Impact and biocompatibility studies on Mg2+-substituted Apatite-derived 3D porous scaffolds for hard tissue engineering

The objective of this study was to develop Mg²⁺-substituted Apatite scaffolds by slip-casting method. The Apatite scaffolds were prepared as engineering constructs with interconnected pore structure with a pore size of 128-194 μm range. The physicochemical properties such as crystalline phase, functional group, microstructure, pore size distribution, and elemental compositions of the scaffolds were characterized. The bioactivity of the developed porous scaffolds was investigated in Simulated Body Fluid (SBF) for various time periods (3 and 7 days). In vitro bioactivity results confirm the hydroxyl Apatite layer formation of the scaffolds and results suggest that the developed microporous scaffold could be used as suitable candidates in bone tissue engineering.     

A simple approach to observe non-conductive hydrated materials with FE-SEM: Case study on porous hydroxyapatite green bodies

The purpose of this study is to develop a simple method to observe the surface morphology of non-conductive material in its hydrated condition. Here porous hydroxyapatite (HAp) green body prepared by gelcasting process was considered as a case study, and, the resultant body was subsequently treated with hydrophilic ionic liquid (IL). The surface morphology and the pore structure of the IL-treated porous HAp green body were successfully observed even in hydrated condition without any charging using field emission scanning electron microscope (FE-SEM). Results showed that the pore diameter from 300 to 600 μm in as-prepared green body was reduced to 100–300 μm in the sample sintered at 1000 °C. Raman results showed that the IL forms weak hydrogen bond with water molecules within the sample and, that prevents from drying in vacuum condition. In addition, the IL acts as a conducting media for HAp ceramics to be observed under FE-SEM.     

Novel gamma irradiated agarose-gelatin-hydroxyapatite nanocomposite scaffolds for skin tissue regeneration

Agarose-gelatin-hydroxyapatite composites prepared by freeze-drying technique were gamma irradiated with various doses (25 kGy, 50 kGy and 100 kGy). X-ray Diffraction (XRD) analysis revealed the pure phase of HAp and the intensity of prominent planes of hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂, HAp) were found to decrease on irradiation. Fourier Transform Infrared spectra (FTIR) showed functional groups of HAp and polymer composites, and higher disorder of the polymer matrix on irradiation. In addition, gamma irradiation led to a drastic reduction in the wettability (62%) and the compressive modulus (76%) of the scaffolds. There was significant enhancement (113%) in pore size of the scaffolds at higher fluence (100 kGy). The swelling and the dissolution studies of the gamma irradiated scaffolds showed that it had an appreciable change in the scaffold's mechanical and biological properties viz., compressive modulus, cell proliferation, hemolysis etc. The irradiated biomaterials exhibited enhanced hemocompatibility, antimicrobial activity and cell viability. The above results clearly reveal that the gamma irradiation is a suitable tool to tailor the multifunctional properties of the composites and could be used for various biomedical applications.     

Application of hydrophilic ionic liquid treatment to the morphological observations of hydrated porous ceramic green bodies

In this study, a simple and convenient method for observing the surface morphology of hydrated porous ceramic green bodies is proposed. The porous hyrdoxyapatite (HAp) green body was prepared by a gelcasting process and was dried in a humid chamber from 90 to 50% relative humidity at 25 °C before subsequent treatment with a hydrophilic ionic liquid (IL). The surface morphology of the IL-treated porous HAp green body was observed using FE-SEM. The results showed that the pore morphology and microstructure of the HAp green body was readily observable without evidence of charging. The as-prepared sample showed pores approximately 300–600 μm in diameter, which gradually contracted to approximately 200–400 μm upon drying in the humid chamber. Following sintering at 1000 °C, the pores had further contracted to approximately 100–300 μm. The IL binds with the surrounding water to prevent the sample from drying in vacuum and acts as a conductive media, allowing the HAp ceramics to be observed in the electron microscope. In comparison to the micro-focused X-ray CT analysis, the fine pore structure (less than 100 μm) could only be observed using FE-SEM when the porous body had also been subjected to the IL treatment.     

Effect of particle size and freezing conditions on freeze-casted scaffold

Freeze casting is one of the emerging and novel manufacturing routes to fabricate porous scaffolds for various applications including orthopedic implants, drug delivery, energy storing devices etc. Thus, it becomes important to understand this process in a deeper sense. Present work was focused to study the effect/influence of basic parameters, particle sizes, and freezing conditions on the mechanical properties and microstructures of porous scaffold fabricated by freeze casting. β-tricalcium phosphate (β-TCP) and hydroxyapatite (HAp) powder with particle sizes of 10?μm and 20?nm were used. Prepared slurries were freeze casted at constant freezing temperature (5?°C) and constant freezing rate (1.86?°C/min) to study the effect of freezing conditions on mechanical and microstructural properties of the porous scaffold. It was observed that porous scaffold fabricated by nanoparticles has given better porosity (63.22–76.16%), than scaffold fabricated by microparticles (13–43.05%) at given solid loading of both freezing conditions. Although, the range of pore size of the scaffold fabricated by nanoparticles (CFR: 2.60–0.84?μm; CFT: 1.66–0.46?μm) was lower than that of scaffold fabricated by microparticles (CFR: 9.45–4.83?μm; CFT: 4.72–2.84?μm). The compressive strength of scaffolds prepared by nanoparticles was in the range of trabecular bone. Moreover, the results of present work will pave the way for the fabrication of porous scaffold with desired pore size and porosity for various implants, energy, and drug delivery applications.     

3D pore-interconnected calcium phosphate bone blocks for bone tissue engineering

Pore size and connectivity of artificial bone scaffolds play key role in regulating cell ingrowth and vascularization during healing. The objective of this study was to develop a novel process for preparing 3D pore-interconnected open-cell bone substitutes with varying pore sizes. This was achieved by thermal-induced expansion, drying, then sintering the mixture of biphasic calcium phosphate (BCP) and a thermal responsive porogen comprising chitosan (CS) and hydroxypropyl methyl cellulose (HPMC). The interpolymer complexes (IPCs) of CS/HPMC were prepared and investigated by FT-IR. The mixtures of IPCs/BCP were heated up to 100 °C for analyzing their thermal expansion properties. This resulted in ~13% and ~42% volume increment for IPC-1/BCP and IPC-2/BCP, respectively, while ~230% volume increased in the case of IPC-3/BCP (therefore chosen for sintering bone blocks). Heating rate-dependent (0.20–0.25 °C/min range) sintering profiles for IPC-3/BCP were utilized to produce BCP bone blocks. Gasification of IPC during sintering resulted in the formation of interconnected porous structures, and the morphology was investigated by SEM, revealing varying sizes ranging from 106 ± 13 μm to 1123 ± 75 μm. The pore size range of bone blocks from 235 ± 46 μm to 459 ± 76 μm portrayed significantly high MC3T3-E1 cell viability with prominent filopodial extensions, and elongated cells, depicting efficient biocompatibility. Therefore, the process for preparing porous interconnected 3D bone blocks were feasible, thereby serving as an alternative for potential bone tissue engineering applications.     

Preparation and characterization of vancomycin releasing PHBV coated 45S5 Bioglass®-based glass–ceramic scaffolds for bone tissue engineering

Porous 45S5 Bioglass^®-based glass–ceramic scaffolds with high porosity (96%) and interconnected pore structure (average pore size 300 μm) were prepared by foam replication method. In order to improve the mechanical properties and to incorporate a drug release function, the scaffolds were coated with a drug loaded solution, consisting of PHBV and vancomycin. The mechanical properties of the scaffolds were significantly improved by the PHBV coating. The bioactivity of scaffolds upon immersion in SBF was maintained in PHBV coated scaffolds although the formation of hydroxyapatite was slightly retarded by the presence of the coating. The encapsulated drug in coated scaffolds was released in a sustained manner (99.9% in 6 days) as compared to the rapid release (99.5% in 3 days) of drug directly adsorbed on the uncoated scaffolds. The obtained drug loaded and bioactive composite scaffolds represent promising candidates for bone tissue engineering applications.     

Characterization of 3D elastic porous polydimethylsiloxane (PDMS) cell scaffolds fabricated by VARTM and particle leaching

In this study, elastic porous polydimethylsiloxane (PDMS) cell scaffolds were fabricated by vacuum‐assisted resin transfer moulding (VARTM) and particle leaching technologies. To control the porous morphology and porosity, different processing parameters, such as compression load, compression time, and NaCl particle size for preparing NaCl preform, were studied. The porous structures of PDMS cell scaffolds were characterized by scanning electron microscopy (SEM). The properties of PDMS cell scaffolds, including porosity, water absorption, interconnectivity, compression modulus, and compression strength were also investigated. The results showed that after the porogen–NaCl particles had been leached, the remaining pores had the sizes of 150–300, 300–450, and 450–600 μm, which matched the sizes of the NaCl particles. The interconnectivity of PDMS cell scaffolds increases with an increase in the size of NaCl particles. It was also found that the smaller the size of the NaCl particles, the higher the porosity and water absorption of PDMS cell scaffolds. The content of residual NaCl in PDMS/NaCl scaffolds reduces under ultrasonic treatment. In addition, PDMS scaffolds with a pore size of 300–450 μm have better mechanical properties compared to those with pore sizes of 150–300 and 450–600 μm. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 42909.     

Low Cost Porous Alumina with Tailored Microstructure and Thermal Conductivity Prepared using Rice Husk and Sucrose

This study demonstrates a cost‐effective way to fabricate porous ceramics with tailored porosity and pore microstructure using 5–40 wt% rice husk (RH) in <75 μm, 75–180 μm, 180–355 μm, 355–420 μm, and 420–600 μm size, as pore former. Sucrose, used as binder, also acted as a pore former. Porous alumina compacts with 20%–66% volume fraction porosity and 50–516 μm pore size (length) were successfully fabricated. Microstructure of samples reveal randomly oriented elongated coarse pores and fine pores (avg. size 4 μm), created during burnout of RH and sucrose, respectively. Samples with isolated and/or interconnected pores were fabricated using this process. Thermal conductivity of the samples prepared was measured using Transient Plane Source (TPS) technique. Thermal conductivity ranges from 1.2 to 24 W/mK. Experimental results agree closely with predictions made based on Effective Medium Theory (EMT) for a two‐phase system.     

Polymer template fabrication of porous hydroxyapatite scaffolds with interconnected spherical pores

Porous hydroxyapatite (HA) scaffolds with interconnected spherical pores were fabricated by slip casting using a polymer template. Templates were produced using polymer beads, NaCl, and adhesive (N100). Effects of the preparation process on the pore structures and mechanical properties of the porous HA scaffolds were investigated. Pore interconnectivity was improved by adding NaCl particles with appropriate diameters to the polymer template. The size of the adhesive area could be controlled by adjusting the concentration of N100. The pore size could be controlled between 200 ± 42 and 400 ± 81 μm, and the porosity between 50.2 and 73.1%, by changing the size of the polymer beads and the volume of the NaCl particles. The compressive strength decreased as the porosity or pore size increased.     

Preparation and characterization of Chitosan and PLGA‐based scaffolds for tissue engineering applications

Three dimensional (3D) biodegradable porous scaffolds play a crucial role in bone tissue repair. In this study, four types of 3D polymer/hydroxyapatite (HAp) composite scaffolds were prepared by freeze drying technique in order to mimic the organic/inorganic nature of the bone. Chitosan (CH) and poly(lactic acid‐co‐glycolic acid) (PLGA) were used as the polymeric part and HAp as the inorganic component. Properties of the resultant scaffolds, such as morphology, porosity, degradation, water uptake, mechanical and thermal stabilities were examined. 3D scaffolds having interconnected macroporous structure and 77–89% porosity were produced. The pore diameters were in the range of 6 and 200 µm. PLGA and HAp containing scaffolds had the highest compressive modulus. PLGA maintained the strength by decreasing water uptake but increased the degradation rate. Scaffolds seeded with SaOs‐2 osteoblast cells showed that all scaffolds were capable of encouraging cell adhesion and proliferation. The presence of HAp particles caused an increase in cell number on CH‐HAp scaffolds compared to CH scaffolds, while cell number decreased when PLGA was incorporated in the structure. CH‐PLGA scaffolds showed highest cell number on days 7 and 14 compared to others. Based on the properties such as interconnected porosity, high mechanical strength, and in vitro cell proliferation, blend scaffolds have the potential to be applied in hard tissue treatments. POLYM. COMPOS., 36:1917–1930, 2015. © 2014 Society of Plastics Engineers     

Fabrication and characterization of alumina and zirconia-toughened alumina porous structures

In the present study, alumina (Al₂O₃) and zirconia-toughened alumina (ZTA) porous structures (foams) were manufactured using the space holder technique. Al₂O₃ and ZTA foams with varying porosities from 20% to 69% were fabricated by adding different sizes (10, 20, and 40 μm) and different volume % of polystyrene beads (space holders) to Al₂O₃ and ZTA powders. All the fabricated foams were investigated under static conditions to assess the compressive behavior. It is observed that the compressive strength of these foams strongly depends on porosity, pore size, pore size distribution and pore wall thickness. Among all fabricated foams, Al₂O₃ foams with 20 vol% beads of 10 μm size showed a higher compressive strength of 700 MPa with low porosity (21%) and a higher pore wall thickness (2.8 μm). It is also observed that the pore wall thickness decreased with the increase in beads size and the volume % of the beads, resulting in a low compressive strength value of 8 MPa with a lower pore wall thickness of 1.75 μm at 80 vol% of 40 μm beads. All the foams, irrespective of pore size, showed a typical ceramic failure phenomenon up to 70 vol% of beads; after that, the failure behavior changed to complete open-cell fracture.     

A comparative study of porous scaffolds with cubic and spherical macropores

Two kinds of polyester porous scaffolds having cubic and spherical macropores were fabricated, and a comparative study of their morphologies and mechanical properties were made in this paper. Poly(d,l-lactic-co-glycolic acid) (PLGA) scaffolds were prepared by room temperature compression molding and particulate leaching method based on cubic NaCl particles and paraffin spheres with a similar size range of 355-450 μm and a series of porosities (77-97%). Scanning electronic microscopy demonstrated that the spherical pore scaffolds exhibited better pore interconnectivity than the cubic pore ones. In compressive tests of both kinds of scaffolds, striking yield peaks were found at relatively low porosities, but just non-linear flexure behavior was observed at high porosities. The power-law relationships of compressive modulus and compressive strength versus porosity were confirmed in both foams. Comparison of the underlying scaling exponents reveals that the scaffolds with spherical pores are, at high porosities, with better compressive properties to a certain degree in contrast to those with cubic pores.     

Preparation of dense β-CaSiO3 ceramic with high mechanical strength and HAp formation ability in simulated body fluid

In present study, dense CaSiO₃ (CS) ceramics have been fabricated through spark plasma sintering (SPS) technique using β-CS powder prepared by chemical precipitation method. The β-CS ceramic sintered at 950 °C has a relative density of about 95% and shows a fine microstructure with an average grain size of 0.6 μm, thus expresses good bending strength of about 294 MPa. The simulated body fluid (SBF) immersion tests show that the dense β-CS ceramic has a high hydroxyapatite (HAp) formation rate on its surface. The HAp layer formed on the CS ceramic surface has a granular structure consisting of silkworm-like HAp grains, and the thickness of HAp and Si-rich layer are 70 and 120 μm, respectively.     

Selective laser sintering of porous Al2O3-based ceramics using both Al2O3 and SiO2 poly-hollow microspheres as raw materials

Porous Al₂O₃-based ceramics with improved mechanical strength and different pore size were fabricated using Al₂O₃ and SiO₂ poly-hollow microspheres (PHMs) as raw materials by selective laser sintering (SLS). The effects of different contents of SiO₂ PHMs on phase compositions, microstructures, mechanical properties and pore size distribution of the prepared ceramics were investigated. It is found that moderate content of SiO₂ PHMs (≤30 wt%) could work as a sintering additive, which could enhance the bonding necks between Al₂O₃ PHMs. When the content of SiO₂ PHMs increased from 0 wt% to 30 wt%, the compressive strength of Al₂O₃-based ceramics increased from 0.3 MPa to 4.0 MPa, and the porosity decreased from 77.0% to 65.0% with open pore size decreased from 52.0 μm to 38.3 μm. However, SiO₂ PHMs could provide pores by keeping its integrity when the content of SiO₂ PHMs increased to 40 wt%, which could result in the porosity increasing to 66.8% and pore size decreasing to 30.1 μm. Selective laser sintering of different kinds of ceramic PHMs is a feasible method to fabricate porous ceramics with complex shape, controllable pore size and improved properties.     

Mechanical properties of porous ceramic scaffolds: Influence of internal dimensions

Highly porous ceramic scaffolds have been fabricated from a 70% SiO₂–30% CaO glass powder using stereolithography and the lost-mould process combined with gel-casting. After sintering at 1200 °C the glass crystallised to a structure of wollastonite and pseudowollastonite grains in a glassy matrix with a bulk porosity of 1.3%. All scaffolds had a simple cubic strut structure with an internal porosity of approximately 42% and internal pore dimensions in the range 300–600 μm. The mean crushing strength of the scaffolds is in the range 10–25 MPa with the largest pore sizes showing the weakest strengths. The variability of scaffold strengths has been characterised using Weibull statistics and each set of scaffolds showed a Weibull modulus of m≈3 independent of pore size. The equivalent strength of the struts within the porous ceramics was estimated to be in the range 40–80 MPa using the models of the Gibson and Ashby. These strengths were found to scale with specimen size consistent with the Weibull modulus obtained from compression tests. Using a Weibull analysis, these strengths are shown to be in accordance with the strength of 3-point bend specimens of the bulk glass material fabricated using identical methods. The strength and Weibull modulus of these scaffolds are comparable to those reported for other porous ceramic scaffold materials of similar porosity made by different fabrication routes.     

Interconnectivity and permeability of supercritical fluid-foamed scaffolds and the effect of their structural properties on cell distribution

This study aims to investigate interconnectivity and permeability of scCO₂-foamed scaffolds and the influence of structural scaffold properties on cell distribution. Supercritical fluid technology was utilized to fabricated scaffolds from 37 kDa, 53 kDa and 109 kDa PLGA (85:15). Pore size, pore size distribution and porosity were quantified by MicroCT, and window sizes were measured using SEM. A novel interconnectivity algorithm allowed the quantification of scaffold interconnectivity in three space dimensions. To determine the permeability of porous materials direct perfusion experiments were performed, where a known flow rate was applied to measure the pressure differential across the scaffolds. The permeability was calculated using Darcy's law. Largest pore sizes, porosities, interconnectivities and permeabilities were obtained for scaffolds fabricated from 37 kDa PLGA. These scaffolds showed a heterogeneous pore structure and distribution, whereas homogeneous pore structure, smallest pore sizes, porosities, interconnectivities and permeabilities were observed for scaffolds fabricated from 109 kDa PLGA. The distribution of 3T3 fibroblasts through scCO₂-foamed scaffolds was investigated by MicroCT and MTT staining. Cells were further visualized by fluorescent imaging. Uniform cell distribution was observed on scaffolds fabricated from 109 kDa PLGA and an average of 10% of the total scaffold volume was covered with cells that had adhered onto them.     

Effectual drug-releasing porous scaffolds from 1,6-diisocyanatohexane-extended poly(1,4-butylene succinate) for bone tissue regeneration

Tooth extraction induces residual ridge resorption which impairs function and aesthetic of dental prostheses. This study aimed at developing new bone scaffolds to be used in a tooth socket for preserving bone mass from the residual ridge resorption. Scaffolds were fabricated from poly(1,4-butylene succinate) extended with 1,6-diisocyanatohexane (PBSu-DCH) by solvent casting and particulate leaching technique. Four different weight ratios of NaCl particles (200-400 μm; used as the porogen species) and the polymer were varied (i.e., 25, 30, 35, and 40% based on the weight of the polymer). Scaffolds were evaluated for their physical (i.e., morphology, porosity, pore volume, and pore size), physico-mechanical (i.e., mechanical properties and water retention ability), and biological properties (i.e., cytotoxicity and bone cell attachment). The potential for use of the as-prepared materials as effectual drug-releasing scaffolds for bone tissue regeneration was assessed by incorporating ipriflavone and studying the release of the drug from the drug-loaded scaffolds.     

Morphological,mechanical, and biological evolution of pure hydroxyapatite and its composites with titanium carbide for biomedical applications

Pure hydroxyapatite (HAp) was synthesized successfully via a wet chemical precipitation method. To study the influence of TiC (weight % of 5, 10, 15) substitution on the mechanical behavior of pure HAp, its composites with TiC were synthesized using a solid-state reaction method. Herein, detailed investigations of pure HAp and its composites using X-ray powder diffraction (XRD), FTIR spectroscopy, Raman spectroscopy, UV-VIS spectroscopy, SEM followed by EDAX and particle size analysis were carried out. XRD study reveals the phase stability of the prepared HAp and composite samples. However, FTIR and Raman spectroscopic studies revealed the bond formation among the various constituents. Mechanical behavior of HAp, and its composites with TiC were studied using numerous parameters like density, Young's modulus, fracture toughness, and load absorption capability. Based on these studies, it was revealed that the addition of 5 wt % substitution of TiC sintered at 1200 °C significantly enhanced the mechanical properties of pure HAp. Hence, 5 wt % of TiC composite 95HAp-5TiC showed the best mechanical characteristics such as density (2.3060 g/cm³), Young's modulus (14.53 MPa), fracture toughness (19.82 MPa m^1/2), maximum compressive strength (186 MPa) respectively. Cytotoxicity and osteogenic activities of the synthesized pure HAp and its composite, 95HAp-5TiC were performed using osteoblast cells (mouse calvarial) at different concentrations of the samples (0.01 μg, - 100 μg). From the above studies, the cell viability and ALP activities of the composite, 95HAp-5TiC found to be excellent than that of pure HAp. Hence, this composite sample may be utilized for bone implant applications.     

Porous β-Ca2SiO4 ceramic scaffolds for bone tissue engineering: In vitro and in vivo characterization

The biodegradable ceramic scaffolds with desirable pore size, porosity and mechanical properties play a crucial role in bone tissue engineering and bone transplantation. A novel porous β-dicalcium silicate (β-Ca₂SiO₄) ceramic scaffold was prepared by sintering the green body consisting of CaCO₃ and SiO₂ at 1300 °C, which generated interconnected pore network with proper pore size of about 300 μm and high compressive strength (28.13±5.37–10.36±0.83 MPa) following the porosity from 53.54±5.37% to 71.44±0.83%. Porous β-Ca₂SiO₄ ceramic scaffolds displayed a good biocompatibility, since human osteoblast-like MG-63 cells and goat bone mesenchymal stem cells (BMSCs) proliferated continuously on the scaffolds after 7 d culture. The porous β-Ca₂SiO₄ ceramic scaffolds revealed well apatite-forming ability when incubated in the simulated body fluid (SBF). According to the histological test, the degradation of porous β-Ca₂SiO₄ ceramic scaffolds and the new bone tissue generation in vivo were observed following 9 weeks implantation in nude mice. These results suggested that the porous β-Ca₂SiO₄ ceramic scaffolds could be potentially applied in bone tissue engineering.