EDTA treatment diminishes the antibacterial and anti-adherence effect of calcium hydroxide on Enterococcus faecalis: an in vitro study

Biofilms ◽

10.1017/s147905050800224x ◽

2008 ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

S. George ◽

A. Kishen

Keyword(s):

Cell Surface ◽

Calcium Hydroxide ◽

Enterococcus Faecalis ◽

Type I Collagen ◽

Membrane Integrity ◽

Cell Surface Hydrophobicity ◽

Prior Exposure ◽

Type I ◽

Bacterial Cells

ABSTRACTThis study sought to understand the cell surface characteristics, viability and biofilm-forming potential ofEnterococcus faecaliscells sequentially exposed to EDTA and calcium hydroxide, as in endodontic treatment. Bacterial cells exposed to EDTA and calcium hydroxide were assayed for cell viability, membrane integrity, cell surface hydrophobicity and surface charge, while alteration in the surface topography ofE. faecaliscells was examined using atomic force microscopy (AFM). The bacterial adherence potential to type I collagen was also examined to assess the biofilm-forming capacity ofE. faecaliscells exposed to EDTA and calcium hydroxide. It was found that calcium hydroxide treatment reduced the viability ofE. faecalis. However, prior exposure to EDTA significantly reduced the antibacterial effect of calcium hydroxide (P< 0.05). Calcium hydroxide treatment resulted in impaired cell wall morphology, observed as increased surface roughness and pore formation under AFM. However, these topographical changes induced by calcium hydroxide were significantly reduced in EDTA pretreated cells (P< 0.05). Calcium hydroxide treatment caused reduction in hydrophobicity and adherence ofE. faecalisto type I collagen. These effects due to calcium hydroxide were also significantly altered in EDTA-pretreated cells (P< 0.001). The findings from this study showed that the antibacterial and anti-adherence effect of calcium hydroxide was diminished by prior exposure ofE. faecaliscells to EDTA.

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MT1-MMP–dependent neovessel formation within the confines of the three-dimensional extracellular matrix

The Journal of Cell Biology ◽

10.1083/jcb.200405001 ◽

2004 ◽

Vol 167 (4) ◽

pp. 757-767 ◽

Cited By ~ 223

Author(s):

Tae-Hwa Chun ◽

Farideh Sabeh ◽

Ichiro Ota ◽

Hedwig Murphy ◽

Kevin T. McDonagh ◽

...

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Cell Surface ◽

Type I Collagen ◽

Ex Vivo ◽

Three Dimensional ◽

Collagenolytic Activity ◽

Type I ◽

Ex Vivo Model

During angiogenesis, endothelial cells initiate a tissue-invasive program within an interstitial matrix comprised largely of type I collagen. Extracellular matrix–degradative enzymes, including the matrix metalloproteinases (MMPs) MMP-2 and MMP-9, are thought to play key roles in angiogenesis by binding to docking sites on the cell surface after activation by plasmin- and/or membrane-type (MT) 1-MMP–dependent processes. To identify proteinases critical to neovessel formation, an ex vivo model of angiogenesis has been established wherein tissue explants from gene-targeted mice are embedded within a three-dimensional, type I collagen matrix. Unexpectedly, neither MMP-2, MMP-9, their cognate cell-surface receptors (i.e., β3 integrin and CD44), nor plasminogen are essential for collagenolytic activity, endothelial cell invasion, or neovessel formation. Instead, the membrane-anchored MMP, MT1-MMP, confers endothelial cells with the ability to express invasive and tubulogenic activity in a collagen-rich milieu, in vitro or in vivo, where it plays an indispensable role in driving neovessel formation.

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Pericellular coat of chick embryo chondrocytes: structural role of hyaluronate.

The Journal of Cell Biology ◽

10.1083/jcb.99.6.2114 ◽

1984 ◽

Vol 99 (6) ◽

pp. 2114-2122 ◽

Cited By ~ 47

Author(s):

R L Goldberg ◽

B P Toole

Keyword(s):

Cell Surface ◽

Chondroitin Sulfate ◽

Type I Collagen ◽

Type Ii Collagen ◽

Enzyme Linked Immunosorbent Assay ◽

Type I ◽

Type Ii ◽

Culture Dish ◽

Tissue Culture Dish

Chondrocytes produce large pericellular coats in vitro that can be visualized by the exclusion of particles, e.g., fixed erythrocytes, and that are removed by treatment with Streptomyces hyaluronidase, which is specific for hyaluronate. In this study, we examined the kinetics of formation of these coats and the relationship of hyaluronate and proteoglycan to coat structure. Chondrocytes were isolated from chick tibia cartilage by collagenase-trypsin digestion and were characterized by their morphology and by their synthesis of both type II collagen and high molecular weight proteoglycans. The degree of spreading of the chondrocytes and the size of the coats were quantitated at various times subsequent to seeding by tracing phase-contrast photomicrographs of the cultures. After seeding, the chondrocytes attached themselves to the tissue culture dish and exhibited coats within 4 h. The coats reached a maximum size after 3-4 d and subsequently decreased over the next 2-3 d. Subcultured chondrocytes produced a large coat only if passaged before 4 d. Both primary and first passage cells, with or without coats, produced type II collagen but not type I collagen as determined by enzyme-linked immunosorbent assay. Treatment with Streptomyces hyaluronidase (1.0 mU/ml, 15 min), which completely removed the coat, released 58% of the chondroitin sulfate but only 9% of the proteins associated with the cell surface. The proteins released by hyaluronidase were not digestible by bacterial collagenase. Monensin and cycloheximide (0.01-10 microM, 48 h) caused a dose-dependent decrease in coat size that was linearly correlated to synthesis of cell surface hyaluronate (r = 0.98) but not chondroitin sulfate (r = 0.2). We conclude that the coat surrounding chondrocytes is dependent on hyaluronate for its structure and that hyaluronate retains a large proportion of the proteoglycan in the coat.

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Collagen fibrillogenesis in vitro: interaction of types I and V collagen

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142670 ◽

1986 ◽

Vol 44 ◽

pp. 210-211

Author(s):

Arthur J. Wasserman ◽

Kathy C. Kloos ◽

David E. Birk

Keyword(s):

Type I Collagen ◽

Corneal Stroma ◽

Minor Component ◽

Homogeneous Distribution ◽

Type I ◽

Type V Collagen ◽

Fibril Morphology ◽

Type V ◽

In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

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Restoration of Osteogenesis by CRISPR/Cas9 Genome Editing of the Mutated COL1A1 Gene in Osteogenesis Imperfecta

Journal of Clinical Medicine ◽

10.3390/jcm10143141 ◽

2021 ◽

Vol 10 (14) ◽

pp. 3141

Author(s):

Hyerin Jung ◽

Yeri Alice Rim ◽

Narae Park ◽

Yoojun Nam ◽

Ji Hyeon Ju

Keyword(s):

Osteogenesis Imperfecta ◽

Type I Collagen ◽

Disease Modeling ◽

Bone Fragility ◽

Osteogenic Potential ◽

Type I ◽

Gene Correction ◽

Col1a1 Gene ◽

Collagen Formation

Osteogenesis imperfecta (OI) is a genetic disease characterized by bone fragility and repeated fractures. The bone fragility associated with OI is caused by a defect in collagen formation due to mutation of COL1A1 or COL1A2. Current strategies for treating OI are not curative. In this study, we generated induced pluripotent stem cells (iPSCs) from OI patient-derived blood cells harboring a mutation in the COL1A1 gene. Osteoblast (OB) differentiated from OI-iPSCs showed abnormally decreased levels of type I collagen and osteogenic differentiation ability. Gene correction of the COL1A1 gene using CRISPR/Cas9 recovered the decreased type I collagen expression in OBs differentiated from OI-iPSCs. The osteogenic potential of OI-iPSCs was also recovered by the gene correction. This study suggests a new possibility of treatment and in vitro disease modeling using patient-derived iPSCs and gene editing with CRISPR/Cas9.

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Effects of chitosan-collagen dressing on wound healing in vitro and in vivo assays

Journal of Applied Biomaterials & Functional Materials ◽

10.1177/2280800021989698 ◽

2021 ◽

Vol 19 ◽

pp. 228080002198969

Author(s):

Min-Xia Zhang ◽

Wan-Yi Zhao ◽

Qing-Qing Fang ◽

Xiao-Feng Wang ◽

Chun-Ye Chen ◽

...

Keyword(s):

Wound Healing ◽

Wound Dressing ◽

Type I Collagen ◽

Inhibition Zone ◽

Negative Control ◽

Type I ◽

Nih3t3 Cells ◽

Post Operation

The present study was designed to fabricate a new chitosan-collagen sponge (CCS) for potential wound dressing applications. CCS was fabricated by a 3.0% chitosan mixture with a 1.0% type I collagen (7:3(w/w)) through freeze-drying. Then the dressing was prepared to evaluate its properties through a series of tests. The new-made dressing demonstrated its safety toward NIH3T3 cells. Furthermore, the CCS showed the significant surround inhibition zone than empty controls inoculated by E. coli and S. aureus. Moreover, the moisture rates of CCS were increased more rapidly than the collagen and blank sponge groups. The results revealed that the CCS had the characteristics of nontoxicity, biocompatibility, good antibacterial activity, and water retention. We used a full-thickness excisional wound healing model to evaluate the in vivo efficacy of the new dressing. The results showed remarkable healing at 14th day post-operation compared with injuries treated with collagen only as a negative control in addition to chitosan only. Our results suggest that the chitosan-collagen wound dressing were identified as a new promising candidate for further wound application.

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In vitro phosphorylation of type I collagen by cyclic AMP-dependent protein kinase.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)57267-2 ◽

1986 ◽

Vol 261 (12) ◽

pp. 5674-5679

Author(s):

D B Glass ◽

J M McPherson

Keyword(s):

Protein Kinase ◽

Cyclic Amp ◽

Type I Collagen ◽

Type I ◽

Dependent Protein Kinase ◽

Dependent Protein

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Relative rates of biosynthesis of collagen type I, type V and type VI in calf cornea

Biochemical Journal ◽

10.1042/bj2740615 ◽

1991 ◽

Vol 274 (2) ◽

pp. 615-617 ◽

Cited By ~ 32

Author(s):

P Kern ◽

M Menasche ◽

L Robert

Keyword(s):

Type I Collagen ◽

Collagen Type ◽

Corneal Stroma ◽

Collagen Type I ◽

Type I ◽

Type Vi Collagen ◽

Sds Page ◽

Proline Incorporation ◽

Type V

The biosynthesis of type I, type V and type VI collagens was studied by incubation of calf corneas in vitro with [3H]proline as a marker. Pepsin-solubilized collagen types were isolated by salt fractionation and quantified by SDS/PAGE. Expressed as proportions of the total hydroxyproline solubilized, corneal stroma comprised 75% type I, 8% type V and 17% type VI collagen. The rates of [3H]proline incorporation, linear up to 24 h for each collagen type, were highest for type VI collagen and lowest for type I collagen. From pulse-chase experiments, the calculated apparent half-lives for types I, V and VI collagens were 36 h, 10 h and 6 h respectively.

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