| 1. |
- Herter, Eva K., et al.
(författare)
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WAKMAR2, a Long Noncoding RNA Downregulated in Human Chronic Wounds, Modulates Keratinocyte Motility and Production of Inflammatory Chemokines
- 2019
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Ingår i: Journal of Investigative Dermatology. - : ELSEVIER SCIENCE INC. - 0022-202X .- 1523-1747. ; 139:6, s. 1373-1384
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Tidskriftsartikel (refereegranskat)abstract
- Chronic wounds represent a major and growing health and economic burden worldwide. A better understanding of molecular mechanisms of normal as well as impaired wound healing is needed to develop effective treatment. Herein we studied the potential role of long noncoding RNA LOC100130476 in skin wound repair. LOC100130476 is an RNA polymerase IIeencoded polyadenylated transcript present in both cytoplasm and nucleus. We found that its expression was lower in wound-edge keratinocytes of human chronic wounds compared to normal wounds of healthy donors and intact skin. In cultured keratinocytes, LOC100130476 expression was induced by TGF-beta signaling. By reducing LOC100130476 expression with antisense oligos or activating its transcription with CRISPR/Cas9 Synergistic Activation Mediator system, we showed that LOC100130476 restricted the production of inflammatory chemokines by keratinocytes, while enhancing cell migration. In line with this, knockdown of LOC100130476 impaired re-epithelization of human ex vivo wounds. Based on these results, we named LOC100130476 wound and keratinocyte migration-associated long noncoding RNA 2 (WAKMAR2). Moreover, we identified a molecular network that may mediate the biological function of WAKMAR2 in keratinocytes using microarray. In summary, our data suggest that WAKMAR2 is an important regulator of skin wound healing and its deficiency may contribute to the pathogenesis of chronic wounds.
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| 2. |
- 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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| 3. |
- Li, Dongqing, et al.
(författare)
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Human skin long noncoding RNA WAKMAR1 regulates wound healing by enhancing keratinocyte migration
- 2019
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 116:19, s. 9443-9452
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Tidskriftsartikel (refereegranskat)abstract
- An increasing number of studies reveal the importance of long noncoding RNAs (lncRNAs) in gene expression control underlying many physiological and pathological processes. However, their role in skin wound healing remains poorly understood. Our study focused on a skin-specific lncRNA, LOC105372576, whose expression was increased during physiological wound healing. In human nonhealing wounds, however, its level was significantly lower compared with normal wounds under reepithelialization. We characterized LOC105372576 as a nuclear-localized, RNAPII-transcribed, and polyadenylated lncRNA. In keratinocytes, its expression was induced by TGF-beta signaling. Knockdown of LOC105372576 and activation of its endogenous transcription, respectively, reduced and increased the motility of keratinocytes and reepithelialization of human ex vivo skin wounds. Therefore, LOC105372576 was termed "wound and keratinocyte migration-associated lncRNA 1" (WAKMAR1). Further study revealed that WAKMAR1 regulated a network of protein-coding genes important for cell migration, most of which were under the control of transcription factor E2F1. Mechanistically, WAKMAR1 enhanced E2F1 expression by interfering with E2F1 promoter methylation through the sequestration of DNA methyltransferases. Collectively, we have identified a lncRNA important for keratinocyte migration, whose deficiency may be involved in the pathogenesis of chronic wounds.
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| 4. |
- Li, Dongqing, et al.
(författare)
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miR-19a/b and miR-20a promote wound healing by regulating the inflammatory response of keratinocytes
- 2021
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Ingår i: Journal of Investigative Dermatology. - : Elsevier. - 0022-202X .- 1523-1747. ; 141:3, s. 659-671
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Tidskriftsartikel (refereegranskat)abstract
- Persistent and impaired inflammation impedes tissue healing and is characteristic of chronic wounds. A better understanding of the mechanisms controlling wound inflammation is needed. Here we show that in human wound-edge keratinocytes, the expression of miR-17, miR-18a, miR-19a, miR-19b, and miR-20a, which all belong to the miR-17∼92 cluster, is upregulated during wound repair. However, their levels are lower in chronic ulcers than acute wounds at the proliferative phase. Conditional knockout of miR-17∼92 in keratinocytes as well as injection of miR-19a/b and miR-20a antisense inhibitors into wound-edges enhanced inflammation and delayed wound closure in mice. In contrast, conditional overexpression of the miR-17∼92 cluster or miR-19b alone in mice keratinocytes accelerated wound closure in vivo. Mechanistically, miR-19a/b and miR-20a decreased TLR3-mediated NF-κB activation by targeting SHCBP1 and SEMA7A, respectively, reducing the production of inflammatory chemokines/cytokines by keratinocytes. Thus, as crucial regulators of wound inflammation, lack of miR-19a/b and miR-20a may contribute to sustained inflammation and impaired healing in chronic wounds. In line with this, we show that a combinatory treatment with miR-19b and miR-20a improved wound healing in a mouse model of type 2 diabetes.
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| 5. |
- Li, Dongqing, et al.
(författare)
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Single-Cell Analysis Reveals Major Histocompatibility Complex II-Expressing Keratinocytes in Pressure Ulcers with Worse Healing Outcomes
- 2022
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Ingår i: Journal of Investigative Dermatology. - : Elsevier. - 0022-202X .- 1523-1747. ; 142:3, s. 705-716
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Tidskriftsartikel (refereegranskat)abstract
- Pressure ulcer (PU) is a chronic wound often seen in patients with spinal cord injury and other bed-bound individuals, particularly in the elderly population. Despite its association with high mortality, the pathophysiology of PU remains poorly understood. In this study, we compared single-cell transcriptomic profiles of human epidermal cells from PU wound edges with those from uninjured skin and acute wounds in healthy donors. We identified significant shifts in the cell composition and gene expression patterns in PU. In particular, we found that major histocompatibility complex class II-expressing keratinocytes were enriched in patients with worse healing outcomes. Furthermore, we showed that the IFN-gamma in PU-derived wound fluid could induce major histocompatibility complex II expression in keratinocytes and that these wound fluid-treated keratinocytes inhibited autologous T-cell activation. In line with this observation, we found that T cells from PUs enriched with major histocompatibility complex II+ keratinocytes produced fewer inflammatory cytokines. Overall, our study provides a high-resolution molecular map of human PU compared with that of acute wounds and intact skin, providing insights into PU pathology and the future development of tailored wound therapy.
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| 6. |
- Rakar, Jonathan, et al.
(författare)
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Evaluating Multi-Lineage Induction of Human Dermal FibroblastsUsing Gene Expression Analysis
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- During the past decades, several adult stem cell populations from a range of tissues have been characterized. Since principally all human cells contain the same genetic material, the specific gene expression profile determines the cell phenotype. The notion of terminally differentiated somatic cells being necessarily restricted to one phenotype has been challenged, and instead an inherent range of plasticity for any given cell type has been suggested. We have in previous work shown that normal human dermal fibroblasts have an inherent plasticity and can be induced to differentiate towards adipogenic, chondrogenic, endotheliogenic and osteogenic lineages when subjected to defined induction media. The aim of the present study was to further study the differentiation of human dermal fibroblasts on a gene expression level. This was achieved by employing genome wide expression analysis using microarray technology. Selected gene expression was also evaluated over time using real-time PCR. Several master regulatory genes important for lineage commitment, as well as phenotypically relevant genes, were found regulated in the respective induced cultures. The results obtained in this study strengthen previously published results showing an inherent ability for controllable phenotype alteration of human dermal fibroblasts in vitro. We conclude that adipogenic, chondrogenic, endotheliogenic and osteogenic induction results in novel phenotypes that show a genetic readiness for lineage-specific biological functionality.
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| 7. |
- Rakar, Jonathan, et al.
(författare)
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Interpreted gene expression of human dermal fibroblasts after adipo-, chondro- and osteogenic phenotype shifts
- 2012
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Ingår i: Differentiation. - : Wiley-Blackwell / Elsevier. - 0301-4681 .- 1432-0436. ; 84:4, s. 305-313
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Tidskriftsartikel (refereegranskat)abstract
- Autologous cell-based therapies promise important developments for reconstructive surgery. In vitro expansion as well as differentiation strategies could provide a substantial benefit to cellular therapies. Human dermal fibroblasts, considered ubiquitous connective tissue cells, can be coaxed towards different cellular fates, are readily available and may altogether be a suitable cell source for tissue engineering strategies. Global gene expression analysis was performed to investigate the changes of the fibroblast phenotype after four-week inductions toward adipocytic, osteoblastic and chondrocytic lineages. Differential gene regulation, interpreted through Gene Set Enrichment Analysis, highlight important similarities and differences of induced fibroblasts compared to control cultures of human fibroblasts, adipocytes, osteoblasts and articular chondrocytes. Fibroblasts show an inherent degree of phenotype plasticity that can be controlled to obtain cells supportive of multiple tissue types.
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| 8. |
- Sellberg, Felix, et al.
(författare)
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Composition of growth factors and cytokines in lysates obtained from fresh versus stored pathogen-inactivated platelet units
- 2016
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Ingår i: Transfusion and apheresis science. - : Elsevier BV. - 1473-0502 .- 1878-1683. ; 55:3, s. 333-337
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Tidskriftsartikel (refereegranskat)abstract
- Background: Platelet lysate is a readily available source of growth factors, and other mediators, which has been used in a variety of clinical applications. However, the product remains poorly standardized and the present investigation evaluates the composition of platelet lysate obtained from either fresh or stored pathogen-inactivated platelet units.Materials and Methods: Platelet pooled units (n = 10) were obtained from healthy blood donors and tested according to standard procedures. All units were pathogen inactivated using amotosalen hydrochloride and UVA exposure. Platelet lysate was subsequently produced at two separate time-points, either from fresh platelet units or after 5 days of storage, by repeated freeze-thaw cycles. The following mediators were determined at each time point: EGF, FGF-2, VEGF, IGF-1, PDGF-AB/BB, BMP-2, PF4, TGF-beta isoform 1, IL-1(i, IL-2, IL-6, IL-10, IL-12p70, 1L-17A, TNF-alpha, and IFN-gamma.Results: The concentration of growth factors and cytokines was affected by time in storage. Notably, TGF-beta, PDGF-AB/BB, and PF4 showed an increase of 27.2% (p < 0.0001), 29.5% (p = 0.04) and 8.2% (p = 0.0004), respectively. A decrease was seen in the levels of IGF-1 and FGF-2 with 22% (p = 0.041) and 11% (p = 0.01), respectively. Cytokines were present only in very low concentrations and all other growth factors remained stable with time in storage.Conclusion: The composition of mediators in platelet lysate obtained from pathogen inactivated platelet units differs when produced from fresh and stored platelet units, respectively. This underscores the need for further standardization and optimization of this important product, which potentially may influence the clinical effects.
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| 9. |
- Sommar, Pehr, et al.
(författare)
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ABC om Brännskador
- 2008
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Ingår i: Läkartidningen. - 0023-7205 .- 1652-7518. ; 105:48-49, s. 3547-3552
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Tidskriftsartikel (refereegranskat)
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| 10. |
- Sommar, Pehr, et al.
(författare)
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[Burns].
- 2008
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Ingår i: Läkartidningen. - 0023-7205 .- 1652-7518. ; 105:48-49, s. 3547-52
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Tidskriftsartikel (refereegranskat)
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