Osteogenic Properties of 3D-Printed Silica-Carbon-Calcite Composite Scaffolds: Novel Approach for Personalized Bone Tissue Regeneration

Abstract

Carbon enriched bioceramic (C-Bio) scaffolds have recently shown exceptional results in terms of their biological and mechanical properties. The present study aims at assessing the ability of the C-Bio scaffolds to affect the commitment of canine adipose-derived mesenchymal stem cells (cAD-MSCs) and investigating the influence of carbon on cell proliferation and osteogenic differentiation of cAD-MSCs in vitro. The commitment of cAD-MSCs to an osteoblastic phenotype has been evaluated by expression of several osteogenic markers using real-time PCR. Biocompatibility analyses through 3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) activity, hemolysis assay, and Ames test demonstrated excellent biocompatibility of both materials. A significant increase in the extracellular alkaline phosphatase (ALP) activity and expression of runt-related transcription factor (RUNX), ALP, osterix (OSX), and receptor activator of nuclear factor kappa-Β ligand (RANKL) genes was observed in C-Bio scaffolds compared to those without carbon (Bio). Scanning electron microscopy (SEM) demonstrated excellent cell attachment on both material surfaces; however, the cellular layer on C-Bio fibers exhibited an apparent secretome activity. Based on our findings, graphene can improve cell adhesion, growth, and osteogenic differentiation of cAD-MSCs in vitro. This study proposed carbon as an additive for a novel three-dimensional (3D)-printable biocompatible scaffold which could become the key structural material for bone tissue reconstruction.

Active Materials for 3D Printing in Small Animals: Current Modalities and Future Directions for Orthopedic Applications

“…The choice of biomaterial type used in 3D printing is determined by the intended application of the final product [19,20]. For instance, a biomaterial for 3D printing in orthopedics should be easily printable and have excellent biocompatibility, controlled long-lasting biodegradation, acceptable mechanical properties, and a well-designed architecture [6,19,20]. For surgical application, the material should be sterilizable [14].…”

“…Recently, different bioceramics have been employed for the construction of 3D-printed scaffolds or implants due to their promising osteoinductive and osteoconductive properties, high stiffness (393 GPa), and similarity to the mineral phase of bone [6,28]. Calcium phosphate ceramics (CaP ceramics) are synthetic materials made of calcium hydroxyapatites (HA) and thus have a composition comparable to the original bone matrix.…”

“…Calcium phosphate ceramics (CaP ceramics) are synthetic materials made of calcium hydroxyapatites (HA) and thus have a composition comparable to the original bone matrix. Tricalcium phosphate (TCP), HA, and biphasic calcium phosphate (BCP) are the most often used CaP ceramics in bone reconstruction [6,29]. Bioactive glasses, often known as bioglasses, are ceramics made from synthetic silicates; they are rapidly resorbed in the first two weeks after implantation, allowing for rapid new bone formation and implant ingrowth.…”

“…CaP ceramics and bioactive glasses have been used to make 3D-printed scaffolds; however, their poor mechanical properties, such as low fracture-toughness and tensile strength, limit their application in load-bearing conditions. This disadvantage can be overcome prior to printing by mixing bioceramics with polymers such as cellulose, poly (D, L-lactic acid-co-glycolic acid), or polycaprolactone (PCL), or by combining with specialized reinforcing materials such as carbon nanotubes, graphene, polyethylene, Al 2 O 3 , and TiO 2 to create ceramic composites with higher mechanical strengths [6,7,28]. Graphene and its derivates showed to be promising materials in the field of material science due to their good biocompatibility and osteogenesis properties [6,[30][31][32][33].…”

“…In spite of this, many orthopedic conditions in animals are still treated with limb amputations and salvage procedures, resulting in reduced or impaired limb function and decreased quality of life [5]. Complex orthopedic disorders such as extensive bone loss, fracture nonunion or malunion, tumors, bone deformities, and large-scale traumatic injuries remain clinical challenges in veterinary practice as conventional surgical techniques usually fail to address these conditions [6][7][8]. The idea of bone tissue engineering (BTE) has been developed to resolve the limitations of conventional approaches in addressing complicated orthopedic circumstances [9][10][11].…”