Three-dimensional printing of polycaprolactone/hydroxyapatite bone tissue engineering scaffolds mechanical properties and biological behavior.

Controlled pore size and desirable internal architecture of bone scaffolds play a significant role in bone regeneration efficiency. In addition to choosing appropriate materials, the manufacturing method is another significant factor in fabricating the ideal scaffold. In this study, scaffolds were designed and fabricated by the fused filament fabrication (FFF) technique. Polycaprolactone (PCL) and composites films with various percentages of hydroxyapatite (HA) (up to 20%wt) were used to fabricate filaments. The influence of (HA) addition on the mechanical properties of filaments and scaffolds was investigated. in vitro biological evaluation was examined as well as the apatite formation in simulated body fluid (SBF). The addition of HA particles increased the compressive strength and Young's modulus of filaments and consequently the scaffolds. Compared to PCL, Young's modulus of PCL/HA20% filament and three-dimensional (3D) printed scaffold has increased by 30% and 50%, respectively. Also, Young's modulus for all scaffolds was in the range of 30-70 MPa, which is appropriate to use in spongy bone. Besides, the MTT assay was utilized to evaluate cell viability on the scaffolds. All the samples had qualified cytocompatibility, and it would be anticipated that addition of HA particles raise the biocompatibility in vivo. Alkaline phosphatase (ALP) evaluation shows that the addition of HA caused higher ALP activity in the PCL/HA scaffolds than PCL. Furthermore, calcium deposition in the PCL/HA specimens is higher than control. In conclusion, the addition of HA particles into the PCL matrix, as well as utilizing an inexpensive commercial FFF device, lead to the fabrication of scaffolds with proper mechanical and biological properties for bone tissue engineering applications. Graphical abstract.

Preparation and characterization of electrospun polycaprolactone/brushite scaffolds to promote osteogenic differentiation of mesenchymal stem cells.

Bone tissue engineering aims to develop effective strategies for repairing or replacing damaged bone tissue. In this study, composite scaffolds consisting of dicalcium phosphate dihydrate (DCDP, brushite) as a bone phase mineral precursor with different weight percentages (0%, 1%, 3%, 5%, and 10%) in combination with polycaprolactone (PCL) were fabricated by electrospinning technique. The morphology and mechanical behavior of scaffolds were characterized using scanning electron microscopy and tensile strength test, respectively. The bioactivity of scaffolds was assessed in simulated body fluid. Adhesion, viability, proliferation, and differentiation of mesenchymal stem cells derived from the human bone marrow on scaffolds were investigated using electron microscopy, MTT assay, live-dead assay, alizarin red staining, alkaline phosphatase activity and, gene expression analysis by real-time PCR. The results showed that the scaffold containing 3 wt. % of DCDP had the highest tensile strength (15.35 MPa). Furthermore, cells seeded on scaffolds showed over 80% viability after 1, 3, 7 days of incubation. Also, the results showed that the addition of DCDP to the PCL significantly increased the alkaline phosphatase activity. The osteocalcin gene expression in the composite scaffold showed a 6.1-fold increase compared to the pure PCL scaffold. It is concluded that electrospun PCL scaffolds containing DCDP with optimum concentration can be a proper candidate for bone tissue engineering applications.

Structure, Properties, and In Vitro Behavior of Heat-Treated Calcium Sulfate Scaffolds Fabricated by 3D Printing.

The ability of inkjet-based 3D printing (3DP) to fabricate biocompatible ceramics has made it one of the most favorable techniques to generate bone tissue engineering (BTE) scaffolds. Calcium sulfates exhibit various beneficial characteristics, and they can be used as a promising biomaterial in BTE. However, low mechanical performance caused by the brittle character of ceramic materials is the main weakness of 3DP calcium sulfate scaffolds. Moreover, the presence of certain organic matters in the starting powder and binder solution causes products to have high toxicity levels. A post-processing treatment is usually employed to improve the physical, chemical, and biological behaviors of the printed scaffolds. In this study, the effects of heat treatment on the structural, mechanical, and physical characteristics of 3DP calcium sulfate prototypes were investigated. Different microscopy and spectroscopy methods were employed to characterize the printed prototypes. The in vitro cytotoxicity of the specimens was also evaluated before and after heat treatment. Results showed that the as-printed scaffolds and specimens heat treated at 300°C exhibited severe toxicity in vitro but had almost adequate strength. By contrast, the specimens heat treated in the 500°C-1000°C temperature range, although non-toxic, had insufficient mechanical strength, which was mainly attributed to the exit of the organic binder before 500°C and the absence of sufficient densification below 1000°C. The sintering process was accelerated at temperatures higher than 1000°C, resulting in higher compressive strength and less cytotoxicity. An anhydrous form of calcium sulfate was the only crystalline phase existing in the samples heated at 500°C-1150°C. The formation of calcium oxide caused by partial decomposition of calcium sulfate was observed in the specimens heat treated at temperatures higher than 1200°C. Although considerable improvements in cell viability of heat-treated scaffolds were observed in this study, the mechanical properties were not significantly improved, requiring further investigations. However, the findings of this study give a better insight into the complex nature of the problem in the fabrication of synthetic bone grafts and scaffolds via post-fabrication treatment of 3DP calcium sulfate prototypes.

Cerebellum-inspired neural network solution of the inverse kinematics problem.

The demand today for more complex robots that have manipulators with higher degrees of freedom is increasing because of technological advances. Obtaining the precise movement for a desired trajectory or a sequence of arm and positions requires the computation of the inverse kinematic (IK) function, which is a major problem in robotics. The solution of the IK problem leads robots to the precise position and orientation of their end-effector. We developed a bioinspired solution comparable with the cerebellar anatomy and function to solve the said problem. The proposed model is stable under all conditions merely by parameter determination, in contrast to recursive model-based solutions, which remain stable only under certain conditions. We modified the proposed model for the simple two-segmented arm to prove the feasibility of the model under a basic condition. A fuzzy neural network through its learning method was used to compute the parameters of the system. Simulation results show the practical feasibility and efficiency of the proposed model in robotics. The main advantage of the proposed model is its generalizability and potential use in any robot.

Epigallocatechin Gallate/Layered Double Hydroxide Nanohybrids: Preparation, Characterization, and In Vitro Anti-Tumor Study.

In recent years, nanotechnology in merging with biotechnology has been employed in the area of cancer management to overcome the challenges of chemopreventive strategies in order to gain promising results. Since most biological processes occur in nano scale, nanoparticles can act as carriers of certain drugs or agents to deliver it to specific cells or targets. In this study, we intercalated Epigallocatechin-3-Gallate (EGCG), the most abundant polyphenol in green tea, into Ca/Al-NO3 Layered double hydroxide (LDH) nanoparticles, and evaluated its efficacy compared to EGCG alone on PC3 cell line. The EGCG loaded LDH nanohybrids were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM) and nanosizer analyses. The anticancer activity of the EGCG-loaded LDH was investigated in prostate cancer cell line (PC3) while the release behavior of EGCG from LDH was observed at pH 7.45 and 4.25. Besides enhancing of apoptotic activity of EGCG, the results showed that intercalation of EGCG into LDH can improve the anti- tumor activity of EGCG over 5-fold dose advantages in in-vitro system. Subsequently, the in-vitro release data showed that EGCG-loaded LDH had longer release duration compared to physical mixture, and the mechanism of diffusion through the particle was rate-limiting step. Acidic attack was responsible for faster release of EGCG molecules from LDH at pH of 4.25 compared to pH of 7.4. The results showed that Ca/Al-LDH nanoparticles could be considered as an effective inorganic host matrix for the delivery of EGCG to PC3 cells with controlled release properties.

Effect of layer thickness and printing orientation on mechanical properties and dimensional accuracy of 3D printed porous samples for bone tissue engineering.

Powder-based inkjet 3D printing method is one of the most attractive solid free form techniques. It involves a sequential layering process through which 3D porous scaffolds can be directly produced from computer-generated models. 3D printed products' quality are controlled by the optimal build parameters. In this study, Calcium Sulfate based powders were used for porous scaffolds fabrication. The printed scaffolds of 0.8 mm pore size, with different layer thickness and printing orientation, were subjected to the depowdering step. The effects of four layer thicknesses and printing orientations, (parallel to X, Y and Z), on the physical and mechanical properties of printed scaffolds were investigated. It was observed that the compressive strength, toughness and Young's modulus of samples with 0.1125 and 0.125 mm layer thickness were more than others. Furthermore, the results of SEM and µCT analyses showed that samples with 0.1125 mm layer thickness printed in X direction have more dimensional accuracy and significantly close to CAD software based designs with predefined pore size, porosity and pore interconnectivity.