| 1. |
- Aulin, Cecilia, et al.
(författare)
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Extracellular matrix-polymer hybrid materials produced in a pulsed-flow bioreactor system
- 2009
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Ingår i: Journal of Tissue Engineering and Regenerative Medicine. - : John Wiley & Sons, Ltd. - 1932-6254 .- 1932-7005. ; 3:3, s. 188-195
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Tidskriftsartikel (refereegranskat)abstract
- Cell adhesion, interaction with material, cell proliferation and the production of an extracellular matrix (ECM) are all important factors determining the successful performance of an engineered scaffold. Scaffold design should aim at creating structures which can guide cells into forming new, functional tissue. In this study, the concept of in situ deposition of ECM by human dermal fibroblasts onto a compliant, knitted poly (ethyleneterephtalate) support is demonstrated, creating in vitro produced ECM polymer hybrid materials for tissue engineering. Comparison of cells cultured under static and dynamic conditions were examined, and the structure and morphology of the materials so formed were evaluated, along with the amount collagen deposited by the seeded cells. In vitro produced ECM polymer hybrid scaffolds could be created in this way, with the dynamic culture conditions increasing ECM deposition. Histological analysis indicated a homogenous distribution of cells in the 1 mm thick scaffold, surrounded by a matrix-like structure. ECM deposition was observed throughout the materials wigh 81.6 µg/cm2 of collagen deposited after 6 weeks. Cell produced bundles of ECM fibres bridged the polymer filaments and anchored cells to the support. These findings open hereto unknown possibilities of producing materials with structure designed by engineering together with biochemical composition given by cells.
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| 2. |
- Bodin, Aase Katarina, 1977, et al.
(författare)
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Bacterial cellulose as a potential meniscus implant
- 2007
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Ingår i: Journal of tissue engineering and regenerative medicine. - : Hindawi Limited. - 1932-6254 .- 1932-7005. ; 1:5, s. 406-8
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Tidskriftsartikel (refereegranskat)abstract
- Traumatic or degenerative meniscal lesions are a frequent problem. The meniscus cannot regenerate after resection. These lesions often progress and lead to osteoarthritis. Collagen meniscal implants have been used in clinical practice to regenerate meniscal tissue after partial meniscectomy. The mechanical properties of bacterial cellulose (BC) gel were compared with a collagen material and the pig meniscus. BC was grown statically in corn steep liquid medium, as described elsewhere. Pig meniscus was harvested from pigs. The collagen implant was packed in sterile conditions until use. The different materials were evaluated under tensile and compression load, using an Instron 5542 with a 500 N load cell. The feasibility for implantation was explored using a pig model. The Young's modulus of bacterial cellulose was measured to be 1 MPa, 100 times less for the collagen material, 0.01 MPa in tensile load. The Young's modulus of bacterial cellulose and meniscus are similar in magnitude under a compression load of 2 kPa and with five times better mechanical properties than the collagen material. At higher compression strain, however, the pig meniscus is clearly stronger. These differences are clearly due to a more ordered and arranged structure of the collagen fibrils in the meniscus. The combination of the facts that BC is inexpensive, can be produced in a meniscus shape, and promotes cell migration makes it an attractive material for consideration as a meniscus implant. Copyright (c) 2007 John Wiley & Sons, Ltd.
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| 3. |
- Bäckdahl, Henrik, 1977, et al.
(författare)
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Engineering microporosity in bacterial cellulose scaffolds
- 2008
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Ingår i: Journal of tissue engineering and regenerative medicine. - : Hindawi Limited. - 1932-6254 .- 1932-7005. ; 2:6, s. 320-330
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Tidskriftsartikel (refereegranskat)abstract
- The scaffold is an essential component in tissue engineering. A novel method to prepare threedimensional (3D) nanofibril network scaffolds with controlled microporosity has been developed. By placing paraffin wax and starch particles of various sizes in a growing culture of Acetobacter xylinum, bacterial cellulose scaffolds of different morphologies and interconnectivity were prepared. Paraffin particles were incorporated throughout the scaffold, while starch particles were found only in the outermost area of the resulting scaffold. The porogens were successfully removed after culture with bacteria and no residues were detected with electron spectroscopy for chemical analysis (ESCA) or Fourier transform infra-red spectroscopy (FT-IR). Resulting scaffolds were seeded with smooth muscle cells (SMCs) and investigated using histology and organ bath techniques. SMC were selected as the cell type since the main purpose of the resulting scaffolds is for tissue engineered blood vessels. SMCs attached to and proliferated on and partly into the scaffolds. Copyright © 2008 John Wiley & Sons, Ltd.
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| 4. |
- Concaro, Sebastian, et al.
(författare)
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Effect of cell seeding concentration on the quality of tissue engineered constructs loaded with adult human articular chondrocytes.
- 2008
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Ingår i: Journal of tissue engineering and regenerative medicine. - : Hindawi Limited. - 1932-6254 .- 1932-7005. ; 2:1, s. 14-21
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Tidskriftsartikel (refereegranskat)abstract
- Many aspects of the process of in vitro differentiation of chondrocytes in three-dimensional (3D) scaffolds need to be further investigated. Chitosan scaffolds were produced by freeze-drying 3% w/v 90% DDA chitosan gels. The effect of the cell seeding concentration was evaluated by culturing human adult chondrocytes in chitosan scaffolds After the first passage, cells were seeded into chitosan scaffolds with a diameter of 8 mm. The final cell seeding concentration per cm3 of chitosan scaffold was: Group A, 3 x 10(6); Group B, 6 x 10(6); Group C, 12 x 10(6); and Group D, 25 x 10(6) cells. After 14 and 28 days in 3D culture, the constructs were assesed for collagen, glucosaminoglycans and DNA content. The mechanical properties of the constructs were determined using a dynamic oscillatory shear test. The histological aspect of the constructs was evaluated using the Bern score. The collagen and GAG concentration increased, varying the cell seeding concentration. There was a significant increase in proteoglycan and hydroxyproline production between groups C and D. The sulphated GAG content increased significantly in the group D as compared to the other groups. The mechanical properties of the different constructs increased over time, from 9.6 G'/kPa at 14 days of 3D culture to 14.6 G'/kPa at 28 days under the same culture conditions. In this study we were able to determine that concentrations of 12-25 million cells/cm2 are needed to increase the matrix production and mechanical properties of human adult chondrocytes under static conditions.
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| 5. |
- Dravida, Subhadra, et al.
(författare)
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A biomimetic scaffold for culturing limbal stem cells: a promising alternative for clinical transplantation
- 2008
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Ingår i: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE. - : John Wiley and Sons. - 1932-6254 .- 1932-7005. ; 2:5, s. 263-271
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Tidskriftsartikel (refereegranskat)abstract
- Limbal tissues can be cultured on various types of scaffolds to create a sheet of limbal-corneal epithelium for research as well as clinical transplantation. An optically clear, biocompatible, biomimetic scaffold would be an ideal replacement graft for transplanting limbal stem cells. in this study, we evaluated the physical and culture characteristics of the recombinant human cross-linked collagen scaffold (RHC-III scaffold) and compared it with denuded human amniotic membrane (HAM). Optical/mechanical properties and microbial susceptibility were measured for the scaffolds. With the approval of the institutional review board, 2 mm. fresh human limbal tissues were cultured on 2.5 x 2.5 cm(2) scaffolds in a medium containing autologous serum in a feeder cell-free submerged system. The cultured cell systems were characterized by morphology and immunohistochemistry for putative stem cells and differentiated cell markers. The refractive index (RI) and tensile strength of the RHC-III scaffold were comparable to human cornea, with delayed in vitro degradation compared to HAM. RHC-III scaffolds were 10-fold less susceptible to microbial growth. Cultures were initiated on day 1, expanded to form a monolayer by day 3 and covered the entire growth surface in 10 days. Stratified epithelium on the scaffolds was visualized by transmission electron microscopy. The cultured cells showed p63 and ABCG2 positivity in the basal layer and were immunoreactive for cytokeratin K3 and K12 in the suprabasal layers. RHC-III scaffold supports and retains the growth and stemness of limbal stem cells, in addition to resembling human cornea; thus, it could be a good replacement scaffold for growing cells for clinical transplantation.
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| 6. |
- Foroughi, Farhad, et al.
(författare)
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Bulk collagen incorporation rates into knitted stiff fibre polymer in tissue-engineered scaffolds : the rate-limiting step
- 2008
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Ingår i: Journal of Tissue Engineering and Regenerative Medicine. - : John Wiley & Sons, Ltd.. - 1932-6254 .- 1932-7005. ; 2:8, s. 507-514
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Tidskriftsartikel (refereegranskat)abstract
- Fabrication of tissue-engineered constructs in vitro relies on sufficient synthesis of extracellular matrix (ECM) by cells to form a material suitable for normal function in vivo. Collagen synthesis by human dermal fibroblasts grown in vitro on two polymers, polyethylene terephthalate (PET) and polyglycolic acid (PGA), was measured by high-performance liquid chromatography (HPLC). Cells were either cultured in a dynamic environment, where meshes were loaded onto a pulsing tube in a bioreactor, or in a static environment without pulsing. Collagen synthesis by cells cultured on a static mesh increased by six-fold compared to monolayer culture, and increased by up to a further 5.4-fold in a pulsed bioreactor. However, little of the collagen synthesized was deposited onto the meshes, almost all being lost to the medium. The amount of collagen deposited onto meshes was highest when cells were cultured dynamically on PET meshes (17.6 µg), but deposition still represented only 1.4% of the total synthesized. Although total collagen synthesis was increased by the use of 3D culture and the introduction of pulsing, the results suggest that the limiting factor for fabrication of a tissue-engineered construct within practical timescales is not the amount of collagen synthesized but the quantity retained (i.e. deposited) within the construct during culture. This may be enhanced by systems which promote or assemble true 3D multi-layers of cells.
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| 7. |
- Hilborn, Jöns, et al.
(författare)
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A new and evolving paradigm for biocompatibitity
- 2007
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Ingår i: Journal of Tissue Engineering and Regenerative Medicine. - : Hindawi Limited. - 1932-6254 .- 1932-7005. ; 1:2, s. 110-119
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Tidskriftsartikel (refereegranskat)abstract
- We propose that the mechanical property of the interface between an implant and its surrounding tissues is critical for the host response and the performance of the device. The interfacial mechanics depends on several different factors related to the physical shape of the device and its surface as well as properties of the host tissue and the loading conditions of the device and surrounding tissue. It seems plausible that the growth of the fibrotic tissue to support mechanical loads is governed by the same priniciples as depicted by Wolfs' Law for bone. Of course, biocompatibility will have different implications depending on which vantage point we look at the host-material interface. Another implication is that only limited aspects of biocompatibility is measurable with current in vitro tests and that the elicited host response in vivo models remains crucial for evaluation of medical devices and tissue engineering constructs.
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| 8. |
- Pettersson, Sofia, et al.
(författare)
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Human articular chondrocytes on macroporous gelatin microcarriers form structurally stable constructs with blood-derived biological glues in vitro.
- 2009
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Ingår i: Journal of tissue engineering and regenerative medicine. - : Hindawi Limited. - 1932-7005 .- 1932-6254. ; 3:6, s. 450-60
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Tidskriftsartikel (refereegranskat)abstract
- Biodegradable macroporous gelatin microcarriers fixed with blood-derived biodegradable glue are proposed as a delivery system for human autologous chondrocytes. Cell-seeded microcarriers were embedded in four biological glues-recalcified citrated whole blood, recalcified citrated plasma with or without platelets, and a commercially available fibrin glue-and cultured in an in vitro model under static conditions for 16 weeks. No differences could be verified between the commercial fibrin glue and the blood-derived alternatives. Five further experiments were conducted with recalcified citrated platelet-rich plasma alone as microcarrier sealant, using two different in vitro culture models and chondrocytes from three additional donors. The microcarriers supported chondrocyte adhesion and expansion as well as extracellular matrix (ECM) synthesis. Matrix formation occurred predominantly at sample surfaces under the static conditions. The presence of microcarriers proved essential for the glues to support the structural takeover of ECM proteins produced by the embedded chondrocytes, as exclusion of the microcarriers resulted in unstable structures that dissolved before matrix formation could occur. Immunohistochemical analysis revealed the presence of SOX-9- and S-100-positive chondrocytes as well as the production of aggrecan and collagen type I, but not of the cartilage-specific collagen type II. These results imply that blood-derived glues are indeed potentially applicable for encapsulation of chondrocyte-seeded microcarriers. However, the static in vitro models used in this study proved incapable of supporting cartilage formation throughout the engineered constructs.
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