liu.seSearch for publications in DiVA
Change search
ReferencesLink to record
Permanent link

Direct link
Human articular chondrocytes on macroporous gelatin microcarriers form structurally stable constructs with blood-derived biological glues in vitro
Linköping University, Department of Clinical and Experimental Medicine, Surgery. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Rheumatology. Linköping University, Faculty of Health Sciences.ORCID iD: 0000-0002-6916-5490
University of Gothenburg.
Linköping University, Department of Clinical and Experimental Medicine, Plastic Surgery, Hand Surgery and Burns. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Reconstruction Centre, Department of Plastic Surgery, Hand surgery UHL.
2009 (English)In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, ISSN 1932-6254, Vol. 35, no 6, 450-460 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2009. Vol. 35, no 6, 450-460 p.
Keyword [en]
cartilage, chondrocyte, microcarrier, tissue engineering, fibrin glue, platelet-rich plasma
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-20597DOI: 10.1002/term.179OAI: diva2:235330
Available from: 2009-09-15 Created: 2009-09-15 Last updated: 2015-06-29
In thesis
1. Biodegradable gelatin microcarriers in tissue engineering: In vitro studies on cartilage and bone
Open this publication in new window or tab >>Biodegradable gelatin microcarriers in tissue engineering: In vitro studies on cartilage and bone
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Tissue engineering is a multidisciplinary field that combines cells, biomaterial scaffolds and environmental factors to achieve functional tissue repair. This thesis focuses on the use of macroporous gelatin microcarriers as scaffolds in tissue engineering applications, with a special focus on cartilage and bone formation by human adult cells in vitro.

In our first study, human articular chondrocytes were seeded on macroporous gelatin microcarriers. The microcarriers were subsequently encapsulated in coagulated blood-derived biological glues and cultured under free-swelling conditions for up to 17 weeks. Even in the absence of recombinant chondrogenic growth factors, the chondrocytes remained viable and metabolically active for the duration of the culture period, as indicated by an increased amount of cell nuclei and extracellular matrix (ECM). The ECM showed several cartilage characteristics, but lacked the cartilage specific collagen type II. Furthermore, ECM formation was seen primarily in a capsule surrounding the tissue-engineered constructs, leading to the conclusion that the used in vitro models were unable to support true cartilage formation.

The capacity of human dermal fibroblasts to produce cartilage- and bone-like tissue in the previously mentioned model was also investigated. Under the influence of chondrogenic induction factors, including TGF-β1 and insulin, the fibroblasts produced cartilage specific molecules, as confirmed by indirect immunohistochemistry, however not collagen type II. Under osteogenic induction, by dexamethasone, ascorbate-2-phosphate and β–glycerophosphate, the fibroblasts formed a calcified matrix with bone specific markers, and an alkaline phosphatase assay corroborated a shift towards an osteoblast like phenotype. The osteogenic induction was enhanced by flow-induced shear stress in a spinner flask system.

In addition, four different types of gelatin microcarriers, differing by their internal pore diameter and their degree of gelatin cross-linking, were evaluated for their ability to support chondrocyte expansion. Chondrocyte densities on the microcarriers were monitored every other day over a twoweek period, and chondrocyte growth was analyzed by piecewise linear regression and analysis of variance (ANOVA). No differences were seen between the different microcarriers during the first week. However, during the second week of culture both microcarrier pore diameter and gelatin crosslinking had significant impacts on chondrocyte density.

Lastly, a dynamic centrifugation regime (f=12.5 mHz for 16 minutes every other day) was administered to chondrocyte-seeded microcarriers, with or without encapsulation in platelet rich plasma (PRP), to study the possible effect of dynamic stimuli on cartilage formation. Presence of PRP enhanced the structural stability of the tissue-engineered constructs, but we were not able to confirm any dose-response pattern between ECM formation and the applied forces. After 12 weeks, distinct gelatin degradation had occurred independent of both dynamic stimuli and presence of PRP.

In summary, this thesis supports a plausible use for gelatin microcarriers in tissue engineering of cartilage and bone. Microcarrier characteristics, specifically gelatin cross-linking and pore diameter, have been shown to affect chondrocyte expansion. In addition, the use of human dermal fibroblasts as an alternative cell source for cartilage and bone formation in vitro was addressed.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2009. 68 p.
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1147
National Category
Medical and Health Sciences
urn:nbn:se:liu:diva-54116 (URN)978-91-7393-557-9 (ISBN)
Public defence
2009-09-25, Berzeliussalen, Hälsouniversitet, Campus US, Linköpings universitet, Linköping, 13:00 (Swedish)
Available from: 2010-02-23 Created: 2010-02-23 Last updated: 2015-06-29Bibliographically approved

Open Access in DiVA

fulltext(1157 kB)137 downloads
File information
File name FULLTEXT01.pdfFile size 1157 kBChecksum SHA-512
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Search in DiVA

By author/editor
Pettersson, SofiaWetterö, JonasKratz, Gunnar
By organisation
SurgeryFaculty of Health SciencesRheumatologyPlastic Surgery, Hand Surgery and BurnsDepartment of Plastic Surgery, Hand surgery UHL
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
Total: 137 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 131 hits
ReferencesLink to record
Permanent link

Direct link