A study on the fabrication of porous chitosan/gelatin network scaffold for tissue engineering

A novel porous chitosan/gelatin scaffold for tissue engineering was prepared via polyelectrolyte complex formation, freeze drying and post‐crosslinking with glutaraldehyde. The porosity and mean pore diameters could be controlled within 30∼100 µm by varying the original water content and the freezing conditions. Dipping the scaffolds in poly(lactic acid) provided good mechanical properties making it a promising candidate towards tissue engineering. © 2000 Society of Chemical Industry     

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     

Control of the structure and mechanical property of porous WS<Subscript>2</Subscript> scaffold during freeze casting

Porous WS₂ scaffolds with aligned lamellar pore were fabricated by freeze casting, and the pore morphology and size can be well controlled by adjusting the processing parameters during freeze casting process. The results indicated that the porosity and the compressive strength of porous WS₂ were greatly affected by the concentration of gelatin. As the gelatin addition increased from 1 to 5 wt%, the porosity of porous WS₂ scaffolds with initial solid content of 3 vol% decreased from 95.12 to 90.23%, and their compressive strength increased from 0.22 to 1.16 MPa. Moreover, the lamellar spacing and wall thickness could be tailored from 90 to 320 and 5 to 30 μm respectively by changing the cooling temperature. And the compressive strength of scaffolds has a slight increase with the decreases of cooling temperature. The porous WS₂ scaffold with fine aligned lamellar structure and proper compressive strength are expected to be used for scaffold materials.     

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.     

Modulating stability and mechanical properties of silica–gelatin hybrid by incorporating epoxy‐terminated polydimethylsiloxane oligomer

Silica‐gelatin hybrids, particularly GT‐G hybrids prepared by crosslinking gelatin (G) with γ‐glycidoxypropyltrimethoxysilane (GT), have attracted much attention in tissue engineering for diverse applications in hard or soft tissue regeneration; however, scaffolds with tunable properties are needed to meet specific requirements. In this work, a silica‐gelatin hybrid (ES/GT‐G) was synthesized by incorporating epoxy‐terminated polydimethylsiloxane oligomer (ES) to modulate the properties of GT‐G hybrid. The ES/GT‐G hybrid sponge presented a 3D network structure with porosity 86.4% ± 0.9%, determined by the liquid displacement method, and average pore size 340 ± 36 μm, determined by SEM observation. Compared with GT‐G hybrid material, the prepared ES/GT‐G hybrid wet film showed a decrease of tensile strength from 2.79 ± 0.04 MPa to 1.87 ± 0.12 MPa, with an increase of elongation at break from 19.96 ± 0.66% to 29.86 ± 0.87%, and the ES/GT‐G hybrid sponge exhibited a decline of compressive yield strength from 1.21 ± 0.04 MPa to 0.72 ± 0.06 MPa, based on the tensile and compression tests respectively. The introduction of ES enhanced the thermal denaturing temperature of GT‐G by 5°C as determined by a DSC study, and increased in vitro biodegradation slightly, without significantly changing surface wettability and swelling behavior. These findings suggest that silica‐gelatin hybrids with tunable properties are promising for applications from hard to soft tissue regeneration. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43059.     

I‐Optimal design of poly(lactic‐co‐glycolic) acid/hydroxyapatite three‐dimensional scaffolds produced by thermally induced phase separation

In bone tissue engineering, three‐dimensional (3D) scaffolds are often designed to have adequate modulus while taking into consideration the requirement for a highly porous network for cell seeding and tissue growth. This article presents the design optimization of 3D scaffolds made of poly(lactic‐co‐glycolic) acid (PLGA) and nanohydroxyapatite (nHA), produced by thermally induced phase separation (TIPS). Slow cooling at a rate of 1°C/min enabled a uniform temperature and produced porous scaffolds with a relatively uniform pore size. An I‐optimal design of experiments (DoE) with 18 experimental runs was used to relate four responses (scaffold thickness, density, porosity, and modulus) to three experimental factors, namely the TIPS temperature (?20, ?10, and 0°C), PLGA concentration (7%, 10%, and 13% w/v), and nHA content (0%, 15%, and 30% w/w). The response surface analysis using JMP® software predicted a temperature of ?18.3°C, a PLGA concentration of 10.3% w/v, and a nHA content of 30% w/w to achieve a thickness of 3 mm, a porosity of 83%, and a modulus of ~4 MPa. The set of validation scaffolds prepared using the predicted factor levels had a thickness of 3.05 ± 0.37 mm, a porosity of 86.8 ± 0.9%, and a modulus of 3.57 ± 2.28 MPa. POLYM. ENG. SCI., 59:1146–1157 2019. © 2019 Society of Plastics Engineers     

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.     

Facile preparation and cytocompatibility of poly(lactic acid)/poly(3‐hydroxybutyrate‐co‐4‐hydroxybutyrate) hybrid fibrous scaffolds

A fibrous scaffold is required to provide three‐dimensional (3D) cell growth microenvironments and appropriate synergistic cell guidance cues. In this study, porous scaffolds with different mass ratio of poly(lactic acid) to poly(3‐hydroxybutyrate‐co‐4‐hydroxybutyrate) (P(3HB‐co‐4HB)) for tissue engineering were prepared by a modified particle leaching method. The effect of the addition of P(3HB‐co‐4HB) on microstructural morphology, compression property, swelling behavior, and enzymatic degradation of hybrid scaffolds was systematically investigated. The results indicated that this method was simple but efficient to prepare highly interconnected biomimetic 3D hybrid scaffolds (PP50/50 and PP33/67) with fibrous pore walls. The cytocompatibility of hybrid scaffolds was evaluated by in vitro culture of mesenchymal stem cells. The cell‐cultured hybrid scaffolds presented a complete 3D porous structure, thus allowing cell proliferation on the surface and infiltration into the inner part of scaffolds. The obtained hybrid scaffolds with pore size ranging from 200 to 450 µm, over 90% porosity, adjustable biodegradability, and water‐uptake capability will be promising for cartilage tissue engineering applications. POLYM. ENG. SCI., 54:2902–2910, 2014. © 2014 Society of Plastics Engineers     

Fabrication of bimodal porous PLGA scaffolds by supercritical CO2 foaming/particle leaching technique

Specific pore structure is a vital essential for scaffolds applied in tissue engineering. In this article, poly(lactide‐co‐glycolide) (PLGA) scaffolds with a bimodal pore structure including macropores and micropores to facilitate nutrient transfer and cell adhesion were fabricated by combining supercritical CO₂ (scCO₂) foaming with particle leaching technique. Three kinds of NaCl particles with different scales (i.e., 100–250, <75, <10 μm) were used as porogens, respectively. In particular, heterogeneous nucleation occurred to modify scCO₂ foaming/particle leaching process when NaCl submicron particles (<10 μm) were used as porogens. The observation of PLGA scaffolds gave a formation of micropores (pore size <10 μm) in the cellular walls of macropores (pore size around 100–300 μm) to present a bimodal pore structure. With different mass fractions of NaCl introduced, the porosity of PLGA scaffolds ranged from 68.4 ± 1.4 to 88.7 ± 0.4% for three NaCl porogens. The results of SEM, EDS, and in vitro cytotoxicity test of PLGA scaffolds showed that they had uniform structures and were compatible for cell proliferation with no toxicity. This novel scCO₂ foaming/particle leaching method was promising in tissue engineering due to its ability to fabricate scaffolds with precise pore structure and high porosity. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43644.     

热致相分离法制备明胶多孔支架及其性能研究

本文对热致相分离法制备明胶多孔支架及其性能进行了系统的研究。采用冷冻致孔真空干燥法制备了明胶基组织工程多孔支架,并对影响其结构与性能的因素(如明胶溶液浓度、交联剂用量、体系pH值等)进行了考察。实验结果表明:真空冷冻干燥法制备的明胶基多孔组织工程支架都具有三维孔洞结构;所制备的支架平均孔径可达100μm。     

Fabrication and characterization of porous β-tricalcium phosphate scaffolds coated with alginate

All porous materials have a common limitation which is lack of strength due to the porosity. In this study, two different methods have been used to produce porous β-tricalcium phosphate (β-TCP) scaffolds: liquid-nitrogen freeze casting and a combination of the direct-foaming and sacrificial-template methods. Among these two methods, porous β-TCP scaffolds with acceptable pore size and compressive strength and defined pore-channel interconnectivity were successfully fabricated by the combined direct-foaming and sacrificial-template method. The average pore size of the scaffolds was in the range of 100–150 µm and the porosity was around 70%. Coating with 4 wt% alginate on porous β-TCP scaffolds led to higher compressive strength and low porosity. In order to make a chemical link between the β-TCP scaffolds and the alginate coating, silane coupling agent was used. Treated β-TCP scaffold showed improvements in compressive strength of up to 38% compared to the pure β-TCP scaffold and 11% compared to coated β-TCP scaffold.     

Effects of formaldehyde solution and nanoparticles on mechanical properties and biodegradation of gelatin/nano β-TCP scaffolds

In this study, gelatin/beta tricalcium phosphate (β-TCP) nanocomposite scaffolds were prepared by solvent casting method. The cross-linking method was carried out by adding formaldehyde to gelatin. The microparticles of sodium chloride were used as porogen agent. Characterization of nano β-TCP was performed using XRD, FTIR, and SEM. Results showed that the size of the particles is about 100 nm with spherical morphology. In addition, the scaffold characterization was carried out using FTIR and SEM techniques. Observations showed a porous texture with pore size between 100 and 400 μm. The biodegradability and bioactivity evaluations of the scaffolds were done by immersing them in a simulated body fluid solution for different time periods. The biodegradability studies demonstrated a reduction in the degradation rate of gelatin/β-TCP nanocomposite scaffolds due to the presence of β-TCP nanoparticles. The obtained results of bioactivity tests confirmed the formation of apatite layer on the surface of the scaffolds. Furthermore, the effects of porosity, cross-linking agent, and β-TCP nanoparticles on the bending and compressive properties of the composite scaffolds were examined. According to the mechanical examinations of the scaffolds, the best bending and compressive properties occurred in the presence of 10 and 20 wt% of β-TCP nanoparticles, respectively. The appropriate mechanical properties and biodegradation rate for tissue engineering applications obtained at 1 g of the formaldehyde solution.     

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.     

Chitosan/gelatin porous bone scaffolds made by crosslinking treatment and freeze‐drying technology: Effects of crosslinking durations on the porous structure,compressive strength,and in vitro cytotoxicity

In this study, freezing was used to separate a solute (polymer) and solvent (deionized water). The polymer in the ice crystals was then crosslinked with solvents, and this diminished the linear pores to form a porous structure. Gelatin and chitosan were blended and frozen, after which crosslinking agents were added, and the whole was frozen again and then freeze‐dried to form chitosan/gelatin porous bone scaffolds. Stereomicroscopy, scanning electron microscopy, compressive strength testing, porosity testing, in vitro biocompatibility, and cytotoxicity were used to evaluate the properties of the bone scaffolds. The test results show that both crosslinking agents, glutaraldehyde (GA) and 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide, were able to form a porous structure. In addition, the compressive strength increased as a result of the increased crosslinking time. However, the porosity and cell viability were not correlated with the crosslinking times. The optimal porous and interconnected pore structure occurred when the bone scaffolds were crosslinked with GA for 20 min. It was proven that crosslinking the frozen polymers successfully resulted in a division of the linear pores, and this resulted in interconnected multiple pores and a compressively strong structure. The 48‐h cytotoxicity did not affect the cell viability. This study successfully produced chitosan/gelatin porous materials for biomaterials application. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41851.     

Bioactive glass based scaffolds incorporating gelatin/manganese doped mesoporous bioactive glass nanoparticle coating

We investigated the bioactivity and cytocompatibility of 45S5 bioactive glass (BG) based scaffolds coated with a composite layer formed by gelatin and manganese doped mesoporous bioactive glass nanoparticles (Mn-MBGNs). The scaffolds were prepared using the foam replica method, and they were further coated with Mn-MBGNs/gelatin via dip coating. The synthesized scaffolds were characterized in relation to morphology, porosity, mechanical stability, bioactivity and cell biology behavior using osteoblast-like (MG-63) cells. The scaffolds were highly porous with interconnected porosity, and a suitable pore structure was maintained even after the Mn-MBGNs/gelatin coating. Energy-dispersive X-ray spectroscopy (EDX) confirmed the presence of Mn-MBGNs in the coatings. Moreover, the presence of gelatin was confirmed by Fourier transform infrared spectroscopy (FTIR). The coated scaffolds exhibited in-vitro bioactivity in simulated body fluid comparable to that of uncoated BG scaffolds. Finally, Mn-MBGNs/gelatin coated scaffolds were shown to be non-cytotoxic to MG-63 cells. Hence, the results presented here confirm that the novel Mn containing scaffolds can be considered in the field of biologically active ion releasing scaffolds for bone tissue engineering applications.     

Injection‐molded porous hydroxyapatite/polyamide‐66 scaffold for bone repair and investigations on the experimental conditions

Hydroxyapatite/polyamide‐66 (HA/PA66) composite scaffolds were prepared using injection‐molding technique and also analyzed by means of scanning electron microscopy, X‐ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and mechanical testing. Compared with common methods to fabricate scaffolds, this process can fabricate composite scaffolds in a rapid and convenient manner by adjusting the experimental conditions of foaming agent and shot size. The interactions between PA66 and HA particles affect the crystallization behavior of PA66 and the pore structure of scaffolds. HA particles can increase the stiffness of composite scaffolds accompanied by the reduction of impact strength, pore size and porosity. The obtained 40 wt% HA/PA66 composite scaffolds with a pore size ranging from 100–500 μm and a porosity more than 65% can simultaneously meet the requirements of porous structure and mechanical performance. POLYM. ENG. SCI., 54:1003–1012, 2014. © 2013 Society of Plastics Engineers     

Electrospun polylactide/silk fibroin–gelatin composite tubular scaffolds for small‐diameter tissue engineering blood vessels

Many synthetic scaffolds have been used as vascular substitutes for clinical use. However, many of these scaffolds may not show suitable properties when they are exposed to physiologic vascular environments, and they may fail eventually because of some unexpected conditions. Electrospinning technology offers the potential for controlling the composition, structure, and mechanical properties of scaffolds. In this study, a tubular scaffold (inner diameter = 4.5 mm) composed of a polylactide (PLA) fiber outside layer and a silk fibroin (SF)–gelatin fiber inner layer (PLA/SF–gelatin) was fabricated by electrospinning. The morphological, biomechanical, and biological properties of the composite scaffold were examined. The PLA/SF–gelatin composite tubular scaffold possessed a porous structure; the porosity of the scaffold reached 82 ± 2%. The composite scaffold achieved the appropriate breaking strength (1.28 ± 0.21 MPa) and adequate pliability (elasticity up to 41.11 ± 2.17% strain) and possessed a fine suture retention strength (1.07 ± 0.07 N). The burst pressure of the composite scaffold was 111.4 ± 2.6 kPa, which was much higher than the native vessels. A mitochondrial metabolic assay and scanning electron microscopy observations indicated that both 3T3 mouse fibroblasts and human umbilical vein endothelial cells grew and proliferated well on the composite scaffold in vitro after they were cultured for some days. The PLA/SF–gelatin composite tubular scaffolds presented appropriate characteristics to be considered as candidate scaffolds for blood vessel tissue engineering. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

仿生型壳聚糖-明胶/溶胶凝胶生物玻璃复合支架的制备及其矿化性能研究

利用冷冻干燥法制备出用于骨和软骨组织工程的壳聚糖-明胶/溶胶凝胶生物玻璃(CS-Gel/SGBG)仿生型复合多孔支架,并进行了孔隙率的测定和显微形貌的观察;探讨了各组分不同用量对CS-Gel/SGBG复合支架显微结构的影响以及复合支架在模拟生理体液中的仿生矿化性能。研究表明,通过调节各组分的不同用量,可以制备出三维连通的复合多孔支架,且孔隙率达到90%以上;在模拟生理体液中浸泡后发现CS-Gel/SGBG支架表面有大量结晶态类骨碳酸羟基磷灰石生成,表明复合支架有良好的生物矿化性能。     

Fabrication of polylactic acid/polyethylene glycol (PLA/PEG) porous scaffold by supercritical CO2 foaming and particle leaching

PLA/PEG/NaCl blends were melt‐blended followed by gas foaming and particle leaching process to fabricate porous scaffold with high porosity and interconnectivity. A home‐made triple‐screw compounding extruder was used to intensify the mixability and dispersion of NaCl and PEG in the PLA matrix. Supercritical carbon dioxide was used as physical blowing agent for the microcellular foaming process. Sodium chloride (NaCl) was used as the porogen to further improve the porosity of PLA scaffold. This study investigated the effects of PEG and NaCl on the structure and properties of the PLA‐based blend, as well as the porosity, pore size, interconnectivity, and hydrophilicity of porous scaffolds. It was found that the incorporation of PEG and NaCl significantly improved the crystallization rate and reduced viscoelasticity of PLA. Moreover, scaffolds obtained from PLA/PEG/NaCl blends had an interconnected bimodal porous structure with the open‐pore content about 86% and the highest porosity of 80%. And the presence of PEG in PLA/NaCl composite improved the extraction ability of NaCl particles during leaching process, which resulted in a well‐interconnected structure. The biocompatibility of the porous scaffolds fabricated was verified by culturing fibroblast cells for 10 days. POLYM. ENG. SCI., 55:1339–1348, 2015. © 2015 Society of Plastics Engineers     

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.