| 1. |
- Brackmann, Christian, 1973, et al.
(författare)
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Coherent anti-Stokes Raman scattering microscopy of human smooth muscle cells in bioengineered tissue scaffolds
- 2011
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Ingår i: Journal of Biomedical Optics. - : SPIE - International Society for Optical Engineering. - 1083-3668 .- 1560-2281. ; 16:2
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Tidskriftsartikel (refereegranskat)abstract
- The integration of living, human smooth muscle cells in biosynthesized cellulose scaffolds was monitored by nonlinear microscopy toward contractile artificial blood vessels. Combined coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy was applied for studies of the cell interaction with the biopolymer network. CARS microscopy probing CH(2)-groups at 2845 cm(-1) permitted three-dimensional imaging of the cells with high contrast for lipid-rich intracellular structures. SHG microscopy visualized the fibers of the cellulose scaffold, together with a small signal obtained from the cytoplasmic myosin of the muscle cells. From the overlay images we conclude a close interaction between cells and cellulose fibers. We followed the cell migration into the three-dimensional structure, illustrating that while the cells submerge into the scaffold they extrude filopodia on top of the surface. A comparison between compact and porous scaffolds reveals a migration depth of <10 μm for the former, whereas the porous type shows cells further submerged into the cellulose. Thus, the scaffold architecture determines the degree of cell integration. We conclude that the unique ability of nonlinear microscopy to visualize the three-dimensional composition of living, soft matter makes it an ideal instrument within tissue engineering.
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| 2. |
- Braide, Magnus, 1955, et al.
(författare)
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Erythrocytes as Volume Markers in Experimental PD Show that Albumin Transport in the Extracellular Space Depends on PD Fluid Osmolarity
- 2016
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Ingår i: Peritoneal Dialysis International. - Toronto, Canada : SAGE Publications. - 0896-8608 .- 1718-4304. ; 36:3, s. 247-256
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Tidskriftsartikel (refereegranskat)abstract
- Background: Macromolecules, when used as intraperitoneal volume markers, have the disadvantage of leaking into the surrounding tissue. Therefore, Cr-51-labeled erythrocytes were evaluated as markers of intraperitoneal volume and used in combination with I-125-labeled bovine serum albumin to study albumin transport into peritoneal tissues in a rat model of peritoneal dialysis (PD). Methods: Single dwells of 20 mL of lactate-buffered filter-sterilized PD fluid at glucose concentrations of 0.5%, 2.5%, and 3.9% were performed for 1 or 4 hours. Tissue biopsies from abdominal muscle, diaphragm, liver, and intestine, and blood and dialysate samples, were analyzed for radioactivity. Results: The dialysate distribution volume of labeled erythrocytes, measured after correction for lymphatic clearance to blood, was strongly correlated with, but constantly 3.3 mL larger than, drained volumes. Erythrocyte activity of rinsed peritoneal tissue biopsies corresponded to only 1 mL of dialysate, supporting our utilization of erythrocytes as markers of intraperitoneal volume. The difference between the distribution volumes of albumin and erythrocytes was analyzed to represent the albumin loss into the peritoneal tissues, which increased rapidly during the first few minutes of the dwell and then leveled out at 2.5 mL. It resumed when osmotic ultrafiltration turned into reabsorption and, at the end of the dwell, it was significantly lower for the highest osmolarity PD fluid (3.9% glucose). Biopsy data showed the lowest albumin accumulation and edema formation in abdominal muscle for the 3.9% fluid. Conclusion: Labeled erythrocytes are acceptable markers of intraperitoneal volume and, combined with labeled albumin, provided novel kinetic data on albumin transport in peritoneal tissues.
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| 3. |
- Bäckdahl, Henrik, 1977, et al.
(författare)
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Engineering microporosity in bacterial cellulose scaffolds
- 2008
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Ingår i: Journal of tissue engineering and regenerative medicine. - : Hindawi Limited. - 1932-6254 .- 1932-7005. ; 2:6, s. 320-330
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Tidskriftsartikel (refereegranskat)abstract
- The scaffold is an essential component in tissue engineering. A novel method to prepare threedimensional (3D) nanofibril network scaffolds with controlled microporosity has been developed. By placing paraffin wax and starch particles of various sizes in a growing culture of Acetobacter xylinum, bacterial cellulose scaffolds of different morphologies and interconnectivity were prepared. Paraffin particles were incorporated throughout the scaffold, while starch particles were found only in the outermost area of the resulting scaffold. The porogens were successfully removed after culture with bacteria and no residues were detected with electron spectroscopy for chemical analysis (ESCA) or Fourier transform infra-red spectroscopy (FT-IR). Resulting scaffolds were seeded with smooth muscle cells (SMCs) and investigated using histology and organ bath techniques. SMC were selected as the cell type since the main purpose of the resulting scaffolds is for tissue engineered blood vessels. SMCs attached to and proliferated on and partly into the scaffolds. Copyright © 2008 John Wiley & Sons, Ltd.
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| 4. |
- Carlsson, Stina K., et al.
(författare)
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Adenosine A2 receptor presence and synergy with cholinergic stimulation in rabbit lacrimal gland.
- 2010
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Ingår i: Current eye research. - : Informa UK Limited. - 1460-2202 .- 0271-3683. ; 35:6, s. 466-74
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Tidskriftsartikel (refereegranskat)abstract
- Secretion from the lacrimal gland is an important part of the well-being of the eye, and a central part in the search for treatment of dry eye syndrome. Adenosine has stimulatory effects on the lacrimal gland, and can potentiate the effect of the cholinergic agonist carbachol (Cch). The aim of the present study is to investigate the presence of the adenosine A(2) receptor subtypes A(2A) and A(2B) in the rabbit lacrimal gland, and to characterize their role in regulated acinar cell secretion.
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| 5. |
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| 6. |
- Delbro, Dick, 1950, et al.
(författare)
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Demonstration of P2Y4 purinergic receptors in the HT-29 human colon cancer cell line
- 2005
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Ingår i: Autonomic & autacoid pharmacology. - 1474-8665. ; 25:4, s. 163-6
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Tidskriftsartikel (refereegranskat)abstract
- 1 The aim of the current study was to investigate the existence of P 2 Y(4) purinergic receptors in the HT-29 human colon cancer cell line. 2 We utilized Western blots and immunocytochemistry for the analysis. 3 Western blotting demonstrated two bands that could not be found after the antibody had been preabsorbed with the control peptide, suggesting that both bands are related to the P 2 Y(4) purinergic receptor. 4 Immunocytochemistry showed immunoreactivity for the P 2 Y(4) purinergic receptor localized in the cytoplasm of the HT-29 cells. 5 This is the first demonstration of the protein expression of P 2 Y(4) purinergic receptors in a human colon cancer cell line.
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| 7. |
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| 8. |
- Delbro, Dick, 1950, et al.
(författare)
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In inflammatory reactive astrocytes co-cultured with brain endothelial cells nicotine-evoked Ca(2+) transients are attenuated due to interleukin-1beta release and rearrangement of actin filaments.
- 2009
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Ingår i: Neuroscience. - : Elsevier BV. - 0306-4522. ; 159:2, s. 770-9
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Tidskriftsartikel (refereegranskat)abstract
- The aim of this study was to investigate whether nicotine acetylcholine receptors (nAChRs) are expressed in a more pronounced way in astrocytes co-cultured with microvascular endothelial cells from adult rat brain, compared with monocultured astrocytes, as a sign of a more developed signal transduction system. Also investigated was whether nicotine plays a role in the control of neuroinflammatory reactivity in astrocytes. Ca(2+) imaging experiments were performed using cells loaded with the Ca(2+) indicator Fura-2/AM. Co-cultured astrocytes responded to lower concentrations of nicotine than did monocultured astrocytes, indicating that they are more sensitive to nicotine. Co-cultured astrocytes also expressed a higher selectivity for alpha7nAChR and alpha4/beta2 subunits and evoked higher Ca(2+) transients compared with monocultured astrocytes. The Ca(2+) transients referred to are activators of Ca(2+)-induced Ca(2+) release from intracellular stores, both IP(3) and ryanodine, triggered by influx through receptor channels. The nicotine-induced Ca(2+) transients were attenuated after incubation with the inflammatory mediator lipopolysaccharide (LPS), but were not attenuated after incubation with the pain-transmitting peptides substance P and calcitonin-gene-related peptide, nor with the infection and inflammation stress mediator, leptin. Furthermore, LPS-induced release of interleukin-1beta (IL-1beta) measured by enzyme-linked immunosorbent assay (ELISA) was more pronounced in co-cultured versus monocultured astrocytes. Incubation with both LPS and IL-1beta further attenuated nicotine-induced Ca(2+) response. We also found that LPS and IL-1beta induced rearrangement of the F-actin filaments, as measured with an Alexa488-conjugated phalloidin probe. The rearrangements consisted of increases in ring formations and a more dispersed appearance of the filaments. These results indicate that there is a connection between a dysfunction of nicotine Ca(2+) signaling in inflammatory reactive astrocytes and upregulation of IL-1beta and the rearrangements of actin filaments in the cells.
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| 9. |
- Delbro, Dick, 1950-, et al.
(författare)
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The extracellular matrix-degrading protein ADAMTS5 is expressed in the nuclei of urothelial cells in healthy rats
- 2018
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Ingår i: Scandinavian Journal of Urology. - : Medical Journals Sweden AB. - 2168-1805 .- 2168-1813. ; 52:2, s. 139-142
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Tidskriftsartikel (refereegranskat)abstract
- Objective: The aim of this study was to investigate whether protein expression of the extracellular matrix-degrading protease ADAMTS5 can be demonstrated in the urinary bladder of healthy rats, and, if so, to determine the localization of this enzyme. Materials and methods: The experiments were conducted with eight inbred male Sprague-Dawley rats. Immunohistochemistry was used to investigate the expression of ADAMTS5 in the urinary bladder. Negative controls were established by either excluding the primary antibody or applying the antibody after it had been preabsorbed with its immunogenic peptide. Confocal microscopy was used to visualize the distribution of ADAMTS5 in the urinary bladder tissue. Results: Immunoreactivity for ADAMTS5 was demonstrated in the urothelium and in the detrusor. This expression was localized not only in the cytoplasm, but also in the nuclei. Confocal microscopy corroborated these findings. Conclusion: Expression of ADAMTS5 was demonstrated in the cytoplasm as well as in the nuclei of the urothelium and detrusor cells, suggesting that it may play a role at the transcriptional level.
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| 10. |
- Edman, Maria C, et al.
(författare)
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Functional expression of the adenosine A1 receptor in rabbit lacrimal gland.
- 2008
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Ingår i: Experimental eye research. - : Elsevier BV. - 0014-4835. ; 86:1, s. 110-7
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Tidskriftsartikel (refereegranskat)abstract
- It has become increasingly clear that purine compounds play a mediator role in exocrine secretion. Therefore, the present study was aimed at examining the presence of the adenosine A1 receptor in rabbit lacrimal gland and to evaluate the role of the A1 receptor in regulated secretion. The expression of the A1 receptor was investigated with reverse transcriptase PCR, cyto- and histochemistry as well as with pharmacological methods. Acinar cells were isolated, cultured in a serum-free medium for 2 days and thereafter treated with purinergic agonists/antagonists and/or carbachol and VIP. Secretory response was assessed by measuring secreted beta-hexosaminidase enzymatic activity. Microscopical evaluation of the immunocyto- and histochemistry specimens indicated that the adenosine A1 receptor is expressed in the rabbit lacrimal gland, which was also supported by reverse transcriptase PCR resulting in a sequence 100% identical with the previously published sequence for the rabbit A1 receptor gene. Incubation of acinar cells with adenosine and the A1 specific agonist N6-cyclopentyladenosine (CPA) resulted in a fourfold increase of secretion at the determined optimal concentrations. Incubation with carbachol alone resulted in a 10- to 15-fold increase. Carbachol combined with either adenosine or CPA increased the secretion 20-fold or more, demonstrating a synergistic effect. Our data provides evidence for the presence of adenosine A1 receptors in both tissue and cultured cells. Even though adenosine and CPA alone had only a moderate effect on secretion, the observed synergistic effect with carbachol suggests a modulatory role for the adenosine A1 receptor in the lacrimal gland.
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