Fabrication of porous scaffolds with a controllable microstructure and mechanical properties by porogen fusion technique

Macroporous scaffolds with controllable pore structure and mechanical properties were fabricated by a porogen fusion technique. Biodegradable material poly (d, l-lactide) (PDLLA) was used as the scaffold matrix. The effects of porogen size, PDLLA concentration and hydroxyapatite (HA) content on the scaffold morphology, porosity and mechanical properties were investigated. High porosity (90% and above) and highly interconnected structures were easily obtained and the pore size could be adjusted by varying the porogen size. With the increasing porogen size and PDLLA concentration, the porosity of scaffolds decreases, while its mechanical properties increase. The introduction of HA greatly increases the impact on pore structure, mechanical properties and water absorption ability of scaffolds, while it has comparatively little influence on its porosity under low HA contents. These results show that by adjusting processing parameters, scaffolds could afford a controllable pore size, exhibit suitable pore structure and high porosity, as well as good mechanical properties, and may serve as an excellent substrate for bone tissue engineering.     

Design and fabrication of 3D‐plotted polymeric scaffolds in functional tissue engineering

Regenerating the load‐bearing tissues requires 3D scaffolds that balance the temporary mechanical function with the biological requirements. In functional tissue engineering, designing scaffolds with biomimetic mechanical properties could promote tissue ingrowth since the cells are sensitive to their local mechanical environment. This work aims to design scaffolds that mimic the mechanical response of the biological tissues under physiological loading conditions. Poly(L ‐lactide) (PLLA) scaffolds with varying porosities and pore sizes were made by the 3D‐plotting technique. The scaffolds were tested under unconfined ramp compression to compare their stress profile under load with that of bovine cartilage. A comparison between the material parameters estimated for the scaffolds and for the bovine cartilage based on the biphasic theory enabled the definition of an optimum window for the porosity and pore size of these constructs. Moreover, the finite element prediction for the stress distribution inside the scaffolds, surrounded by the host cartilaginous tissue, demonstrated a negligible perturbation of the stress field at the site of implantation. The finite element modeling tools in combination with the developed methodology for optimal porosity/pore size determination can be used to improve the design of biomimetic scaffolds. POLYM. ENG. SCI., 47:608–618, 2007. © 2007 Society of Plastics Engineers.     

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.     

Fabrication and characterization of honeycomb β-tricalcium phosphate scaffolds through an extrusion technique

In this study, for the first time honeycomb β-tricalcium phosphate (β-TCP) scaffolds were fabricated through an extrusion technique. The physicochemical properties and cell behaviors of the honeycomb β-TCP scaffolds were investigated. The results showed that scaffolds were characterized by ordered channel-like macropores and unidirectional interconnection. The pore structure and mechanical strength could be tailored by changing the parameters of extrusion molds. The pore size of scaffolds was in the range of 400–800 µm approximately, while their compressive strength parallel to the pore direction and porosity ranged from 14 to 20 MPa and 60–70%, respectively. The in vitro cell behavior demonstrated that cells could well attach on the surfaces and grow into the inner channel-like pores of thescaffolds; the scaffolds with higher porosity showed better cell proliferation but poorer cell differentiation. The honeycomb scaffolds fabricated by extrusion technique are potential candidate for bone tissue engineering.     

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.     

Improvement of mechanical properties of macroporous β-tricalcium phosphate bioceramic scaffolds with uniform and interconnected pore structures

The biocompatible and degradable macroporous bioceramic scaffolds with high mechanical properties and interconnected porous structures play an important role in hard tissue regeneration and bone tissue engineering applications. In this study, the improvement of mechanical properties of macroporous β-tricalcium phosphate [β-Ca₃(PO₄)₂, β-TCP] bioceramic scaffolds with uniform macropore size and interconnected pores were fabricated by impregnation of the synthesized β-TCP nano-powder slurry into polymeric frames. The microstructures, mechanical properties and in vitro degradation of the fabricated samples were investigated. For a comparison, β-TCP scaffolds were also fabricated from commercial micro-size powders under the same conditions. The resultant scaffolds showed porosities ∼65% with uniform macropore size ranging from 400 to 550 μm and interconnected pore size ∼100 μm. The compressive strength of the samples fabricated from nano-size powders reached 10.87 MPa, which was almost twice as high as those fabricated from commercial micro-size powders, and was comparable to the high-end value (2–10 MPa) of human cancellous bone. Furthermore, the degradation of the β-TCP bioceramics fabricated from nano-size powders was apparently lower than those fabricated from commercial micro-size powders, suggesting the possible control of the degradation of the scaffolds by regulating initial powder size. Regarding the excellent mechanical properties and porous structures, the obtained macroporous β-TCP bioceramic scaffolds can be used in hard tissue regeneration and bone tissue engineering applications.     

Solvent-Free Fabrication of Tissue Engineering Scaffolds With Immiscible Polymer Blends

A completely organic solvent-free fabrication method is developed for tissue engineering scaffolds by gas foaming of immiscible polylactic acid (PLA) and sucrose blends, followed by water leaching. PLA scaffolds with above 90% porosity and 25–200 µm pore size were fabricated. The pore size and porosity was controlled with process parameters including extrusion temperature and foaming process parameters. Dynamic mechanical analysis showed that the extrusion temperature could be used to control the scaffold strength. Both unfoamed and foamed scaffolds were used to culture glioblastoma (GBM) cells M059 K. The results showed that the cells grew better in the foamed PLA scaffolds. The method presented in the paper is versatile and can be used to fabricate tissue engineering scaffolds without any residual organic solvents.     

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.     

Morphology,wettability, and mechanical properties of polycaprolactone/hydroxyapatite composite scaffolds with interconnected pore structures fabricated by a mini‐deposition system

A new mini‐deposition system (MDS) was developed to fabricate scaffolds with interconnected pore structures and anatomical geometry for bone tissue engineering. Polycaprolactone/hydroxyapatite (PCL/HA) composites with varying hydroxyapatite (HA) content were adopted to manufacture scaffolds by using MDS with a porosity of 54.6%, a pore size of 716 μm in the x‐y plane, and 116 μm in the z direction. The water uptake ratio and compressive modulus of PCL/HA composite scaffold increase from 8 to 39% and from 26.5 to 49.8 MPa, respectively, as the HA content increases from 0 to 40%. PCL/HA composite scaffolds have better wettability and mechanical properties than pure PCL scaffold. A PCL/HA composite scaffold for mandible bone repair was successfully fabricated with both interconnected pore structures and anatomical shape to demonstrate the versatility of MDS. POLYM. ENG. SCI., 2012. © 2012 Society of Plastics Engineers     

Novel fabrication of hierarchically porous hydroxyapatite scaffolds with refined porosity and suitable strength

Based on extrusion deposition and foaming technique, a novel method for biological hydroxyapatite (HA) scaffolds was introduced in this paper. The scaffolds were primarily characterised by interconnected and hierarchically porous structures with high porosity, adjustable distribution of pore sizes, as well as considerable mechanical strength. In order to confirm that fine control of bulk porosity and mechanical strength was possible and feasible, further analysis of obtained scaffolds was carried out by field emission scanning electron microscope (FESEM), compressive test and calculation of volumetric shrinkage; in particular, the additional porosity resulting from the introduction of pore former was evaluated. The results indicated that this method can have a great potential to construct HA scaffolds of suitable quality for spongy bone in bone tissue engineering.     

Evaluation of poly(L-lactide) and chitosan composite scaffolds for cartilage tissue regeneration

The present study delineates the development of chitosan and poly(L-lactide) (PLLA) scaffolds cross-linked using a mixture of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), n-hydroxysuccinimide (NHS), and chondroitin sulfate (CS) for cartilage tissue engineering applications. Chitosan and PLLA were varied in concentration for developing scaffolds and prepared by freeze-drying method. The various scaffolds were studied using scanning electron microscopy (SEM), porosity by mercury intrusion porosimeter, and the molecular interactions among polymers using Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) studies. Differential scanning calorimetry was used to predict the thermal properties of the scaffolds. The mechanical properties of the scaffolds were studied using static mechanical tester. The ability of the scaffolds to support chondrocyte proliferation was also studied. The microscopy suggests that the pore size of the scaffolds varied with the composition in the range of 38–172 μm and the porosities in the range of 73–93%. The XRD and the FTIR studies suggested that an alternation in the composition of the scaffolds altered the molecular interactions among the scaffold components. An increase in the chitosan content enhanced the swelling property. The degradation of the scaffolds was least when the proportion of chitosan and PLLA was in the ratio of 70:30. The in vitro cell proliferation study suggested that the developed scaffolds were able to support chondrogenesis, the glycosaminoglycan (GAG) content of the mature chondrocyte was 40 μg/ml and the viability was approximately 90%. Hence, the so designed scaffolds may be tried for cartilage tissue engineering applications.     

Macroporous Bioglass-derived scaffolds for bone tissue regeneration

Since it was introduced at the end of the ‘60s, the 45S5 Bioglass^® has played a fundamental role among the materials for orthopedic applications because of its ability to build a stable bond with the surrounding bone. The recent development of bone tissue engineering has led the interest of many scientists in the design of Bioglass^®-based scaffolds, i.e. porous systems able to drive and foster the bone tissue regrowth. Among the available techniques to realize scaffolds, the polymer burning out method, which employs organic particles as pore generating agents in a ceramic matrix, combines versatility and low cost. In spite of the advantages of the polymer burning out method, this technique has been rarely applied to 45S5 Bioglass^® and a systematic feasibility study has not been carried out on this issue yet. In order to fill this gap, in the present contribution the polymer burning out method was employed to design macroporous scaffolds based on 45S5 Bioglass^®. Different amounts of organic phase were used to obtain samples with different porosity. The samples were characterized from a microstructural point of view, in order to evaluate the pore morphology, dimension and degree of interconnectivity. Such findings proved that a proper setting of the processing parameters made it possible to achieve very high porosity values, among the best ones obtained in the literature with the same technique, together with an appreciable mechanical behaviour, according to compression tests. Finally, the scaffolds bioactivity was assessed by means of in vitro tests in a simulated body fluid (SBF) solution. Moreover, in the view of a potential application for bone tissue engineering, a preliminary biological evaluation of the obtained scaffolds to sustain cell proliferation was carried out.     

Characterisation of Bioglass based foams developed via replication of natural marine sponges

A comparative characterisation of Bioglass based scaffolds for bone tissue engineering applications developed via a replication technique of natural marine sponges as sacrificial template is presented, focusing on their architecture and mechanical properties. The use of these sponges presents several advantages, including the possibility of attaining higher mechanical properties than those scaffolds made by foam replica method (up to 4?MPa) due to a decrease in porosity (68–76%) without affecting the pore interconnectivity (higher than 99%). The obtained pore structure possesses not only pores with a diameter in the range 150–500?μm, necessary to induce bone ingrowth, but also pores in the range of 0–200?μm, which are requested for complete integration of the scaffold and for neovascularisation. In this way, it is possible to combine the main properties that a three-dimensional scaffold should have for bone regeneration: interconnected and high porosity, adequate mechanical properties and bioactivity.     

Bioactive and Biocompatible Macroporous Scaffolds with Tunable Performances Prepared Based on 3D Printing of the Pre‐Crosslinked Sodium Alginate/Hydroxyapatite Hydrogel Ink

Bioactive and biocompatible porous scaffold materials with adjustable pore structures and drug delivery capability are one of the key elements in bone tissue engineering. In this work, bioactive and biocompatible sodium alginate (SA)/hydroxyapatite (HAP) macroporous scaffolds are facilely and effectively fabricated based on 3D printing of the pre‐crosslinked SA/HAP hydrogels followed by further crosslinking to improve the mechanical properties of scaffolds. The pore structures and porosity (>80%) of the porous scaffolds can be readily tailored by varying the formation conditions. Furthermore, the in vitro biomineralization tests show that the bioactivity of the porous scaffolds is effectively enhanced by the addition of HAP nanoparticles into the scaffold matrix. Furthermore, the anti‐inflammatory drug curcumin is loaded into the porous scaffolds and the in vitro release study shows the sustainable drug release function of the porous scaffolds. Moreover, mouse bone mesenchymal stem cells (mBMSCs) are cultured on the porous scaffolds, and the results of the in vitro biocompatibility experiment show that the mBMSCs can be adhered well on the porous scaffolds. All of the results suggest that the bioactive and biocompatible SA/HAP porous scaffolds have great application potential in bone tissue engineering.     

Fabrication of cancellous-bone-mimicking β-tricalcium phosphate bioceramic scaffolds with tunable architecture and mechanical strength by stereolithography 3D printing

It is highly challenging to fabricate bioceramic scaffolds mimicking architecture and mechanical strength of cancellous bone. Gyroid structure, which is based on triply periodic minimal surface, highly resembles the architecture of cancellous bone. Herein, β-tricalcium phosphate (β-TCP) bioceramic scaffolds with gyroid structure were fabricated by stereolithography (SLA) 3D printing. The SLA 3D printing ensured high precision of ceramic part. The porosity (51–87%), pore size (250 – 2400 µm), pore wall thickness (< 300 µm) and compressive strength (0.6 – 16.8 MPa) of gyroid bioceramic scaffolds were readily adjusted to match various sites of cancellous bone. The gyroid bioceramic scaffolds were more favorable for cell proliferation than the grid-like bioceramic scaffolds. The cancellous-bone-mimicking gyroid bioceramic scaffolds with tunable architecture and mechanical strength were expected to efficiently repair the target bone defects.     

In vitro and in vivo properties of graphene-incorporated scaffolds for bone defect repair

The employment of graphene and its derivatives, graphene oxide and reduced graphene oxide, is extending from bioimaging and fabrications of biosensors to drug delivery and tissue engineering in the biomedical area. Graphene family-incorporated scaffolds, used in bone tissue engineering and bone regenerative medicine, profit superior properties of these materials, such as enhanced mechanical properties, large surface area, and the existence of functional groups. At the same time, problems related to cytotoxicity and adverse immune response of graphene family are solved when they are applied to produce 3-dimensional scaffolds. The objective of this review is to focus on in vitro properties of scaffolds consisting of graphene or its derivatives, especially osteogenic and antibacterial properties, as well as the influence of graphene and its derivatives on in vivo performances of implanted bone scaffolds. The positive effect of graphene and its two derivatives on attachment, and cell proliferation, as well as in vitro osteogenic differentiation of different cells was undeniable. Besides, the synergetic outcome of using graphene family on the antibacterial feature of scaffolds, especially incorporation with the silver element, was effective. Moreover, successful treatment of critical-sized bone defects was reported during in vivo preclinical tests when graphene or its derivatives-incorporated scaffolds were used. However, the limited number of in vivo studies should be considered as one of the main shortcomings to use graphene as a promising candidate for treating bone defects. It is anticipated that the increased number of well-designed preclinical studies could improve the applications of graphene incorporated scaffolds in bone tissue engineering/regeneration, and find out explanations and appropriate solutions to possible long-term toxicity and nonbiodegradability of these materials.     

3D打印骨组织支架孔隙结构对支架性能影响的研究进展

从骨组织工程支架孔隙形状、孔径大小、孔隙率、表面粗糙度、连接通路5方面对支架孔的微观结构的研究现状进行了综述。发现梯度结构的支架孔隙形状更接近天然骨,粗糙的表面可以改善细胞的黏附和增殖,非正交连接通路的内部结构可以为后期骨再生提供一个动态的生长空间。     

Numerical simulations of bioextruded polymer scaffolds for tissue engineering applications

Scaffolds provide a temporary mechanical and vascular support for tissue regeneration while shaping the in‐growth tissues. These scaffolds must be biocompatible, biodegradable, enclose appropriate porosity, pore structure and pore distribution, and have optimal structural and vascular performance, with both surface and structural compatibility. Surface compatibility means a chemical, biological and physical suitability to the host tissue. Structural compatibility corresponds to an optimal adaptation to the mechanical behaviour of the host tissue. Recent advances in the design of tissue engineering scaffolds are increasingly relying on computer‐aided design modelling and numerical simulations. The design of optimized scaffolds based on fundamental knowledge of their macro microstructure is a relevant topic of research. This research work presents a comparison between experimental compressive data and numerical simulations of bioextruded polymer scaffolds with different pore sizes for the elastic and plastic domain. Constitutive behaviour models of cellular structures are used in numerical simulations to compare numerical data with the experimental compressive data. Vascular simulation is also used in the design process of the extrusion‐based scaffolds in order to define an optimized scaffold design. © 2013 Society of Chemical Industry     

Direct ink writing of silica-carbon-calcite composite scaffolds from a silicone resin and fillers

Calcite-based composite scaffolds have been successfully 3D-printed by direct ink writing, starting from a paste comprising a silicone polymer and calcite (CaCO₃) powders. The firing in nitrogen, at 600?°C, after preliminary cross-linking step at 350?°C, determined the transformation of the polymer matrix into a silica-carbon nano-composite, embedding unreacted calcite particles. Compared to previously developed silica-calcite scaffolds, obtained after firing in air, the new composites exhibited a significant strength improvement (up to ～10?MPa, for a total open porosity of 56%). The new formulation did not compromise the in vitro bioactivity and the biocompatibility of the scaffolds, as shown by dissolution studies in SBF and preliminary cell culture tests, with human fibroblasts. Due to the simplicity of the processing and the outstanding mechanical performances, the developed scaffolds are promising candidates for bone tissue engineering applications.     

Compressive strength of β-TCP scaffolds fabricated via lithography-based manufacturing for bone tissue engineering

Bone is a vital organ that is responsible for the support and movement of body as well as the storage and transportation of cells and nutrients. Disease, along with traumatic events, can leave regions of bone with large voids and/or defects. Related surgical procedures, such as allografts, autografts, and arthroplasty, are reported to amount to roughly €9.6bn annually, emphasising the need for bone repair/replacement globally. Tricalcium phosphate (TCP) is a bioactive ceramic that has been identified as a suitable material for bone tissue engineering applications due to its excellent bioresorbability and overall biocompatibility. Through lithography-based ceramic manufacturing (LCM), β -TCP scaffolds were fabricated across nine different designs in this work. Pore size, unit cell size, and unit cell geometry were altered to vary the porosity of these scaffolds. Following fabrication, the material composition, topography, macrostructure, and microstructure of the β-TCP scaffolds were characterised. The effects of porosity and unit cell geometry on the compressive strengths of β -TCP scaffolds were analysed in detail. Compressive strengths of the scaffolds were measured between 1.4 ± 0.5 MPa and 67.6 ± 13.3 MPa across a porosity range of 5.58 ± 0.09% to 59.36 ± 0.18%. The strength of these scaffolds was considerably lower than that of the compressive strength of cortical bone (100–200 MPa), but mimic the compressive strength of cancellous bone well (0.1–16 MPa). While scaffolds of β-TCP alone may not be suitable for load-bearing applications, they demonstrate enough mechanical stability for bone regeneration/tissue engineering applications. They hold more potential in the regeneration of small bone defects/voids or in composite materials.