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

Article type: Research Article

Authors: Ayala-Ham, Alfredo^{a; b} | Aguilar-Medina, Maribel^c | León-Félix, Josefina^d | Romero-Quintana, José Geovanni^c | Bermúdez, Mercedes^e | López-Gutierrez, Jorge^a | Jiménez-Gastélum, German^a | Avendaño-Félix, Mariana^c | Lizárraga-Verdugo, Erik^c | Castillo-Ureta, Hipólito^a | López-Camarillo, César^f | Ramos-Payan, Rosalio^c;

Affiliations: [a] Faculty of Biology, Autonomous University of Sinaloa, Culiacan, Mexico | [b] Faculty of Odontology, Autonomous University of Sinaloa, Culiacan, Mexico | [c] Faculty of Biological and Chemical Sciences, Autonomous University of Sinaloa, Culiacan, Mexico | [d] Center for Research in Food and Development (CIAD), Culiacan, Mexico | [e] Faculty of Odontology, Autonomous University of Chihuahua, Chihuahua, Mexico | [f] Autonomous University of Mexico City, Mexico City, Mexico

Correspondence: [*] Corresponding author: Rosalio Ramos-Payan, PhD, Faculty of Chemical and Biological Sciences, Autonomous University of Sinaloa, Josefa Ortiz de Domínguez s/n y Avenida de las Américas, 80010 Culiacán, Sinaloa, Mexico. E-mail: [email protected]

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, demineralized bone matrix, biocompatibility, regeneration, biomaterial

DOI: 10.3233/BME-211387

Journal: Bio-Medical Materials and Engineering, vol. 33, no. 6, pp. 491-504, 2022