Abstract:
:The efficacy of connective tissue explants is difficult to determine, particularly where autopsy material is required for research or clinical applications. We report here an optimised protocol using 5-chloromethylfluorescein diacetate (CMFDA) and ethidium homodimer-1 to distinguish viable and non-viable cells in a range of connective tissue explants. Biopsies and explants of corneae, arteries, cartilage and skin were loaded with fluoroprobes for extended periods (< or = 24h) at low temperatures (4 degrees C), fixed in paraformaldehyde, and processed using a variety of embedding, sectioning, autoradiographic, and immunohistochemical procedures. Detection of fluorescent green CMFDA and red ethidium homodimer was achieved using epi-illuminated light or dual channel confocal microscopy, and clearly differentiated live from dead cells throughout the explants. Furthermore, the intracellular distribution of CMFDA provided superior images of cell shape and morphology not previously available using conventional histochemical techniques. Adaptations of this protocol could prove valuable in a variety of research and clinical applications.
journal_name
Connect Tissue Resjournal_title
Connective tissue researchauthors
Poole CA,Brookes NH,Gilbert RT,Beaumont BW,Crowther A,Scott L,Merrilees MJdoi
10.3109/03008209609028880subject
Has Abstractpub_date
1996-01-01 00:00:00pages
233-41issue
4eissn
0300-8207issn
1607-8438journal_volume
33pub_type
杂志文章abstract::Part of the hyaluronic acid (HA) synthesized in peripheral tissues enters the blood circulation through the lymph. It is rapidly taken up by the endothelial cells in the liver (half-life in blood is 2.5-5.5 minutes) and degraded. Pure primary cultures of liver endothelial cells were obtained by a newly developed techn...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208609001971
更新日期:1986-01-01 00:00:00
abstract::Keratocan is an extracellular matrix protein that belongs to the small leucine-rich proteoglycan family that also includes lumican, biglycan, decorin, mimecan, and fibromodulin. Members of this family are known to play a role in regulating cellular processes such as proliferation and modulation of osteoprogenitor line...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207.2010.546536
更新日期:2011-10-01 00:00:00
abstract::Heparin treated and aldehyde crosslinked rat aorta segments were implanted in infrarenal aorta of homologous rats. One year following aortic replacement, the subendothelial scar and the prosthetic remnants were excised. The scar and the host intima-media were incubated with 3H-valine for 4 h and extracted with 5 M gua...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208309005609
更新日期:1983-01-01 00:00:00
abstract::The fibrous collagens form the structural basis of all mammalian connective tissues, including the vasculature, dermis, bones, tendons, cartilage, and those tissues that support organs such as the heart, kidneys, liver, and lungs. The helical structure of collagen has been extensively studied but in addition to its he...
journal_title:Connective tissue research
pub_type: 杂志文章,评审
doi:10.3109/03008207.2010.511353
更新日期:2011-02-01 00:00:00
abstract::Although bone is a nanocomposite of mineral and collagen, mineral has been the more elusive component of study. A standard for bone mineral is clearly needed. We hypothesized that the most natural, least-processed bone mineral could be retrieved from the most highly mineralized bone. We therefore studied the rostrum o...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207.2013.769973
更新日期:2013-01-01 00:00:00
abstract::Osteogenesis imperfecta is a congenital disease commonly characterized by brittle bones and caused by mutations in the genes encoding Type I collagen, the single most abundant protein produced by the body. The oim model has a natural collagen mutation, converting its heterotrimeric structure (two α1 and one α2 chains)...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207.2014.923860
更新日期:2014-08-01 00:00:00
abstract::Depolymerization of Ch-4-S, Ch-6-S, HA and HP was observed by gel filtration after treatment with 2-10 N NaOH or Ba(OH)2 in the presence of NaBH4 at 80-83 degrees for 1-5 hr. Depolymerized products were isolated after these treatments. 4-Deoxy-alpha-L-threo-4-enohexopyranosyluronic acid residue as the nonreducing end ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207509152343
更新日期:1975-01-01 00:00:00
abstract::Conditions were established to stimulate human gingival fibroblast explant cultures to synthesize milligram quantities of the metalloproteinase proenzymes, prostromelysin and procollagenase. To stimulate enzyme production, cells were treated with 1 nM recombinant human IL-1 beta for approximately 7 days under serum fr...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008209009009812
更新日期:1990-01-01 00:00:00
abstract::Multiple hereditary exostoses (MHE) is an autosomal dominant disorder that affects about 1 in 50,000 children worldwide. MHE, also known as hereditary multiple exostoses (HME) or multiple osteochondromas (MO), is characterized by cartilage-capped outgrowths called osteochondromas that develop adjacent to the growth pl...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008207.2017.1394295
更新日期:2018-01-01 00:00:00
abstract:AIM OF THE STUDY:To further elucidate the pathogenesis of systemic sclerosis (SSc) an experimental avian model was used. The University of California at Davis line 200 (UCD-200) chickens spontaneously develop a SSc-like disease that has most features of human SSc with vascular effects, inflammation, autoimmunity, and f...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008207.2016.1182997
更新日期:2016-09-01 00:00:00
abstract::The eggshell is an highly ordered structure deposited in the distal oviduct and composed of calcium carbonate and an organic matrix which is believed to influence its fabric. We have identified ovotransferrin as an 80 kDa matrix protein observed at high concentration in the uterine fluid at the initial stage of shell ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008200109016840
更新日期:2001-01-01 00:00:00
abstract::Fras1 and the structurally related proteins Frem1, Frem2, and Frem3, comprise a novel family of extracellular matrix proteins, which localize in a similar fashion underneath the lamina densa of epithelial basement membranes. They are involved in the structural adhesion of the skin epithelium to its underlying mesenchy...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008200802148025
更新日期:2008-01-01 00:00:00
abstract::Activation of protein kinase C (PKC) can upregulate tooth eruption molecules such as osteoprotegerin (OPG) and vascular endothelial growth factor (VEGF). In this study, we examined the in vivo gene expression of classic isoforms of PKC in the dental follicle of postnatal rats. The expression level of PKC-alpha was sig...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008200490882628
更新日期:2004-01-01 00:00:00
abstract::To elucidate the mechanism of accumulation and fragmentation of hyaluronic acid (HA) under inflammatory conditions, we investigated the effect of proinflammatory cytokines on hyaluronic acid synthetase (HAS) mRNA expression using cultured rabbit synovial membrane cells. HASs mRNA levels were determined by real-time PC...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008200109005649
更新日期:2001-01-01 00:00:00
abstract::Idiopathic pulmonary fibrosis (IPF) is associated with aberrant repair, persistence of collagen deposition, and the development of vascular remodeling. However, the role of angiogenesis in the pathogenesis of IPF is still undetermined. The aim of this study was to evaluate the combined mRNA expression of vascular endo...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008200903056150
更新日期:2010-01-01 00:00:00
abstract::The connective tissue protein elastin is largely responsible for maintaining the elasticity of major blood vessels and lung tissue. Comparatively few studies have been made of the immunologic properties of elastin because of its high degree of insolubility and because of its apparent limited capacity to elicit precipi...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208109152131
更新日期:1981-01-01 00:00:00
abstract::Since cartilage is mainly an avascular tissue, chondrocytes exist in a low-level oxygen environment in vivo. In the present study, we investigated the effect of oxygen tension (20%, 5% and 1% gas phase oxygen concentrations) over a 20-day period on the extracellular matrix accumulation of bovine articular chondrocytes...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008200109014251
更新日期:2001-10-01 00:00:00
abstract::Quantitative levels of mRNAs coding for elastin, types I and III procollagen and gamma-actin were measured in porcine vascular material following balloon catheterization. A balloon catheter was introduced into the thoracic aorta and jugular vein of 3-6 week old pigs; following distention and six days of postoperative ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208809008059
更新日期:1988-01-01 00:00:00
abstract:UNLABELLED:Keratocystic odontogenic tumors (KCOTs) are locally invasive, rapidly proliferating cystic lesions of the jaw. The bone-invasive nature of these tumors has been previously associated with the expression of matrix metalloproteinases (MMPs), which degrade the extracellular matrix. The purpose of this study was...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207.2014.923875
更新日期:2014-08-01 00:00:00
abstract::The in-vitro fluoride treatment technique has been introduced to investigate the composite behavior of bone tissue. Bone tissue with different mechanical properties can be obtained by varying the concentration, pH and immersion time in fluoride ion solutions. The chemical and physical changes in intact pieces of bone ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:
更新日期:2002-01-01 00:00:00
abstract::A method is described for accurate and reproducible determination of wet and dry weights of small articular cartilage samples. The water content of the articular cartilage samples before and after treatment with selective degradative enzymes and "artificial fibrillation" was determined. It was found that the water con...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208209160266
更新日期:1982-01-01 00:00:00
abstract::The novel method of high voltage electron microscopic tomography (3D) has been applied for the first time to examine ultrastructural features and spatial relations between collagen fibrils and mineral crystals in a mouse mutant (oim/oim) which replicates a moderate to severe form of osteogenesis imperfecta. The animal...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008209509010825
更新日期:1995-01-01 00:00:00
abstract::Muscle-derived cells can differentiate into chondrogenic cells. In our present study, we investigated the pattern of expression of Sox9, a transcription factor known to play a key role in chondrogenesis, in a rat myoblastic cell line, L6. In addition, we evaluated expression of type II collagen and myogenic regulatory...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008200490514130
更新日期:2004-01-01 00:00:00
abstract::This study was designed to determine the effects of pulsed electromagnetic fields (PEMF) on the mRNA expression of the receptor activator of NF-kappa-B (RANK) and carbonic anhydrase II (CA II) in ovariectomized rat osteoclast-like cells. Marrow cells were harvested from femora and tibiae of rats, from which the ovarie...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008200902855917
更新日期:2010-01-01 00:00:00
abstract::Molecular distributions of the intermolecular cross-links in fetal bovine bone type I collagen fibrils were quantitatively determined and compared with those of periodontal ligament. Results indicated that Hyl and Lys residues in the COOH-terminal nonhelical peptide portions (residues 16C) of both alpha 1 chains were ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208909050006
更新日期:1989-01-01 00:00:00
abstract::The pharmacokinetics of desmosine following a 5 mg intravenous dose was determined in five sheep. The biological half-life (t 1/2 beta) of desmosine varied between 81-489 min. Urinary recovery of unchanged desmosine indicated that the compound is not extensively metabolized and that the primary route of elimination is...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208409013681
更新日期:1984-01-01 00:00:00
abstract::The thermal stability and solubility profiles of human hypertrophic scar collagen have been measured and compared with those of the collagens present in human normal scar, human normal skin and a lathyritic dermal tissue. The thermal stability profiles of the hypertrophic scar collagens were found to be very similar t...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008208409152138
更新日期:1984-01-01 00:00:00
abstract::Scanning electron microscopy and high angle X-ray diffraction were used to define the relationship between collagen and elastin of bovine aortic wall. The diffraction pattern shows on one hand that the broad rings at 4.5 A and 9 A, due to elastin, do not orient on stretching and on the other hand, that the collagen ri...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.3109/03008207709152610
更新日期:1977-01-01 00:00:00
abstract::To address the molecular mechanisms of cementogenesis, we have isolated dental follicle cells and examined them to see if they contain cementoblast progenitors. Dental follicle tissue was dissected from the root surface of bovine tooth germ and cells were released by digestion with bacterial collagenase. The released ...
journal_title:Connective tissue research
pub_type: 杂志文章
doi:10.1080/03008200290001023
更新日期:2002-01-01 00:00:00
abstract::Genetic diseases affecting tooth structure have been classified by the tissue affected enamel versus dentin, and their pattern of inheritance autosomal dominant, autosomal recessive, or X-linked. Advances in molecular genetics and the Human Genome Project have provided substantial progress regarding the identification...
journal_title:Connective tissue research
pub_type: 杂志文章,评审
doi:
更新日期:2003-01-01 00:00:00