In vitro and in vivo study of microporous ceramics using MC3T3 cells, CAM assay and a pig animal model

Abstract

Bone tissue engineering combines biomaterials with biologically active factors and cells to hold promise for reconstructing craniofacial defects. In this study the biological activity of biphasic hydroxyapatite ceramics (HA; a bone substitute that is a mixture of hydroxyapatite and β-tricalcium phosphate in fixed ratios) was characterized (1) in vitro by assessing the growth of MC3T3 mouse osteoblast lineage cells, (2) in ovo by using the chick chorioallantoic membrane (CAM) assay and (3) in an in vivo pig animal model. Biocompatibility, bioactivity, bone formation and biomaterial degradation were detected microscopically and by radiology and histology. HA ceramics alone demonstrated great biocompatibility on the CAM as well as bioactivity by increased proliferation and alkaline phosphatase secretion of mouse osteoblasts. The in vivo implantation of HA ceramics with bone marrow mesenchymal stem cells (MMSCs) showed de novo intramembranous bone healing of critical-size bone defects in the right lateral side of pig mandibular bodies after 3 and 9 weeks post-implantation. Compared with the HA ceramics without MMSCs, the progress of bone formation was slower with less-developed features. This article highlights the clinical use of microporous biphasic HA ceramics despite the unusually shaped elongated micropores with a high length/width aspect ratio (up to 20) and absence of preferable macropores (>100 µm) in bone regenerative medicine.

Evolving applications of the egg: chorioallantoic membrane assay andex vivoorganotypic culture of materials for bone tissue engineering

“…For example, nanohydroxyapatite rod cores surrounded by a silica sheath incorporated into a gelatin matrix were found to be angiogenic, 101 as hydroxyapatite is a native component of bone, and silicon has effects on bone formation and remodelling. 102 Hydroxyapatite-based scaffolds were tested by Tomco et al 103 in the CAM and a pig mandibular defect model with the bone marrow stromal cell seeded-scaffold replaced by bone in 9 weeks, although it appeared that the study lacked a ‘no-scaffold’ control to compare normal bone healing rate and morphology in this anatomical region potentially limiting interpretation.…”

Tissue engineering applications in otolaryngology—The state of translation

“…Heatmap representing the proportion of publications by year that utilized specific translational research methodologies from construct characterization to human trial to investigate craniofacial tissue engineering 49‐72,74‐77,79,81‐84,145‐197 …”

Modification of silicone elastomers with Bioglass 45S5® increases in ovo tissue biointegration

“…The CAM assay is an easy and fast assay often used in the field of tissue engineering and regenerative medicine . It allows short‐term determination of a biomaterial's biocompatibility and integration into the chorioallantoic membrane of the chicken embryo . First, we tested biointegration of non‐porous silicone and composites during the 7‐day incubation in the CAM assay.…”