The role of apoptosis associated speck-like protein containing a caspase-1 recruitment domain (ASC) in response to bone substitutes

Author(s)

Sartoretto, Suelen C. ^{[1, 2]} ; Calasans-Maia, Monica D. ^[3] ; Alves, Adriana T. N. N. ^[4] ; Resende, Rodrigo F. B. ^{[1, 2]} ; da Costa Fernandes, Celio Junior ^[5] ; Padilha, Pedro de Magalhaes ^[5] ; Rossi, Alexandre M. ^[6] ; Teti, Anna ^[7] ; Granjeiro, Jose M. ^{[8, 9]} ; Zambuzzi, Willian F. ^[5]

Affiliation

^[1] Univ Veiga Almeida, Oral Surg Dept, Rio De Janeiro, RJ - Brazil

^[2] Univ Iguacu, Oral Surg Dept, Nova Iguacu, RJ - Brazil

^[3] Univ Fed Fluminense, Oral Surg Dept & Clin Res Lab Dent, Niteroi, RJ - Brazil

^[4] Univ Fed Fluminense, Oral Diag Dept, Niteroi, RJ - Brazil

^[5] UNESP Sao Paulo State Univ, Inst Biosci Botucatu, Dept Chem & Biochem, Botucatu, SP - Brazil

^[6] Ctr Brasileiro Pesquisas Fis CBPF, Dept Appl Phys, Rio De Janeiro, RJ - Brazil

^[7] Univ Aquila, Dept Biotechnol & Appl Clin Sci, Laquila - Italy

^[8] Univ Fed Fluminense, Clin Res Lab Dent, Niteroi, RJ - Brazil

^[9] Inst Nacl Metrol Qualidade & Tecnol INMETRO, Directory Life Sci Appl Metrol, Duque De Caxias, RJ - Brazil

• Document type: Journal article
• Source: Materials Science & Engineering C-Materials for Biological Applications; v. 112, JUL 2020.
• Web of Science Citations: 0

Abstract

The apoptosis-associated Speck-like protein containing a caspase-1 recruitment domain (ASC), present in inflammasomes, regulates inflammation events and is involved in osteogenic phenotype. Nevertheless, its function in bone repair induced by bone substitute biomaterials is unclear. This study aimed to unveil the role of ASC on osteoprogenitor and tissue response to stoichiometric-hydroxyapatite (HA), nanostructured carbonated-hydroxyapatite (CHA), and CHA containing 5% Strontium (SrCHA), characterized previously by XRD, uXRF-SR, and FTIR spectroscopy implants. Thereafter, conditioned media by the biomaterials were used later to treat preosteoblasts and an osteogenic stimulus was shown in response to the materials, with higher expression of Runx2, Osterix, ALP, and Collagen 1a1 genes, with significant involvement of inflammatory-related genes. Thus, to better address the involvement of inflammasome, primary cells obtained from both genotypes {[}Wild-Type (WT) and ASC Knockout (ASC-KO) mice] were subjected to conditioned media up to 7 days, and our data reinforces both HA and CHA induces lower levels of alkaline phosphatase (ALP) than SrCHA, considering both genotypes (p < 0.01), and ASC seems contribute with osteogenic stimulus promoted by SrCHA. Complimentarily, the biomaterials were implanted into both subcutaneous and bone defects in tibia. Histological analysis on 28 days after implantation of biomaterials into mice's subcutaneous tissue revealed moderate inflammatory response to them. Both histomorphometry and mu CT analysis of tibias indicated that the biomaterials did not reverse the delay in bone repair of ASC KO, reinforcing the involvement of ASC on bone regeneration and bone de novo deposition. Also, the bone density in CHA was > 2-fold higher in WT than ASC-KO samples. HA was virtually not resorbed throughout the experimental periods, in opposition to CHA in the WT group. CHA reduced to half-area after 28 days, and the bone deposition was higher in CHA for WT mice than HA. Taken together, our results show that biomaterials did not interfere with the healing pattern of the ASC KO, but CHA promoted higher bone deposition in the WT group, probably due to its greater biodegradability. These results reinforce the importance of ASC during bone de novo deposition and healing. (AU)

• FAPESP's process: 14/22689-3 - Microvesicle/proteins-mediated paracrine signaling among bone and endothelial cells during bone development and regeneration
• Grantee: Willian Fernando Zambuzzi
• Support Opportunities: Research Grants - Young Investigators Grants