Production and Characterization of Chitooligosaccharides: Evaluation of Acute Toxicity, Healing, and Anti-Inflammatory Actions

Int J Mol Sci. 2021 Sep 30;22(19):10631. doi: 10.3390/ijms221910631.

Abstract

The search for promising biomolecules such as chitooligosaccharides (COS) has increased due to the need for healing products that act efficiently, avoiding complications resulting from exacerbated inflammation. Therefore, this study aimed to produce COS in two stages of hydrolysis using chitosanases derived from Bacillus toyonensis. Additionally, this study aimed to structurally characterize the COS via mass spectrometry, to analyze their biocompatibility in acute toxicity models in vivo, to evaluate their healing action in a cell migration model in vitro, to analyze the anti-inflammatory activity in in vivo models of xylol-induced ear edema and zymosan-induced air pouch, and to assess the wound repair action in vivo. The structural characterization process pointed out the presence of hexamers. The in vitro and in vivo biocompatibility of COS was reaffirmed. The COS stimulated the fibroblast migration. In the in vivo inflammatory assays, COS showed an antiedematogenic response and significant reductions in leukocyte migration, cytokine release, and protein exudate. The COS healing effect in vivo was confirmed by the significant wound reduction after seven days of the experiment. These results indicated that the presence of hexamers influences the COS biological properties, which have potential uses in the pharmaceutical field due to their healing and anti-inflammatory action.

Keywords: biocompatibility; chitosan; enzymatic hydrolysis; in vivo tests; oligosaccharides.

MeSH terms

  • 3T3 Cells
  • Animals
  • Anti-Inflammatory Agents / administration & dosage*
  • Anti-Inflammatory Agents / chemistry
  • Bacillus / enzymology
  • Biocompatible Materials / administration & dosage*
  • Biocompatible Materials / chemistry
  • Cell Movement / drug effects
  • Cell Survival / drug effects
  • Chitosan / administration & dosage*
  • Chitosan / chemistry
  • Cytokines / metabolism
  • Disease Models, Animal
  • Ear Diseases / chemically induced
  • Ear Diseases / drug therapy*
  • Edema / chemically induced
  • Edema / drug therapy*
  • Female
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Glycoside Hydrolases / chemistry
  • Hydrolysis
  • Inflammation / drug therapy
  • Inflammation / metabolism
  • Leukocytes / drug effects
  • Leukocytes / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Oligosaccharides / administration & dosage*
  • Oligosaccharides / chemistry
  • Wound Healing / drug effects*

Substances

  • Anti-Inflammatory Agents
  • Biocompatible Materials
  • Cytokines
  • Oligosaccharides
  • oligochitosan
  • Chitosan
  • Glycoside Hydrolases
  • chitosanase

Supplementary concepts

  • Bacillus toyonensis