• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2013.05a
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• pp.38-38
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• 2013

The mechanical cues that adherent cells derive from the extracellular matrix (ECM) can effect dramatic changes in cell migration, proliferation, and differentiation. Using a thin film of Type I collagen fibrils comprised of 100 nm to 200 nm collagen fibrils overlaying a bed of smaller fibrils, changes in cellular response to systematically controlled changes in mechanical properties of collagen was investigated. Further, an experimental and modeling approaches to calculate the elastic modulus of individual collagen fibrils, and thereby the effective stiffness of the entire collagen thin film matrix, from atomic force microscopy force spectroscopy data was performed. These results demonstrate an approach to analysis of fundamental properties of thin, heterogeneous, organic films, and add further insights into the mechanical properties of collagen fibrils that are of relevance to cell response to the ECM.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.6
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• pp.1332-1338
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• 1999

Intermolecular collagen cross links stabilize collagen fibrils and are necessary for normal tensile strength in collagen fibrils. Once the fibrils are aligned, hydroxyllysine, hydroxylysine derived aldehyde modified enzymatically, reacts with hydroxylysine to form the dehydrodihydroxylysinonorleucine (DHLNL), an immature crosslink. Pyridinoline, one of matured cross links is presumably formed nonenzymatically through condensation of DHLNL and hydroxylysine residue. It is widely distributed in hard connective tissues such as cartilage, bone and tendon. L ascorbic acid(AsA) is well known to be required for the enzymatic hydroxylation of proline and lysine in collagen fibrils. The purpose of this study is to clarify the role of AsA on the biosynthesis of DHLNL in vitro. We examined the effect of AsA on the formation of hydroxylysine and DHLNL in collagen. Pyridinoline and DHLNL were measured as a function of time. The contents of DHLNL was increased, reached maximum within 2 hr and was held until 24 hr, then it decreased slowly. On the contrary, pyridinoline increased gradually after 24 hr and continued to increase for 2 weeks. Moreover, the contents of DHLNL remarkably decreased at 60 min after incubation, the contents of DHLNL was decreased by addition of AsA or dehydroascorbic acid(DHA). These results suggest that the supplementation of AsA causes decrease in DHLNL formation and pyridinoline formed by nonenzymatic reaction of DHLNL.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.2
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• pp.305-312
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• 1998

Normal tensile strength in collagen fibrils is due to intermolecular and intramolecular crosslinks which are known to be altered in aging. Pyridinoline, a mature crosslink which is stable and nonreducible, is derived from two hydroxyallysine and one hydroxylysine residues of collagen fibrils. The excess formation of pyridinoline in collagen is associated with making the tissue stiffer, less soluble and less digestible by enzymes. Lysyl oxidase is the enzyme that initiates the biosynthesis or crosslinks in collagen by catalyzing the oxidative deamination of the lysyl and hydroxylysyl residues in these molecules, and its activity is inhibited by $\beta$-aminopropionitrile(BAPN). Our previous work demonstrated that the pyridinoline content of bone collagen significantly was increased during incubation for 5 weeks at 37$^{\circ}C$ invitro, but it was diecrased by the addition of ascorbic acdi(AsA). In this study, we clarified the specific action of AsA in aging process in vitro enzymatic reaction.

• Applied Microscopy
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• v.26 no.2
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• pp.157-175
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• 1996

The development of flexor digital tendon of the hand was studied by electron microscopy in human fetuses ranging from 9 mm to 260 mm crown rump length. The primordium of tendons was first identified as discrete collection of mesenchymal cells at 25 mm fetus. Synovial sheath formation had commenced by 40 mm fetus and was complete by 70 mm fetus. Cell junction or adhesion sites at all ages were noted between the tendon cells. When dilatation of the synovial cavity occurred, two types of synovial cells were observed. A-type cells had numerous vesicles and large vacuoles. In contrast, B-type cells were characterized by abundant rough endoplasmic reticulum and well-developed Golgi complex. By $150mm{\sim}260mm$ fetuses, a mojority of the synovial cells were type B. The most remarkable difference between the synovial cells of full-term fetus and adult was the larger amount of collagen fibers in the latter. The vascular buds were first observed between the individual fibril bundles in the interfascicular space at 150 mm fetus. At 25 mm fetus, collagen fibrils were first noted within narrow cytoplasmic recesses which were continued with the extracellular space. Collagen fibrils were filled in almost entire extracellular space at 150 mm fetus. Besides collagen fibrils in the extracellular space small elastic fibers were also identified and followed in their development.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.3
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• pp.475-479
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• 1997

A decrease in the circulating levels of estrogen, occuring as a consequence of post menopausal decline or from surgical ovariectomy, results in an accelerated loss of bone. Estrogen has been shown to stimulate lysyl oxidase activity, and the treatment with estrogen increased the pyridinium content of cortical bone. a trivalent mature cross-links collagen fibrils named pyridinoline, which is especially abundant in collagen of cartilage and bone, markedly increases with growth in humans and rats. The main aim of this study was to examine the increased bone loss caused by ovariectomy through monitoring the concentrations of the collagen and the pyridinium cross-links of collagen, pyridinoline. The ovariectomized rats, 4 weeks old, were divided at random into two or three groups of 5. Ovariectomies were carried out on both of the saline-treated group(OVX(NH)) and the estrogen-treated group(OVX(H)) using the dorsal approach and sham operations were performed on the sham-operated group(sham). They were maintained under identical conditions for 4 or 8 weeks and were allowed free access to food and water. it was observed that there was no significant difference between the control group and the sham-operated group, however, the control group had a higher content of collagen than the saline-treated group after 4 weeks and 8 weeks. Based on these results, iot is supposed that estrogen can enhance collagen synthesis and affects the pyridinoline formation in collagen fibrils through stimulating lysyl oxidase activity.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.609-619
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• 2000

Cyclosporin A(CsA) is now widely used to treat organ transplant recipients. But CsA has various short-and long-term side effects. Especially, gingival hyperplasia is not easy to resolve since its nature is still unknown. This study discusses the pathogenesis of CsA-induced gingival hyperplasia on the basis of data obtained from light and electron microscopic studies of biopsis from patients on CsA treatment after kidney transplantation. Light microscopically, the multilayered squamous epithelium showed an irregular surface of parakeratosis and deep invaginations in the subepithelial tissue. At lamina propria, we observed bundles of irregularly arranged collagen fiber, some fibroblasts, numerous capillary vessels and a large diffuse infiltration of plasma cells. Ultrastructurally, many fibroblasts, collagen fibers, collagen fibrils were present in lamina propria. On the basis of the data collected, we propose that the morphological features of the dimensional increase in gingival tissue associated with CsA treatment in kidney transplant patients may be considered proliferative fibroblasts, collagen fibers, collagen fibrils in lamina propria.

• Tissue Engineering and Regenerative Medicine
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• v.15 no.6
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• pp.711-719
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• 2018

BACKGROUND: Collagen organization within tissues has a critical role in wound regeneration. Collagen fibril diameter, arrangements and maturity between connective tissue growth factor (CTGF) small interfering RNA (siRNA) and mismatch scrambled siRNA-treated wound were compared to evaluate the efficacy of CTGF siRNA as a future implement for scar preventive medicine. METHODS: Nanocomplexes of CTGF small interfering RNA (CTGF siRNA) with cell penetrating peptides (KALA and $MPG^{{\Delta}NLS}$) were formulated and their effects on CTGF downregulation, collagen fibril diameter and arrangement were investigated. Various ratios of CTGF siRNA and peptide complexes were prepared and down-regulation were evaluated by immunoblot analysis. Control and CTGF siRNA modified cells-populated collagen lattices were prepared and rates of contraction measured. Collagen organization in rabbit ear 8 mm biopsy punch wound at 1 day to 8 wks post injury time were investigated by transmission electron microscopy and histology was investigated with Olympus System and TS-Auto software. CONCLUSION: CTGF expression was down-regulated to 40% of control by CTGF siRNA/KALA (1:24) complexes (p<0.01) and collagen lattice contraction was inhibited. However, down-regulated of CTGF by CTGF $siRNA/MPG^{{\Delta}NLS}$ complexes was not statistically significant. CTGF KALA-treated wound appeared with well formed-basket weave pattern of collagen fibrils with mean diameter of $128{\pm}22nm$ (n = 821). Mismatch siRNA/KALA-treated wound showed a high frequency of parallel small diameter fibrils (mean $90{\pm}20nm$, n = 563). CONCLUSION: Controlling over-expression of CTGF by peptide-mediated siRNA delivery could improve the collagen orientation and tissue remodeling in full thickness rabbit ear wound.

• Applied Microscopy
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• v.21 no.2
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• pp.1-13
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• 1991

In order to investigate the factors of the control mechanism of somitogenesis, early chick embryos (H-H stage $8{\sim}13$) were treated with heat shock, ${\alpha}$-amanitin and cycloheximide and morphological changes of somite were examined by light microscopy, transmission and scanning electron microscopy. In normal chick embryo, somites were formed from the somitomere which preexisted in segmental plate. Somites were wrapped with extracellular collagen fibrils and connected with neural tube, notochord and ectoderm. And then, somites were differentiated to sclerotome, dermatome and myotome by the interaction of nervous tissue. Abnormal somites were observed after formation of six or seven so mites in heat shock treated group. Amounts of collagen fibrils were obviously decreased in this group. In cycloheximide treated group, most so mites were smaller and neural tube formation was incomplete. Chromatins were condenced and formed several heterochromatins in the nucleus of somite cells. Lipid like cytoplasmic dense mass and lipid droplets were also observed. Segmentation of somites seemed to be normal progress in ${\alpha}$-amanitin treated group. Center of somite, however, hollowed in longitudinal sectioned samples. These results suggested that so mites were already existed in the segmental plate as the form of somitomere. Segmented somites were contacted with neural tube or notochord and the somites were tightly connected with each other by the extracellular collagen fibrils which were secreted from neuroepithelium and somite cells. Somites are thought to differentiate into sclerotome, dermatome and myotome by these interactions.

• Journal of Periodontal and Implant Science
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• v.23 no.2
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• pp.203-218
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• 1993

This study was designed to examine the effect of tetracycline application to the planed periodontal surgery of the experimentally induced periodontal disease in dogs. Modified Widman flap surgery was done and aqueous tetracycline (1%) was applied to the root surface for 5min, after which the wound was rinsed with saline, and flaps were coronally repositioned. Root surface ntoches were used as reference points. The animals were sacrificed 1 week, 2 weeks, 4 weeks, and 8 weeks after surgery, and block sections of tooth and surround tissue were processed for conventional light and electron microscopy. The results were as follows : 1. A more coronal position of junctional epithelium was observed in the area treated with tetracycline. 2. In the most of the tetracycline - treated teeth, the new collagen fibrils of connective tissue were oriented vertical/or oblique and parallel to the root surface. The vertical or oblique fibers were inserted into the denuded dentin matrix and contacted with exposed dentin collagen fibrils. 3. In the tetracycline - treated root, new cementum apposition, most of acellular extrinsic fiber cementum, was seen with bundles of oriented collagen fibrils incoporating into the cementum. 4. In the control and tetracyclin - treated teeth, bone resorption was observed at the alveolar crest in the 1 week groups.

• Journal of Food Hygiene and Safety
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• v.14 no.1
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• pp.45-54
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• 1999

The purpose of this study was to examine the increased bone loss caused by ovariectomy through monitoring the concentrations of the collagen and the pyridinoline crosslinks of collagen. The ovariectomized rats treated for 8 weeks, were divided at random into two or three groups of 10. Ovariectomies were carried out from the saline-treated group (Ovx), the estrogentreated group (Ovx+ES) and chondroitin sulfate-treated group (Ovx+CS). Sham operations were performed on the sham-operated group (Sham). Ovx+ES and Ovx+CS groups showed the remarkably increased collagen and pyridinoline amount in the bone and cartilage compared to Ovx group. And as the result of the measurement of SOD, Catalase and GPx which are antioxidant enzyme, SOD and Catalase activities in Ovx group were much higher than in Sham group. But they were significantly decreased in Ovx+CS group. Based on these results, it is supposed that estrogen and condroitin sulfate can enhance collagen synthesis and affect the pyridinoline formation in collagen fibrils through stimulating lysyl oxidase activity. And it is also thought that chondroitin sulfate can inhibit aging by reducing antioxidant enzyme.