In Vitro Study of Human Immune Responses to Hyaluronic Acid Hydrogels, Recombinant Spidroins and Human Neural Progenitor Cells of Relevance to Spinal Cord Injury Repair

Cells. 2021 Jul 6;10(7):1713. doi: 10.3390/cells10071713.

Abstract

Scaffolds of recombinant spider silk protein (spidroin) and hyaluronic acid (HA) hydrogel hold promise in combination with cell therapy for spinal cord injury. However, little is known concerning the human immune response to these biomaterials and grafted human neural stem/progenitor cells (hNPCs). Here, we analyzed short- and long-term in vitro activation of immune cells in human peripheral blood mononuclear cells (hPBMCs) cultured with/without recombinant spidroins, HA hydrogels, and/or allogeneic hNPCs to assess potential host-donor interactions. Viability, proliferation and phenotype of hPBMCs were analyzed using NucleoCounter and flow cytometry. hPBMC viability was confirmed after exposure to the different biomaterials. Short-term (15 h) co-cultures of hPBMCs with spidroins, but not with HA hydrogel, resulted in a significant increase in the proportion of activated CD69+ CD4+ T cells, CD8+ T cells, B cells and NK cells, which likely was caused by residual endotoxins from the Escherichia coli expression system. The observed spidroin-induced hPBMC activation was not altered by hNPCs. It is resource-effective to evaluate human compatibility of novel biomaterials early in development of the production process to, when necessary, make alterations to minimize rejection risk. Here, we present a method to evaluate biomaterials and hPBMC compatibility in conjunction with allogeneic human cells.

Keywords: artificial spidroin; human immune response; human neural progenitor cell; hyaluronic acid hydrogel; spinal cord injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abortion, Legal
  • CD4-Positive T-Lymphocytes / cytology
  • CD4-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / cytology
  • CD8-Positive T-Lymphocytes / immunology
  • Cell Encapsulation / methods
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Coculture Techniques
  • Female
  • Fetus
  • Fibroins / chemistry
  • Fibroins / pharmacology*
  • Humans
  • Hyaluronic Acid / chemistry
  • Hyaluronic Acid / pharmacology*
  • Hydrogels / chemistry
  • Hydrogels / pharmacology*
  • Killer Cells, Natural / cytology
  • Killer Cells, Natural / immunology
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / immunology
  • Lymphocyte Activation
  • Models, Biological
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects*
  • Neural Stem Cells / immunology
  • Pregnancy
  • Pregnancy Trimester, First
  • Primary Cell Culture
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / pharmacology
  • Spinal Cord / cytology
  • Spinal Cord / drug effects*
  • Spinal Cord / immunology
  • Spinal Cord Injuries / immunology
  • Spinal Cord Injuries / pathology

Substances

  • Hydrogels
  • Recombinant Proteins
  • Hyaluronic Acid
  • Fibroins