Extracellular matrix hydrogel derived from bovine bone is biocompatible in vitro and in vivo

Biomed Mater Eng. 2022;33(6):491-504. doi: 10.3233/BME-211387.

Abstract

Background: Nowadays, biomaterials used as a scaffold must be easy to deliver in the bone defect area. Extracellular matrix (ECM) hydrogels are highly hydrated polymers that can fill irregular shapes and act as bioactive materials.

Objective: This work aims to show the effects of ECM hydrogels derived from bovine bone (bECMh) on proliferation, cytotoxicity and expression of pro-inflammatory cytokines in three cells types involved in tissue regeneration, as well as biocompatibility in vivo.

Methods: In vitro, we used an extract of bECMh to test it on macrophages, fibroblasts, and adipose-derived mesenchymal stem cells (AD-MCSs). Cell proliferation was measured using the MTT assay, cytotoxicity was measured by quantifying lactate dehydrogenase release and the Live/Dead Cell Imaging assays. Concentrations of IL-6, IL-10, IL-12p70, MCP-1 and TNF-α were quantified in the supernatants using a microsphere-based cytometric bead array. For in vivo analysis, Wistar rats were inoculated into the dorsal sub-dermis with bECMh, taking as reference the midline of the back. The specimens were sacrificed at 24 h for histological study.

Results: In vitro, this hydrogel behaves as a dynamic biomaterial that increases fibroblast proliferation, induces the production of pro-inflammatory cytokines in macrophages, among which MCP-1 and TNF-α stand out. In vivo, bECMh allows the colonization of host fibroblast-like and polymorphonuclear cells, without tissue damage or inflammation.

Conclusions: The results indicate that bECMh is a biocompatible material that could be used as a scaffold, alone or in conjunction with cells or functional biomolecules, enhancing proliferation and allowing the filling of bone defects to its further regeneration.

Keywords: Bovine extracellular matrix hydrogel; biocompatibility; biomaterial; demineralized bone matrix; regeneration.

MeSH terms

  • Animals
  • Biocompatible Materials / pharmacology
  • Cattle
  • Extracellular Matrix
  • Hydrogels* / pharmacology
  • Rats
  • Rats, Wistar
  • Tissue Scaffolds*
  • Tumor Necrosis Factor-alpha

Substances

  • Hydrogels
  • Tumor Necrosis Factor-alpha
  • Biocompatible Materials