and desmosome similar to human skin were observed in result of an electron micrograph. Conclusion: As a result of research, the structure seen in normal skin such as rete ridge, is found in reproduced artificial skin. This type of artificial skin can be used as a useful model for investigating skin disease and for clinical application also.
Kim, Yong-Hun;Yun, Pil-Young;Myoung, Hoon;Kim, Myung-Jin
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.28
no.6
/
pp.434-439
/
2002
Oral squamous cell carcinoma (OSCC) is one of the most common head and neck cancers. OSCC generally has a poor prognosis due to its tendency towards a local invasion and subsequent metastasis, which is mediated by multiple proteolytic enzymes and angiogenesis. Soy products contain high levels of isoflavonoids, including the tyrosine kinase inhibitor, genistein, which has been identified as a potent inhibitor of cell proliferation and in vitro angiogenesis. The purpose of this in vitro study is to evaluate the anti-cancer effect of genistein with respect to the angiogenesis and basement membrane invasion in OSCC. The highly invasive OSCC cell line, HSC-3 cells were cultured in the presence of $10{\mu}M$ genistein for 24h. To evaluate the effects of genistein on the invasiveness and the gelatinolytic activity, in vitro invasion assay and zymography were performed. In order to evaluate the effect on the VEGF and bFGF mRNA expression, RT-PCR and northern hybridization reaction, and chemiluminescence detection were applied. The in vitro invasion assay showed that the genistein treatment reduced the cellular invasion through the artificial basement membrane and significant difference between the control group and the genistein treated group was shown in MMP-2 activity. Especially, the 62 kDa activated form of MMP-2 in the control group was 1.8 times higher than that in the genistein treated group. The results of the northern blot analyses indicated that VEGF mRNA expression in the genistein treated group was significantly down regulated. This study showed that genistein inhibits angiogenesis and reduces basement membrane invasion in OSCC. It seems to support the possibility of genistein as an anti-cancer agent.
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