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Home / Equine Research Bank / PeerJ / Culture of equine fibroblast-like synoviocytes on synthetic ...
PeerJ2014; 2; e353; doi: 10.7717/peerj.353

Culture of equine fibroblast-like synoviocytes on synthetic tissue scaffolds towards meniscal tissue engineering: a preliminary cell-seeding study.

Abstract: Introduction. Tissue engineering is a new methodology for addressing meniscal injury or loss. Synovium may be an ideal source of cells for in vitro meniscal fibrocartilage formation, however, favorable in vitro culture conditions for synovium must be established in order to achieve this goal. The objective of this study was to determine cellularity, cell distribution, and extracellular matrix (ECM) formation of equine fibroblast-like synoviocytes (FLS) cultured on synthetic scaffolds, for potential application in synovium-based meniscal tissue engineering. Scaffolds included open-cell poly-L-lactic acid (OPLA) sponges and polyglycolic acid (PGA) scaffolds cultured in static and dynamic culture conditions, and PGA scaffolds coated in poly-L-lactic (PLLA) in dynamic culture conditions. Materials and Methods. Equine FLS were seeded on OPLA and PGA scaffolds, and cultured in a static environment or in a rotating bioreactor for 12 days. Equine FLS were also seeded on PGA scaffolds coated in 2% or 4% PLLA and cultured in a rotating bioreactor for 14 and 21 days. Three scaffolds from each group were fixed, sectioned and stained with Masson's Trichrome, Safranin-O, and Hematoxylin and Eosin, and cell numbers and distribution were analyzed using computer image analysis. Three PGA and OPLA scaffolds from each culture condition were also analyzed for extracellular matrix (ECM) production via dimethylmethylene blue (sulfated glycosaminoglycan) assay and hydroxyproline (collagen) assay. PLLA coated PGA scaffolds were analyzed using double stranded DNA quantification as areflection of cellularity and confocal laser microscopy in a fluorescent cell viability assay. Results. The highest cellularity occurred in PGA constructs cultured in a rotating bioreactor, which also had a mean sulfated glycosaminoglycan content of 22.3 µg per scaffold. PGA constructs cultured in static conditions had the lowest cellularity. Cells had difficulty adhering to OPLA and the PLLA coating of PGA scaffolds; cellularity was inversely proportional to the concentration of PLLA used. PLLA coating did not prevent dissolution of the PGA scaffolds. All cell scaffold types and culture conditions produced non-uniform cellular distribution. Discussion/Conclusion. FLS-seeding of PGA scaffolds cultured in a rotating bioreactor resulted in the most optimal cell and matrix characteristics seen in this study. Cells grew only in the pores of the OPLA sponge, and could not adhere to the PLLA coating of PGA scaffold, due to the hydrophobic property of PLA. While PGA culture in a bioreactor produced measureable GAG, no culture technique produced visible collagen. For this reason, and due to the dissolution of PGA scaffolds, the culture conditions and scaffolds described here are not recommended for inducing fibrochondrogenesis in equine FLS for meniscal tissue engineering.
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Publication Date: 2014-04-17 PubMed ID: 24765587PubMed Central: PMC3994628DOI: 10.7717/peerj.353Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study focuses on exploring the effectiveness of different synthetic scaffolds for growing equine fibroblast-like synoviocytes (FLS) for potential use in meniscal tissue engineering. The results suggest that polyglycolic acid (PGA) scaffolds in dynamic culture conditions show the best growth characteristics; however, none of the tested scaffolds or conditions could successfully induce fibrochondrogenesis, thereby highlighting their unsuitability for meniscal tissue engineering.

Research Objective and Methodology

  • The main objective of this study was to assess the growth characteristics of equine fibroblast-like synoviocytes (cells found in the membrane lining joints) on different synthetic scaffolds— specifically, poly-L-lactic acid (OPLA) sponges and polyglycolic acid (PGA) scaffolds. Researchers wanted to evaluate the potential of these cells for use in meniscal tissue engineering.
  • The FLS were seeded on the different scaffolds and cultured either in a stationary environment or a rotating bioreactor for up to 21 days. They were also seeded on PGA scaffolds coated with a substance called poly-L-lactic (PLLA) and cultured in the rotating bioreactor.
  • After culture, the scaffolds were analyzed for cell numbers, cell distribution, and production of extracellular matrix (ECM), the three-dimensional network of proteins and other biomolecules that cells produce and live within.

Research Findings

  • The PGA scaffolds cultured in the rotating bioreactor showed the highest cell numbers and the highest concentration of sulfated glycosaminoglycan, an important component of the ECM. However, these scaffolds had poor adherence properties, as cells struggled to attach to the OPLA and the PLLA-coated PGA scaffolds.
  • Cell numbers were inversely proportional to the concentration of PLLA used, suggesting that this coating was not beneficial to cell growth. Furthermore, the PLLA coating did not prevent the PGA scaffolds from dissolving over time.
  • All of the tested scaffolds and culture conditions resulted in a non-uniform cell distribution, suggesting that the cells did not grow evenly across the scaffold structure.

Conclusions and Implications

  • Of all the tested scaffold-culture combinations, the FLS-seeded PGA scaffolds in a rotating bioreactor showed the most promising results in terms of cell numbers and ECM production. However, the issues with cell attachment and scaffold dissolution make them unsuitable for their intended purpose— to induce fibrochondrogenesis, the formation of fibrocartilage tissue, in meniscal tissue engineering.
  • This preliminary study reveals the challenges related to culture conditions and scaffold properties in meniscal tissue engineering and points to the need for further research in this area.

Cite This Article

APA
Warnock JJ, Fox DB, Stoker AM, Beatty M, Cockrell M, Janicek JC, Cook JL. (2014). Culture of equine fibroblast-like synoviocytes on synthetic tissue scaffolds towards meniscal tissue engineering: a preliminary cell-seeding study. PeerJ, 2, e353. https://doi.org/10.7717/peerj.353

Publication

PeerJ
ISSN: 2167-8359
NlmUniqueID: 101603425
Country: United States
Language: English
Volume: 2
Pages: e353
PII: e353

Researcher Affiliations

Warnock, Jennifer J
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.
Fox, Derek B
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.
Stoker, Aaron M
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.
Beatty, Mark
  • VA Nebraska-Western Iowa Health Care System and University of Nebraska Medical Center College of Dentistry , Lincoln, NE , USA.
Cockrell, Mary
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.
Janicek, John C
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.
Cook, James L
  • Comparative Orthopaedic Laboratory, University of Missouri , Columbia, MO , USA.

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