| 1. |
- Backesjo, CM, et al.
(författare)
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Activation of Sirt1 decreases adipocyte formation during osteoblast differentiation of mesenchymal stem cells
- 2009
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 189:1-4, s. 93-97
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Tidskriftsartikel (refereegranskat)abstract
- Mesenchymal stem cells (MSC) can differentiate into osteoblasts, adipocytes, chondrocytes and myoblasts. It has been suggested that a reciprocal relationship exists between the differentiation of MSC into osteoblasts and adipocytes. Peroxisome proliferator-activated receptor γ2 (PPARγ2) is a key element for the differentiation into adipocytes. Activation of the nuclear protein deacetylase Sirt1 has recently been shown to decrease adipocyte development from preadipocytes via inhibition of PPARγ2. In vitro, MSC differentiate to osteoblasts when exposed to bone-inducing medium. However, adipocytes are also developed. In the present study we have targeted Sirt1 to control adipocyte development during differentiation of MSC into osteoblasts. The finding that resveratrol and isonicotinamide markedly inhibited adipocyte and promoted osteoblast differentiation demonstrates an interesting alternative to PPARγ antagonists. These results are important for the evolving field of cell-based tissue engineering, but may also be relevant in the search for new treatments of osteoporosis.
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| 2. |
- Barreto Henriksson, Helena, et al.
(författare)
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Development and Regeneration Potential of the Mammalian Intervertebral Disc
- 2013
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Ingår i: Cells Tissues and Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 197:1
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Forskningsöversikt (refereegranskat)abstract
- At the present time, the normal cell proliferation rate and regeneration processes in the intervertebral disc (IVD) are not fully known. Historically, the IVD has been considered an organ with little or no regenerative capacity. However, several studies have identified the presence of cells expressing progenitor/stem cell markers in adult cartilage tissue and recent data suggest that adult mammalian IVDs have regenerative capacity, albeit slow. The aim of this review is to give an overview of the present knowledge regarding IVD development, regeneration and repair mechanisms in mammals, with a special focus on human discs. At a time when regenerative medicine is making progress and biological treatment options, such as stem cell therapy, are suggested for patients with degenerated discs causing chronic low back pain, basic knowledge about disc cells and their regenerative capacity form a useful basis for the exploration of new treatment options.
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| 3. |
- Bloecker, K., et al.
(författare)
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Morphometric Differences between the Medial and Lateral Meniscus in Healthy Men - A Three-Dimensional Analysis Using Magnetic Resonance Imaging
- 2012
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 195:4, s. 353-364
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Tidskriftsartikel (refereegranskat)abstract
- The objective of this work was to characterize tibial plateau coverage and morphometric differences of the medial (MM) and lateral meniscus (LM) in a male reference cohort using three-dimensional imaging. Coronal multiplanar reconstructions of a sagittal double-echo steady state with water excitation magnetic resonance sequence (slice thickness: 1.5 mm, and in-plane resolution: 0.37 x 0.70 mm) were analyzed in 47 male participants without symptoms, signs or risk factors of knee osteoarthritis of the reference cohort of the Osteoarthritis Initiative. The medial and lateral tibial (LT) plateau cartilage area and the tibial, femoral and external surfaces of the MM and LM were manually segmented throughout the entire knee. This process was assisted by parallel inspection of a coronal intermediately weighted turbo spin echo sequence. Measures of tibial coverage, meniscus size, and meniscus position were computed three-dimensionally for the total menisci, the body, and the anterior and the posterior horn. The LM was found to cover a significantly greater (p < 0.001) proportion of the LT plateau (59 +/- 6.8%) than the MM of the medial plateau (50 +/- 5.5%). Whereas the volume of both menisci was similar (2.444 vs. 2.438 ml; p = 0.92), the LM displayed larger tibial and femoral surface areas (p < 0.05) and a smaller maximal (7.2 +/- 1.0 vs. 7.7 +/- 1.1 mm; p < 0.01) and mean thickness (2.7 +/- 0.3 vs. 2.8 +/- 0.3 mm; p < 0.001) than the medial one. Also, the LM displayed less (physiological) extrusion than the medial one. These data may guide strategies for meniscal tissue engineering and transplantation aiming to restore normal joint conditions. Copyright 2011 (C) S. Karger AG, Basel
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| 4. |
- Brisby, Helena, 1965, et al.
(författare)
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Moderate Physical Exercise Results in Increased Cell Activity in Articular Cartilage of the Knee Joint in Rats.
- 2013
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 198:3, s. 237-248
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Tidskriftsartikel (refereegranskat)abstract
- Background: Moderate exercise regimens have shown minor positive effects on matrix turnover in articular cartilage (AC), while effects at cellular level, e.g. proliferation, are scarcely described. Aim: The aim of this study was to investigate the effects of moderate exercise on cell proliferation and recruitment of cells possibly active in regeneration in different regions of cartilage in the rat knee joint. Methods: Eighteen rats were orally given 5-bromo-2-deoxyuridine (BrdU) for 14 days for in vivo DNA labeling. Nine rats underwent treadmill training for 50 min/day, 5 days/week (exercise group), and 9 rats served as controls (no exercise). Animals were sacrificed after 14, 56 and 105 days, and knee joints were harvested. BrdU+ cells were visualized immunohistochemically (IHC) and counted in AC, posterior stem cell niche (PN), potential migration route (PMR; area between PN and the AC border), potential migration area (PMA; region between PN and AC including PN) and epiphyseal cartilage plate (EP) of the tibia and femur. Results: Compared to controls, in the exercise group BrdU+ cells/mm(2) were increased on days 14 (p = 0.022) and 105 (p = 0.045) in AC of the tibia and on day 105 (p = 0.014) in AC of the femur. BrdU+ cell numbers were increased in the PMR region of the tibia on days 14 (p = 0.023) and 105 (p = 0.0018) and in the PMR region of the femur on day 105 (p = 0.0099) as well as in the PMA region of the tibia (p = 0.0008) and femur (p = 0.0080) on day 105. No significant differences in BrdU+ cells/mm(2) were seen in PN or EP between the groups at any time point. Regarding collagen 2A1 expression and proteoglycan accumulation, no significant differences between groups were detected. Conclusions: The results indicate increased cell activity in AC in response to physical exercise and may help to understand the complexity of AC regeneration in the normal mammal knee joint. © 2013 S. Karger AG, Basel.
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| 5. |
- Dare, Emma V., et al.
(författare)
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Genipin Cross-Linked Fibrin Hydrogels for in vitro Human Articular Cartilage Tissue-Engineered Regeneration
- 2009
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Ingår i: Cells Tissues Organs. - : Karger. - 1422-6405 .- 1422-6421. ; 190:6, s. 313-325
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Tidskriftsartikel (refereegranskat)abstract
- Our objective was to examine the potential of a genipin cross-linked human fibrin hydrogel system as a scaffold for articular cartilage tissue engineering. Human articular chondrocytes were incorporated into modified human fibrin gels and evaluated for mechanical properties, cell viability, gene expression, extracellular matrix production and subcutaneous biodegradation. Genipin, a naturally occurring compound used in the treatment of inflammation, was used as a cross-linker. Genipin cross-linking did not significantly affect cell viability, but significantly increased the dynamic compression and shear moduli of the hydrogel. The ratio of the change in collagen II versus collagen I expression increased more than 8-fold over 5 weeks as detected with real-time RT-PCR. Accumulation of collagen II and aggrecan in hydrogel extracellular matrix was observed after 5 weeks in cell culture. Overall, our results indicate that genipin appeared to inhibit the inflammatory reaction observed 3 weeks after subcutaneous implantation of the fibrin into rats. Therefore, genipin cross-linked fibrin hydrogels can be used as cell-compatible tissue engineering scaffolds for articular cartilage regeneration, for utility in autologous treatments that eliminate the risk of tissue rejection and viral infection.
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| 6. |
- Fossum, Magdalena, et al.
(författare)
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Long-term culture of human urothelial cells : a qualitative analysis
- 2005
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 181:1, s. 11-22
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Tidskriftsartikel (refereegranskat)abstract
- Today, in vitro culturing of autologous cells is an established method in the field of tissue reconstruction. It can be applied to urothelial cells and could have many clinical implications in urological reconstructive surgery. This development calls for quality controls concerning cells used for clinical treatment when cells are autotransplanted back to the patient. We have studied cultured cells in order to detect whether genetic or morphologic changes occur. Urothelial cells isolated from bladder lavage were cultured according to different protocols based on the presence or absence of feeder cells. Genetic studies were performed by means of karyotyping with standard G-banding and interphase fluorescent in situ hybridization (FISH) analyses. The morphology of these epithelial cells was judged as well as immunostaining for epithelial cell markers. In addition, to minimize the risk of feeder cell contamination, proliferation studies were performed on cultures including feeder cells that had been pretreated with different doses of mitomycin or radiation. In initial studies, when using feeder cells in each passage according to standard protocols, urothelial cells proliferated unfavourably after the fourth passage with increasing numbers of mouse cells as well as urothelial tetraploid cells. We could also show that urothelial cells from bladder lavage need feeder cells in order to establish primary cultures. Further propagation up to 14 passages was performed without feeder cells and the urothelial cells retained normal karyotypes. We also found that mitomycin treatment had its main effect on feeder cells during the first 2 h. When feeder cells were irradiated, 20 Gy was effective and no feeder cell contamination was seen. In conclusion, we found that a high standard of quality in urothelial cell culturing can be achieved with a careful culturing technique.
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| 7. |
- Gaida, James Edmund, et al.
(författare)
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Evidence of the TNF-α system in the human Achilles tendon : expression of TNF-α and TNF receptor at both protein and mRNA levels in the tenocytes
- 2012
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Ingår i: Cells Tissues Organs. - : S. Karger. - 1422-6405 .- 1422-6421. ; 196:4, s. 339-352
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Tidskriftsartikel (refereegranskat)abstract
- Understanding adaption to load is essential for prevention and treatment of tendinopathy/tendinosis. Cytokine release in response to load is one mechanism involved in mechanotransduction. The cytokine tumor necrosis factor alpha (TNF-α) is implicated in tendinosis and can induce apoptotic effects via tumor necrosis factor receptor 1 (TNFR1). The complete absence of information concerning the TNF-α system in Achilles tendon is a limitation as mid-portion Achilles tendinosis is very frequent. Purpose: To examine expression patterns of TNF-α and its two receptors (TNFR1 and TNFR2) in human Achilles tendinosis and control tissue and to biochemically confirm the presence of TNF-α in tendinosis tissue. Methods: TNF-α and TNFR1 mRNA were detected via in situ hybridization. TNF-α, TNFR1, and TNFR2 were demonstrated immunohistochemically. Apoptosis markers were utilized. ELISA was used to detect TNF-α. Results: TNF-α and TNFR1 mRNA was detected in tenocytes of both tendinosis and control tendons. Tenocytes from both groups displayed specific immunoreactions for TNF-α, TNFR1, and TNFR2. The widened/rounded tenocytes of tendinosis samples exhibited the most intense immunoreactions. Apoptosis was detected in only a subpopulation of the tenocytes in tendinosis tissue. TNF-α was measurable in tendinosis tissue. Inflammatory cells were not seen. Conclusion: This is the first evidence of the existence of the TNF-α system in the human Achilles tendon. Findings are confirmed at mRNA and protein levels as well as biochemically. The TNF-α system was in principle confined to the tenocytes. The connection between tenocyte morphology and the expression pattern of TNF-α, TNFR1, and TNFR2 suggests that the TNF-α system may be involved in tenocyte activation in Achilles tendinosis.
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| 8. |
- Gormus, U, et al.
(författare)
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Comparative Proteome Analyses of Ureteropelvic Junction Obstruction and Surrounding Ureteral Tissue
- 2020
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 209:1, s. 2-12
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Tidskriftsartikel (refereegranskat)abstract
- Ureteropelvic junction (UPJ) obstruction is a common problem in children, but its etiology remains unclear. In this study, the proteome profiles of the obstructed segment and its surrounding distal and proximal parts were comparatively evaluated. Twelve children younger than 2 years of age with unilateral intrinsic UPJ obstruction were included. The excised operational tissue was divided into three parts immediately after resection: the obstructed part (Obst), the distal normal ureteral part (Dist), and the proximal part of the obstructed segment (Prox). Proteins extracted from the tissue samples were subjected to two-dimensional gel electrophoresis analysis to identify differentially regulated proteins. Spot analysis revealed that four proteins, namely tropomyosin beta and alpha-1 chains, actin and desmin, were upregulated in Obst in comparison to Dist. A similar analysis between Obst and Prox showed that heat shock protein beta-1 and carbonic anhydrase-1 were upregulated in Obst, while tropomyosin alpha 3 chain and ATP synthase beta were upregulated in Prox. The last comparative analysis between Dist and Prox revealed upregulation of annexin-A5 and annexin-A1 in Dist and vimentin, mitochondrial ATP synthase subunit-beta, peroxiredoxin-2, and apolipoprotein-A1 in Prox. Bioinformatics analysis using the STRING server indicated that the differentially regulated proteins, altogether, point to the changes occurring in muscle filament sliding pathway. When regulations occurring in each group were mutually compared, a change in lipase inhibition activity was detected by STRING. This is the first study scrutinizing changes occurring in protein profiles in UPJ.
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| 9. |
- Gradin, P, et al.
(författare)
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Transgenic overexpression of tartrate-resistant acid phosphatase is associated with induction of osteoblast gene expression and increased cortical bone mineral content and density
- 2012
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 196:1, s. 68-81
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Tidskriftsartikel (refereegranskat)abstract
- Bone remodeling is a central event in the maintenance of skeletal tissue, and involves cycles of resorption followed by the formation of bone tissue. The activity of osteoclasts and osteoblasts during these cycles is tightly regulated by systemic and local factors coupling the action of these cells. Tartrate-resistant acid phosphatase (TRAP) is predominantly expressed in bone by osteoclasts but has also been detected in osteoblasts and osteocytes. Moreover, TRAP can stimulate the differentiation of mesenchymal lineage cells, i.e. progenitors of osteoblasts and adipocytes. In order to further explore the effects of TRAP on bone turnover, the structural and molecular phenotypes of osteoclasts and osteoblasts were assessed in TRAP-overexpressing transgenic mice. Transgenic mice of both sexes display increased cortical bone mineral content and density, which cannot be accounted for by decreased bone resorption since osteoclast numbers and resorptive activity do not differ from wild-type mice. Examination of the osteoblast phenotype revealed that markers of bone formation, i.e. procollagen type I N-terminal propeptides, and osteoblast lineage markers as well as the TRAP 1B mRNA transcript are increased in TRAP-overexpressing mice. Expression of the osteoclast-selective TRAP 1C mRNA is not increased in TRAP transgenic mice. Elevated expression of TRAP mRNA and protein were detected in osteoblasts, osteocytes and in the bone matrix of TRAP transgenic mice, suggesting that TRAP overexpression in osteoblast lineage cells is associated with increased cortical bone mineral content and density. The data presented here support the hypothesis that TRAP overexpression in the osteoblastic cell lineage stimulates the differentiation and/or activation of these cells.
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| 10. |
- Granberg, I, et al.
(författare)
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Capillary supply in relation to myosin heavy chain fibre composition of human intrinsic tongue muscles
- 2010
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 192:5, s. 303-313
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Tidskriftsartikel (refereegranskat)abstract
- The capillary supply and myosin heavy chain (MyHC) composition of three different intrinsic tongue muscles was analysed in the anterior and posterior regions of the human tongue with biochemical and immunohistochemical techniques. Mean capillary density for the whole tongue was 796 ± 82 cap/mm², without regional differences. The overall number of capillaries around each fibre (CAF) was higher in the posterior than in the anterior region (2.5 vs. 2.1, p = 0.009). However, correcting for regional differences in fibre size, CAF per fibre area was higher in the anterior region (4.3 vs. 3.0, p < 0.001). Muscle fibres containing fast MyHCs predominated in the anterior region (78.7%), consisting of MyHCIIa (58.5%), MyHCIIx (1.0%), MyHCIIa+MyHCIIx (11.3%) and MyHCI+MyHCIIa (7.9%). Fibres containing slow MyHC predominated in the posterior region (65.2%), consisting of MyHCI (45.5%) and MyHCI+MyHCIIa (19.7%). A minor fibre population (<2%) contained unusual MyHC isoforms, namely MyHC foetal, MyHC slow-tonic, MyHC α-cardiac or MyHC embryonic. The microvascularization of the human tongue was twice as high as in human limb muscles. Regional similarities in capillary supply, but differences in fibre phenotype composition, suggest that human tongue muscle fibres are fatigue resistant independently of MyHC content. High frequency of hybrid fibres, that is fibres co-expressing two or more MyHC isoforms, indicates a wider spectrum of fibre contractile properties than in limb muscles. In conclusion, human intrinsic tongue muscles showed internal specialization in distribution of MyHC isoforms and capillary supply, but not in the expression of unusual MyHCs.
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| 11. |
- Granéli, Cecilia, 1981, et al.
(författare)
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Virtual Ligand-Based Screening Reveals Purmorphamine Analogs with the Capacity to Induce the Osteogenic Differentiation of Human Mesenchymal Stem Cells.
- 2012
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 197:2, s. 89-102
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Tidskriftsartikel (refereegranskat)abstract
- Human mesenchymal stem cells (hMSCs) have extensive proliferative capacity, are able to self-renew and have the potential to differentiate into cells of the connective tissue lineages. These properties make them a putative cell type for tissue engineering applications, as well as a possible in vivo target for the pharmaceutical modulation of the differentiation processes. The aim of this study was to find one or more small-molecule substances that would enhance the osteogenic differentiation of hMSCs in vitro. The strategy used here was ligand-based virtual screening for substances similar to the previously suggested osteoinductive purmorphamine followed by an in vitro screening of the selected analogs in hMSCs isolated from bone marrow. We investigated the osteoinductive capacity of several purmorphamine analogs by determining the protein and gene expression of markers for osteogenic differentiation as well as the extracellular matrix (ECM) mineralization of these cells. Treatment with two candidate substances or purmorphamine resulted in increased levels of alkaline phosphatase (ALP) activity compared to the control. Other purmorphamine analogs demonstrated higher calcium deposition in the ECM after 5 weeks of osteogenic differentiation, compared to both purmorphamine and the control condition. The resulting substances, which had positive effects on the osteogenic differentiation, are promising as possible modes of treatment for bone-related diseases or defects that target and enhance the osteogenic differentiation of MSCs, in vitro or in vivo. Furthermore, the concept of combining the virtual ligand-based screening method with in vitro screening, using human adult stem cells as a possible strategy for drug discovery, is demonstrated.
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| 12. |
- Grimsholm, Ola, et al.
(författare)
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Expression patterns of neurotrophins and neurotrophin receptors in articular chondrocytes and inflammatory infiltrates in knee joint arthritis.
- 2008
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 188:3, s. 299-309
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: It is likely that neurotrophins (NTs) are of great importance for the articular cartilage and the inflammation process in arthritis. METHODS: The immunohistochemical expression of the NTs nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) and the associated receptors p75, TrkA and TrkB was examined in the knee joint of arthritic and healthy mice. RESULTS: Immunoreactions for NGF and BDNF were detected in cells and nerve fiber varicosities in the inflammatory infiltrates of the synovial tissue of arthritic joints but not in synovial tissue of controls. p75-immunoreactive nerve fiber-like strands were detected in inflammatory infiltrates. Immunostaining for NGF, BDNF, p75, TrkA and TrkB was noted in articular chondrocytes. There was a statistically significant decrease in reactions for NGF (p < 0.001), TrkA (p = 0.001) and p75 (p < 0.001) in articular chondrocytes in joints exhibiting severe arthritis. CONCLUSION: The findings show that an NT system develops in inflammatory infiltrates of the synovial tissue. Furthermore, most interestingly, autocrine/paracrine effects appear to exist concerning NTs for the articular chondrocytes. The downregulated expression of NGF and NT receptors in articular chondrocytes in arthritis is a new aspect concerning the involvement of NTs in cartilage.
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| 13. |
- Hagert, Elisabet, et al.
(författare)
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General innervation pattern and sensory corpuscles in the scapholunate interosseous ligament.
- 2004
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 177:1, s. 47-54
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Tidskriftsartikel (refereegranskat)abstract
- The scapholunate interosseous ligament (SLIL) is biomechanically important in maintaining wrist motion and grip strength in the hand, but its possible sensory role in the dynamic muscular stability of the wrist joint has not been examined. The aim of this study was to use immunohistochemical methods to analyze the general innervation and the possible existence of sensory corpuscles in the SLIL. The ligament was excised in its entirety from 9 patients. Antibodies against the low-affinity p75 neurotrophic receptor (p75) were used to reveal sensory corpuscles as well as general innervation. Furthermore, antibodies against the general nerve marker protein gene product 9.5 (PGP 9.5) and the glial marker S-100 were used to additionally depict innervation and corpuscular structures. Blood vessels occurred in areas interspersed throughout the homogeneous collagenous structure. In these vascularized areas, the SLIL was found to be supplied with nerve fascicles and sensory corpuscles of both the Ruffini and lamellated type. p75 immunoreactivity (IR) was detected in association with the nerve fascicles and the corpuscles, particularly in their capsule. S-100 IR was found in the Schwann cells in the central regions of the corpuscle, and PGP 9.5 IR marked the axonal structures in the corpuscles. New information on neurotrophin receptor distribution in ligaments has been obtained here. The presence of nerve fascicles and particularly sensory corpuscles in the SLIL suggests that the ligament has a proprioceptive role in the stability of the wrist. The marked p75 IR further indicates that neurotrophins play a part in a proprioceptive system in the ligament, given the importance of neurotrophins in maintaining sensory function.
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| 14. |
- Hellström, Martin, et al.
(författare)
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Expression of the CD44 receptor in the blood vessel system : an experimental study in rat.
- 2005
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 179:3, s. 102-108
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Tidskriftsartikel (refereegranskat)abstract
- The CD44 receptor is a transmembrane glycoprotein expressed on a variety of cells like endothelial, epithelial and smooth muscle cells. This molecule has many important functions, e.g. in cell-cell and cell-matrix interactions and signal transduction. The main ligand for CD44 is hyaluronan (HYA). HYA is a glycosaminoglycan with structural and cell biological properties. The localization of HYA in the vessel wall of arteries and veins in the healthy adult and newborn rat has been described earlier. In this study the occurrence of the CD44 receptor was investigated in the same vessels and compared to the localization of HYA. Both CD44 and its ligand showed an increased expression in the vessel wall of newborn rats compared to that of adult rats. Although HYA is abundant in the adventitia of adult rats, virtually no expression of CD44 was observed. Our results indicate that the CD44 receptor expression is increased during the stage of maturation of the vessel tree whereas the CD44 receptor is less needed by HYA in the healthy vessel wall.
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| 15. |
- Hellström, Martin, et al.
(författare)
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Hyaluronan is differently located in arteries and veins : an immunohistochemical study in the rat
- 2003
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 173:4, s. 227-233
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Tidskriftsartikel (refereegranskat)abstract
- The matrix components of the vessel wall are of great importance for the function of the vessel system of which both endurance and elasticity are prerequisites. One component of the vessel wall is the glycosaminoglycan hyaluronan (HYA) with its unique physico-chemical properties, e.g. viscoelasticity and barrier function. The present study aimed to map and compare the normal localisation and distribution of hyaluronan in the arteries and veins of both adult and newborn rats, using a specific staining method utilizing a hyaluronan-binding protein. The hyaluronan stained clearly different in veins and arteries both in newborn and adult rats. In the veins, the tunica intima stained intensely whereas the corresponding area in the arteries only showed a weak and scattered staining pattern. In the adult rats, the matrix between the smooth muscle cells of the tunica media of the veins had a clearly positive staining pattern compared to the media in the arteries which showed only a few scattered areas of positive staining. In the newborn rats, the media of the arteries stained more intensely. In both newborn and adult rats, the adventitia stained intensely both in veins and arteries. Moreover, the HYA-staining pattern differed in veins in different regions of the body. Since HYA is known to be involved in many different biological processes, the results are of importance to understand physiological properties of the vessel tree and to explain the development of vascular diseases.
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| 16. |
- HILLIGES, M, et al.
(författare)
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Innervation of the human vaginal mucosa as revealed by PGP 9.5 immunohistochemistry
- 1995
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Ingår i: Acta anatomica. - : S. Karger AG. - 0001-5180. ; 153:2, s. 119-126
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Tidskriftsartikel (refereegranskat)abstract
- In order to obtain a description of the innervation of the vaginal wall we employed an antiserum against the general neuronal marker, protein gene product 9.5, on normal human vaginal mucosa. Specimens were taken from the anterior and posterior fornices, from the anterior vaginal wall at the bladder neck level and from the introitus vaginae region, and then processed for indirect immunohistochemistry. All regions studied revealed a profound innervation, although regional differences were noted. The more distal areas of the vaginal wall had more nerve fibers compared to the more proximal parts. Also, biopsies from the anterior wall generally were more densely innervated than the posterior wall. Some large nerve coils were observed in lamina propria of the anterior wall as well as gatherings of thick-walled medium-sized blood vessels. Free intraepithelial nerve endings were only detected in the introitus vaginae region. These fibers were very thin, always varicose and could be observed just a few cell layers from the surface. In this part of the vagina, protein gene product 9.5 anti¬bodies also stained cells within the basal parts of the epithelium. These cells were also neurone-specific enolase positive and resembled, from a morphological point of view, Merkel cells.
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| 17. |
- Hingert, Daphne, et al.
(författare)
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Human Levels of MMP-1 in Degenerated Disks Can Be Mitigated by Signaling Peptides from Mesenchymal Stem Cells
- 2020
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 209:2-3, s. 144-153
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Tidskriftsartikel (refereegranskat)abstract
- Degradation of extracellular matrix (ECM) in intervertebral disks (IVDs) during IVD degeneration plays a vital role in low back pain (LBP). In healthy IVDs, synthesis and degradation of ECM are kept in balance by matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs. MMPs are enzymes responsible for ECM degradation, and their expression levels are known to increase in degenerated disks. However, the exact pathophysiological concentration of MMP-1 in the degenerated disks of patients with chronic LBP has not been reported previously. Factors secreted by human mesenchymal stem cells (hMSCs) have shown positive results in cell therapy of degenerated disks. The aim of this study was to investigate the pathophysiological MMP-1 concentration (in ng/mL) in degenerated disk tissue and to evaluate if conditioned media (CM) from hMSCs could mitigate the effects of MMP-1 at the detected levels in a 3D in vitro disk cell (DC) pellet model. Tissue levels of MMP-1 were quantified in disk tissue collected from 6 chronic LBP patients undergoing surgery. DC pellet cultures were performed to investigate the effects of MMP-1 alone and the effects of conditioned media (CM) in the presence of MMP-1. MMP-1 was introduced in the pellets on day 14 at concentrations of 5, 50, or 100 ng/mL. The pellets were harvested on day 28 and evaluated for cell viability, proliferation, and ECM production. The mean concentration of MMP-1 in disk tissue was 151 ng/mL. Results from pellet cultures demonstrated a higher number of viable cells, glycosaminoglycan production, and ECM accumulation in the CM group even in the presence of MMP-1 compared to the controls. However, the level decreased with increasing MMP-1 concentration. The results demonstrated that CM has the ability to mitigate matrix degradation property of MMP-1 up to 50 ng/mL suggesting that CM could potentially be used to treat early stages of disk degeneration.
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| 18. |
- Hingert, Daphne, et al.
(författare)
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Investigation of the Effect of Secreted Factors from Mesenchymal Stem Cells on Disc Cells from Degenerated Discs
- 2020
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 208:1-2, s. 76-88
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Tidskriftsartikel (refereegranskat)abstract
- Low back pain is experienced by a large number of people in western countries and may be caused and influenced by many different pathologies and psychosocial factors including disc degeneration. Disc degeneration involves the increased expression of proinflammatory cytokines and matrix metalloproteinases (MMPs) in the disc environment, which leads to the loss of extracellular matrix (ECM) and the viability of the native disc cells (DCs). Treatment approaches using growth factors and cell therapy have been proposed due to the compelling results that growth factors and mesenchymal stem cells (MSCs) can influence the degenerated discs. The aim of this study was to investigate the effects of conditioned media (CM) from human MSCs (hMSCs) and connective tissue growth factor (CTGF) and TGF-β on disc cells, and hMSCs isolated from patients with degenerative discs and severe low back pain. The aim was also to examine the constituents of CM in order to study the peptides that could bring about intervertebral disc (IVD) regeneration. DCs and hMSC pellets (approx.. 200,000 cells) were cultured and stimulated with hMSC-derived CM or CTGF and TGF-β over 28 days. The effects of CM and CTGF on DCs and hMSCs were assessed via cell viability, proteoglycan production, the expression of ECM proteins, and chondrogenesis in 3D pellet culture. To identify the constituents of CM, CM was analyzed with tandem mass spectrometry. The findings indicate that CM enhanced the cellular viability and ECM production of DCs while CTGF and the control exhibited nonsignificant differences. The same was observed in the hMSC group. Mass spectrometry analysis of CM identified >700 peptides, 129 of which showed a relative abundance of ≥2 (CTGF among them). The results suggest that CM holds potential to counter the progression of disc degeneration, likely resulting from the combination of all the substances released by the hMSCs. The soluble factors released belong to different peptide families. The precise mechanism underlying the regenerative effect needs to be investigated further, prior to incorporating peptides in the development of new treatment strategies for low back pain that is potentially caused by IVD degeneration.
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| 19. |
- Hingert, Daphne, et al.
(författare)
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Pathological Effects of Cortisol on Intervertebral Disc Cells and Mesenchymal Stem Cells from Lower Back Pain Patients
- 2019
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 207:1, s. 34-45
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Tidskriftsartikel (refereegranskat)abstract
- In western countries, lower back pain (LBP) is one of the most common disorders, experienced by more than 80% of the population. Chronic LBP due to disc degeneration has been linked to ongoing inflammatory processes in the disc and endplates. Pain effects the body in different ways, inducing a general stress response in which the body responds by releasing the stress hormone cortisol. Little is known about the impact of pain-induced stress on the progression of disc degeneration. Thus, the effects of cortisol on disc cells (DCs) and human mesenchymal stem cells (hMSCs) were explored in vitro with the objective of investigating the repercussions of cortisol on these cell types involved in de- and regenerative mechanisms of the disc. DC and hMSC pellet cultures were exposed to cortisol at two concentrations (150 and 300 ng/mL) for 28 days to simulate pain-induced stress. Cell viability, histological staining, and GAG DNA, along with apo-ptotic assays were conducted. Detection of OCT4, SOX9, IL-1R, and CXCR2 expressions was performed by immunohistochemistry. With cortisol treatment, restricted cell proliferation and less GAG production in both DCs and hMSCs were observed. Suppression of the differentiation and immunomodulatory efficacy of hMSCs was also detected. Moreover, elevated expressions of IL-1R and CXCR2 were detected in both cell types. To conclude, constant exposure to cortisol even at a physiological level enhanced pathological cellular processes in both DCs and hMSCs, which further jeopardized chondrogenesis. This suggests that cortisol resulting from pain-induced stress is a contributing component of intervertebral disc degeneration and may negatively affect regenerative attempts of the disc.
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| 20. |
- Hinkula, J, et al.
(författare)
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Genetic immunization with multiple HIV-1 genes provides protection against HIV-1/MuLV pseudovirus challenge in vivo
- 2004
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 177:3, s. 169-184
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Tidskriftsartikel (refereegranskat)abstract
- Superinfection by HIV-1 of a cell line containing the complete murine leukemia virus (MuLV) genome was shown to give rise to pseudotyped HIV-1/MuLV. Such superinfection was successful with certain strains of HIV-1 subtypes A–D. Primary spleen cells and cells of the peritoneal cavity of immunocompetent mice of the C57Bl/6 strain were infectable with the pseudotype HIV-1/MuLV and secreted HIV-1 in vitro and in vivo. In contrast, the murine cell lines, NIH 3T3, myeloma cell line Sp2/0, and two murine hybridoma cell lines were relatively resistant to infection and produced no or little HIV. After primary murine spleen cells had been infected with pseudotyped HIV-1 and transferred to C57Bl/6 mice, replication-competent HIV-1 was obtained from the peritoneal cavity for at least 10–14 days. High amounts (>10<sup>5</sup> vRNA copies/ml) of HIV-1 vRNA could be measured in the peritoneal fluid. Presence of HIV-1 proviral DNA was detectable in cells from the peritoneal cavity for up to 24 days after infected cell transfer. Active reverse transcriptase representing both HIV-1 and C-type murine retroviruses was detected in the peritoneal washes. The HIV-infected spleen cells injected into the peritoneal cavity elicited HIV-1-specific cellular immune responses to p24gag, gp160Env, Nef, Tat and Rev. Mice immunized with HIV-1 DNA, but not with HIV-1 protein, cleared their HIV-1-infected cells within 10–14 days after challenge with HIV-1/MuLV-infected syngeneic spleen cells. This novel model system of primarily cellular reactivity to HIV-1-infected cells in vivo may become useful for assaying experimental HIV-1 immunization schedules.
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| 21. |
- Huss, Fredrik R.M., et al.
(författare)
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Mammary epithelial cell and adipocyte co-culture in a 3-D matrix : The first step towards tissue-engineered human breast tissue
- 2001
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 169:4, s. 361-367
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Tidskriftsartikel (refereegranskat)abstract
- Reconstruction of the female breast after cancer surgery is a demanding task where the methods used today suffer from several disadvantages. In the present study we have investigated the possibility to use tissue engineering methods to regenerate human autologous breast tissue. Human mammary epithelial cells and preadipocytes were derived from breast tissue biopsies from healthy women undergoing reduction mammoplasty, and the two celltypes were co-cultured with conventional cell culture methods as well as in 3-D matrices. The study shows that it is possible to harvest both human mammary epithelial cells and preadipocytes in a single session, propagate several subcultures, and that the cells maintain a normal intercellular distribution and growth-pattern when co-cultured in a 3-D collagen gel. We propose that growth and formation of a tissue closely resembling normal human breast tissue be readily obtained in the described in vitro cell culture set-up using basic tissue engineering principles. This concept may be of great importance in the development of new methods for reconstruction of the human breast.
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| 22. |
- Junker, Johan, 1980-, et al.
(författare)
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Adipogenic, Chondrogenic and Osteogenic Differentiation of ClonallyDerived Human Dermal Fibroblasts
- 2010
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Ingår i: Cells Tissues Organs. - Basel : Karger AG. - 1422-6405 .- 1422-6421. ; 191:2, s. 105-118
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Tidskriftsartikel (refereegranskat)abstract
- The apparent need of an autologous cell source for tissueengineering applications has led researchers to explore thepresence of cells with stem cell plasticity in several humantissues. Dermal fibroblasts (FBs) are easy to harvest, expandin vitro and store, rendering them plausible candidates forcell-based therapies. The aim of the present study was toobserve the effects of adipogenic, chondrogenic and osteogenicinduction media on the phenotype of human FBs.Human preadipocytes obtained from fat tissue have beenproposed as an adult stem cell source with suitable characteristics,and were used as control cells in regard to their differentiationpotential. Routine staining, immunohistochemicalanalysis and alkaline phosphatase assay were employed,in order to study the phenotypic shift. FBs were shown topossess multilineage potential, giving rise to fat-, cartilageandbone-like cells. To exclude contaminant progenitor cellsor cell fusion giving rise to tissue with adipocyte-, chondrocyte-and osteoblast-like cells, single-cell cloning was performed.Single-cell-cloned FBs (sccFBs) displayed a similardifferentiation potential as primary-culture FBs. The pres-ence of ‘stem-cell-specific’ surface antigens was analyzedusing flow cytometry. The results reveal that sccFBs haveseveral of the markers associated with cells exhibiting stemcell plasticity. The findings presented here are corroboratedby the findings of other groups, and suggest the use of humandermal FBs in cell-based therapies for the reconstructionof fat, cartilage and bone.
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| 23. |
- Juul-Kristensen, Birgit, et al.
(författare)
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Muscle Sizes and Moment Arms of Rotator Cuff Muscles Determined by Magnetic Resonance Imaging
- 2000
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 167, s. 214-222
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Tidskriftsartikel (refereegranskat)abstract
- Biomechanical models which require information on, e.g., joint torque and muscle force are useful in the estimation of when and how mechanical overload of the musculoskeletal system may lead to disorders. The aim was to study the reliability and validity of magnetic resonance imaging (MRI) to quantify muscle sizes and moment arms by MRI and to test selected anthropometric measures as predictors of muscle sizes and moment arms. A total of 20 healthy Scandinavian women (age 22–58 years) participated in an MRI scanning of their dominant shoulder. With a PC-based program the reliability and the validity of the MRI measurements was estimated to be high, and mean anatomical cross-sectional areas (ACSA) and muscle lengths were measured to be 4.0, 9.8 and 12.1 cm2 and 12.0, 12.6 and 12.8 cm for m. supraspinatus, m. infraspinatus and m. subscapularis, respectively. Volumes were calculated to be 48.8, 125.1 and 153.6 cm3. Moment arms were measured with the upper arm in a neutral position and in a functional position of 34° abduction for m. supraspinatus only, and were 2.4 and 2.6 cm. Physiological cross-sectional area (PCSA) and its fiber force component were estimated from dissected fiber length and pennation angle. MRI volume and PCSA were 1.4–1.7 times higher than dissection data, primarily because of age differences. No external anthropometric measures were found to be predictors of volumes or moment arms.
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| 24. |
- Karlsson, Camilla, 1977, et al.
(författare)
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Neither Notch1 expression nor cellular size correlate with mesenchymal stem cell properties of adult articular chondrocytes.
- 2008
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 187:4, s. 275-85
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Tissue repair is thought to be regulated by progenitor cells, which in other tissues are characterized by their Notch1 expression or small cellular size. Here we studied if these traits affect the chondrogenic potential and are markers for multipotent progenitor cell populations in adult articular cartilage. METHODS: Directly isolated articular chondrocytes were sorted with regard to their Notch1 expression or cellular size. Their colony forming efficiency (CFE) and their potential to differentiate towards adipogenic, osteogenic and chondrogenic lineages were investigated. The different sorted populations were also expanded in monolayer and analyzed in the same manner as the directly isolated cells. RESULTS: No differences in CFE or adipogenic, osteogenic and chondrogenic potentials were detected among the sorted populations. Expanded cells displayed a higher osteochondral potential than directly isolated cells. CONCLUSION: Cellular size or Notch1 expression is not per se a specific marker for mesenchymal progenitor cells in adult articular cartilage. Monolayer-expanded adult chondrocytes contain a larger mesenchymal progenitor cell-like population than directly isolated cells, highly likely as a result of dedifferentiation. If there are resident Notch1-positive cells or cells of a specific size in adult articular cartilage with functional features of progenitor cells, the population consists of only a very small number of cells.
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| 25. |
- Karlsson, Camilla, 1977, et al.
(författare)
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Notch1, Jagged1, and HES5 are abundantly expressed in osteoarthritis.
- 2008
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 188:3, s. 287-98
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Notch signalling controls differentiation and proliferation in various cell types and is associated with several diseases. We investigated the localization and regulation of several Notch markers in human osteoarthritic (OA) cartilage as well as identified genes controlled by Notch signalling. METHODS: Immunolocalization and real-time PCR analysis of Notch markers in healthy and OA articular cartilage were performed. Genes regulated by Notch signalling were studied using microarray. Cytokine-induced transcription of Notch markers was analyzed using real-time PCR and its effect on cellular localization of the intracellular domain of Notch1 (NICD1) was investigated using immunohistochemistry, subcellular fractionation, and transfection. The effect of NFkappaB activation on HES5 transcription was studied using the NFkappaB inhibitor pyrrolidine dithiocarbamate. RESULTS: Notch signalling was activated in OA cartilage and Notch1, Jagged1, and HES5 were abundantly expressed compared to healthy cartilage. Notch signalling regulated the expression of several genes associated with OA, like interleukin-8, lubricin, CD10, matrix metalloproteinase-9, and bone morphogenetic protein-2. Cytokines significantly affected the expression of several Notch markers and repressed expression of HES5, but did not affect the cellular localization of NICD1. CONCLUSION: Notch signalling is dysregulated in OA, inducing and repressing transcription of genes that could potentially partly contribute to the OA phenotype.
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| 26. |
- Kingham, Paul J., et al.
(författare)
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Adipose-Derived Stem Cells for Nerve Repair : Hype or Reality?
- 2014
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 200:1, s. 23-30
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Forskningsöversikt (refereegranskat)abstract
- Peripheral nerve injury is a relatively commonly occurring trauma which seriously compromises the quality of life for many individuals. There is a major need to devise new treatment strategies, and one possible approach is to develop cellular therapies to bioengineer new nerve tissue and/or modulate the endogenous regenerative mechanisms within the peripheral nervous system. In this short review we describe how stem cells isolated from adipose tissue could be a suitable element of this approach. We describe the possible mechanisms through which the stem cells might exert a positive influence on peripheral nerve regeneration. These include their ability to differentiate into cells resembling Schwann cells and their secretion of a plethora of neurotrophic growth factors. We also review the literature describing the effects of these cells when tested using in vivo peripheral nerve injury models.
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| 27. |
- Kopp, S, et al.
(författare)
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Reduction of temporomandibular joint pain after treatment with a combination of methotrexate and infliximab is associated with changes in synovial fluid and plasma cytokines in rheumatoid arthritis
- 2005
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 180:1, s. 22-30
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Tidskriftsartikel (refereegranskat)abstract
- The aims were to investigate the effect of intravenous infusions of the tumor necrosis factor-α (TNF-α) antibody infliximab on symptoms and signs of temporomandibular joint (TMJ) involvement in relation to effects on synovial fluid and plasma proinflammatory TNF-α, interleukin-1β (IL-1β) and interleukin-6 as well as antiinflam matory soluble TNF receptor II (TNF-sRII), interleukin-1 receptor antagonist (IL-1ra), soluble IL-1 receptor II (IL-1sRII) and interleukin-10 (IL-10) in patients with active rheumatoid arthritis (RA). Nineteen patients with TMJ involvement taking methotrexate were included in the study. TMJ and general joint pain intensity as well as pain on mandibular movements, tenderness to digital palpation, pressure pain threshold and maximum mouth-opening capacity were assessed in a clinical examination. The effect of infliximab was assessed after 2 and 14 or 22 weeks. TMJ synovial fluid and venous blood were collected for cytokine analysis at all occasions while determination of erythrocyte sedimentation rate and C-reactive protein were performed at baseline and at long-term follow-up only. Reduction of TMJ pain was associated with raised levels of synovial fluid TNF-sRII and IL-1sRII as well as raised plasma levels of IL-1ra and IL-10. Decreased erythrocyte sedimentation rate was associated with decreased tenderness to digital palpation. Reduced general joint pain intensity was associated with reduced plasma levels of IL-6 and C-reactive protein. In conclusion, systemic treatment with a combination of infliximab and methotrexate reduces TMJ pain in RA in association with an increase in anti-inflammatory cytokines and receptors in synovial fluid and plasma.
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| 28. |
- Larsson, Karin, 1955, et al.
(författare)
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Electron Microscopy Analysis of Neurites Extending from Dorsal Root Ganglia in vitro following Exposure to Intervertebral Disc Cells.
- 2012
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 196:1, s. 82-89
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Tidskriftsartikel (refereegranskat)abstract
- Nucleus pulposus cells from the intervertebral disc have been shown to have inhibiting effects on neurite outgrowth in vitro. The nucleus pulposus consists of at least 2 cell populations, notochordal cells and chondrocyte-like cells. The aim of this study was to analyze the morphology of the neurites, from rat dorsal root ganglia (DRG) in a culture system, after exposure of these 2 cell populations. DRG from perinatal rats was harvested and placed in culture dishes for 24 h. Nucleus pulposus cells from donor rats were separated into 2 populations and applied to the DRG and neurite culture for a further 24 h and compared to control cultures exposed to culture medium without cells. The DRG and neurites were thereafter prepared for scanning or transmission electron microscopy (SEM/TEM). Descriptive SEM and TEM analyses and calculations of the neurite diameter were performed. The visual appearance after SEM and TEM preparation was similar in the three different culture conditions. However, there was a statistically significant reduction of the neurite diameter for the cultures exposed to notochordal cells compared to the cultures exposed to medium and chondrocyte-like cells (TEM preparation). Prominent and frequent pathologic abnormalities in peripheral nerve diseases have been observed with changes in axonal caliber. This study may suggest that a preserved small amount of notochordal cells, as seen in human adults, may play a role in clinical situations where nerve tissue is exposed to disc material, i.e. in disc herniation or degeneration.
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| 29. |
- Lee, J, et al.
(författare)
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Immunofluorescent triple-staining technique to identify sensory nerve endings in human thumb ligaments
- 2012
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 195:5, s. 456-464
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Tidskriftsartikel (refereegranskat)abstract
- Ligament innervation purportedly plays a critical role in stability, proprioception and pathology of joints with minimal bony constraints. The human thumb carpometacarpal (CMC) joint is such a joint: with a complex saddle configuration and wide circumduction, its constraint is primarily ligamentous and it is prone to osteoarthritis. CMC reconstruction is the most commonly performed arthritis surgery in the upper extremity. Little, however, is known about CMC ligament innervation. We describe a novel triple-staining immunofluorescence technique using the markers for low-affinity neurotrophin receptor p75, the pan-neuronal marker protein gene product (PGP) 9.5 and 4′,6′-diamidino-2-phenylindole (DAPI) to simultaneously detect and differentiate between specific sensory nerve endings: the Pacini corpuscles, the Ruffini endings and nerve fascicles. Five primary CMC ligaments (dorsal radial, dorsal central, posterior oblique, anterior oblique and ulnar collateral ligaments) were harvested from 10 fresh-frozen human cadaver hands. Following paraffin sectioning, each ligament was stained using a triple-stain technique and imaged with fluorescence microscopy. Multidimensional acquisition permitted simultaneous capture of images at different wavelengths. Pacini corpuscles were distinguished by their distinct p75 immunoreactive capsules, and Ruffini endings by their overlapping p75 and PGP9.5 immunoreactive dendritic nerve endings. Simultaneous use of PGP9.5, p75 and DAPI immunofluorescence to analyze innervation patterns in human ligaments provides descriptive analysis of staining patterns and receptor structure as well as clues as to the proprioceptive function of CMC ligaments and the joint as a whole. Our novel findings of CMC ligament innervation augment the study of normal and pathological joint mechanics in this joint so prone to osteoarthritis.
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| 30. |
- Långsjö, Teemu, et al.
(författare)
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Quantitative analysis of collagen network structure and fibril dimensions in cartilage repair with autologous chondrocyte transplantation.
- 2010
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 192:6, s. 351-360
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: The aim of this study was to undertake a stereological analysis to quantify the dimensions of the collagen network in the repair tissue of porcine joints after they had been subjected to autologous chondrocyte transplantation (ACT).METHOD: ACT was used to repair cartilage lesions in knee joints of pigs. Electron-microscopic stereology, immunostaining for type II collagen, and quantitative polarized-light microscopy were utilized to study the collagen fibrils in the repair tissue 3 and 12 months after the operation.RESULTS: The collagen volume density (V(V)) was lower in the repair tissue than in normal cartilage at 3 months (20.4 vs. 23.7%) after the operation. The collagen surface density (S(V), 1.5·10(-2) vs. 3.1·10(-2) nm(2)/nm(3)) and V(V) increased with time in the repair tissue (20.4 vs. 44.7%). Quantitative polarized-light microscopy detected a higher degree of collagen parallelism in the repair tissue at 3 months after the operation (55.7 vs. 49.7%). In contrast, 1 year after the operation, fibril parallelism was lower in the repair tissue than in the control cartilage (47.5 vs. 69.8%).CONCLUSION: Following ACT, V(V) and S(V) increased in the repair tissue with time, reflecting maturation of the tissue. One year after the operation, there was a lower level of fibril organization in the repair tissue than in the control cartilage. Thus, the newly synthesized collagen fibrils in the repair tissue appeared to form a denser network than in the control cartilage, but the fibrils remained more randomly oriented.
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| 31. |
- Melnick, M., et al.
(författare)
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Meckel's cartilage differentiation is dependent on hedgehog signaling
- 2005
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 179:4, s. 146-157
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Tidskriftsartikel (refereegranskat)abstract
- The hedgehog (Hh) signaling pathway has been shown to be essential for craniofacial development. Although mandibular arch derivatives are largely absent in Shh null mice, little is known about the role of Hh signaling during Meckel's cartilage development per se. Mandible development is dependent on the morphogenesis of Meckel's cartilage, which then serves as a template for subsequent skeletal differentiation. In this study, we examine the biological function of Hh signaling during Meckel's cartilage development in vivo and in vitro. E13.5 Shh null mice present a small mesenchymal condensation in the region of a presumptive Meckel's cartilage in the hypoplastic mandibular arch. By E15.5, the Shh mutant exhibits a mere remnant of the mandibular arch, without evidence of Meckel's cartilage differentiation. Further, wild-type embryonic (E11 or E12) mandibular explants cultured for up to 5 days in the presence of cyclopamine, a steroidal alkaloid that specifically disrupts the Hh signaling pathway, exhibit a stage-dependent inhibition of Meckel's cartilage chondroblast differentiation to mature chondrocytes. This phenotype can be rescued by exogenous FGF8, a downstream effector of Hh signaling. Taken together, our results indicate that the Hh signaling pathway is critical to Meckel's cartilage ontogenesis and the rate of chondrogenesis, but not to initial primordium formation. The reliance on Hh signaling is stage dependent. Copyright (C) 2005 S. Karger AG, Basel.
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| 32. |
- Rein, S, et al.
(författare)
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Immunohistochemical analysis of sensory nerve endings in ankle ligaments: a cadaver study
- 2013
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 197:1, s. 64-76
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Tidskriftsartikel (refereegranskat)abstract
- <b><i>Background:</i></b> The aim of this study was to analyze the pattern and types of sensory nerve endings in ankle ligaments using immunohistochemical techniques, in order to gain more insight into functional ankle stability. <b><i>Methods:</i></b> One hundred forty ligaments from 10 cadaver feet were included: the calcaneofibular and anterior/posterior talofibular ligaments from the lateral complex; inferior extensor retinaculum complex, talocalcaneal oblique and canalis tarsi ligaments from the sinus tarsi; deltoid ligament with its individual portions from the medial complex, and anterior tibiofibular ligament (ATiFL) from the syndesmosis. Mechanoreceptors were classified according to Freeman and Wyke [Acta Anat (Basel) 1967;68:321–333] after staining with hematoxylin-eosin, low-affinity neurotrophin receptor p75, protein gene product 9.5, and S-100 protein. <b><i>Results:</i></b> Free nerve endings were the predominant sensory endings in all four complexes, with the greatest density in the lateral and medial complexes; followed by Ruffini endings, unclassifiable corpuscles, Pacini corpuscles, and Golgi-like endings. Ruffini endings were significantly more prevalent in the ATiFL than in the medial complex, and more common than Pacini corpuscles and Golgi-like endings in the lateral, medial, and sinus tarsi complexes. A greater number of blood vessels correlated with a greater number of free nerve endings. There was a negative correlation between the number of Ruffini endings, unclassifiable corpuscles, and age. <b><i>Conclusions:</i></b> Free nerve endings are the dominant mechanoreceptor type in the ankle ligaments, followed by Ruffini endings. The ligaments of the lateral and medial ankle complexes are more innervated than the sinus tarsi ligaments.
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| 33. |
- Sandstedt, Joakim, et al.
(författare)
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SSEA-4+ CD34- Cells in the Adult Human Heart Show the Molecular Characteristics of a Novel Cardiomyocyte Progenitor Population.
- 2014
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 199:2-3, s. 103-116
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Tidskriftsartikel (refereegranskat)abstract
- Stage-specific embryonic antigen (SSEA) expression is used to describe the differentiation state of an embryonic stem cell (ESC). In human ESCs, SSEA-3 and SSEA-4 are highly expressed in undifferentiated cells and downregulated upon differentiation. SSEA-4 has also been described as a marker for adult stem cells in various tissues, including human neonatal cardiac tissue. However, there is currently little data on the expression of SSEAs in human adult cardiac tissue. We obtained right and left atrial biopsies from patients undergoing cardiac surgery. These were dissociated, stained for SSEAs and other cardiac stem cell markers and analyzed by flow cytometry. Directly isolated cells expressed variable levels of SSEA-1, SSEA-3 and SSEA-4. The SSEA-1+ population was established as contaminating hematopoietic cells. The SSEA-4+ population, on the other hand, could be subdivided based on the endothelial progenitor marker CD34. The SSEA-4+ CD34- population in the right atrium had a high gene expression of both early (TBX5, NKX2.5) and late (TNNT2) cardiomyocyte markers. The SSEA-4+ CD34+ population, on the other hand, overlapped with previously described C-kit+ CD45- cardiac stem cells. Primary monolayer-cultured cells retained expression of SSEAs while the cardiomyogenic specification in the SSEA-4+ CD34- population was lost. In tissue sections, SSEA-4+ cells could be identified both within and outside the myocardium. Within the myocardium, some SSEA-4+ cells coexpressed cardiomyogenic markers. In conclusion, the results show that the adult human heart expresses SSEAs and that there is a subpopulation of SSEA-4+ CD34- cells that show features of a cardiomyocyte progenitor population. © 2014 S. Karger AG, Basel.
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| 34. |
- Shah, Furqan A., et al.
(författare)
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Mapping Bone Surface Composition Using Real-Time Surface Tracked Micro-Raman Spectroscopy.
- 2021
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Ingår i: Cells tissues organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 209:4-6, s. 266-276
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Tidskriftsartikel (refereegranskat)abstract
- The surface of bone tells a story - one that is worth a thousand words - of how it is built and how it is repaired. Chemical (i.e., composition) and physical (i.e., morphology) characteristics of the bone surface are analogous to a historical record of osteogenesis and provide key insights into bone quality. Analysis of bone chemistry is of particular relevance to the advancement of human health, cell biology, anthropology/archaeology, and biomedical engineering. Although scanning electron microscopy remains a popular and versatile technique to image bone across multiple length scales, limited chemical information can be obtained. Micro-Raman spectroscopy is a valuable tool for nondestructive chemical/compositional analysis of bone. However, signal integrity losses occur frequently during wide-field mapping of non-planar surfaces. Samples for conventional Raman imaging are, therefore, rendered planar through polishing or sectioning to ensure uniform signal quality. Here, we demonstrate ν1 PO43- and ν1 CO32- peak intensity losses where the sample surface and the plane of focus are offset by over 1-2 μm when underfocused and 2-3 μm when overfocused at 0.5-1 s integration time (15 mW, 633 nm laser). A technique is described for mapping the composition of the inherently irregular/non-planar surface of bone. The challenge posed by the native topology characteristic of this unique biological system is circumvented via real-time focus-tracking based on laser focus optimization by continuous closed-loop feedback. At the surface of deproteinized and decellularized/defatted sheep tibial cortical bone, regions of interest up to 1 mm2 were scanned at micrometer and submicrometer resolution. Despite surface height deviations exceeding 100 μm, it is possible to seamlessly probe local gradients in organic and inorganic constituents of the extracellular matrix as markers of bone metabolism and bone turnover, blood vessels and osteocyte lacunae, and the rope-like mineralized bundles that comprise the mineral phase at the bone surface.
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| 35. |
- Shore, R. C., et al.
(författare)
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The Structure and Composition of Deciduous Enamel Affected by Local Hypoplastic Autosomal Dominant Amelogenesis Imperfecta Resulting from an ENAM Mutation
- 2010
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Ingår i: Cells Tissues Organs. - : S. Karger. - 1422-6405 .- 1422-6421. ; 191:4, s. 301-306
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Tidskriftsartikel (refereegranskat)abstract
- In a group of families in northern Sweden, a mutation in the ENAM gene (predicted to produce a highly truncated protein) results in the local hypoplastic form of autosomal dominant amelogenesis imperfecta. In this study, sections of deciduous teeth from members of 3 of these families were examined by scanning electron microscopy (SEM) and the enamel mineral was analysed by energy dispersive X-ray spectroscopy (EDX). The sections were also probed with antibodies raised to a conserved sequence of the enamelin protein. Selected intact teeth were first analysed by digital imaging and ascribed with an 'Enamel Defects Index' (EDI) score. SEM of tooth sections revealed disrupted prism morphology and the prisms had a glass-like appearance in some areas. These areas of dysplasia were sometimes irregular but formed regular arrays in others. Comparison of EDI scores with SEM indicated that in one tooth the surface had no measurable defects but significant defects were present in the underlying enamel microstructure. SEM immunohistochemistry with the antibody raised to a fragment of the enamelin protein produced positive, but light, labelling throughout normal enamel. In dysplastic areas, however, the labelling intensity appeared to be reduced. The results indicate that the presence of functional enamelin in the correct amounts is necessary for correct prism morphogenesis. In addition, a combination of EDI and structural analysis indicate that defects in enamel microstructure are not necessarily visible as defects on the surface of the tooth, suggesting the possibility, at least, that some instances of under-diagnosis may occur. Copyright (C) 2009 S. Karger AG, Basel
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| 36. |
- Stål, Per, et al.
(författare)
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Fibre composition of human intrinsic tongue muscles.
- 2003
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 173:3, s. 147-161
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Tidskriftsartikel (refereegranskat)abstract
- The muscle fibre composition of three human intrinsic tongue muscles, the longitudinalis, verticalis and transversus, was investigated in four anterior to posterior regions of the tongue using morphological and enzyme- and immunohistochemical techniques. All three muscles typically contained type I, IIA and IM/IIC fibres. Type I fibres expressed slow myosin heavy chain (MyHC), type II fibres fast MyHC, mainly fast A MyHC, whereas type IM/IIC coexpressed slow and fast MyHCs. Type II fibres were in the majority (60%), but regional differences in proportion and diameter of fibre types were obvious. The anterior region of the tongue contained a predominance of relatively small type II fibres (71%), in contrast to the posterior region which instead showed a majority of larger type I and type IM/IIC fibres (66%). In general, the fibre diameter was larger in the posterior region. This muscle fibre composition of the tongue differs from those of limb, orofacial and masticatory muscles, probably reflecting genotypic as well as phenotypic functional specialization in oral function. The predominance of type II fibres and the regional differences in fibre composition, together with intricate muscle structure, suggest generally fast and flexible actions in positioning and shaping the tongue, during vital tasks such as mastication, swallowing, respiration and speech. Copyright 2003 S. Karger AG, Basel
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| 37. |
- Svare, Frida, et al.
(författare)
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It’s About Time: Time-Dependent Tissue Damage in the Adult Porcine Retina After Enucleation
- 2021
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Ingår i: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; , s. 1-8
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Tidskriftsartikel (refereegranskat)abstract
- The ex vivo large animal retina is extensively used in research ranging from discovery of disease mechanisms to future treatment paradigms. Due to limited standardization when harvesting the tissue, the time after enucleation is often ex- tended for several hours, a factor that so far has not yet been fully characterized. The purpose of this study was to investi- gate the relationship between time after enucleation and retinal tissue damage. Adult, porcine retinal explants were dissected and fixed 90 or 240 min after enucleation. In a sep- arate experiment, explants were cultured for 48 h, following dissection either 90 or 240 min after enucleation. Retinas were analyzed morphologically using hematoxylin and eo- sin for overall tissue damage, TUNEL staining for detection of apoptosis, and RBPMS immunohistochemistry for evalua- tion of ganglion cell survival. In addition, medium from the cultured explants was sampled after 2, 24, and 48 h of culture and assessed for the cell damage marker lactate dehydroge- nase (LDH). Retinas examined 240 min after enucleation dis- played a significant increase in overall tissue damage, in- creased apoptosis, and decreased ganglion cell survivalcompared with 90-min counterparts. In the culture experi- ment, no significant difference in overall tissue damage was found between the 2 groups, however, apoptosis was sig- nificantly increased, and ganglion cell survival decreased in the cultured 240-min group. In addition, a significantly in- creased LDH medium activity was found in the 240-min group compared with the 90-min counterpart at all time points. The adult porcine retina is relatively resistant to tis- sue damage 90 min after enucleation but displays distinct signs of injury after 240 min. The importance of these time points is further highlighted when retinal explants are cul- tured. Our results strongly suggest that time after enucle- ation is a crucial factor that should be considered in experi- ments involving the ex vivo adult porcine retina.
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| 38. |
- Tallheden, Tommi, 1972, et al.
(författare)
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Human articular chondrocytes--plasticity and differentiation potential.
- 2006
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6421 .- 1422-6405. ; 184:2, s. 55-67
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Tidskriftsartikel (refereegranskat)abstract
- Articular cartilage has no or very low ability of self-repair, and untreated lesions may lead to the development of osteoarthritis. One method which has been proven to result in long-term repair of isolated lesions is autologous chondrocyte transplantation. In this method, culture-expanded chondrocytes isolated from full-thickness biopsies, taken from a non-weight-bearing area at the supromedial edge of the femoral condyle, are transplanted back to the patient under a cover of periosteum. The treatment is able to regenerate hyaline cartilage with long-term durability. Although the repair mechanism behind this treatment has not been fully elucidated, emerging data generated by microarray technologies reveal an interesting regeneration process involving cellular and molecular mechanisms found during fetal development. In hyaline cartilage, the human chondrocyte population is generally considered a homogenous cell population, but recently several investigators have demonstrated that cells isolated from human articular cartilage have stem cell properties and that the superficial layer contains such cells. This paper will discuss these recent data and their implications for future treatment strategies aiming to induce regeneration in articular cartilage surfaces.
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| 39. |
- Thornemo, Maria, 1968, et al.
(författare)
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Clonal populations of chondrocytes with progenitor properties identified within human articular cartilage.
- 2005
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Ingår i: Cells, tissues, organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 180:3, s. 141-50
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Tidskriftsartikel (refereegranskat)abstract
- The aim of the present study was to identify and characterize progenitor properties of human articular chondrocytes selected by using agarose suspension culture. In this chondrogenic selective culture condition, about 3.6% of seeded surplus chondrocytes from patients undergoing articular chondrocyte transplantation proliferated and formed cell clusters after 6 weeks. Phase-contrast microscopy and transmission electron microscopy revealed four different types of cell clusters differing in cellular content and matrix production. Based on their morphological features, they were named the homogenous (H), the homogenous matrix (HM), the differentiated matrix (DM) and the differentiated (D) cell clusters. All cell clusters showed positive safranin O staining, and matrix was positive for antibodies detecting type II collagen and aggrecan. The clusters were further demonstrated to express the genes for fibroblast growth factor receptor 3, type IIA collagen and type IIB collagen, while type X collagen was not expressed. After subcloning, the H and HM clusters demonstrated the best proliferative capacity. Chondrocytes from these two cell clusters also showed phenotypic plasticity in chondrogenic, adipogenic as well as osteogenic assays. This study demonstrates that existing subpopulations of cells with chondroprogenitor properties can be isolated from human adult articular cartilage using agarose suspension cultures.
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| 40. |
- Svanvik, Teresia, et al.
(författare)
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Human Disk Cells from Degenerated Disks and Mesenchymal Stem Cells in Co-Culture Result in Increased Matrix Production.
- 2009
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Ingår i: Cell Tissues and Organs. - 1422-6405. ; 191:1, s. 2-11
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Tidskriftsartikel (refereegranskat)abstract
- Transplantation of mesenchymal stem cells (MSCs) has been suggested for disk degeneration, which is characterized by dysfunctional cells and low proteoglycan production. The aim of this study was to examine the effects of a 3D co-culture system using human disk cells (DCs) and MSCs on collagen and proteoglycan production. DCs and MSCs were expanded in monolayer and grown in pellet cultures for 7, 14 and 28 days and analyzed for hydroxyproline (HP), reflecting total collagen production, and glycosaminoglycan (GAG) accumulation. DCs and MSCs co-cultured at different ratios (25/75, 50/50 and 75%/25%) were examined for GAG accumulation. Collagen type II expression was analyzed immunohistochemically. In a second series, conditioned media were added to pellet cultures of degenerated DCs or MSCs. DCs from degenerated disks and MSCs demonstrated lower total collagen production than non-degenerated DC pellets. GAG production was comparable in DCs and MSCs, except in the youngest donor, with MSC producing about 10 times higher GAG/DNA. Co-cultures resulted in approximately 1.5 times higher GAG/DNA production than DCs. Increased collagen type II expression was seen in co-cultures compared to DC or MSC culture alone, except in the case with highly active MSCs. No positive effect of conditioned media was seen. In conclusion, co-culture of MSCs with degenerated DCs increased proteoglycan and collagen-type ceII production, indicating that in future clinical therapy MSCs can be transplanted without pre-differentiation in vitro. The lack of effect of conditioned media suggests that the positive effect of co-culture on matrix production is not due to soluble factors.
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