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PubMed2006

A three-dimensional in vitro model of angiogenesis in the airway mucosa.

Bronchial asthma is an inflammatory disease characterized by chronic intermittent bronchoconstriction. A key feature of the disease is structural changes in the airway wall (airway remodeling) consistent with tissue growth and chronic wound healing including angiogenesis. The epithelium directs mesenchymal processes during both embryogenesis and wound healing, and thus we hypothesized that the bronchial epithelium plays a critical role in directing angiogenesis. To study angiogenesis in the airways, we have developed a three-dimensional (3-D) in vitro model of the airway mucosa that consists of normal differentiated human bronchial epithelial cells (NHBE), normal human lung fibroblasts (NHLF), and human umbilical vein endothelial cells (HUVEC). The HUVEC are coated on dextran beads and suspended in a fibrin gel approximately 2mm beneath a confluent monolayer of NHLF which are just beneath the confluent monolayer of differentiated NHBE. In the presence of fibroblasts, visible capillaries reaching lengths of up to 1mm sprout from the HUVEC-coated beads. Over 11 days in culture, the bronchial epithelium produces transforming growth factor-beta2 (TGFbeta2, 60pg/ml), significantly increases vascular endothelial growth factor (VEGF) more than 6-fold to a concentration of 1.85ng/ml, but does not significantly impact total network formation. Exogenous TGFbeta2 stimulates VEGF production in a dose-dependent fashion (0-400pg/ml) through a MAPK-dependent pathway, but also inhibits capillary network formation. We conclude that the bronchial epithelium produces biologically relevant concentrations of VEGF and TGFbeta2 in a 3-D model of the airway mucosa that may be useful in probing mechanisms of angiogenesis in asthma.

Bronchi↗
PubMed1996

[Culture of endothelial cells as an in vitro model for experimental and clinical research].

Culture of endothelial cell is gotten from human umbilical cord by enzymatic digestion. For the growth of cells in culture, medial RPMI 1640 with 20% mixed human serum (NHS) or 20% fetal calf serum (FCS), endothelial cell growth factor (ECGF) and heparin have been used. Plastic, 0.1% gelatin and fibronectin have been used as a fundament. Immune identification of endothelial cells was culture is performed by monoclonal antibodies on vWF:Ag. Homogeneous cell line in culture might be used as in vitro model in both experimental and practice work.

Cell Culture Techniques↗
PubMed1996

Comparison of in vitro models for the study of Mycobacterium tuberculosis invasion and intracellular replication.

We recently evaluated several tissue culture model systems for the study of invasion and intracellular multiplication of Mycobacterium tuberculosis. These model systems include a human alveolar pneumocyte epithelial cell line, a murine macrophage cell line (J774), and fresh human peripheral blood-derived macrophages. Our data indicated that the initial level of association of M. tuberculosis with human alveolar pneumocyte cells (2%) was less than that observed with fresh human peripheral blood macrophages (9%) or J774 murine macrophages (13%) within 6 h of the addition of the bacteria. M. tuberculosis replicated in association with the pneumocyte cells by more than 55-fold by day 7 postinfection. In contrast, total bacteria] growth in the J774 cells and human macrophages was considerably less, with increases of only fourfold and threefold, respectively, over the same 7-day period. Amikacin, an aminoglycoside antimicrobial agent, was added to inhibit the growth of extracellular bacteria after the initial 6-h infection period. Decreases in viable counts were observed in all three cell cultures within the first 3 days after infection. However, unlike the case with either macrophage culture, intracellular bacterial CFU obtained from the infected pneumocytes increased by fourfold by day 7 after the addition of amikacin. These data indicate that M. tuberculosis infects and multiplies intracellularly in human lung epithelial cells and that these cells may be an alternative in vitro model for the study of intracellular multiplication of M. tuberculosis in the human lung.

Animals↗
PubMed2005

Comparison of optical resolution with digital and standard fiberoptic cystoscopes in an in vitro model.

OBJECTIVES: To compare two different distal sensor flexible endoscopes with standard fiberoptic cystoscopes for optical resolution. Distal video sensor flexible endoscopy may represent the cusp of a new endoscopic revolution. METHODS: Twenty-three participants evaluated the optical resolution capabilities of six flexible cystoscopes. The two presently available distal sensor flexible cystoscopes, ACMI DCN-2010 digital flexible cystoscope and Olympus CYF-V EndoEYE flexible video cystoscope, were compared with the four leading fiberoptic cystoscopes, ACMI ACN-2 fiberoptic cystoscope, Olympus CYF-4 Cystofiberscope, Karl Storz 11272CU1 Flexible Cystoscope, and Richard Wolf Flexible Fibre Urethro-Cystoscope, in an in vitro model. The participants recorded the numerical digits they were able to visualize through five solutions with progressively increasing concentrations of hematoxylin dye (C1 to C5). The cystoscopic images were viewed on a 13-in. medical grade Olympus monitor with an Olympus Visera camera system. RESULTS: No differences were detected among the cystoscopes at the lowest concentrations of hematoxylin dye (C1 and C2). At C3 to C5, the mean percentage of visualized numbers for the ACMI and Olympus distal sensor flexible cystoscopes was significantly greater than for all fiberoptic cystoscopes (P <0.01). At the highest concentration (C5), the ACMI distal sensor cystoscope performed significantly better than did the Olympus distal sensor flexible cystoscopes (P <0.01), although at all other concentrations, the two digital video cystoscopes performed equivalently. CONCLUSIONS: The results of this in vitro study suggest that cystoscopes with distal sensor technology improve visibility in a simulated challenging working environment compared with fiberoptic cystoscopes.

Cystoscopes↗
PubMed1999

Fluoride release from orthodontic bonding agents: a comparison of three in vitro models.

The aim of this in vitro study was to compare fluoride release from two cement disc models (partially varnished and unvarnished) of three orthodontic bonding materials with fluoride release from the same materials when used to bond an orthodontic bracket onto a tooth surface. A resin-modified glass ionomer cement (Vitremer) and a compomer material (Dyract Ortho) were compared with a conventional resin adhesive (Right-On). Ten discs (3.0 mm diameter and 1.5 mm thick) of each material were manufactured in a silicone mould and ten premolar teeth were bonded with orthodontic brackets using each material. All the premolar teeth were covered with nail varnish up to the bracket periphery and five of the discs for each material were coated with nail varnish on both upper and lower surfaces which halved the surface area. The fluoride released into 2 ml of deionised water, from each tooth or disc, was measured at regular intervals over 60 days. At 60 days, cumulative fluoride release was highest from the unvarnished discs, less for the varnished discs and least from the tooth-bracket model for both test materials. Despite having twice the surface area, the unvarnished disc model released only 1.2 to 1.5 times more fluoride than the varnished disc model. Compared with the tooth-bracket model, the unvarnished discs released 3.0 to 4.5 times more fluoride, whereas the varnished discs released only 2.2 to 3.7 times more fluoride. For the materials tested in this study, halving the surface of the discs did not reduce the cumulative fluoride release by half. The tooth/bracket model with minimal bonding material demonstrated substantial cumulative fluoride release. The pattern of fluoride release was similar for all models tested.

Analysis of Variance↗
PubMed1987

Correlation between bactericidal activity of fosfomycin trometamol in an in vitro model of the urinary bladder and susceptibility testing.

The present study was undertaken to define an interpretative guideline for disk diffusion susceptibility testing with fosfomycin trometamol, a new antimicrobial agent which has been developed for the treatment of urinary tract infections. Two potencies of fosfomycin disk were used: 50 and 200 micrograms, prepared in the presence and absence of glucose-6-phosphate. To verify the reliability of the results obtained in susceptibility testing, we have also evaluated the bactericidal activity of fosfomycin trometamol versus sensitive and resistant strains in an 'in vitro' model simulating the hydrokinetic aspects involved in the treatment of bacterial cystitis. The data obtained evidenced the role of glucose-6-phosphate in antimicrobial susceptibility tests as well as the importance of the urinary antibiotic concentrations to define sensitive and resistant bacteria. On the basis of our results, we recommend that a 200-microgram disk of fosfomycin containing 50 micrograms of glucose-6-phosphate should be used in antimicrobial susceptibility testing with fosfomycin trometamol.

Computer Simulation↗
PubMedDate not supplied

Evaluation of setpoint temperature and pullback speed on vein adventitial temperature during endovenous radiofrequency energy delivery in an in-vitro model.

Endovenous occlusion of the greater saphenous vein with radiofrequency (RF) energy has been shown to be a safe and effective treatment for saphenofemoral reflux. Operating parameters initially developed for the VNUS Closure system were selected in order to provide sufficient energy to controllably and transmurally heat the vein wall to cause collagen contraction and destruction of the vein wall while limiting the degree of perivascular heating. Procedural advances, such as ultrasound-guided tumescent infiltration along the course of the vein to be treated, have provided an added level of thermal protection to the perivenous tissue during the application of RF energy. The purpose of this study was to evaluate the effect of raising the setpoint temperature from 85 degrees C to 90 degrees C while doubling the pullback speed from 3 to 6 cm/minute on vein adventitial temperature. An in vitro simulation of a peripheral vein surrounded by body tissue was created by fixing freshly excised bovine veins in beefsteak kept moist by normal saline. A VNUS RF generator, in conjunction with a VNUS Closure catheter, delivered temperature-controlled RF energy to the bovine vein intima. Adventitial temperatures were recorded as the catheter was withdrawn and peak temperatures from 4 treatment groups were analyzed. The mean peak adventitial temperatures for the groups in which the adventitia was lightly bathed in saline were 64.4 degrees C and 64.9 degrees C with a setpoint temperature of 85 degrees C at a pullback speed of 3 cm/minute and 90 degrees C at a pullback speed of 6 cm/minute, respectively. The mean peak adventitial temperatures for the groups in which the adventitia was bathed in a 2.0 mm layer of saline were 51.3 degrees C and 47.7 degrees C with a setpoint temperature of 85 degrees C at a pullback speed of 3 cm/minute and 90 degrees C at a pullback speed of 6 cm/minute, respectively. Temperature-controlled radiofrequency occlusion of the saphenous vein, performed using an intimal setpoint temperature of 90 degrees C and double the pullback speed of the historical VNUS Closure treatment (6 vs 3 cm/minute), produced comparable temperatures at the adventitial surface in an in vitro model. The addition of a 2.0 mm layer of saline significantly reduced the peak adventitial temperatures.

Animals↗
PubMed1994

Interleukin-6 does not stimulate rat myometrial contractions in an in vitro model.

PROBLEM: Interleukin-6 (IL-6) increases in culture-positive amniotic fluid in women with preterm labor. IL-6 stimulates the production of prostaglandins leading to increased uterine activity. METHODS: We tested the hypothesis that IL-6 increases myometrial activity through release of uterotonic mediators. We studied the effect of IL-6 on uterine contractions in the absence and presence of fetal membranes to determine if the effect was on myometrium alone or was mediated through fetal membranes/decidua. IL-6 in concentrations of 100, 10, 0.1 or 0 ng/ml was added to the maternal side of the dual chamber-fetal membrane-uterine muscle in vitro model. RESULTS: We found that 10 ng/ml of IL-6 alone, without fetal membranes, caused a significant decrease in uterine contractions over time (P < or = 0.01). This decrease was not observed with the addition of term, nonlabored fetal membranes. CONCLUSIONS: IL-6 in the presence or absence of membranes, over a four log fold dose range, did not stimulate uterine contractions.

Animals↗
PubMed2003

A novel in vitro model of trophoblast-mediated decidual blood vessel remodeling.

In vivo the extravillous trophoblasts (EVTs) penetrate the decidua and the first third of the myometrium to remodel the uterine spiral arteries and achieve the high-flow, low-resistance circulation characteristic of the intervillous space of the term placenta. Much of our understanding of these processes comes from histologic analysis of placental bed biopsies, a limited tissue source and one that can provide only a snapshot of a dynamic process. To better characterize these cellular interactions, we have developed an in vitro co-culture system in which first trimester villous explants are cultured at low oxygen tension in contact with 2-mm(2) sections of decidua parietalis from the same patient. Hematoxylin eosin counterstaining of paraffin sections shows that EVT columns form at the tips of the placental villi and adhere and penetrate the decidual surface. The decidual blood vessels in the path of the EVT show morphologic disruption. Immunohistochemical analysis of the co-cultures using both an endothelial specific anti-CD31 and an anti-smooth muscle actin antibody show a disruption of the integrity of the vessel lining together with a complete loss of organized smooth muscle actin surrounding the blood vessels. In contrast control decidua samples in the absence of placental villi exhibit blood vessels with a complete endothelial lining and an organized muscular sheath. Using both an anti-cytokeratin-7 and anti-Cdx-2 antibody specific to trophoblasts, we show that these changes coincide with invasion of the vessels by endovascular trophoblasts and penetration of the decidua by interstitial EVTs. No EVTs were found in the control decidua. Thus we conclude that this in vitro model mimics the physiologic change observed in vivo during trophoblast invasion into maternal decidual tissues, and as such it may provide useful information concerning the interactions between EVTs and decidual cells and vessels during early gestation.

Actins↗
PubMed1996

Functional characterization of the spontaneously transformed human umbilical vein endothelial cell line ECV304: use in an in vitro model of angiogenesis.

To gain insight into the role of the endothelial cell during the pathophysiology of the angiogenic response, investigators have isolated micro- and macrovascular endothelial cells from a wide range of both animal and human vessels, including the umbilical vein. Human umbilical vein endothelial cells (HUVECs) isolated from umbilical cords remain a readily available and popular source of endothelial cell. However, the isolation and culture of these cells have several disadvantages, including the risk of infection, exogenous growth factor requirement, and low proliferative capacity. The heterogeneity of endothelial cells from different vascular beds as well as the heterogeneity between HUVEC isolates from different cords can make the critical interpretation of results difficult. ECV304 is a unique spontaneously transformed human umbilical vein endothelial cell line. In this report, the novel use of ECV304 cells as an alternative to HUVECs in an in vitro model of angiogenesis using the reconstituted basement membrane extract (Matrigel) was investigated. ECV304 cells were characterized immunohistochemically and their angiogenic behavior on Matrigel was analyzed functionally by phase-contrast, electron, and time-lapse video microscopy. ECV304 cells had several practical advantages over HUVEC culture and in contrast to HUVECs, ECV304 cells exhibited an enhanced and highly reproducible capacity for in vitro angiogenesis. However, several differentiated functions were lost or reduced in the ECV304 cell line which also exhibited anomalous cytokeratin expression. ECV304 cells may provide novel insights into the mechanisms governing angiogenesis under both normal physiological and pathological conditions.

Animals↗
PubMed2006

An optimized in vitro model of the respiratory tract wall to study particle cell interactions.

As a part of the respiratory tissue barrier, lung epithelial cells play an important role against the penetration of the body by inhaled particulate foreign materials. In most cell culture models, which are designed to study particle-cell interactions, the cells are immersed in medium. This does not reflect the physiological condition of lung epithelial cells which are exposed to air, separated from it only by a very thin liquid lining layer with a surfactant film at the air-liquid interface. In this study, A549 epithelial cells were grown on microporous membranes in a two chamber system. After the formation of a confluent monolayer the cells were exposed to air. The morphology of the cells and the expression of tight junction proteins were studied with confocal laser scanning and transmission electron microscopy. Air-exposed cells maintained monolayer structure for 2 days, expressed tight junctions and developed transepithelial electrical resistance. Surfactant was produced and released at the apical side of the air-exposed epithelial cells. In order to study particle-cell interactions fluorescent 1 microm polystyrene particles were sprayed over the epithelial surface. After 4 h, 8.8% of particles were found inside the epithelium. This fraction increased to 38% after 24 h. During all observations, particles were always found in the cells but never between them. In this study, we present an in vitro model of the respiratory tract wall consisting of air-exposed lung epithelial cells covered by a liquid lining layer with a surfactant film to study particle-cell interactions.

Air↗
PubMed1992

An in vitro model for studying growth and effect of trauma and external agents on the cricoid at the cellular level.

Over the past several years it has become evident that expansion laryngotracheal surgery is effective in the treatment of laryngotracheal stenosis. Several clinical and animal studies have been performed to study the process of laryngotracheal stenosis and its treatment. However, there are still many questions that cannot be addressed by currently used clinical and animal research. Further indepth study of the behavior of the subglottis at the cellular level is necessary. We present an in vitro model for studying chondrocyte metabolism of the bovine cricoid. Cartilage was successfully grown in an explant culture system, and it was shown that the chondrocytes were metabolically active and responded to external agents. This model will serve to study the mechanism of growth and effects of trauma and external agents on the cricoid at the cellular level.

Animals↗
PubMed1999

Work of breathing-tidal volume relationship: analysis on an in vitro model and clinical implications.

OBJECTIVE: Work of breathing (WoB) is currently employed to assess the afterload on the respiratory muscles and to estimate the energy expenditure for breathing. Since WoB depends on the ventilated tidal volume (TV), WoB*L(-1), the indicized form of WoB has been employed as a measure of WoB which is independent of TV. Actually, the independence of WoB*L(-1) from the ventilated TV has never been demonstrated. The aim of this study was to verify the predicted TV-independence of WoB*L(-1) on an in vitro model. METHODS: Our experimental model was constituted as follows: two endotracheal tubes, with internal diameter measuring respectively 6.5 and 8.5 mm, were alternatively connected with two rubber balloons whose compliance was respectively 0.02 and 0.06 L/hPa; the system was mechanically ventilated at ten different tidal volumes, ranging from 0.3-1 l. Flow rate was kept constant (35 l/m) during the whole experiment. RESULTS: Both elastic components of the model showed a static volume-pressure relationship which was linear in the experimental range of TV. In all combinations of resistance and compliance WoB increased quadratically whereas WoB*L(-1) increased linearly with the growing TV (p < 0.001). CONCLUSIONS: These results demonstrate the TV-dependence of WoB*L(-1) and suggest that WoB*L(-1), if TV changes, cannot be considered as an index of respiratory muscle afterload and should not be used as a guide for weaning patients from the mechanical ventilation. Finally, we introduced a new parameter (WoB1L) which seems to be a more TV-independent measure of respiratory work.

Humans↗
PubMed2004

Interleukin-1beta attenuates endothelin B receptor-mediated airway contractions in a murine in vitro model of asthma: roles of endothelin converting enzyme and mitogen-activated protein kinase pathways.

BACKGROUND: Asthma is a chronic airway disease, known to involve several inflammatory mediators. Little is known about how these mediators interact in order to produce or attenuate even basic features of the disease, like airway hyper-reactivity and remodelling. Endothelin-1 (ET-1) and IL-1beta are two mediators suggested to play important roles in the induction of airway inflammation. OBJECTIVE: To investigate the interactions between ET-1 and IL-1beta, using a novel in vitro model of asthma, focusing on airway smooth muscle contractility. METHODS: Isolated murine tracheal segments were cultured from 1 to 8 days in the absence and presence of IL-1beta. The subsequent contractile responses to sarafotoxin 6c (S6c) (selective agonist for ETB receptor) and sarafotoxin 6b (S6b) (ETA and ETB receptor agonist) were recorded by a myographs system. In all experiments, ETB receptors were desensitized before the contractile response to S6b was recorded. Thus, the response to S6b is only mediated by ETA receptors in the present study. The mRNA expressions for ET-1 and endothelin (ET) receptors were quantified by real-time PCR. RESULTS: Organ culture in the presence of IL-1beta attenuated the maximal contraction induced by S6c, but not S6b. This reduction was concentration-dependent and was significant after 2, 4 and 8 days of culture. To investigate the mechanisms behind this, inhibitors for endothelin converting enzyme (ECE) phosphoramidon, c-JUN N-terminal kinase (JNK) SP600125, extracellular-signal-regulated kinase 1/2(ERK 1/2) PD98059 and p38 pathway SB203580 were used. Individually, SP600125 and PD98059, but not SB203580, could partly reverse the reduction induced by IL-1beta. An additional effect was obtained when SP600125 and PD98059 were combined. The mRNA expressions for ET-1 and ETB receptor were up- and down-regulated, respectively, by IL-1beta. CONCLUSION: Presence of IL-1beta in the airways attenuate the contractile response mediated via ETB receptors, an effect dependent on ECE, JNK and ERK 1/2 pathways.

Animals↗
PubMed2005

An in vitro model to measure the effect of a silver fluoride and potassium iodide treatment on the permeability of demineralized dentine to Streptococcus mutans.

BACKGROUND: Diamine silver fluoride (Ag(NH3)2F), referred to as AgF, has been used to reduce the incidence of caries in primary dentitions but has been limited by the associated staining of both teeth and restorative materials. The application of potassium iodide (KI), following AgF prevents staining but its effects on the ability of AgF to reduce caries are not known. The aim of this study was to develop an in vitro model that would provide an indication of the permeability of demineralized dentine to Streptococcus mutans after treatment of the dentine with AgF followed by KI. METHODS: Forty dentine discs were bonded to the base of forty 5mL polycarbonate screw top vials (that had had their bases removed), filled with nutrient medium, sterilized and placed into a continuous culture of S. mutans. Samples were divided into four groups as follows: 10 samples of demineralized dentine as a control, 10 samples of demineralized dentine treated with AgF/KI, 10 samples of demineralized dentine treated with KI and 10 samples of demineralized dentine treated with AgF. After two weeks the optical density of the growth medium chambers was measured to determine bacterial penetration and growth. Cultures were plated out to determine migration through the discs by S. mutans. RESULTS: S. mutans migrated through all dentine discs. However, the samples treated with AgF and AgF/KI had significantly lower optical densities than the corresponding controls. The range of optical densities was least amongst demineralized samples treated with AgF/KI. CONCLUSIONS: Under the conditions of this study, treatment of demineralized dentine discs with AgF followed by KI allowed the penetration of S. mutans. Based on optical density measurements, the treatment resulted in significantly fewer microorganisms being present subjacent to the discs treated with AgF and KI than the control discs at the end of the experimental period.

Bacteriological Techniques↗
PubMed1998

The hemodynamic effects of mechanical prosthetic valve type and orientation on fluid mechanical energy loss and pressure drop in in vitro models of ventricular hypertrophy.

BACKGROUND AND AIMS OF THE STUDY: When choosing a prosthetic replacement for a natural heart valve, one objective should be to minimize the workload placed on the heart. This workload can be raised by fluid mechanical energy losses imposed by the valve. For a patient with left ventricular hypertrophy, certain aortic valve types and orientations could be hemodynamically superior to others. METHODS: This study used a control volume analysis to investigate the effects of prosthetic mechanical aortic valve type and orientation on fluid mechanical energy losses in four in vitro models of the left ventricular outflow/aortic inflow tract in various degrees of hypertrophy. Flow visualization studies were performed to qualitatively validate this analysis. The two most commonly used mechanical valve designs were studied: the St. Jude Medical (SJM) bileaflet valve and the Medtronic Hall (MH) tilting disk valve. Experiments were performed in pulsatile flow at a constant heart rate of 60 beats per min for five valve type/orientation combinations. The stroke volume was varied between 40 and 120 ml in five increments for each model and valve/orientation studied. RESULTS: Valve type and orientation was found to have a significant effect on energy losses in these models (p < 0.05). Valve/orientation combinations with leaflets or disks approximately parallel to the proximal flow direction created lower energy losses than others. The MH valve in the 180 degrees orientation caused significantly less energy losses and pressure drops (orifice and recovered) than any of the SJM valve/orientations studied (p < 0.05). The SJM and MH valves in the 0 degree orientation were responsible for significantly more energy loss than other valve/orientations studied (p < 0.05). An aortic inflow tract model with severe (45 degrees) curvature created significantly more energy loss (p < 0.05) than those with less curvature (15 and 30 degrees). However, the insertion of an obstruction simulating a hypertrophic tissue outgrowth caused much more energy loss than increasing the severity of outflow tract curvature from 15 to 45 degrees. Both orifice pressure drop and recovered pressure drop had excellent linear correlations with energy losses found in these models. CONCLUSIONS: These results imply that: (i) prosthetic valve type and orientation should be considered when replacing the aortic valve of a hypertropic patient; (ii) removal of obstructions within the aortic inflow tract will decrease ventricular workload; and (iii) the Doppler-estimated pressure gradients commonly use by cardiologists to assess the performance of a prosthetic valve, correlate very well with left ventricular energy loss and work load.

Aortic Valve↗
PubMed1996

In vitro models to study wound healing fibroblasts.

Phenotypic and contractile properties of human fibroblasts from dermis and from an experimental wound model were studied in vitro. When cultured in monolayer, dermal fibroblasts had an elongated spindle shape, were small in diameter and grew at a high rate. Wound fibroblasts grew slowly and were large, star shaped and had cytoplasmic stress fibres. Smooth muscle alpha actin was detected in 10 percent of dermal cells, whereas 20-80 per cent of wound fibroblasts contained this protein in their cytoplasm. The contractile property of cells was evaluated using a three-dimensional cell culture model. Our results show that wound fibroblasts contract collagen gels during the first days more strongly than dermal fibroblasts. These results show that, in vitro, wound fibroblasts have greater contractile capacity than dermal cells. The significant proportion of wound fibroblasts containing alpha-smooth muscle actin suggests that alpha-smooth muscle actin ratio may be related to wound contraction.

Cells, Cultured↗
PubMed1995

Ultraviolet radiation activates the human elastin promoter in transgenic mice: a novel in vivo and in vitro model of cutaneous photoaging.

The major alteration in photoaged skin is the deposition of massive amounts of abnormal elastic material, termed solar elastosis. In previous work, it has been shown that solar elastosis is accompanied by increased abundance of elastin and fibrillin mRNAs and upregulation of elastin promoter activity. Using a transgenic mouse line, which expresses the human elastin promoter, linked to a chloramphenicol acetyltransferase reporter gene, in a tissue-specific and developmentally regulated manner, we investigated the effects of ultraviolet A radiation and ultraviolet B radiation on human elastin promoter activity in vivo and in vitro. Irradiation of mice with a single dose of ultraviolet B radiation (491.4 mJ/cm2) resulted in an increase up to 8.5-fold in promoter activity, whereas a more modest increase of 1.8-fold was measured with ultraviolet A radiation (38.2 J/cm2). In addition, in vitro studies revealed over a thirtyfold increase in elastin promoter activity in response to ultraviolet B radiation (5.5 mJ/cm2), whereas no change was measured in response to ultraviolet A radiation (2.2 J/cm2). These results confirm the role of ultraviolet B radiation in elastin promoter activation in photoaging, and identify ultraviolet A radiation as a contributing factor. This system should serve as a useful in vivo and in vitro model to study cutaneous photoaging, and for testing compounds that may protect against cutaneous photodamage.

Animals↗