Sulfated Hydrogels as Primary Intervertebral Disc Cell Culture Systems

Author:

Bermudez-Lekerika Paola12ORCID,Crump Katherine B.12ORCID,Wuertz-Kozak Karin34ORCID,Le Maitre Christine L.5ORCID,Gantenbein Benjamin16ORCID

Affiliation:

1. Tissue Engineering for Orthopaedics and Mechanobiology, Bone & Joint Program, Department for BioMedical Research (DBMR), Medical Faculty, University of Bern, 3008 Bern, Switzerland

2. Graduate School for Cellular and Biomedical Sciences (GCB), University of Bern, 3012 Bern, Switzerland

3. Department of Biomedical Engineering, Rochester Institute of Technology, Rochester, NY 14623, USA

4. Spine Center, Schön Klinik München Harlaching Academic Teaching Hospital, Spine Research Institute, Paracelsus Private Medical University Salzburg (Austria), 81547 Munich, Germany

5. Division of Clinical Medicine, School of Medicine and Population Health, University of Sheffield, Sheffield S10 2TN, UK

6. Inselspital, Department of Orthopedic Surgery & Traumatology, Medical Faculty, University of Bern, 3010 Bern, Switzerland

Abstract

The negatively charged extracellular matrix plays a vital role in intervertebral disc tissues, providing specific cues for cell maintenance and tissue hydration. Unfortunately, suitable biomimetics for intervertebral disc regeneration are lacking. Here, sulfated alginate was investigated as a 3D culture material due to its similarity to the charged matrix of the intervertebral disc. Precursor solutions of standard alginate, or alginate with 0.1% or 0.2% degrees of sulfation, were mixed with primary human nucleus pulposus cells, cast, and cultured for 14 days. A 0.2% degree of sulfation resulted in significantly decreased cell density and viability after 7 days of culture. Furthermore, a sulfation-dependent decrease in DNA content and metabolic activity was evident after 14 days. Interestingly, no significant differences in cell density and viability were observed between surface and core regions for sulfated alginate, unlike in standard alginate, where the cell number was significantly higher in the core than in the surface region. Due to low cell numbers, phenotypic evaluation was not achieved in sulfated alginate biomaterial. Overall, standard alginate supported human NP cell growth and viability superior to sulfated alginate; however, future research on phenotypic properties is required to decipher the biological properties of sulfated alginate in intervertebral disc cells.

Funder

Marie Skłodowska Curie International Training Network (ITN) “disc4all”

Publisher

MDPI AG

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