A review on bovine hydroxyapatite; extraction and characterization

High rate of bone grafting surgeries emphasizes the need for optimal bone substitutes. Biomaterials mimicking the interconnected porous structure of the original bone with osteoconductive and osteoinductive capabilities have long been considered. Hydroxyapatite (HA), as the main inorganic part of natural bone, has exhibited excellent regenerative properties in bone tissue engineering. This manuscript reviews the HA extraction methods from bovine bone, as one of the principal biosources. Essential points in the extraction process have also been highlighted. Characterization of the produced HA through gold standard methods such as XRD, FTIR, electron microscopies (SEM and TEM), mechanical/thermodynamic tests, and bioactivity analysis has been explained in detail. Finally, future perspectives for development of HA constructs are mentioned.

Spatiotemporal regulation of angiogenesis/osteogenesis emulating natural bone healing cascade for vascularized bone formation

Engineering approaches for growth factor delivery have been considerably advanced for tissue regeneration, yet most of them fail to provide a complex combination of signals emulating a natural healing cascade, which substantially limits their clinical successes. Herein, we aimed to emulate the natural bone healing cascades by coupling the processes of angiogenesis and osteogenesis with a hybrid dual growth factor delivery system to achieve vascularized bone formation. Basic fibroblast growth factor (bFGF) was loaded into methacrylate gelatin (GelMA) to mimic angiogenic signalling during the inflammation and soft callus phases of the bone healing process, while bone morphogenetic protein-2 (BMP-2) was bound onto mineral coated microparticles (MCM) to mimics osteogenic signalling in the hard callus and bone remodelling phases. An Initial high concentration of bFGF accompanied by a sustainable release of BMP-2 and inorganic ions was realized to orchestrate well-coupled osteogenic and angiogenic effects for bone regeneration. In vitro experiments indicated that the hybrid hydrogel markedly enhanced the formation of vasculature in human umbilical vein endothelial cells (HUVECs), as well as the osteogenic differentiation of mesenchymal stem cells (BMSCs). In vivo results confirmed the optimal osteogenic performance of our F/G-B/M hydrogel, which was primarily attributed to the FGF-induced vascularization. This research presents a facile and potent alternative for treating bone defects by emulating natural cascades of bone healing. Graphical Abstract

Enzymatically Crosslinked In Situ Synthesized Silk/Gelatin/Calcium Phosphate Hydrogels for Drug Delivery

Our research focuses on combining the valuable properties of silk fibroin (SF) and calcium phosphate (CaP). SF is a natural protein with an easily modifiable structure; CaP is a mineral found in the human body. Most of the new age biocomposites lack interaction between organic/inorganic phase, thus SF/CaP composite could not only mimic the natural bone, but could also be used to make drug delivery systems as well, which can ensure both healing and regeneration. CaP was synthesized in situ in SF at different pH values, and then crosslinked with gelatin (G), horseradish peroxide (HRP), and hydrogen peroxide (H2O2). In addition, dexamethasone phosphate (DEX) was incorporated in the hydrogel and drug delivery kinetics was studied. Hydrogel made at pH 10.0 was found to have the highest gel fraction 110.24%, swelling degree 956.32%, and sustained drug delivery for 72 h. The highest cell viability was observed for the hydrogel, which contained brushite (pH 6)—512.43%.

Fractal Design Boosts Extrusion-Based 3D Printing of Bone-Mimicking Radial-Gradient Scaffolds

Although extrusion-based three-dimensional (EB-3D) printing technique has been widely used in the complex fabrication of bone tissue-engineered scaffolds, a natural bone-like radial-gradient scaffold by this processing method is of huge challenge and still unmet. Inspired by a typical fractal structure of Koch snowflake, for the first time, a fractal-like porous scaffold with a controllable hierarchical gradient in the radial direction is presented via fractal design and then implemented by EB-3D printing. This radial-gradient structure successfully mimics the radially gradual decrease in porosity of natural bone from cancellous bone to cortical bone. First, we create a design-to-fabrication workflow with embedding the graded data on basis of fractal design into digital processing to instruct the extrusion process of fractal-like scaffolds. Further, by a combination of suitable extruded inks, a series of bone-mimicking scaffolds with a 3-iteration fractal-like structure are fabricated to demonstrate their superiority, including radial porosity, mechanical property, and permeability. This study showcases a robust strategy to overcome the limitations of conventional EB-3D printers for the design and fabrication of functionally graded scaffolds, showing great potential in bone tissue engineering.

Calcium substituted with magnesium, silver and zinc in hydroxyapatite: a review

Hydroxyapatite with the chemical formula Ca10(-PO4)6(OH)2 is an important bioceramic well known for its high osteoconductive properties, non-toxic nature, high bioactivity and good biocompatibility. Moreover, it is known to mimic natural bone. Inspite of the above mentioned advantages, it has certain disadvantages such as having poor mechanical properties, being brittle, not showing an inhibitory effect on microbes and taking a long time to resorb in the body. These disadvantages can be covered up by the addition of dopants which include cations, anions and polymers. These are sufficiently known to improve the properties of hydroxyapatite. This review focuses on the substitution of hydroxyapatite with silver, zinc, magnesium ions and alloys to show the changes in the morphology and biocompatible properties of hydroxyapatite after substitution of the ions in the matrix.

Evaluating the Prospects of Ti-Base Lattice Infiltrated with Biodegradable Zn–2%Fe Alloy as a Structural Material for Osseointegrated Implants—In Vitro Study

The term “osseointegrated implants” mainly relates to structural systems that contain open spaces, which enable osteoblasts and connecting tissue to migrate during natural bone growth. Consequently, the coherency and bonding strength between the implant and natural bone can be significantly increased, for example in operations related to dental and orthopedic applications. The present study aims to evaluate the prospects of a Ti–6Al–4V lattice, produced by selective laser melting (SLM) and infiltrated with biodegradable Zn2%Fe alloy, as an OI–TiZn system implant in in vitro conditions. This combined material structure is designated by this study as an osseointegrated implant (OI–TiZn) system. The microstructure of the tested alloys was examined both optically and using scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis. The mechanical properties were assessed in terms of compression strength, as is commonly acceptable in cases of lattice-based structures. The corrosion performance was evaluated by immersion tests and electrochemical analysis in terms of potentiodynamic polarization and electrochemical impedance spectroscopy (EIS), all in simulated physiological environments in the form of phosphate buffered saline (PBS) solution. The cytotoxicity was evaluated in terms of indirect cell viability. The results obtained demonstrate the adequate performance of the OI–TiZn system as a non-cytotoxic structural material that can maintain its mechanical integrity under compression, while presenting acceptable corrosion rate degradation.

Microarchitecture of titanium cylinders obtained by additive manufacturing does not influence osseointegration in the sheep

Large bone defects are a challenge for orthopedic surgery. Natural (bone grafts) and synthetic biomaterials have been proposed but several problems arise such as biomechanical resistance or viral/bacterial safety. The use of metallic foams could be a solution to improve mechanical resistance and promote osseointegration of large porous metal devices. Titanium cylinders have been prepared by additive manufacturing (3D printing/rapid prototyping) with a geometric or trabecular microarchitecture. They were implanted in the femoral condyles of aged ewes; the animals were left in stabling for 90 and 270 days. A double calcein labeling was done before sacrifice; bones were analyzed by histomorphometry. Neither bone volume, bone/titanium interface nor mineralization rate were influenced by the cylinder’s microarchitecture; the morphometric parameters did not significantly increase over time. Bone anchoring occurred on the margins of the cylinders and some trabeculae extended in the core of the cylinders but the amount of bone inside the cylinders remained low. The rigid titanium cylinders preserved bone cells from strains in the core of the cylinders. Additive manufacturing is an interesting tool to prepare 3D metallic scaffolds, but microarchitecture does not seem as crucial as expected and anchoring seems limited to the first millimeters of the graft.

Design and Mechanical Properties Verification of Gradient Voronoi Scaffold for Bone Tissue Engineering

In order to obtain scaffold that can meet the therapeutic effect, researchers have carried out research on irregular porous structures. However, there are deficiencies in the design method of accurately controlling the apparent elastic modulus of the structure at present. Natural bone has a gradient porous structure. However, there are few studies on the mechanical property advantages of gradient bionic bone scaffold. In this paper, an improved method based on Voronoi-tessellation is proposed. The method can get controllable gradient scaffolds to fit the modulus of natural bone, and accurately control the apparent elastic modulus of porous structure, which is conducive to improving the stress shielding. To verify the designed structure can be fabricated by additive manufacturing, several designed models are obtained by SLM and EBM. Through finite element analysis (FEA), it is verified that the irregular porous structure based on Voronoi-tessellation is more stable than the traditional regular porous structure of the same structure volume, the same pore number and the same material. Furthermore, it is verified that the gradient irregular structure has a better stability than the non-gradient structure. An experiment is conducted successfully to verify the stability performance got by FEA. In addition, a dynamic impact FEA is also performed to simulate impact resistance. The result shows that the impact resistance of the regular porous structure, the irregular porous structure and the gradient irregular porous structure becomes better in turn. The mechanical property verification provides a theoretical basis for the structural design of gradient irregular porous bone tissue engineering scaffolds.

INVESTIGATION OF PROPERTIES OF SYNTHETIC BONE SUBSTITUTES

The article compares different materials of bone substitutes – bioceramics: hydroxyapatite (HA), tricalcium phosphate (CaP) and polymer: polylactide (PLA). In the paper determines which of the substitutes is mechanically similar to the natural bone. Universal testing machine for tensile, compression was used for research. The properties of the test substances were determined by a compression and hardness test. Comparative tests are conducted with HA, CaP, PLA which were kept for 3 weeks in physiological saline and with natural pig bone. The mechanical properties of PLA specimens produced by 3D printers have been found to be similar to natural bone. When held in saline, PLA does not change its properties and dissolves less quickly than tricalcium phosphate.

Preparation and Characterization of Sustained-Release Naringin Coating on Magnesium Surface

Given the three-dimensional multi-level structure of natural bone and the multi-factor time-shifting effect in the healing process after bone trauma, there are plans to introduce drug-controlled release systems into the treatment of orthopedic diseases. To achieve multi-level loading and controlled release of biologically active substances, it is necessary to create synergistic behavior between biological factors, thereby improving the bone regeneration ability of artificial bone replacement materials. A naringin-loaded (NG) coating was prepared, compared with ultrasonic micro-arc oxidation (UMAO). The coating was characterized by X-ray diffraction, infrared spectroscopy, and scanning electron microscopy. The corrosion resistance of the coating was studied through the wetting angle and polarization curve. The high-performance liquid chromatography method was used to test the release of the drug. It can be seen from the experimental results that the NG coating has a larger wetting angle and better corrosion resistance. In addition, the NG coating produces more apatite substances and has good biological activity. The NG coatings can stimulate the natural bone regeneration and repair process by releasing drugs during the process, which can effectively promote bone regeneration and repair after implantation in the body.