TY - JOUR
T1 - Impact of Endotoxins in Gelatine Hydrogels on Chondrogenic Differentiation and Inflammatory Cytokine Secretion In Vitro
AU - Groen, Wilhelmina M G A C
AU - Utomo, Lizette
AU - Castilho, Miguel
AU - Gawlitta, Debby
AU - Malda, Jos
AU - Weeren, P René van
AU - Levato, Riccardo
AU - Korthagen, Nicoline M
N1 - Funding Information:
Funding: This research was funded by the European Research Council under grant agreement n◦647426 (3D-JOINT), and the Dutch Arthritis Society (LLP-12, LLP-22). The APC was funded by the Department of Clinical Sciences from the faculty of Veterinary Medicine of Utrecht University.
Publisher Copyright:
© 2020, MDPI AG. All rights reserved.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2020/11/2
Y1 - 2020/11/2
N2 - Gelatine methacryloyl (GelMA) hydrogels are widely used in studies aimed at cartilage regeneration. However, the endotoxin content of commercially available GelMAs and gelatines used in these studies is often overlooked, even though endotoxins may influence several cellular functions. Moreover, regulations for clinical use of biomaterials dictate a stringent endotoxin limit. We determined the endotoxin level of five different GelMAs and evaluated the effect on the chondrogenic differentiation of equine mesenchymal stromal cells (MSCs). Cartilage-like matrix production was evaluated by biochemical assays and immunohistochemistry. Furthermore, equine peripheral blood mononuclear cells (PBMCs) were cultured on the hydrogels for 24 h, followed by the assessment of tumour necrosis factor (TNF)-α and C-C motif chemokine ligand (CCL)2 as inflammatory markers. The GelMAs were found to have widely varying endotoxin content (two with >1000 EU/mL and three with <10 EU/mL), however, this was not a critical factor determining in vitro cartilage-like matrix production of embedded MSCs. PBMCs did produce significantly higher TNF-α and CCL2 in response to the GelMA with the highest endotoxin level compared to the other GelMAs. Although limited effects on chondrogenic differentiation were found in this study, caution with the use of commercial hydrogels is warranted in the translation from in vitro to in vivo studies because of regulatory constraints and potential inflammatory effects of the content of these hydrogels.
AB - Gelatine methacryloyl (GelMA) hydrogels are widely used in studies aimed at cartilage regeneration. However, the endotoxin content of commercially available GelMAs and gelatines used in these studies is often overlooked, even though endotoxins may influence several cellular functions. Moreover, regulations for clinical use of biomaterials dictate a stringent endotoxin limit. We determined the endotoxin level of five different GelMAs and evaluated the effect on the chondrogenic differentiation of equine mesenchymal stromal cells (MSCs). Cartilage-like matrix production was evaluated by biochemical assays and immunohistochemistry. Furthermore, equine peripheral blood mononuclear cells (PBMCs) were cultured on the hydrogels for 24 h, followed by the assessment of tumour necrosis factor (TNF)-α and C-C motif chemokine ligand (CCL)2 as inflammatory markers. The GelMAs were found to have widely varying endotoxin content (two with >1000 EU/mL and three with <10 EU/mL), however, this was not a critical factor determining in vitro cartilage-like matrix production of embedded MSCs. PBMCs did produce significantly higher TNF-α and CCL2 in response to the GelMA with the highest endotoxin level compared to the other GelMAs. Although limited effects on chondrogenic differentiation were found in this study, caution with the use of commercial hydrogels is warranted in the translation from in vitro to in vivo studies because of regulatory constraints and potential inflammatory effects of the content of these hydrogels.
KW - Articular cartilage regeneration
KW - Gelatine Methacryloyl (GelMA)
KW - Inflammatory mediator
KW - Lipopolysaccharide (LPS)
KW - Mesenchymal stromal cell (MSC)
KW - Peripheral blood mononuclear cell (PBMC)
KW - Regenerative medicine
KW - Type A and type B gelatine
UR - http://www.scopus.com/inward/record.url?scp=85096239552&partnerID=8YFLogxK
U2 - 10.3390/ijms21228571
DO - 10.3390/ijms21228571
M3 - Article
C2 - 33202964
SN - 1422-0067
VL - 21
SP - 1
EP - 17
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 22
M1 - 8571
ER -