Tissue engineering-based bone graft is an emerging viable treatment modality to repair and regenerate tissues damaged as a result of diseases or injuries. The structure and composition of scaffolds should modulate the classical osteogenic pathways in human stem cells. The osteoinductivity properties of the hydroxylapatite-collagen hybrid scaffold named Coll/Pro Osteon 200 were investigated in an in vitro model of human adipose mesenchymal stem cells (hASCs), whereas the clinical evaluation was carried out in maxillofacial patients. Differentially expressed genes (DEGs) induced by the scaffold were analyzed using the Osteogenesis RT2 PCR Array. The osteoinductivity potential of the scaffold was also investigated by studying the alkaline phosphatase (ALP) activity, matrix mineralization, osteocalcin (OCN), and CLEC3B expression protein. Fifty patients who underwent zygomatic augmentation and bimaxillary osteotomy were evaluated clinically, radiologically, and histologically during a 3-year follow-up. Among DEGs, osteogenesis-related genes, including BMP1/2, ALP, BGLAP, SP7, RUNX2, SPP1, and EGFR, which play important roles in osteogenesis, were found to be upregulated. The genes to cartilage condensation SOX9, BMPR1B, and osteoclast cells TNFSF11 were detected upregulated at every time point of the investigation. This scaffold has a high osteoinductivity revealed by the matrix mineralization, ALP activity, OCN, and CLEC3B expression proteins. Clinical evaluation evidences that the biomaterial promotes bone regrowth. Histological results of biopsy specimens from patients showed prominent ossification. Experimental data using the Coll/Pro Osteon 200 indicate that clinical evaluation of bone regrowth in patients, after scaffold implantation, was supported by DEGs implicated in skeletal development as shown in "in vitro" experiments with hASCs.

Hydroxylapatite-collagen hybrid scaffold induces human adipose-derived mesenchymal stem cells to osteogenic differentiation in vitro and bone regrowth in patients.

Mechanoadaptive strain and functional osseointegration of dental implants in rats.

Many cellular models have been used to investigate bone substitutes including coralderived hydroxylapatite (HA). The aim of this study was to verify whether a new cellular model represented by Saos-eGFP cells can be used to test biomaterials by in vitro assays. SaoseGFP cells which express the enhanced Green Fluorescent Protein (eGFP), were derived from the osteoblast-like cell line Saos-2. To this purpose Saos-eGFP cells were employed to investigate the in-vitro bioactivity of a well characterized coral-derived HA biomaterial, in block and granule forms. This engineered cell line, by evaluating the emitted fluorescence, allowed us to assay (i) cell adhesion, (ii) cell proliferation and (iii) colony capability. Electron microscopy analysis was employed to evaluate the (iv) morphology of cells seeded on the biomaterial surface. (v) Histological analysis of the bone grown after scaffold implantation was carried out in specimens from two clinical cases. Saos-eGFP cells indicate, as established before, that the coralline-HA biomaterials has a good in vitro cytocompatibility when tested with human osteoblast-like cells. Some differences in proliferation activity was detected for the two different forms assayed. Cytocompatibility data from in vitro analyses were confirmed by in vivo behaviour of biomaterials. Our tests suggest that the engineered cell line Saos-eGFP represents a suitable in vitro mode for studying the biocompatibility, the cell adhesion, spreading and proliferation on biomaterials developed for clinical applications. The main advantages of this cellular model are (i) less time consuming and (ii) a reduced cost of the experiments.

/pdf/novel-engineered-human-fluorescent-osteoblasts-for-scaffolds-2njv1lsgar.pdf

Novel Engineered Human Fluorescent Osteoblasts for Scaffolds Bioassays

MC3T3-E1 Hücre Dizisinin Uygulandığı Biyoseramiklerin Taramalı Elektron Mikroskobu ve Konfokal Mikroskop ile İncelenmesi

Bioactive scaffolds for potential bone regenerative medical applications

Bone grafts are used to treat various disorders, including delayed union and nonunion of fractures, congenital pseudoarthrosis, and osseous defects from trauma, infection, and tumors. Bone graft substitutes provide surgeons a wide range of materials, structures, and potential delivery systems to use in bone grafting procedures. In addition, the materials may be used as scaffolds to deliver patient derived bone cells. The goal of this study was to seed MG63 osteoblast cells onto three different material constructs--TCP, ZnCaP and ProOsteon--and to compare the growth characteristics of the cells in reaction to the materials. Cellular response to the material was evident as early as 24 hours following the addition of cells onto the constructs. Cells were found adjacent to and along the TCP and ZnCaP carrier, while in the ProOsteon group, the cells appeared non-adhered and more round than spindle shaped. Type I collagen synthesis was highly evident in the TCP and ZnCaP groups at the end of 16 days, whereas little cellular activity was noted around the ProOsteon construct. The overall results suggest a favorable environment for bone cell growth on TCP and ZnCaP materials. The materials may serve as delivery systems for cells in tissue engineering fields in the future.

Comparison of osteoconductive materials on MG63 osteoblast cell function.

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Comparison of osteoconductive materials on MG63 osteoblast cell function.