3D-printed PLA/Gel hybrid in liver tissue engineering: Effects of architecture on biological functions.

The liver is one of the vital organs in the body, and the gold standard of treatment for liver function impairment is liver transplantation, which poses many challenges. The specific three-dimensional (3D) structure of liver, which significantly impacts the growth and function of its cells, has made biofabrication with the 3D printing of scaffolds suitable for this approach. In this study, to investigate the effect of scaffold geometry on the performance of HepG2 cells, poly-lactic acid (PLA) polymer was used as the input of the fused deposition modeling (FDM) 3D-printing machine. Samples with simple square and bioinspired hexagonal cross-sectional designs were printed. One percent and 2% of gelatin coating were applied to the 3D printed PLA to improve the wettability and surface properties of the scaffold. Scanning electron microscopy pictures were used to analyze the structural properties of PLA-Gel hybrid scaffolds, energy dispersive spectroscopy to investigate the presence of gelatin, water contact angle measurement for wettability, and weight loss for degradation. In vitro tests were performed by culturing HepG2 cells on the scaffold to evaluate the cell adhesion, viability, cytotoxicity, and specific liver functions. Then, high-precision scaffolds were printed and the presence of gelatin was detected. Also, the effect of geometry on cell function was confirmed in viability, adhesion, and functional tests. The albumin and urea production of the Hexagonal PLA scaffold was about 1.22 ± 0.02-fold higher than the square design in 3 days. This study will hopefully advance our understanding of liver tissue engineering toward a promising perspective for liver regeneration.

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.

Bone tissue engineering gelatin-hydroxyapatite/graphene oxide scaffolds with the ability to release vitamin D: fabrication, characterization, and in vitro study.

Developing smart scaffolds with drug release capability is one of the main approaches to bone tissue engineering. The current study involves the fabrication of novel gelatin (G)-hydroxyapatite (HA)-/vitamin D (VD)-loaded graphene oxide (GO) scaffolds with different concentrations through solvent-casting method. Characterizations confirmed the successful synthesis of HA and GO, and VD was loaded in GO with 36.87 ± 4.87% encapsulation efficiency. Physicochemical characterizations showed that the scaffold containing 1% VD-loaded GO had the best mechanical properties and its porosity percentage and density was in the range of natural spongy bone. All scaffolds were degraded after 1-month, subjecting to phosphate buffer saline. The release profile of VD did not match any mathematical kinetics model, porosities and the degradation rate of the scaffolds were dominant controlling factors of release behavior. Studies on the bioactivity of scaffolds immersed in simulated body fluid indicated that VD and HA could encourage the formation of secondary apatite crystals in vitro. Buccal fat pad-derived stem cells (BFPSCs) were seeded on the scaffolds, MTT assay, alkaline phosphatase activity as an indicator of osteoconductivity, and cell adhesion were conducted in order to evaluate in vitro biological responses. All scaffolds highly supported cell adhesion, MTT assay indicated better cell viability in 0.5% VD-loaded GO containing scaffold, and the scaffold enriched with 2% VD-loaded GO performed the most ALP activity. The results demonstrated the potential of these scaffolds to induce bone regeneration. Developing smart scaffolds with drug release capability is one of the main approaches to bone tissue engineering. The current study involves the fabrication of novel gelatin (G)-hydroxyapatite (HA)-/vitamin D (VD)-loaded graphene oxide (GO) scaffolds with different concentrations through solvent-casting method. Characterizations confirmed the successful synthesis of HA and GO, and VD was loaded in GO with 36.87 ± 4.87% encapsulation efficiency. Physicochemical characterizations showed that the scaffold containing 1% VD-loaded GO had the best mechanical properties and its porosity percentage and density was in the range of natural spongy bone. All scaffolds were degraded after 1-month, subjecting to phosphate buffer saline. The release profile of VD did not match any mathematical kinetics model, porosities and the degradation rate of the scaffolds were dominant controlling factors of release behavior. Studies on the bioactivity of scaffolds immersed in simulated body fluid indicated that VD and HA could encourage the formation of secondary apatite crystals in vitro. Buccal fat pad-derived stem cells (BFPSCs) were seeded on the scaffolds, MTT assay, alkaline phosphatase activity as an indicator of osteoconductivity, and cell adhesion were conducted in order to evaluate in vitro biological responses. All scaffolds highly supported cell adhesion, MTT assay indicated better cell viability in 0.5% VD-loaded GO containing scaffold, and the scaffold enriched with 2% VD-loaded GO performed the most ALP activity. The results demonstrated the potential of these scaffolds to induce bone regeneration.

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.

The effects of preservation procedures on amniotic membrane's ability to serve as a substrate for cultivation of endothelial cells.

Amniotic membrane (AM) has been used as a scaffold for the ex vivo expansion of different types of cells and a cell delivery matrix in regenerative medicine. Since the preservation procedures can influence the AM properties for experimental and clinical purposes, this study was established to investigate the feasibility of using the AM after different preservation methods to serve as substrates for endothelial cell expansion ex vivo. The effects of cryopreservation and lyophilization were evaluated on mechanical and histological characteristics of the AM, and the results were compared with the fresh AM. The ECM components of the basement membrane were well conserved in all groups. Although lyophilization resulted in more histological changes and lower level of physical variables including thickness, F(max), elongation at break and suture retention than the fresh and cryopreserved AM, endothelial cells grown on the lyophilized AM were better attached to the basement membrane. Cytotoxicity assay by MTT showed that the lyophilized AM is a compatible substrate for endothelial cells cultivation. The findings of this study suggest that the lyophilized AM is a suitable matrix for cultivation of endothelial cells due to this fact that lyophilization led to exposure of basement membrane of the AM.

Microfluidic organ-on-a-chip models of human liver tissue.

The liver is the largest internal organ of the body with complex microarchitecture and function that plays critical roles in drug metabolism. Hepatotoxicity and drug-induced liver injury (DILI) caused by various drugs is the main reason for late-stage drug failures. Moreover, liver diseases are among the leading causes of death in the world, with the number of new cases arising each year. Although animal models have been used to understand human drug metabolism and toxicity before clinical trials, tridimensional microphysiological systems, such as liver-on-a-chip (Liver Chip) platforms, could better recapitulate features of human liver physiology and pathophysiology and thus, are often more predictive of human outcome. Liver Chip devices have shown promising results in mimicking in vivo condition by recapitulating the sinusoidal structure of the liver, maintaining high cell viability and cellular phenotypes, and emulating native liver functions. Here, we first review the cellular constituents and physiology of the liver and then critically discuss the state-of-the-art chip-based liver models and their applications in drug screening, disease modeling, and regenerative medicine. We finally address the pending issues of existing platforms and touch upon future directions for developing new, advanced on-chip models.

Signaling molecules orchestrating liver regenerative medicine.

The liver is in charge of more than 500 functions in the human body, which any damage and failure to the liver can significantly compromise human life. Numerous studies are being carried out in regenerative medicine, as a potential driving force, toward alleviating the need for liver donors and fabrication of a 3D-engineered transplantable hepatic tissue. Liver tissue engineering brings three main factors of cells, extracellular matrix (ECM), and signaling molecules together, while each of these three factors tries to mimic the physiological state of the tissue to direct tissue regeneration. Signaling molecules play a crucial role in directing tissue fabrication in liver tissue engineering. When mimicking the natural in vivo process of regeneration, it is tightly associated with three main phases of differentiation, proliferation (progression), and tissue maturation through vascularization while directing each of these phases is highly regulated by the specific signaling molecules. The understanding of how these signaling molecules guide the dynamic behavior of regeneration would be a tool for further tailoring of bioengineered systems to help the liver regeneration with many cellular, molecular, and tissue-level functions. Hence, the signaling molecules come to aid all these phases for further improvements toward the clinical use of liver tissue engineering as the goal.

Novel calcium phosphate coated calcium silicate-based cement: in vitro evaluation.

Calcium silicate-based cements are known for their wide applications in dentistry and orthopedics. The alkaline pH (up to 12) of these cements limits their application in other orthopedic areas. In this study, the effect of dicalcium phosphate dihydrate (DCPD) coating on set cement on pH reduction and biocompatibility improvement was examined. Samples with 0 and 10 weight ratio DCPD were prepared and characterized by XRD, FTIR, and SEM. The DCPD coating on the set cement was performed by a 7 d immersion in 1% monocalcium phosphate (MCP) solution and characterized by XRD, FTIR, SEM, and EDX. Also, the compressive strength and cytotoxicity of the samples were tested. The results showed that DCPD coating did not significantly change the compressive strength of the cement, but by decreasing the pH of the culture medium to the physiological range, it led to enhance adhesion, spreading and proliferation of human osteosarcoma cell line (Saos-2). The novel DCPD coated calcium silicate-based cement could be served as a bulk or porous bone substitute and scaffold.

Liver Tissue Engineering as an Emerging Alternative for Liver Disease Treatment.

Chronic liver diseases affect thousands of lives throughout the world every year. The shortage of liver donors for transplantation has been the main driving force to employ alternative methods such as liver tissue engineering (LTE) in fabricating a three-dimensional transplantable liver tissue or enhancing cell delivery techniques alleviating the need for liver donors. LTE consists of three components, cells, ECM (extracellular matrix), and signaling molecules, which we discuss the first and second. The three most common cell sources used in LTE are human and animal primary hepatocytes, and stem cells for different applications. Two major categories of ECM are used to mimic the microenvironment of these cells, named scaffolds and microbeads. Scaffolds have been made by numerous methods with a wide range of synthetic and natural biomaterials. Cell encapsulation has also been utilized by many polymeric biomaterials. To investigate their functions, many properties have been discussed in the literature, such as biochemical, geometrical, and mechanical properties, in both of these categories. Overall, LTE shows excellent potential in assisting hepatic disorders. However, some challenges exist that prevent the practical use of it clinically, making LTE an ongoing research subject in the scientific society.

Release behavior and signaling effect of vitamin D3 in layered double hydroxides-hydroxyapatite/gelatin bone tissue engineering scaffold: An in vitro evaluation.

Incorporating the controlled release of vitamin D3 (VD3) into biodegradable porous scaffolds is a new approach to equipping multifunctional therapeutics for osteoporosis. The current investigation involves the encapsulation of VD3 into gelatin through the one-step desolvation method. The layered double hydroxides-hydroxyapatite nanocomposite (LDH-HAp) and pure LDH were combined with the gelatin-VD3 complex to reinforce the porous biodegradable structure and enhance the biological response. Afterwards, glutaraldehyde was used to form crosslinks within the gelatin chains. The encapsulation efficiency and loading capacity showed approximately 40% and 50% reduction after crosslinking, respectively. The particle size, zeta potential, contact angle, Young's modulus and porosity were measured to find the effect of VD3 on the scaffolds' physiochemical properties. To explore the bioactivity and degradation behavior, the scaffolds were immersed in simulated body fluid. The VD3 release kinetics followed the Korsmeyer-Peppas model and non-Fickian release pattern. The greater osteblastic expression was observed in VD3-containing scaffolds due to the higher alkaline phosphatase activity which was excited more by HAp (P<0.05). Alizarin red staining illustrated that VD3 induced more calcium deposition, which indicates the signaling role of VD3 on osteoconductivity and biomineralization. The findings provide new insights on the VD3 encapsulation within hydrophilic matrices to protect VD3 and enable the signaling ability for bone tissue engineering scaffolds, which could improve the bone healing efficiency.

Novel layered double hydroxides-hydroxyapatite/gelatin bone tissue engineering scaffolds: Fabrication, characterization, and in vivo study.

Developing porous biodegradable scaffolds through simple methods is one of the main approaches of bone tissue engineering (BTE). In this work, a novel BTE composite containing layered double hydroxides (LDH), hydroxyapatite (HA) and gelatin (GEL) was fabricated using co-precipitation and solvent-casting methods. Physiochemical characterizations showed that the chemical composition and microstructure of the scaffolds were similar to the natural spongy bone. Interconnected macropores ranging over 100 to 600µm were observed for both scaffolds while the porosity of 90±0.12% and 92.11±0.15%, as well as, Young's modulus of 19.8±0.41 and 12.5±0.35GPa were reported for LDH/GEL and LDH-HA/GEL scaffolds, respectively. The scaffolds were degraded in deionized water after a month. The SEM images revealed that between two scaffolds, the LDH-HA/GEL with needle-like secondary HA crystals showed better bioactivity. According to the alkaline phosphatase activity and Alizarin red staining results, LDH-HA/GEL scaffolds demonstrated better bone-specific activities comparing to LDH/Gel scaffold as well as control sample (P<0.05). The rabbit adipose stem cells (ASCs) were extracted and cultured, then seeded on the LDH-HA/GEL scaffolds after confluence. Three groups of six adult rabbits were prepared: the scaffold+ASCs group, the empty scaffold group and the control group. The critical defects were made on the left radius and the scaffolds with or without ASCs were implanted there while the control group was left without any treatment. All animals were sacrificed after 12weeks. Histomorphometric results showed that the regeneration of defects was accelerated by scaffold implantation but ASC-seeding significantly improved the quality of new bone formation (P<0.05). The results confirmed the good performance of LDH-HA/GEL scaffold to induce bone regeneration.

Phase evaluation of an effervescent-added apatitic calcium phosphate bone cement.

Development of macroporosity during setting would allow fast bone ingrowth and good osteointegration of the implant. The interconnected macropores could be created in calcium phosphate cements (CPCs) through the addition of an effervescent porogen mixture to the component of the cements. But this addition could also affect other characteristics of CPCs, such as setting time, mechanical strength, extent of conversion of reactant to apatite phase, crystallinity, and chemical composition of apatite lattice. In this study, these properties were investigated in an effervescent-added calcium phosphate bone cement. From 0 to 20 wt % of an effervescent mixture was added to calcium phosphate cement (CPC) components and phase evaluations were performed after 24 h incubation at 37 degrees C and 28% relative humidity and 1, 3, 7, and 14 days immersion in a specific simulated body fluid. XRD and FTIR techniques were used to characterize the cement composition, crystallinity, and chemical groups in final CPCs. The results showed that addition of effervescent porogen affects the extent of conversion of reactant to apatite phase and crystallinity. In other words, using the effervescent porogen in CPCs could accelerate the rate of conversion of TTCP/DCPA reactant to apatite phase with smaller crystallites, so that it was the predominant phase (about 67%) after only 3 days soaking in SBF solution. The content of carbonate groups substituted for phosphate groups in apatite lattice increased when the effervescent additive was further added. The compressive strength of the set calcium phosphate cement decreased significantly with the addition of the effervescent agent and reached from 8 MPa for additive-free CPC to 1.3 MPa for 20% effervescent-added CPC. The compressive strength was improved after 3 days immersing of CPC in the simulated body fluid solution.

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.

Polycaprolactone/starch composite: Fabrication, structure, properties, and applications.

Interests in the use of biodegradable polymers as biomaterials have grown. Among the different polymeric composites currently available, the blend of starch and polycaprolactone (PCL) has received the most attention since the 1980s. Novamont is the first company that manufactured a PCL/starch (SPCL) composite under the trademark Mater-Bi®. The properties of PCL (a synthetic, hydrophobic, flexible, expensive polymer with a low degradation rate) and starch (a natural, hydrophilic, stiff, abundant polymer with a high degradation rate) blends are interesting because of the composite components have completely different structures and characteristics. PCL can adjust humidity sensitivity of starch as a biomaterial; while starch can enhance the low biodegradation rate of PCL. Thus, by appropriate blending, SPCL can overcome important limitations of both PCL and starch components and promote controllable behavior in terms of mechanical properties and degradation which make it suitable for many biomedical applications. This article reviewed the different fabrication and modification methods of the SPCL composite; different properties such as structural, physical, and chemical as well as degradation behavior; and different applications as biomaterials.

Facile fabrication of egg white macroporous sponges for tissue regeneration.

The availability of 3D sponges combining proper biochemical, biophysical, and biomechanical properties with enhanced capacity of in vivo engraftment and vascularization is crucial in regenerative medicine. A simple process is developed to generate macroporous scaffolds with a well-defined architecture of interconnected pores from chicken egg white (EW), a material with protein- and growth factor-binding features which has not yet been employed in regenerative medicine. The physicomechanical properties and degradation rates of the scaffold are finely tuned by using varying concentrations of the cross-linker, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, without alteration of the biochemical traits. In vitro, EW scaffolds supported active metabolism, proliferation, and migration of human dermal fibroblasts, thereby generating uniform cellular constructs. In vivo, subcutaneous implantation in mice reveals negligible immune reaction and efficient cell and tissue ingrowth. Angiogenesis into EW scaffolds is enhanced as compared to standard collagen type I sponges used as reference material, likely due to significantly higher adsorption of the proangiogenic factor vascular endothelial growth factor. In summary, a material is presented derived by facile processing of a highly abundant natural product. Due to the efficient subcutaneous engraftment capacity, the sponges can find utilization for soft tissue regeneration.

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.

Enhancing glass ionomer cement features by using the HA/YSZ nanocomposite: a feed forward neural network modelling.

Despite brilliant properties of glass ionomer cement (GIC), its weak mechanical property poses an obstacle for its use in medical applications. The present research aims to formulate hydroxyapatite/yttria-stabilized zirconia (HA/YSZ) in the composition of GIC to enhance mechanical properties and to improve fluoride release of GIC. HA/YSZ was synthesized via a sol-gel method and characterized by applying X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photo-emission spectroscopy (XPS) and simultaneous thermal analysis (STA) along with transmission electron microscopy (TEM) methods. The synthesized nanocomposite was mixed with GIC at a fixed composition of 5wt%. The effect of different weight percentages of YSZ:HA on GIC was investigated by measuring the compressive strength, diametral tensile strength, microhardness and fluoride release. The results showed that, after 1 and 7 days of setting, the 20wt% nanohydroxyapatite/80wt% stabilized zirconia cement exhibited higher compressive strength (1857-245MPa), higher diametral tensile strength (11-14MPa) and greater microhardness (104-106MPa) as compared with the pure GIC (65-88MPa in compressive strength, 5-9.5MPa in diametral tensile strength and 70-89MPa in microhardness). The reinforced cement, also, exhibited higher fluoride release compared with pure GIC. The artificial neural network (ANN) was trained for modeling the system. Results obtained by ANN have proved to be completely in accordance with expectations.

Fabrication and characterization of regenerated silk scaffolds reinforced with natural silk fibers for bone tissue engineering.

We introduce a novel Bombyx mori silk-based composite material developed for bone tissue engineering. Three-dimensional scaffolds were fabricated by embedding of natural degummed silk fibers in a matrix of regenerated fibroin, followed by freeze-drying. Different ratios of fibers to fibroin were investigated with respect to their influence on mechanical and biological properties. For all scaffold types, an interconnected porous structure suitable for cell penetration was proven by scanning electron microscopy. Compressive tests, carried out in static and cyclic mode under dry as well as wet conditions, revealed a strong impact of fiber reinforcement on compressive modulus and compressive stress. Cell culture experiments with human mesenchymal stem cells demonstrated that the fiber/fibroin composite scaffolds support cell attachment, proliferation, as well as differentiation along the osteoblastic lineage. Considering the excellent mechanical and biological properties, novel fiber/fibroin scaffolds appear to be an interesting structure for prospect studies in bone tissue engineering.

The effect of crystallographic orientation of titanium substrate on the structure and bioperformance of hydroxyapatite coatings.

This study investigates the effects of crystallographic orientation of titanium substrates on the atomic structure and biological characteristics of hydroxyapatite (HA) coatings. Samples are prepared from extruded rod and rolled sheet of commercially pure titanium having distinct distribution of crystallographic planes. Electrophoresis is used to coat titanium substrates having different microstructures. The biological performance of both HA-coated and non-coated samples is assessed by osteoblast cell attachment, proliferation, differentiation and morphological studies. X-ray diffraction (XRD) analysis of the HA-coated samples indicates the predominant orientation of (002) for HA-coated sheets compared to (211) for the HA-coated rod samples. The numbers of attached and grown cells are higher on the surface of the HA-coated sheet samples. There is also a significant difference in alkaline phosphatase activity on the HA-coated sheet samples. Scanning electron microscopy (SEM) analysis of osteoblast cells grown on HA-coated and non-coated samples demonstrates differences in morphology with respect to spreading and attachment patterns. We believe that the specific atomic structure that is induced in the HA coating by the crystallographic orientation of the sheet substrate causes orientation-dependent coordination with biomolecules and improves cellular interactions. This suggests that crystal orientation of the substrate can be used to both influence the structure of the coating material and improve and control cell-substrate interactions.