Investigating the effect of sterilisation methods on the physical properties and cytocompatibility of methyl cellulose used in combination with alginate for 3D-bioplotting of chondrocytes

Ella Hodder, Sarah Duin, David Kilian, Tilman Ahlfeld, Julia Seidel, Carsten Nachtigall, Peter Bush, Derek Covill, Michael Gelinsky, Anja Lode

Research output: Contribution to journalArticleResearchpeer-review

Abstract

For both the incorporation of cells and future therapeutic applications the sterility of a biomaterial must be ensured. However, common sterilisation techniques are intense and often negatively impact on material physicochemical attributes, which can affect its suitability for tissue engineering and 3D printing. In the present study four sterilisation methods, autoclave, supercritical CO2 (scCO2) treatment, UV- and gamma (γ) irradiation were evaluated regarding their impact on material properties and cellular responses. The investigations were performed on methyl cellulose (MC) as a component of an alginate/methyl cellulose (alg/MC) bioink, used for bioprinting embedded bovine primary chondrocytes (BPCs). In contrast to the autoclave, scCO2 and UV-treatments, the γ-irradiated MC resulted in a strong reduction in alg/MC viscosity and stability after extrusion which made this method unsuitable for precise bioprinting. Gel permeation chromatography analysis revealed a significant reduction in MC molecular mass only after γ-irradiation, which influenced MC chain mobility in the Ca2+-crosslinked alginate network as well as gel composition and microstructure. With regard to cell survival and proteoglycan matrix production, the results determined UV-irradiation and autoclaving as the best candidates for sterilisation. The scCO2-treatment of MC resulted in an unfavourable cell response indicating that this method needs careful optimisation prior to application for cell encapsulation. As proven by consistent FT-IR spectra, chemical alterations could be excluded as a cause for the differences seen between MC treatments on alg/MC behaviour. This investigation provides knowledge for the development of a clinically appropriate 3D-printing-based fabrication process to produce bioengineered tissue for cartilage regeneration.
Original languageEnglish
Article number10
JournalJournal of Materials Science: Materials in Medicine
Volume30
Issue number1
DOIs
Publication statusPublished - 4 Jan 2019

Fingerprint

Methylcellulose
Physical properties
Autoclaves
Irradiation
Printing
alginic acid
Cartilage
Gel permeation chromatography
Molecular mass
Biocompatible Materials
Proteoglycans
Encapsulation
Tissue engineering
Extrusion
Materials properties
Gels
Cells
Viscosity
Tissue
Fabrication

Bibliographical note

This is a post-peer-review, pre-copyedit version of an article published in Journal of Materials Science: Materials in Medicine. The final authenticated version is available online at: http://dx.doi.org/10.1007/s10856-018-6211-9

Keywords

  • autoclaving
  • UV-irradiation
  • gamma irradiation
  • supercritical CO2
  • bioprinting
  • methyl cellulose
  • hydrogels
  • bioink
  • biomaterial
  • additive manufacturing

Cite this

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title = "Investigating the effect of sterilisation methods on the physical properties and cytocompatibility of methyl cellulose used in combination with alginate for 3D-bioplotting of chondrocytes",
abstract = "For both the incorporation of cells and future therapeutic applications the sterility of a biomaterial must be ensured. However, common sterilisation techniques are intense and often negatively impact on material physicochemical attributes, which can affect its suitability for tissue engineering and 3D printing. In the present study four sterilisation methods, autoclave, supercritical CO2 (scCO2) treatment, UV- and gamma (γ) irradiation were evaluated regarding their impact on material properties and cellular responses. The investigations were performed on methyl cellulose (MC) as a component of an alginate/methyl cellulose (alg/MC) bioink, used for bioprinting embedded bovine primary chondrocytes (BPCs). In contrast to the autoclave, scCO2 and UV-treatments, the γ-irradiated MC resulted in a strong reduction in alg/MC viscosity and stability after extrusion which made this method unsuitable for precise bioprinting. Gel permeation chromatography analysis revealed a significant reduction in MC molecular mass only after γ-irradiation, which influenced MC chain mobility in the Ca2+-crosslinked alginate network as well as gel composition and microstructure. With regard to cell survival and proteoglycan matrix production, the results determined UV-irradiation and autoclaving as the best candidates for sterilisation. The scCO2-treatment of MC resulted in an unfavourable cell response indicating that this method needs careful optimisation prior to application for cell encapsulation. As proven by consistent FT-IR spectra, chemical alterations could be excluded as a cause for the differences seen between MC treatments on alg/MC behaviour. This investigation provides knowledge for the development of a clinically appropriate 3D-printing-based fabrication process to produce bioengineered tissue for cartilage regeneration.",
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Investigating the effect of sterilisation methods on the physical properties and cytocompatibility of methyl cellulose used in combination with alginate for 3D-bioplotting of chondrocytes. / Hodder, Ella; Duin, Sarah; Kilian, David; Ahlfeld, Tilman; Seidel, Julia; Nachtigall, Carsten; Bush, Peter; Covill, Derek; Gelinsky, Michael; Lode, Anja.

In: Journal of Materials Science: Materials in Medicine, Vol. 30, No. 1, 10, 04.01.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Investigating the effect of sterilisation methods on the physical properties and cytocompatibility of methyl cellulose used in combination with alginate for 3D-bioplotting of chondrocytes

AU - Hodder, Ella

AU - Duin, Sarah

AU - Kilian, David

AU - Ahlfeld, Tilman

AU - Seidel, Julia

AU - Nachtigall, Carsten

AU - Bush, Peter

AU - Covill, Derek

AU - Gelinsky, Michael

AU - Lode, Anja

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AB - For both the incorporation of cells and future therapeutic applications the sterility of a biomaterial must be ensured. However, common sterilisation techniques are intense and often negatively impact on material physicochemical attributes, which can affect its suitability for tissue engineering and 3D printing. In the present study four sterilisation methods, autoclave, supercritical CO2 (scCO2) treatment, UV- and gamma (γ) irradiation were evaluated regarding their impact on material properties and cellular responses. The investigations were performed on methyl cellulose (MC) as a component of an alginate/methyl cellulose (alg/MC) bioink, used for bioprinting embedded bovine primary chondrocytes (BPCs). In contrast to the autoclave, scCO2 and UV-treatments, the γ-irradiated MC resulted in a strong reduction in alg/MC viscosity and stability after extrusion which made this method unsuitable for precise bioprinting. Gel permeation chromatography analysis revealed a significant reduction in MC molecular mass only after γ-irradiation, which influenced MC chain mobility in the Ca2+-crosslinked alginate network as well as gel composition and microstructure. With regard to cell survival and proteoglycan matrix production, the results determined UV-irradiation and autoclaving as the best candidates for sterilisation. The scCO2-treatment of MC resulted in an unfavourable cell response indicating that this method needs careful optimisation prior to application for cell encapsulation. As proven by consistent FT-IR spectra, chemical alterations could be excluded as a cause for the differences seen between MC treatments on alg/MC behaviour. This investigation provides knowledge for the development of a clinically appropriate 3D-printing-based fabrication process to produce bioengineered tissue for cartilage regeneration.

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KW - gamma irradiation

KW - supercritical CO2

KW - bioprinting

KW - methyl cellulose

KW - hydrogels

KW - bioink

KW - biomaterial

KW - additive manufacturing

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