| 1. |
- De Santis, Martina M., et al.
(författare)
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Lung tissue bioengineering for transplantation and modelling of development, disease and regeneration
- 2021
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Ingår i: ERS Monograph. - 2312-508X. ; 2021:91, s. 248-272
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Tidskriftsartikel (refereegranskat)abstract
- The lung is a vital and dynamic organ that serves as a critical first barrier to environmental stimuli and facilitates gas exchange from birth until death. Developmental or genetic defects that impair lung function severely impact quality of life. Minor defects present in early life increase the likelihood that patients will develop chronic lung diseases in adulthood that have no cure. Current therapies aim to slow disease progression, with lung transplantation remaining the only option at end-stage disease. While a number of discoveries have been made using conventional cell culture and in vivo animal studies, new approaches are needed to develop effective therapies. Recent advances using bioengineering have created new models that more closely recapitulate human development and disease. In parallel, progress has been made towards generating lung tissue in the laboratory with the ultimate aim of transplantation. This chapter covers the progress and recent advances in applying bioengineering approaches towards improving our understanding of lung development, disease and regeneration.
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| 2. |
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| 3. |
- Ota, Chiharu, et al.
(författare)
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Dynamic expression of HOPX in alveolar epithelial cells reflects injury and repair during the progression of pulmonary fibrosis
- 2018
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 8
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Tidskriftsartikel (refereegranskat)abstract
- Mechanisms of injury and repair in alveolar epithelial cells (AECs) are critically involved in the progression of various lung diseases including idiopathic pulmonary fibrosis (IPF). Homeobox only protein x (HOPX) contributes to the formation of distal lung during development. In adult lung, alveolar epithelial type (AT) I cells express HOPX and lineage-labeled Hopx+ cells give rise to both ATI and ATII cells after pneumonectomy. However, the cell function of HOPX-expressing cells in adult fibrotic lung diseases has not been investigated. In this study, we have established a flow cytometry-based method to evaluate HOPX-expressing cells in the lung. HOPX expression in cultured ATII cells increased over culture time, which was accompanied by a decrease of proSP-C, an ATII marker. Moreover, HOPX expression was increased in AECs from bleomycin-instilled mouse lungs in vivo. Small interfering RNA-based knockdown of Hopx resulted in suppressing ATII-ATI trans-differentiation and activating cellular proliferation in vitro. In IPF lungs, HOPX expression was decreased in whole lungs and significantly correlated to a decline in lung function and progression of IPF. In conclusion, HOPX is upregulated during early alveolar injury and repair process in the lung. Decreased HOPX expression might contribute to failed regenerative processes in end-stage IPF lungs.
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| 4. |
- Bölükbas, Deniz A., et al.
(författare)
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The Preparation of Decellularized Mouse Lung Matrix Scaffolds for Analysis of Lung Regenerative Cell Potential
- 2019
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Ingår i: Methods in molecular biology (Clifton, N.J.). - New York, NY : Springer New York. - 1940-6029. ; 1940, s. 275-295
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Tidskriftsartikel (refereegranskat)abstract
- Lung transplantation is the only option for patients with end-stage lung disease, but there is a shortage of available lung donors. Furthermore, efficiency of lung transplantation has been limited due to primary graft dysfunction. Recent mouse models mimicking lung disease in humans have allowed for deepening our understanding of disease pathomechanisms. Moreover, new techniques such as decellularization and recellularization have opened up new possibilities to contribute to our understanding of the regenerative mechanisms involved in the lung. Stripping the lung of its native cells allows for unprecedented analyses of extracellular matrix and sets a physiologic platform to study the regenerative potential of seeded cells. A comprehensive understanding of the molecular pathways involved for lung development and regeneration in mouse models can be translated to regeneration strategies in higher organisms, including humans. Here we describe and discuss several techniques used for murine lung de- and recellularization, methods for evaluation of efficacy including histology, protein/RNA isolation at the whole lung, as well as lung slices level.
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| 5. |
- Costa, Rita, et al.
(författare)
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A drug screen with approved compounds identifies amlexanox as a novel Wnt/β-catenin activator inducing lung epithelial organoid formation
- 2021
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Ingår i: British Journal of Pharmacology. - : Wiley. - 0007-1188 .- 1476-5381. ; 178:19, s. 4026-4041
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Tidskriftsartikel (refereegranskat)abstract
- Background and Purpose: Emphysema is an incurable disease characterized by loss of lung tissue leading to impaired gas exchange. Wnt/β-catenin signalling is reduced in emphysema, and exogenous activation of the pathway in experimental models in vivo and in human ex vivo lung tissue improves lung function and structure. We sought to identify a pharmaceutical able to activate Wnt/β-catenin signalling and assess its potential to activate lung epithelial cells and repair. Experimental Approach: We screened 1216 human-approved compounds for Wnt/β-catenin signalling activation using luciferase reporter cells and selected candidates based on their computationally predicted protein targets. We further performed confirmatory luciferase reporter and metabolic activity assays. Finally, we studied the regenerative potential in murine adult epithelial cell-derived lung organoids and in vivo using a murine elastase-induced emphysema model. Key Results: The primary screen identified 16 compounds that significantly induced Wnt/β-catenin-dependent luciferase activity. Selected compounds activated Wnt/β-catenin signalling without inducing cell toxicity or proliferation. Two compounds were able to promote organoid formation, which was reversed by pharmacological Wnt/β-catenin inhibition, confirming the Wnt/β-catenin-dependent mechanism of action. Amlexanox was used for in vivo evaluation, and preventive treatment resulted in improved lung function and structure in emphysematous mouse lungs. Moreover, gene expression of Hgf, an important alveolar repair marker, was increased, whereas disease marker Eln was decreased, indicating that amlexanox induces pro-regenerative signalling in emphysema. Conclusion and Implications: Using a drug screen based on Wnt/β-catenin activity, organoid assays and a murine emphysema model, amlexanox was identified as a novel potential therapeutic agent for emphysema.
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| 6. |
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| 7. |
- De Santis, Martina M, et al.
(författare)
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Extracellular-Matrix-Reinforced Bioinks for 3D Bioprinting Human Tissue
- 2021
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Ingår i: Advanced Materials. - : Wiley. - 1521-4095 .- 0935-9648. ; 33:3
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Tidskriftsartikel (refereegranskat)abstract
- Recent advances in 3D bioprinting allow for generating intricate structures with dimensions relevant for human tissue, but suitable bioinks for producing translationally relevant tissue with complex geometries remain unidentified. Here, a tissue-specific hybrid bioink is described, composed of a natural polymer, alginate, reinforced with extracellular matrix derived from decellularized tissue (rECM). rECM has rheological and gelation properties beneficial for 3D bioprinting while retaining biologically inductive properties supporting tissue maturation ex vivo and in vivo. These bioinks are shear thinning, resist cell sedimentation, improve viability of multiple cell types, and enhance mechanical stability in hydrogels derived from them. 3D printed constructs generated from rECM bioinks suppress the foreign body response, are pro-angiogenic and support recipient-derived de novo blood vessel formation across the entire graft thickness in a murine model of transplant immunosuppression. Their proof-of-principle for generating human tissue is demonstrated by 3D bioprinting human airways composed of regionally specified primary human airway epithelial progenitor and smooth muscle cells. Airway lumens remained patent with viable cells for one month in vitro with evidence of differentiation into mature epithelial cell types found in native human airways. rECM bioinks are a promising new approach for generating functional human tissue using 3D bioprinting.
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| 8. |
- De Santis, Martina M, et al.
(författare)
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How to build a lung : latest advances and emerging themes in lung bioengineering
- 2018
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Ingår i: European Respiratory Journal. - : European Respiratory Society (ERS). - 1399-3003 .- 0903-1936. ; 52, s. 1-19
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Forskningsöversikt (refereegranskat)abstract
- Chronic respiratory diseases remain a major cause of morbidity and mortality worldwide. The only option at end-stage disease is lung transplantation, but there are not enough donor lungs to meet clinical demand. Alternative options to increase tissue availability for lung transplantation are urgently required to close the gap on this unmet clinical need. A growing number of tissue engineering approaches are exploring the potential to generate lung tissue ex vivo for transplantation. Both biologically derived and manufactured scaffolds seeded with cells and grown ex vivo have been explored in pre-clinical studies, with the eventual goal of generating functional pulmonary tissue for transplantation. Recently, there have been significant efforts to scale-up cell culture methods to generate adequate cell numbers for human-scale bioengineering approaches. Concomitantly, there have been exciting efforts in designing bioreactors that allow for appropriate cell seeding and development of functional lung tissue over time. This review aims to present the current state-of-the-art progress for each of these areas and to discuss promising new ideas within the field of lung bioengineering.
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| 9. |
- De Santis, Martina M.
(författare)
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Next generation bioengineering of lung tissue for transplantation
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Lung transplantation is the only option for end-stage lung diseases, but organ shortage remains problematic. Generating lungs ex vivo could overcome shortages with current approaches being explored for lung tissue engineering utilizing a biologically derived, synthetic or hybrid scaffold which is seeded with cells and cultured ex vivo. Ideally, cells could be sourced from the transplant recipient and thus are conceptualized to reduce the long-term requirements for immunosuppressive drugs and the risk for rejection. Progenitor cell populations can be controlled more easily than induced pluripotent stem cells (IPSCs), with lower risk of tumour formation. However, as presented in this thesis, progenitor cells can be deranged in diseased lungs such as for example idiopathic pulmonary fibrosis (IPF) and are therefore unlikely candidates to generate healthy tissue. In addition to healthy cells, scaffolds with attributes known to be pro-regenerative are required to generate healthy tissues. In the thesis it is shown that both scaffold and cell age plays a role in the regenerative capacity of a tissue. From this it is clear that to generate a healthy tissue or organ, it is critical to find the appropriate scaffold and cell type. Additionally, bioengineering manufacturing methods that generate reproducible, custom-made, high resolution constructs using cytocompatible materials are ideal for tissue engineering approaches. One such method which is compatible with the criteria above and that has emerged in recent years is 3D printing. 3D printing or bioprinting (when cells are printed) can generate custom structures relevant for human lungs. In this thesis, potential bioinks for bioprinting lung tissue are investigated. A tissue-specific hybrid bioink consisting of alginate, reinforced with extracellular matrix from decellularized lung tissue (rECM) was used to 3D bioprint human airways comprised of regionally specified primary cells which remained patent over time. The biocompatibility and vascularisation of rECM hydrogels was investigated in both T-cell immunodeficient mice mimicking the clinical scenario and immunocompetent mice. Bioprinted rECM hydrogels support the formation of an intact vascular network throughout the full thickness of the graft, comprised of both large and small size blood vessels and integrate well in the surrounding tissue.
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| 10. |
- Jiang, Dongsheng, et al.
(författare)
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Two succeeding fibroblastic lineages drive dermal development and the transition from regeneration to scarring
- 2018
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Ingår i: Nature Cell Biology. - : Springer Science and Business Media LLC. - 1465-7392 .- 1476-4679. ; 20:4, s. 422-431
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Tidskriftsartikel (refereegranskat)abstract
- During fetal development, mammalian back-skin undergoes a natural transition in response to injury, from scarless regeneration to skin scarring. Here, we characterize dermal morphogenesis and follow two distinct embryonic fibroblast lineages, based on their history of expression of the engrailed 1 gene. We use single-cell fate-mapping, live three dimensional confocal imaging and in silico analysis coupled with immunolabelling to reveal unanticipated structural and regional complexity and dynamics within the dermis. We show that dermal development and regeneration are driven by engrailed 1-history-naive fibroblasts, whose numbers subsequently decline. Conversely, engrailed 1-history-positive fibroblasts possess scarring abilities at this early stage and their expansion later on drives scar emergence. The transition can be reversed, locally, by transplanting engrailed 1-naive cells. Thus, fibroblastic lineage replacement couples the decline of regeneration with the emergence of scarring and creates potential clinical avenues to reduce scarring.
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