Bone Regeneration Using PEVAV/β-Tricalcium Phosphate Composite Scaffolds in Standardized Calvarial Defects: Micro-Computed Tomographic Experiment in Rats

Bone regeneration using beta-tricalcium phosphate (β-TCP) can be practiced using a biocomposite scaffold. Poly(ethylene-co-vinylalcohol)/poly(δ-valerolactone)/β-tricalcium phosphate (PEVAV/β-TCP) composite scaffolds showed promising in vitro results. This study evaluated the bone regenerative potential of PEVAV/β-TCP biocomposite scaffolds in standardized calvarial defects in a rat model over 4 and 10 weeks. Bilateral calvarial defects (5 mm in diameter and about 1.5 mm thick, equivalent to the thickness of the calvaria) were created in 40 male Wistar albino rats. The defects were grafted with either commercially available β-TCP (positive control), PEVAV/β-TCP 70, or PEVAV/β-TCP 50, or left empty (negative control), depending on the group to which the animal was randomly assigned, to be covered before flap closure with resorbable collagen membrane (RCM). At 4 and 10 weeks post-surgery, the collected rat calvaria were evaluated using micro computed tomography (micro-CT) analysis, to assess the newly formed bone volume (NFBV), newly formed bone mineral density (NFBMD), and remaining graft volume (RGV). The results showed that calvarial defects grafted with the PEVAV/β-TCP biocomposite exhibited higher NFBV than did control defects, both at 4 and 10 weeks post-surgery. Furthermore, calvarial defects grafted with PEVAV/β-TCP 70 showed the highest NFBV among all grafting conditions, with a statistically significant difference recorded at 10 weeks post-surgery. The PEVAV/β-TCP composite scaffold showed potentiality for the regeneration of critical-sized calvarial bone defects in a rat model.     

The Effect of Whitlockite as an Osteoconductive Synthetic Bone Substitute Material in Animal Bony Defect Model

This study aimed to evaluate the biomechanical properties in vitro and the bone regeneration of whitlockite (WH) compared with hydroxyapatite (HA) or β-tricalcium phosphate (β-TCP)-based material. We investigated the morphology and phase composition of the bone grafts using a scanning electron microscope and X-ray diffractometer patterns and tested the compressive strength. Four circular defects of 8 mm in diameter were created on the calvaria of twelve rabbits. One defect was left empty, and each of the other defects was filled with WH, HA, and β-TCP. At 4 and 8 weeks, the specimens were harvested to evaluate for the new bone formation and the remaining bone grafts. Regarding the biomechanical properties, the three grafts had a similar micropore size, and WH showed nanopores. The compressive strength of WH was higher than HA and β-TCP without statistical significance. The radiological and histomorphometric analyses demonstrated that the new bone formation was similar among the groups. The remaining bone graft of the WH group was greater than that of the HA and β-TCP groups at 4 weeks (p < 0.05), and the total bone area of the WH, HA, and β-TCP groups was greater than that of the other (p < 0.01). WH has excellent volumetric stability and osteoconductivity compared with HA and β-TCP.     

A Novel Porcine Graft for Regeneration of Bone Defects

Bone regeneration procedures require alternative graft biomaterials to those for autogenous bone. Therefore, we developed a novel porcine graft using particle sizes of 250–500 μm and 500–1000 μm in rabbit calvarial bone defects and compared the graft properties with those of commercial hydroxyapatite (HA)/beta-tricalcium phosphate (β-TCP) over eight weeks. Surgery was performed in 20 adult male New Zealand white rabbits. During a standardized surgical procedure, four calvarial critical-size defects of 5 mm diameter and 3 mm depth were prepared. The defects were filled with HA/β-TCP, 250–500 μm or 500–1000 μm porcine graft, and control defects were not filled. The animals were grouped for sacrifice at 1, 2, 4, and 8 weeks post-surgery. Subsequently, sample blocks were prepared for micro-computed tomography (micro-CT) scanning and histological sectioning. Similar bone formations were observed in all three treatment groups, although the 250–500 μm porcine graft performed slightly better. Rabbit calvarial bone tissue positively responded to porcine grafts and commercial HA/β-TCP, structural analyses showed similar crystallinity and porosity of the porcine and HA/β-TCP grafts, which facilitated bone formation through osteoconduction. These porcine grafts can be considered as graft substitutes, although further development is required for clinical applications.     

Amine Plasma-Polymerization of 3D Polycaprolactone/β-Tricalcium Phosphate Scaffold to Improving Osteogenic Differentiation In Vitro

This study aims to investigate the surface characterization and pre-osteoblast biological behaviors on the three-dimensional (3D) poly(ε-caprolactone)/β-tricalcium phosphate (β-TCP) scaffold modified by amine plasma-polymerization. The 3D PCL scaffolds were fabricated using fused deposition modeling (FDM) 3D printing. To improve the pre-osteoblast bioactivity, the 3D PCL scaffold was modified by adding β-TCP nanoparticles, and then scaffold surfaces were modified by amine plasma-polymerization using monomer allylamine (AA) and 1,2-diaminocyclohexane (DACH). After the plasma-polymerization of PCL/β-TCP, surface characterizations such as contact angle, AFM, XRD, and FTIR were evaluated. In addition, mechanical strength was measured by UTM. The pre-osteoblast bioactivities were evaluated by focal adhesion and cell proliferation. Osteogenic differentiation was investigated by ALP activity, Alizarin red staining, and Western blot. Plasma-polymerization induced the increase in hydrophilicity of the surface of the 3D PCL/β-TCP scaffold due to the deposition of amine polymeric thin film on the scaffold surface. Focal adhesion and proliferation of pre-osteoblast improved, and osteogenic differentiation was increased. These results indicated that 3D PCL/β-TCP scaffolds treated with DACH plasma-polymerization showed the highest bioactivity compared to the other samples. We suggest that 3D PCL/β-TCP scaffolds treated with DACH and AA plasma-polymerization can be used as a promising candidate for osteoblast differentiation of pre-osteoblast.     

Long-Term Effect of Honeycomb β-Tricalcium Phosphate on Zygomatic Bone Regeneration in Rats

In recent years, artificial bones with high biocompatibility have been developed for hard tissue reconstruction. However, current bone replacement methods are inadequate for large defects, causing infection, exposure, and damage. We have developed a new honeycomb β-tricalcium phosphate (TCP) material, which achieved good bone regeneration after implantation in a rat complete zygomatic bone defect. In this study, we further investigated the ability of honeycomb β- TCP for remodeling after bone regeneration as a long-term result. Bone morphogenic protein (BMP)-2-free honeycomb β-TCP (TCP group) and honeycomb β-TCP with BMP-2 (BMP group) were implanted in the zygomatic bone of rats. Micro-computed tomography was performed to track the zygomatic bone morphology, and specimens were histologically examined for osteogenesis and remodeling. In the TCP group, no bone formation was observed at 1 month, but it was observed at 6 months. Bone formation was observed in the BMP group at 1 month, and β-TCP absorption reproducing the zygomatic bone morphology was observed at 6 months. This honeycomb β-TCP with BMP-2 may provide appropriate remodeling that reproduces good bone formation in the early stage and good morphology in the long term, offering an alternative bone reconstruction material to vascularized bone grafts.     

Three-Dimensional Printing of a Hybrid Bioceramic and Biopolymer Porous Scaffold for Promoting Bone Regeneration Potential

In this study, we proposed a three-dimensional (3D) printed porous (termed as 3DPP) scaffold composed of bioceramic (beta-tricalcium phosphate (β-TCP)) and thermoreversible biopolymer (pluronic F-127 (PF127)) that may provide bone tissue ingrowth and loading support for bone defect treatment. The investigated scaffolds were printed in three different ranges of pore sizes for comparison (3DPP-1: 150–200 μm, 3DPP-2: 250–300 μm, and 3DPP-3: 300–350 μm). The material properties and biocompatibility of the 3DPP scaffolds were characterized using scanning electron microscopy, X-ray diffractometry, contact angle goniometry, compression testing, and cell viability assay. In addition, micro-computed tomography was applied to investigate bone regeneration behavior of the 3DPP scaffolds in the mini-pig model. Analytical results showed that the 3DPP scaffolds exhibited well-defined porosity, excellent microstructural interconnectivity, and acceptable wettability (θ < 90°). Among all groups, the 3DPP-1 possessed a significantly highest compressive force 273 ± 20.8 Kgf (* p < 0.05). In vitro experiment results also revealed good cell viability and cell attachment behavior in all 3DPP scaffolds. Furthermore, the 3DPP-3 scaffold showed a significantly higher percentage of bone formation volume than the 3DPP-1 scaffold at week 8 (* p < 0.05) and week 12 (* p < 0.05). Hence, the 3DPP scaffold composed of β-TCP and F-127 is a promising candidate to promote bone tissue ingrowth into the porous scaffold with decent biocompatibility. This scaffold particularly fabricated with a pore size of around 350 μm (i.e., 3DPP-3 scaffold) can provide proper loading support and promote bone regeneration in bone defects when applied in dental and orthopedic fields.     

Biomimetic Ceramic Composite: Characterization,Cell Response,and In Vivo Biocompatibility

The present study aimed to synthesize biphasic calcium phosphate ceramics (CaPs) composed of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HAp) from the propagated Scleractinian coral and dicalcium phosphate anhydrous using a solid-state reaction followed by heat treatment at a temperature of 1100 °C for 1 h to 7 days. The as-prepared coral and coral-derived biphasic CaPs samples were characterized through scanning electron microscopy, X-ray diffractometry, Fourier transform infrared spectroscopy, and Raman spectroscopy. The cell response of the biphasic CaPs was evaluated by in vitro cytotoxicity assessment using mouse fibroblast (L929) cells. The bilateral femoral defect rabbit model was used to assess the early local reaction of the coral-derived biphasic CaPs bone graft on tissue. The results confirmed that the co-existence of β-TCP and HAp was formed at 1100 °C for 1 h. The ratio of HA/β-TCP increased as the heat-treatment time increased. The coral-derived biphasic CaPs comprising 61% HAp and 39% β-TCP (defined as HT-3) were not cytotoxic. Furthermore, no significant differences in local tissue reaction were observed between the HT-3 sample and autogenous bone. Therefore, the synthesized coral-derived biphasic CaPs is a candidate for bone grafting due to its good biocompatibility.     

Simultaneous Substitution of Fe and Sr in Beta-Tricalcium Phosphate: Synthesis,Structural, Magnetic,Degradation, and Cell Adhesion Properties

β-tricalcium phosphate is a promising bone graft substitute material with biocompatibility and high osteoinductivity. However, research on the ideal degradation and absorption for better clinical application remains a challenge. Now, we focus on modifying physicochemical properties and improving biological properties through essential ion co-substitution (Fe and Sr) in β-TCPs. Fe- and Sr-substituted and Fe/Sr co-substituted β-TCP were synthesized by aqueous co-precipitation with substitution levels ranging from 0.2 to 1.0 mol%. The β-TCP phase was detected by X-ray diffraction and Fourier transform infrared spectroscopy. Changes in Ca–O and P–O bond lengths of the co-substituted samples were observed through X-ray photoelectron spectroscopy. The results of VSM represent the M-H graph having a combination of diamagnetic and ferromagnetic properties. A TRIS–HCl solution immersion test showed that the degradation and resorption functions act synergistically on the surface of the co-substituted sample. Cell adhesion tests demonstrated that Fe enhances the initial adhesion and proliferation behavior of hDPSCs. The present work suggests that Fe and Sr co-substitution in β-TCP can be a candidate for promising bone graft materials in tissue engineering fields. In addition, the possibility of application of hyperthermia for cancer treatment can be expected.     

Tissue Level Changes after Maxillary Sinus Floor Elevation with Three Types of Calcium Phosphate Ceramics: A Radiological Study with a 5-Year Follow-Up

This study evaluates the radiological changes in tissue height after maxillary sinus floor elevation (MSFE) using three types of calcium phosphate ceramics over a period of up to 5 years after dental implant placement. In 163 patients, MSFE was performed. Three groups of patients were distinguished and treated based on the type of calcium phosphate ceramic used and radiologically evaluated: 40 patients with β-tricalcium phosphate (β-TCP), 76 patients with biphasic calcium phosphate (BCP) 20% hydroxyapatite (HA)-80% β-TCP, and 47 patients with BCP 60% HA-40% β-TCP. Radiological measurements were performed on panoramic radiographs at several time points up to 5 years after dental implant placement. After MSFE, a slow decrease in tissue height measured over time was seen in all three study groups. Resorption of the grafted bone substitutes was more prominent in β-TCP than in BCP ceramics with an HA component (60/40 and 20/80). Loss of tissue height after 5 years was lowest in BCP 60/40 and highest in β-TCP. This radiological study shows a predictable and comparable behavior of the slow decrease in tissue height over time for all three types of calcium phosphate ceramics used in MSFE. The fraction of HA in calcium phosphate ceramics and dental implant loading seems to be beneficial for tissue height maintenance after MSFE.     

Synthesis and Characterization of Calcium Phosphate Materials Derived from Eggshells from Different Poultry with and without the Eggshell Membrane

Calcium phosphate materials such as hydroxyapatite (HA) or tricalcium phosphate (β-TCP) are highly attractive due to their multitude of applications in bone replacement as well as their environmental and ecological credentials. In this research, quail, hen, duck, and pigeon eggshells were used as a calcium source to obtain calcium phosphate materials via the environmentally friendly wet synthesis. Using the eggshells with the organic membrane, the biphasic calcium phosphate materials composed mainly of HA were obtained. The second mineral phase was β-TCP in the case of using quail, hen, and pigeon eggshells and octacalcium phosphate (OCP) in the case of duck eggshells. The HA content in the obtained materials depended on the amount of membrane in the eggshells and decreased in the order of pigeon, duck, hen, and quail eggshells. The eggshell membrane removal from the eggshells caused the reduced content of HA and the presence of the more soluble β-TCP or OCP phase in the obtained materials. The calcium ions release profile in the PBS buffer indicates the potential biomedical application of these materials.     

Enhancing Stability of High-Concentration β-Tricalcium Phosphate Suspension for Biomedical Application

We propose a novel process to efficiently prepare highly dispersed and stable Tricalcium Phosphate (β-TCP) suspensions. TCP is coupled with a polymer to enhance its brittleness to be used as an artificial hard tissue. A high solid fraction of β-TCP is mixed with the polymer in order to improve the mechanical strength of the prepared material. The high solid fractions led to fast particle aggregation due to Van der Waals forces, and sediments appeared quickly in the suspension. As a result, we used a dispersant, dispex AA4040 (A40), to boost the surface potential and steric hindrance of particles to make a stable suspension. However, the particle size of β-TCP is too large to form a suspension, as the gravity effect is much more dominant than Brownian motion. Hence, β-TCP was subjected to wet ball milling to break the aggregated particles, and particle size was reduced to ~300 nm. Further, to decrease sedimentation velocity, cellulose nanocrystals (CNCs) are added as a thickening agent to increase the overall viscosity of suspension. Besides the viscosity enhancement, CNCs were also wrapped with A40 micelles and increase the stability of the suspension. These CNC/A40 micelles further facilitated stable suspension of β-TCP particles with an average hydration radius of 244.5 nm. Finally, β-TCP bone cement was formulated with the suspension, and the related cytotoxicity was estimated to demonstrate its applicability for hard tissue applications.     

Active Nanofibrous Membrane Effects on Gingival Cell Inflammatory Response

Alpha-melanocyte stimulating hormone (α-MSH) is involved in normal skin wound healing and also has anti-inflammatory properties. The association of α-MSH to polyelectrolyte layers with various supports has been shown to improve these anti-inflammatory properties. This study aimed to evaluate the effects of nanofibrous membrane functionalized with α-MSH linked to polyelectrolyte layers on gingival cell inflammatory response. Human oral epithelial cells (EC) and fibroblasts (FB) were cultured on plastic or electrospun Poly-ε-caprolactone (PCL) membranes with α-MSH covalently coupled to Poly-L-glutamic acid (PGA-α-MSH), for 6 to 24 h. Cells were incubated with or without Porphyromonas gingivalis lipopolysaccharide (Pg-LPS). Cell proliferation and migration were determined using AlamarBlue test and scratch assay. Expression of interleukin-6 (IL-6), tumor necrosis factor (TNF-α), and transforming growth factor-beta (TGF-β) was evaluated using RT-qPCR method. Cell cultures on plastic showed that PGA-α-MSH reduced EC and FB migration and decreased IL-6 and TGF-β expression in Pg-LPS stimulated EC. PGA-α-MSH functionalized PCL membranes reduced proliferation of Pg-LPS stimulated EC and FB. A significant decrease of IL-6, TNF-α, and TGF-β expression was also observed in Pg-LPS stimulated EC and FB. This study showed that the functionalization of nanofibrous PCL membranes efficiently amplified the anti-inflammatory effect of PGA-α-MSH on gingival cells.     

Impact of Cross-Linking of Collagen Matrices on Tissue Regeneration in a Rabbit Calvarial Bone Defect

The cross-linking of collagen matrices (Cl_CM) may provide volume-stable enhanced defect regeneration when compared to non-cross-linked matrices (Ncl_CM). The aim of the present study was to investigate the bone forming potential of collagen matrices (CMs) and the effects of cross-linking CMs in a rabbit calvaria defect model. (1) Empty controls (n = 6), (2) Ncl_CM (n = 8), and (3) Cl_CM (n = 8) were selected to be observed for the healing in 10 mm critical-sized calvarial bone defects. The potential for the bone as well as the connective tissue formation were evaluated by micro-CT and histomorphometry at three months post-surgery. There were no statistically significant differences in terms of new bone volume in the defects between the groups. However, the Cl_CM induced significantly greater fibrous tissue regeneration (5.29 ± 1.57 mm²) when compared to the controls (3.51 ± 0.93 mm²) by histomorphometry. The remnants of collagen fibers with immune cells, including macrophages and giant cells, were occasionally observed in the Cl_CM group but not in the Ncl_CM group. In conclusion, the cross-linking of collagen did not influence the potential for bone formation. Nevertheless, Cl_CM might be advantageous for the maintenance of fibrous tissue volume without disturbing bone formation in the defects.     

Hydroxyapatite Film Coating by Er:YAG Pulsed Laser Deposition Method for the Repair of Enamel Defects

There are treatments available for enamel demineralization or acid erosion, but they have limitations. We aimed to manufacture a device that could directly form a hydroxyapatite (HAp) film coating on the enamel with a chairside erbium-doped yttrium aluminum garnet (Er:YAG) laser using the pulsed laser deposition (PLD) method for repairing enamel defects. We used decalcified bovine enamel specimens and compacted α-tricalcium phosphate (α-TCP) as targets of Er:YAG-PLD. With irradiation, an α-TCP coating layer was immediately deposited on the specimen surface. The morphological, mechanical, and chemical characteristics of the coatings were evaluated using scanning electron microscopy (SEM), scanning probe microscopy (SPM), X-ray diffractometry (XRD), and a micro-Vickers hardness tester. Wear resistance, cell attachment of the HAp coatings, and temperature changes during the Er:YAG-PLD procedure were also observed. SEM demonstrated that the α-TCP powder turned into microparticles by irradiation. XRD peaks revealed that the coatings were almost hydrolyzed into HAp within 2 days. Micro-Vickers hardness indicated that the hardness lost by decalcification was almost recovered by the coatings. The results suggest that the Er:YAG-PLD technique is useful for repairing enamel defects and has great potential for future clinical applications.     

Medium-Term Function of a 3D Printed TCP/HA Structure as a New Osteoconductive Scaffold for Vertical Bone Augmentation: A Simulation by BMP-2 Activation

Introduction: A 3D-printed construct made of orthogonally layered strands of tricalcium phosphate (TCP) and hydroxyapatite has recently become available. The material provides excellent osteoconductivity. We simulated a medium-term experiment in a sheep calvarial model by priming the blocks with BMP-2. Vertical bone growth/maturation and material resorption were evaluated. Materials and methods: Titanium hemispherical caps were filled with either bare- or BMP-2 primed constructs and placed onto the calvaria of adult sheep (n = 8). Histomorphometry was performed after 8 and 16 weeks. Results: After 8 weeks, relative to bare constructs, BMP-2 stimulation led to a two-fold increase in bone volume (Bare: 22% ± 2.1%; BMP-2 primed: 50% ± 3%) and a 3-fold decrease in substitute volume (Bare: 47% ± 5%; BMP-2 primed: 18% ± 2%). These rates were still observed at 16 weeks. The new bone grew and matured to a haversian-like structure while the substitute material resorbed via cell- and chemical-mediation. Conclusion: By priming the 3D construct with BMP-2, bone metabolism was physiologically accelerated, that is, enhancing vertical bone growth and maturation as well as material bioresorption. The scaffolding function of the block was maintained, leaving time for the bone to grow and mature to a haversian-like structure. In parallel, the material resorbed via cell-mediated and chemical processes. These promising results must be confirmed in clinical tests.     

Orientation of Poly(ε-caprolactone) in Its Poly(vinyl chloride) Blends Crystallized under Strain: The Role of Strain Rate

The blends of high and low molecular weights poly(ε-caprolactone) (PCL) with poly(vinyl chloride (PVC) were prepared. The samples before and after the crystallization of PCL were uniaxially stretched to different draw ratios. The orientation features of PCL in a stretched crystalline PCL/PVC blend and crystallized from the amorphous PCL/PVC blends under varied strains were studied by wide-angle X-ray diffraction (WAXD). It was found that a uniaxial stretching of crystalline PCL/PVC blend with high molecular weight PCL results in the c-axis orientation along the stretching direction, as is usually done for the PCL bulk sample. For the stretched amorphous PCL/PVC blend samples, the crystallization of high molecular weight PCL in the blends under a draw ratio of λ = 3 with a strain rate of 6 mm/min leads to a ring-fiber orientation. In the samples with draw ratios of λ = 4 and 5, the uniaxial orientation of a-, b-, and c-axes along the strain direction coexist after crystallization of high molecular weight PCL. With a draw ratio of λ = 6, mainly the b-axis orientation of high molecular weight PCL is identified. For the low molecular weight PCL, on the contrary, the ring-fiber and a-axis orientations coexist under a draw ratio of λ = 3. The a-axis orientation decreases with the increase of draw ratio. When the λ reaches 5, only a poorly oriented ring-fiber pattern has been recognized. These results are different from the similar samples stretched at a higher strain rate as reported in the literatures and demonstrate the important role of strain rate on the crystallization behavior of PCL in its blend with PVC under strain.     

Histological and Histomorphometric Analyses of Bone Regeneration in Osteoporotic Rats Using a Xenograft Material

We evaluated the effect of osteoporotic induction after eight weeks of initial healing of bone defects grafted with a xenograft material in a rat model. Bone defects were created in the femoral condyles of 16 female Wistar rats (one defect per rat). The defects were filled with bovine bone (Inter-Oss) granules. After eight weeks of bone healing, rats were randomly ovariectomized (OVX) or sham-operated (SHAM). At 14 weeks of bone healing, all animals were euthanized. Bone specimens were harvested and processed for histological and histomorphometric analyses to assess new bone formation (N-BF%), remaining bone graft (RBG%) and trabecular bone space (Tb.Sp%) within the defect area. After 14 weeks of bone healing, histological evaluation revealed a significant alteration in trabecular bone in OVX rats compared to SHAM rats. There was lower N-BF% in OVX rats (22.5% ± 3.0%) compared to SHAM rats (37.7% ± 7.9%; p < 0.05). Additionally, the RBG% was significantly lower in OVX (23.7% ± 5.8%) compared to SHAM (34.8% ± 9.6%; p < 0.05) rats. Finally, the Tb.Sp% was higher in OVX (53.8% ± 7.7%) compared to SHAM (27.5% ± 14.3%; p < 0.05) rats. In conclusion, within the limitations of this study, inducing an osteoporotic condition in a rat model negatively influenced bone regeneration in the created bone defect and grafted with a xenograft material.     

PCL/Si-Doped Multi-Phase Calcium Phosphate Scaffolds Derived from Cuttlefish Bone

Increasing attention is focused on developing biomaterials as temporary scaffolds that provide a specific environment and microstructure for bone tissue regeneration. The aim of the present work was to synthesize silicon-doped biomimetic multi-phase composite scaffolds based on bioactive inorganic phases and biocompatible polymers (poly(ε-caprolactone), PCL) using simple and inexpensive methods. Porous multi-phase composite scaffolds from cuttlefish bone were synthesized using a hydrothermal method and were further impregnated with (3-aminopropyl)triethoxysilane 1–4 times, heat-treated (1000 °C) and coated with PCL. The effect of silicon doping and the PCL coating on the microstructure and mechanical and biological properties of the scaffolds has been investigated. Multi-phase scaffolds based on calcium phosphate (hydroxyapatite, α-tricalcium phosphate, β-tricalcium phosphate) and calcium silicate (wollastonite, larnite, dicalcium silicate) phases were obtained. Elemental mapping revealed homogeneously dispersed silicon throughout the scaffolds, whereas silicon doping increased bovine serum albumin protein adsorption. The highly porous structure of cuttlefish bone was preserved with a composite scaffold porosity of ~78%. A compressive strength of ~1.4 MPa makes the obtained composite scaffolds appropriate for non-load-bearing applications. Cytocompatibility assessment by an MTT assay of human mesenchymal stem cells revealed the non-cytotoxicity of the obtained scaffolds.     

Design of 3D Additively Manufactured Hybrid Structures for Cranioplasty

A wide range of materials has been considered to repair cranial defects. In the field of cranioplasty, poly(methyl methacrylate) (PMMA)-based bone cements and modifications through the inclusion of copper doped tricalcium phosphate (Cu-TCP) particles have been already investigated. On the other hand, aliphatic polyesters such as poly(ε-caprolactone) (PCL) and polylactic acid (PLA) have been frequently investigated to make scaffolds for cranial bone regeneration. Accordingly, the aim of the current research was to design and fabricate customized hybrid devices for the repair of large cranial defects integrating the reverse engineering approach with additive manufacturing, The hybrid device consisted of a 3D additive manufactured polyester porous structures infiltrated with PMMA/Cu-TCP (97.5/2.5 w/w) bone cement. Temperature profiles were first evaluated for 3D hybrid devices (PCL/PMMA, PLA/PMMA, PCL/PMMA/Cu-TCP and PLA/PMMA/Cu-TCP). Peak temperatures recorded for hybrid PCL/PMMA and PCL/PMMA/Cu-TCP were significantly lower than those found for the PLA-based ones. Virtual and physical models of customized devices for large cranial defect were developed to assess the feasibility of the proposed technical solutions. A theoretical analysis was preliminarily performed on the entire head model trying to simulate severe impact conditions for people with the customized hybrid device (PCL/PMMA/Cu-TCP) (i.e., a rigid sphere impacting the implant region of the head). Results from finite element analysis (FEA) provided information on the different components of the model.     

Influence of Natural Polysaccharides on Properties of the Biomicroconcrete-Type Bioceramics

In this paper, novel hybrid biomicroconcrete-type composites were developed and investigated. The solid phase of materials consisted of a highly reactive α -tricalcium phosphate (α-TCP) powder, hybrid hydroxyapatite-chitosan (HAp-CTS) material in the form of powder and granules (as aggregates), and the polysaccharides sodium alginate (SA) or hydroxypropyl methylcellulose (HPMC). The liquid/gel phase in the studied materials constituted a citrus pectin gel. The influence of SA or HPMC on the setting reaction, microstructure, mechanical as well as biological properties of biomicroconcretes was investigated. Studies revealed that manufactured cement pastes were characterized by high plasticity and cohesion. The dual setting system of developed biomicroconcretes, achieved through α-TCP setting reaction and polymer crosslinking, resulted in a higher compressive strength. Material with the highest content of sodium alginate possessed the highest mechanical strength (~17 MPa), whereas the addition of hydroxypropyl methylcellulose led to a subtle compressive strength decrease. The obtained biomicroconcretes were chemically stable and characterized by a high bioactive potential. The novel biomaterials with favorable physicochemical and biological properties can be prosperous materials for filling bone tissue defects of any shape and size.