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.     

Ultraporous poly(lactic acid) scaffolds with improved mechanical performance using high‐pressure molding and salt leaching

A novel processing technique, i.e. high‐pressure compression molding/salt leaching, was developed to fabricate ultraporous poly(lactic acid) (PLA) scaffolds. The optimized composition was studied in relation to the porosity, pore morphology, thermal property, and mechanical performance of the PLA scaffolds. At a porogen (CaCO₃) content of 90 wt %, the scaffolds have an interconnected open pore structure and a porosity above 80%. It was truly interesting that the structural stability of high‐pressure molded scaffolds was remarkably improved based on the fact that its glass transition temperature (83.5°C) increased about 20°C, as compared to that of the conventional compression‐molded PLA (60°C), which is not far from physiological temperature (～37°C) at the risk of structural relaxation or physical aging. More importantly, the mechanical performance of PLA scaffolds was drastically enhanced under optimized processing conditions. At pressure and temperature of 1000 MPa and 190°C, the porous PLA scaffolds attained a storage modulus of 283.7 MPa, comparable to the high‐end value of trabecular bone (250 MPa) ever reported. In addition, our prepared PLA scaffolds showed excellent cellular compatibility and biocompatibility in vitro tests, further suggesting that the high‐pressure molded PLA scaffolds have high potential for bone tissue engineering applications. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 3509–3520, 2013     

Fabrication of a poly(ɛ‐caprolactone)/starch nanocomposite scaffold with a solvent‐casting/salt‐leaching technique for bone tissue engineering applications

A series of nanocomposite scaffolds of poly(?‐caprolactone) (PCL) and starch with a range of porosity from 50 to 90% were fabricated with a solvent‐casting/salt‐leaching technique, and their physical and mechanical properties were investigated. X‐ray diffraction patterns and Fourier transform infrared spectra confirmed the presence of the characteristic peaks of PCL in the fabricated scaffolds. Microstructure studies of the scaffolds revealed a uniform pore morphology and structure in all of the samples. The experimental measurements showed that the average contact angle of the PCL/starch composite was 88.05 ± 1.77°. All of the samples exhibited compressive stress/strain curves similar to those of polymeric foams. The samples with 50, 60, 70, and 80 wt % salt showed compressive‐load‐resisting capabilities in the range of human cancellous bone. With increasing porosity, a significant decrease in the mechanical properties of the scaffolds was observed. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43523.     

Study on development and in vitro biodegradation behaviour of poly(D,L-lactide-co-glycolide) 75/25 bioscaffolds

Abstract

Three-dimensional porous scaffolds based on biodegradable polymers are widely researched for applications to replace and restore the functions of diseased or damaged organs. The requirements for the scaffolds include: highly interconnected pore structures to facilitate cell adhesion for tissue regeneration, maintenance of mechanical properties and structural integrity until cells adapt to its environment and biodegradability with a controlled degradation rate. This paper focuses on the development and in vitro biodegradation behaviour of poly (D,L-lactide-co-glycolide acid) (PLGA) 75/25 and changes on pore morphology affected by initial pore sizes and degradation media. The pore morphology, mechanical properties, and geometric transformation were examined over the course of 13 weeks. It is concluded that the PLGA 75/25 scaffolds degraded after seven weeks and completely degraded after 13 weeks. The degradation time of scaffolds with small pores and in distilled water was comparatively shorter due to poorer interconnectivity of the pores and a more aggressive environment.     

Evaluation of mechanical behavior,bioactivity, and cytotoxicity of chitosan/akermanite-TiO2 3D-printed scaffolds for bone tissue applications

This report aimed to evaluate the mechanical behavior, bioactivity, and cytotoxicity of novel chitosan/akermanite-TiO₂ (CS/AK/Ti) composite scaffolds fabricated using the 3D-printing method. The morphological and structural properties of these scaffolds were characterized by Fourier transform spectroscopy (FTIR) and scanning electron microscopy (SEM). The mechanical behavior was examined by measuring the compressive strength, while the bioactivity was estimated in the simulated body fluid (SBF), and also the cytotoxicity of the scaffolds was assessed by conducting cell culturing experiments in vitro. It was found that the mechanical properties were considerably affected by the amount of TiO_2. The scaffolds had the possessed bone-like apatite forming ability, which indicated high bioactivity. Furthermore, L929 cells spread well on the surface, proliferated, and had good viability regarding the cell behaviors. The outcomes confirmed that the morphological, biological, and mechanical properties of developed 3D-composite scaffolds nearly mimicked the features of natural bone tissue. In summary, these findings showed that the 3D-printed scaffolds with an interconnected pore structure and improved mechanical properties were a potential candidate for bone tissue applications.     

Robocasting of 45S5 bioactive glass scaffolds for bone tissue engineering

Three dimensional scaffolds with controlled pore architecture were prepared from 45S5 Bioglass^® powders by robocasting (direct-writing) using carboxymethyl cellulose (CMC) as the single processing additive. A comprehensive sintering study of the resulting structures was performed within the temperature range 500–1050 °C. Robocast scaffolds with interconnected porosities ranging from 60 to 80% were obtained for a fixed scaffold design. All scaffolds exhibited compressive strengths comparable to that of cancellous bone (2–13 MPa), including those sintered at temperatures below the crystallization temperature of 45S5 bioactive glass. These strength values are substantially higher than any previously reported data for 45S5 Bioglass^® scaffolds and imply that robocasting is the first technique which can be considered suitable for producing vitreous 45S5 scaffolds with a sufficient mechanical integrity for any practical application. Moreover, this process will enable the development of 45S5 Bioglass^® scaffolds with customized external geometry, and optimized pore architecture.     

Microstructure and mechanical properties of poly(L‐lactide) scaffolds fabricated by gelatin particle leaching method

Biodegradable poly(L ‐lactide) (PLLA) scaffolds with well‐controlled interconnected irregular pores were fabricated by a porogen leaching technique using gelatin particles as the porogen. The gelatin particles (280–450 μm) were bonded together through a treatment in a saturated water vapor condition at 70°C to form a 3‐dimensional assembly in a mold. PLLA was dissolved in dioxane and was cast onto the gelatin assembly. The mixtures were then freeze‐dried or dried at room temperature, followed by removal of the gelatin particles to yield the porous scaffolds. The microstructure of the scaffolds was characterized by scanning electron microscopy with respect to the pore shape, interpore connectivity, and pore wall morphology. Compression measurements revealed that scaffolds fabricated by freeze‐drying exhibited better mechanical performance than those by room temperature dying. Along with the increase of the polymer concentration, the porosity of the scaffolds decreased whereas the compressive modulus increased. When the scaffolds were in a hydrated state, the compressive modulus decreased dramatically. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 1373–1379, 2005     

The effect of pore morphology and agarose coating on mechanical properties of tricalcium phosphate scaffolds

Three-dimensional biocompatible porous structures can be fabricated using different methods. However, the biological and mechanical behaviors of scaffolds are the center of focus in bone tissue engineering. In this study, tricalcium phosphate scaffolds with similar porosity contents but different pore morphologies were fabricated using two different techniques, namely, the replica method and the pore-forming agent method. The samples fabricated using the pore-forming agent showed more than two times higher compressive and bending strengths and more than three times higher compressive moduli. Furthermore, a thin layer of agarose coating improved the compressive and bending strength of both types of ceramic scaffolds. Subsequently, the samples’ capability to guide biomineralization was evaluated by immersion into a simulated body fluid that developed Ca-P nano-platelets formation and enhanced the compressive strength. Finally, the tetrazolium-based colorimetric (MTT) assay was used to evaluate L929 cell viability and proliferation on all the samples and confirmed that cell behavior was not affected by pore morphology or agarose coating. In summary, samples produced by the use of the pore-forming agent showed higher potential to be applied as bone scaffolds in tissue engineering applications.     

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     

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.     

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     

Biocompatible scaffolds composed of chemically crosslinked chitosan and gelatin for tissue engineering

Chitosan‐based scaffolds are widely studied in tissue regeneration because of their biocompatibility and biodegradability. Scaffolds are obtained by different techniques and can be modified with other polymers allowing controlling their properties. This article discusses the assembling of three‐dimensional chitosan porous scaffolds blended with gelatin. Gelatin was used to enhance cells attachment due to the presence of cell adhesion motifs, while improving mechanical strength. 2,5‐dimethoxy‐2,5‐dihydrofurane (DHF) was used as the crosslinking agent, because it allowed to control the reaction kinetics through temperature, time and DHF concentration. The results indicate that scaffolds morphology, pore sizes and distribution, compressive moduli and biodegradation in vitro with lysozyme, can be customized with variations of gelatin content and crosslinking degree. Scaffolds were neither cytotoxic nor genotoxic for human keratinocytes, exhibiting cell–substrate interactions. Our findings demonstrated that chitosan–gelatin scaffolds crosslinked with DHF, as a new crosslinking agent, are suitable in tissue engineering applications. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43814.     

Bioactive monetite‐containing whisker‐like fibers reinforced chitosan scaffolds

The aim of this work was to develop bioactive chitosan scaffolds reinforced with monetite‐containing whisker‐like fibers. The fibers synthesized by homogeneous precipitation were characterized as monetite/hydroxyapatite short fibers (MAFs), using XRD, FTIR and SEM. The pure chitosan and MAFs/chitosan composite scaffolds were produced by freeze‐drying, and characterized with respect to porosity, pore size, swelling behavior, compressive strength and modulus, and in vitro bioactivity. The incorporation of MAFs in chitosan matrices led to increase the pore size, according to the evaluation by FE‐SEM, and decrease the porosity of composite scaffolds. The swelling ratio decreased as MAFs content of scaffolds increased. The compressive strength and modulus of scaffolds were improved by an increase in MAFs content. The noncross‐linked scaffolds with a chitosan: MAFs weight ratio of 1:1 (CW3) showed a porosity of 75.5%, and the strength and modulus of 259 kPa and 2.8 MPa in dry state, respectively. The crosslinking by glutaraldehyde resulted in improved mechanical properties. The strength and modulus of cross‐linked CW3 scaffolds in wet state reached to 345 kPa and 1.8 MPa, respectively. The in vitro bioactivity of the reinforced scaffolds, evaluated by FE‐SEM/EDS, XRD, and ATR‐FTIR, was confirmed by the formation of a carbonated apatite layer on their surfaces when they soaked in simulated body fluid (SBF). The results of this initial study indicate that the monetite‐containing whisker‐like fibers may be an appropriate reinforcement of chitosan scaffolds.     

Preparation and characterization of aliphatic polyurethane and hydroxyapatite composite scaffold

Novel porous composite scaffolds for tissue engineering were prepared from aliphatic biodegradable polyurethane (PU) elastomer and hydroxyapatite (HA). It was found that the aliphatic PU was possible to load up to 50 wt % HA. The morphology and properties of the scaffolds were characterized by scanning electron microscope, X‐ray diffraction, infrared absorption spectra, mechanical testing, dynamic mechanical analysis, and in vitro degradation measurement. The results indicated that the HA/PU scaffolds had an interconnected porous structure with a pore size mainly ranging from 300 to 900 μm, and 50–200 μm micropores existed on the pores' walls. The average pore size of macropores and micropores are 510 and 100 μm, respectively. The compressive strength of the composite scaffolds showed higher enhancement with increasing HA content. In addition, the polymer matrix was completely composed of aliphatic component and exhibited progressive mass loss in vitro degradation, and the degradation rate depended on the HA content in PU matrix. The porous HA/PU composite may have a good prospect to be used as scaffold for tissue engineering. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Bending strength of porous ceramics tiles: Bounds and estimates of effective properties of an Intermingled Fractal Units’ model

In this work, a semi-analytical model for the determination of effective mechanical properties of porous ceramic tiles obtained by pore forming agent is proposed. MIP tests allow measuring porosity and pore size distribution of experimental systems. These data are used for developing an Intermingled Fractal Units’ model (IFU) as approximant of porous microstructures. IFU model is then combined with classical structural mechanics theory for the analytical computation of the bending strength of brittle ceramic beams. Bounds and estimates are given in full form and the detailed algorithm can be easily implemented in a numerical package. A preliminary comparison with experimental data shows the capability of the proposed model to reproduce the effective mechanical behaviour of ceramic tiles.     

Improvement of mechanical properties of polycaprolactone scaffolds by applying piezoelectric vibration system based on a dispensing process: A new concept

The mechanical properties of biomedical scaffolds are important for applications in tissue regeneration. The dispensing system described herein, which is based on a solid free‐form fabrication technique, enables the production of design‐based scaffolds with controllable pore structures. Although current plotting systems can easily fabricate a variety of three‐dimensional scaffolds, the mechanical properties of these constructs are difficult to control because of low processing speed. To overcome this limitation, a new dispensing method, which uses a piezoelectric vibration system to improve mechanical properties, has been developed. Polycaprolactone (PCL) strands fabricated using this technique were roughly 70% stronger than normal PCL strands. To explain this increase in mechanical strength, the combined effects of the piezoelectric system and the melt‐dispensing process on the crystalline morphology and molecular orientation of PCL strands were investigated. POLYM. ENG. SCI., 2011. © 2010 Society of Plastics Engineers     

Supercritical CO2 deposition and foaming process for fabrication of biopolyester–ZnO bone scaffolds

Subsequent supercritical CO₂‐assisted deposition and foaming process followed by in situ synthesis was used to fabricate functional polylactide (PLA) and polylactide–poly(?‐caprolactone) (PLA–PCL) bone scaffolds. Deposition of zinc bis(2‐thenoyltrifluoroacetonate) as a ZnO precursor onto biopolyester substrates (30 MPa; 110 °C) was followed by fast depressurization to create cellular structure. Contact time was optimized regarding the deposition yield (2 h), while PCL content in PLA was varied (1–10 wt %). Scaffolds impregnated with the precursor were treated with hydrazine alcoholic solution to obtain biopolyester–ZnO composites. Precursor synthesis and deposition onto the scaffolds was confirmed by Fourier‐transform infrared. Processed scaffolds had micron‐sized pores (d₅₀ ～ 20 μm). High open porosity (69–77%) and compressive strength values (2.8–8.3 MPa) corresponded to those reported for trabecular bone. PLA blending with PCL positively affected precursor deposition, crystallization rate, and compressive strength of the scaffolds. It also improved PLA surface roughness and wettability which are relevant for cell adhesion. ZnO improved compressive strength of the PLA scaffolds without significant effect on thermal stability. Analysis of structural, thermal, and mechanical properties of biopolyester–ZnO scaffolds testified a great potential of the obtained platforms as bone scaffolds. Proposed processing route is straightforward and ecofriendly, fast, easy to control, and suitable for processing of thermosensitive polymers. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 45824.     

Potential application of gelatin scaffolds prepared through in situ gas foaming in skin tissue engineering

Gelatin’s excellent foaming ability allows the application of in situ gas foaming as a preparation technique for porous scaffold development. Here, a new iterative experimental design for in situ gas foaming method is reported. The prepared scaffolds were studied for applying the findings to the future skin tissue engineering scaffolds. The thermal stability, mechanical properties, and pore structure of the scaffolds are reported and their degradation resistance by using collagenase enzyme and their cytotoxicity by using fibroblasts were studied. The results of this study demonstrated that gas foaming method can be modified to produce an interconnected porous structure with enhanced mechanical properties.     

A review of bioceramic porous scaffolds for hard tissue applications: Effects of structural features

Tissue engineering has acquired remarkable attention as an alternative strategy to treat and restore bone defects during recent years. A scaffold is a fundamental component for tissue engineering, on which cells attach, proliferate and differentiate to form new desirable functional tissue. The composition, and structural features of scaffolds, including porosity and pore size, play a fundamental role in the success of tissue-engineered construct. This review summarizes the effect of porosity and pore size of bioceramic-based scaffolds on their mechanical properties and biological performances. The focus of this review is on scaffolds with porosities 40% and above. From the mechanical point of view, the degree of porosity is a more important factor than pore size and scaffolds with porosities greater than 40% were more likely to substitute trabecular bones. While for in vitro and in vivo performances, pore size appeared more influential feature and co-existence of macropores and micropores led to better bone formation.     

High performance shape memory foams with isocyanate-modified hydroxyapatite nanoparticles for minimally invasive bone regeneration

This study conducts a comprehensive assessment of shape memory polyurethane (SMPU) composite foams with isocyanate-modified hydroxyapatite (imHA) nanoparticles in terms of their pore structures, mechanical properties, shape memory effects and biocompatibility in vitro. The results obtained in the research reveal the effectiveness of imHA nanoparticles in SMPU foams as inorganic cross-linking fillers, which contribute to the enhancement of mechanical properties and shape memory performance. Pore structures and compressive properties are simultaneously optimized when imHA content increases. The imHA enhanced SMPU foam could be adopted as a promising alternative for overcoming the disadvantages of traditional polymer scaffolds, such as insufficient mechanical properties, inadequate pore structure, low bioactivity and inconvenience in operation for bone regeneration.