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Chondrocyte behavior on nanostructured micropillar polypropylene and polystyrene surfaces
Department of Applied Physics, University of Eastern Finland, Kuopio, Finland. (Chondrogenic and Osteogenic Differentiation Group)
Department of Chemistry, University of Eastern Finland, Joensuu, Finland.
Department of Applied Physics, University of Eastern Finland, Kuopio, Finland. (Chondrogenic and Osteogenic Differentiation Group)ORCID-id: 0000-0002-9294-7431
Department of Chemistry, University of Eastern Finland, Joensuu, Finland.
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2014 (Engelska)Ingår i: Materials Science and Engineering. C, Materials for Biological Applications, ISSN 0928-4931, Vol. 43, s. 424-431Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

This study was aimed to investigate whether patterned polypropylene (PP) or polystyrene (PS) could enhance the chondrocytes' extracellular matrix (ECM) production and phenotype maintenance. Bovine primary chondrocytes were cultured on smooth PP and PS, as well as on nanostructured micropillar PP (patterned PP) and PS (patterned PS) for 2 weeks. Subsequently, the samples were collected for fluorescein diacetate-based cell viability tests, for immunocytochemical assays of types I and II collagen, actin and vinculin, for scanning electronic microscopic analysis of cell morphology and distribution, and for gene expression assays of Sox9, aggrecan, procollagen α1(II), procollagen α1(X), and procollagen α2(I) using quantitative RT-PCR assays. After two weeks of culture, the bovine primary chondrocytes had attached on both patterned PP and PS, while practically no adhesion was observed on smooth PP. However, the best adhesion of the cells was on smooth PS. The cells, which attached on patterned PP and PS surfaces synthesized types I and II collagen. The chondrocytes' morphology was extended, and an abundant ECM network formed around the attached chondrocytes on both patterned PP and PS. Upon passaging, no significant differences on the chondrocyte-specific gene expression were observed, although the highest expression level of aggrecan was observed on the patterned PS in passage 1 chondrocytes, and the expression level of procollagen α1(II) appeared to decrease in passaged chondrocytes. However, the expressions of procollagen α2(I) were increased in all passaged cell cultures. In conclusion, the bovine primary chondrocytes could be grown on patterned PS and PP surfaces, and they produced extracellular matrix network around the adhered cells. However, neither the patterned PS nor PP could prevent the dedifferentiation of chondrocytes.

Ort, förlag, år, upplaga, sidor
Elsevier, 2014. Vol. 43, s. 424-431
Nyckelord [en]
Chondrocyte, polypropylene, polystyrene, micropillar, nanostructure
Nationell ämneskategori
Materialkemi Cell- och molekylärbiologi
Forskningsämne
materialvetenskap; cellforskning
Identifikatorer
URN: urn:nbn:se:umu:diva-103870DOI: 10.1016/j.msec.2014.07.045ISI: 000342529000052PubMedID: 25175232OAI: oai:DiVA.org:umu-103870DiVA, id: diva2:816068
Tillgänglig från: 2015-06-02 Skapad: 2015-06-02 Senast uppdaterad: 2018-06-07Bibliografiskt granskad
Ingår i avhandling
1. Studies on various culture systems for chondrocytes and osteoblasts
Öppna denna publikation i ny flik eller fönster >>Studies on various culture systems for chondrocytes and osteoblasts
2017 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Osteoarthritis and osteochondral defects are ailments that are increasing in frequency as the lifespan of the population increases and sedentary lifestyle becomes more common. Osteoarthritis is an inflammatory disease that causes the progressive degeneration of articular surfaces and the underlying bone. Accidents and injuries can cause osteochondral defects similar to osteoarthritis. In both cases the structure of the articular cartilage fails, leading to pain and disability. Articular cartilage has a naturally poor ability to regenerate since there is no vasculature and it is aneural. The sparse chondrocytes mainly act to maintain the healthy extracellular matrix. Once the defect is severe enough, a surgical intervention becomes necessary. For small defects and young patients, a cell-based treatment can be used, whereas for larger defects and severe osteoarthritis a partial or whole joint arthroplasty is performed. Methods to repair osteochondral defects have been improving over the years as the inter-disciplinary understanding of joints, and what is required to repair them, has increased. However, there are still issues to solve in order to achieve consistently good results in both joint replacement and repair of cartilage. The main issue faced with current techniques used for joint replacement is poor integration of the artificial joint, leading to loosening at the bone interface over time, while cartilage repair techniques face the problem of generating mechanically inferior fibrocartilage. It is known that surface chemistry and structures at micro- and nanoscale influence cell behaviour, which can be utilised to guide their attachment, proliferation and phenotype. Scaffold-free approaches and mechanical stimulation have previously given promising results in generating articular neocartilage.

This thesis aims at exploring tools and solutions to the problems involved in implant integration, chondrocyte expansion and neocartilage tissue engineering. We hypothesised that 1) ultra-short pulsed laser deposition can be used to create biocompatible coatings; 2) micropillars with nanoscale features can improve the maintenance of the chondrocyte phenotype in culture and 3) hypergravity can aid in the production of more native-like neocartilage constructs.

Our studies showed that ultra-short pulsed laser ablation can be used to create various surfaces for studying cell behaviour. Cell viability was slightly higher on a rough titanium oxide, whereas the cell area was significantly smaller on rough titanium oxide, indicating a lower amount of focal adhesions. Nanopatterned microstructures were not capable of maintaining the chondrocyte phenotype in culture, but they were not disadvantageous either. Hypergravity might help in creating a native-like distribution of collagen and proteoglycans. The constructs were more uniform in shape, but biomechanically the constructs were not different from non-centrifuged controls.

Ort, förlag, år, upplaga, sidor
Umeå: Umeå University, 2017. s. 76
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 1911
Nyckelord
Cell biology, chondrocyte, osteoarthritis, surface materials, topography, tissue engineering, mechanical stimulation
Nationell ämneskategori
Cell- och molekylärbiologi
Forskningsämne
medicinsk cellbiologi
Identifikatorer
urn:nbn:se:umu:diva-138954 (URN)978-91-7601-757-9 (ISBN)
Disputation
2017-09-27, Hörsal E, Humanisthuset, Umeå, 09:00 (Engelska)
Opponent
Handledare
Tillgänglig från: 2017-09-06 Skapad: 2017-09-04 Senast uppdaterad: 2018-06-09Bibliografiskt granskad

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