Hyaluronic Acid/Bone Substitute Complex Implanted on Chick Embryo Chorioallantoic Membrane Induces Osteoblastic Differentiation and Angiogenesis, but not Inflammation

• Published in: International journal of molecular sciences 2018-12, Vol.19 (12), p.4119
• Main Authors: Cirligeriu, Laura, Cimpean, Anca Maria, Calniceanu, Horia, Vladau, Mircea, Sarb, Simona, Raica, Marius, Nica, Luminita
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; Augmentation; Biomedical materials; Biopsy; Blood vessels; BMP4; Bone biomaterials; Bone matrix; Bone morphogenetic protein 4; Bone Morphogenetic Protein 4 - metabolism; bone substitutes; Bone Substitutes - pharmacology; Cbfa-1 protein; Cell differentiation; Cell Differentiation - drug effects; Chick Embryo; Chorioallantoic membrane; Chorioallantoic Membrane - cytology; Chorioallantoic Membrane - drug effects; Chorioallantoic Membrane - metabolism; Core Binding Factor Alpha 1 Subunit - metabolism; Differentiation; Embryos; Experiments; Hyaluronic acid; Hyaluronic Acid - pharmacology; Immune system; In vivo methods and tests; Inflammation; Inflammation - pathology; Inflammatory response; Membranes; Mesenchyme; Mineralization; Molecular modelling; Morphology; Neovascularization, Physiologic - drug effects; Orthopaedic implants; Osteoblastogenesis; Osteoblasts; Osteoblasts - cytology; Osteoblasts - drug effects; Osteonectin; Osteonectin - metabolism; Osteoprogenitor cells; Phenotypes; RUNX 2; Saline solutions; SPARC; Stem cells; Substitute bone; Transplants & implants
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms19124119

Microscopic and molecular events related to alveolar ridge augmentation are less known because of the lack of experimental models and limited molecular markers used to evaluate this process. We propose here the chick embryo chorioallantoic membrane (CAM) as an in vivo model to study the interaction between CAM and bone substitutes (B) combined with hyaluronic acid (BH), saline solution (BHS and BS, respectively), or both, aiming to point out the microscopic and molecular events assessed by Runt-related transcription factor 2 (RUNX 2), osteonectin (SPARC), and Bone Morphogenic Protein 4 (BMP4). The BH complex induced osteoprogenitor and osteoblastic differentiation of CAM mesenchymal cells, certified by the RUNX2 +, BMP4 +, and SPARC + phenotypes capable of bone matrix synthesis and mineralization. A strong angiogenic response without inflammation was detected on microscopic specimens of the BH combination compared with an inflammatory induced angiogenesis for the BS and BHS combinations. A multilayered organization of the BH complex grafted on CAM was detected with a differential expression of RUNX2, BMP4, and SPARC. The BH complex induced CAM mesenchymal cells differentiation through osteoblastic lineage with a sustained angiogenic response not related with inflammation. Thus, bone granules resuspended in hyaluronic acid seem to be the best combination for a proper non-inflammatory response in alveolar ridge augmentation. The CAM model allows us to assess the early events of the bone substitutes⁻mesenchymal cells interaction related to osteoblastic differentiation, an important step in alveolar ridge augmentation.