Sustained release of dexamethasone from drug‐loading PLGA scaffolds with specific pore structure fabricated by supercritical CO2 foaming

Inducing differentiation of bone marrow stem cells to generate new bone tissue is highly desirable by controlling the release of some osteoinductive or osteoconductive factors from porous scaffolds. In this study, dexamethasone was selected as a representative of small molecule drugs and dexamethasone‐loading porous poly(lactide‐co‐glycolide) (PLGA) scaffolds were successfully fabricated by supercritical CO₂ foaming. Scanning electron microscopy images showed that scaffolds had rough and relatively interconnected pores facilitating cells adhesion and growth. Specially, dexamethasone which was incorporated into PLGA matrix in a molecularly dispersed state could serve as a nucleation agent to be helpful for the formation of interconnected pores. Dexamethasone‐loading porous PLGA scaffolds exhibited sustained release profile, and the delivery of dexamethasone from porous scaffolds could last for up to 2 months. The cumulative released amount of dexamethasone was relevant with drug loading capacity (1.66%–2.95%) and pore structure of scaffolds; while the release behavior was anomalous (non‐Fickian) transport by fitting with the simple exponential equation, which had a diffusional exponent n higher than 0.5. It is feasible to fabricate drug‐loading porous scaffolds by supercritical CO₂ foaming with specific pore structure and sustained release profile, which can be well applied in bone tissue engineering. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46207.     

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.     

可注射nHA/PU复合多孔骨修复支架制备及表征

兼具良好孔隙率和原位任意塑形固化的可注射复合多孔骨修复材料在临床不规则骨缺损的治疗方面显示出巨大的优势。本研究通过优化双组分设计,以水为发泡剂制备可注射纳米羟基磷灰石/聚氨酯（nHA/PU）复合多孔支架。利用扫描电子显微镜（SEM）、傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）、力学测试及Gillmore针测试等手段对制备的支架进行结构形貌、化学组成、力学性能和固化时间表征。结果表明,本研究制备的可注射nHA/PU复合多孔支架孔隙率高、孔隙贯通性好,孔径分布在100~700μm,适宜细胞和组织向孔内生长;添加20% nHA显著提高了PU支架的力学强度,但降低了支架的孔隙率;可注射支架在8h固化,适宜临床操作。本研究制备的可注射nHA/PU复合多孔支架在不规则骨缺损修复领域具有较大的应用潜力。     

3D gel-printing of hierarchically porous BCP scaffolds for bone tissue engineering

In this study, a conventional technique of porous preparation was used to improve the constructive capability of direct ink writing on microstructures, and the hierarchically porous scaffolds were successfully prepared by 3D gel printing (3DGP). Micron-sized hydroxyapatite (HA) was coated with tricalcium phosphate (TCP) nanopowders synthesized by chemical co-precipitation to form biphasic calcium phosphate (BCP). The random structure of concave micropores was achieved by filling the BCP slurry with PMMA microspheres while successfully controlling the internal porosity of printed filaments. The results showed that the three-stage porous structure was successfully constructed, i.e., macroscopic pores of 1.50–2.00 mm, spherical micropores of 100–200 µm, and inter-powder interstices of 1.00–10.00 µm. Nano-TCP coated micron-HA powders improved the sintering activity of BCP particles. The compressive strength and porosity of the scaffolds sintered at 1400 °C were 2.78 MPa and 84.98%. The hierarchically porous BCP scaffolds had bright applications in bone tissue engineering.     

Formation of porous PLGA scaffolds by a combining method of thermally induced phase separation and porogen leaching

A novel method for the fabrication of porous poly(L -lactide-co-glycolide) (PLGA) scaffolds by combining thermally induced phase separation and porogen leaching is presented in this article. Big pores with about 75–400 μm diameters in the obtained scaffolds were generated by the porogen, sucrose particles, while small pores with diameters less than 20 μm induced via phase separation. Extraction of the solvent, chloroform by ethanol at cool temperatures could reduce the scaffold toxicity. Effects of PLGA concentration, freezing temperature, volume fraction of porogen, and introduction of β-tricalcium phosphate (β-TCP) on morphology, porosity, and compressive properties of the scaffolds were systematically discussed. Results showed that the size of small pores decreased by decreasing the polymer concentration and reducing the freezing temperature, whereas the interconnectivity of the scaffolds was improved by increasing the porogen fraction. The compressive modulus and strength were significantly lowered by increasing the scaffold porosity, that is, by increasing porogen fraction, or decreasing the polymer concentration, or reducing the freezing temperature. Addition of β-TCP into the scaffolds did not influence the compressive modulus significantly but tended to decrease the compressive strength. The obtained scaffolds with diverse pore sizes would be potentially used in bone tissue engineering. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Subtractive manufacturing of customized hydroxyapatite scaffolds for bone regeneration

An approach to the preparation of customized hydroxyapatite scaffolds for bone regeneration based on subtractive manufacturing has been developed. The developed method is intended as a simple alternative to rapid prototyping based on additive manufacturing methods. The method for the preparation of customized scaffolds consists in computer numerical controlled (CNC) milling of porous hydroxyapatite foam. The machinable foam for customized scaffolds was prepared by direct foaming of a colloidal hydroxyapatite suspension and the foam structure was consolidated by the gelcasting method. The optimal foam after sintering reached a high porosity of 83.3% and the cellular-like structure contained spherical pores with an average diameter of 613 µm, which were connected through windows with an average diameter of 161 µm. The compressive strength of the sintered foam reached an average value of 2 MPa. CNC milling was tested at different stages of the production cycle of the porous foam and after the reinforcing of the foam by impregnation. The best results were obtained by milling the foam presintered at 1100 °C. The milling process was demonstrated and scaffold examples were successfully manufactured.     

Highly interconnected macroporous MBG/PLGA scaffolds with enhanced mechanical and biological properties via green foaming strategy

In this study, mesoporous bioactive glass particles (MBGs) are incorporated into poly(lactic-co-glycolic acid) (PLGA) to fabricate highly interconnected macroporous composite scaffolds with enhanced mechanical and biological properties via a developed supercritical carbon dioxide (scCO₂) foaming method. Scaffolds show favorable highly interconnected and macroporous structure through a high foaming pressure and long venting time foaming strategy. Specifically, scaffolds with porosity from 73% to 85%, pore size from 120 μm to 320 μm and interconnectivity of over 95% are controllably fabricated at MBG content from 0 wt% to 20 wt%. In comparison with neat PLGA scaffolds, composite scaffolds perform improved strength (up to 1.5 folds) and Young's modulus (up to 3 folds). The interconnected macroporous structure is beneficial to the ingrowth of cells. More importantly, composite scaffolds also provide a more promising microenvironment for cellular proliferation and adhesion with the release of bioactive ions. Hopefully, MBG/PLGA scaffolds developed by the green foaming strategy in this work show promising morphological, mechanical and biological features for tissue regeneration.     

骨组织工程用PLGA多孔支架的制备及细胞毒性研究

制备能在骨组织工程研究中应用,并具有良好孔隙结构的块状聚（D,L-乳酸-CO-乙醇酸）（PLGA）多孔支架,探索出以冰粒子作为致孔剂,采用粒子滤出方法结合冷冻干燥工艺制备多孔支架的方法.首先将冰颗粒加入预冻的PLGA氯仿溶液中混合均匀,然后把混合物置于液氮中深度冷冻后冷冻干燥,制得多孔支架.对支架孔隙结构分析表明,该工艺制备的多孔支架无致孔剂残留、三维结构良好、孔径与孔隙可通过改变冰粒子的粒径和质量分数来控制;细胞毒性实验表明该多孔支架毒性在0～1级,可作为骨组织工程研究用多孔支架.     

3D porous HA/TCP composite scaffolds for bone tissue engineering

Calcium phosphates (apatites) are considered as a research frontier for bone regeneration applications by virtue of similarity to the mineral constituent of bone, suitable biocompatibility and remarkable osteogenesis ability. In this regard, the biodegradability and mechanical properties of monophasic apatites, typically hydroxyapatite (HA) and tricalcium phosphate (TCP), are imperfect and do not fulfill some requirements. To overcome these drawbacks, 3D porous HA/TCP composite scaffolds prepared by conventional and more recently, 3D printing techniques have shown to be promising since their bioperformance is adjustable by the HA/TCP ratio and pores. Despite the publication of several reviews on either 3D porous scaffolds or biphasic calcium phosphates (BCPs), no review paper has to our knowledge focused on 3D porous BCP scaffolds. This paper comprehensively reviews the production methods, properties, applications and modification approaches of 3D porous HA/TCP composite scaffolds for the first time. In addition, new insights are introduced towards developing HA/TCP scaffolds with more impressive bioperformance for further tissue engineering applications, including those with different interior and exterior frameworks, patient-specific specifications and drugs (or other biological factors) loading.     

Foaming of polymers with supercritical CO2: An experimental and theoretical study

Microcellular polystyrene (PS) foams and porous structures of the biodegradable poly(d,l-lactic acid) (P_d,lLA) were prepared with the batch foaming technique (pressure quench) using supercritical CO₂ as blowing agent. The effect of pressure, temperature and depressurization rate on the final porous structure was investigated. The results revealed that the size of the pores decreases and their population density increases with pressure increase, or decrease of temperature, and/or increase of the depressurization rate. The results were correlated by combining nucleation theory with NRHB model in order to account for and emphasize the physical mechanism related to nucleation of bubbles inside the supersaturated polymer matrix. A satisfactory agreement between correlations and experimental data was obtained indicating that the nucleation theory yields quantitative correlations when variables such as sorption, degree of plasticization, and surface tension of the system polymer-supercritical fluid are accurately described.     

The use of poly (ε‐caprolactone) to enhance the mechanical strength of porous Si‐substituted carbonate apatite

Poly(ε‐caprolactone) (PCL)/silicon‐substituted carbonate apatite (Si‐CO3Ap) composite derived from the interconnected porous Si‐CO3Ap reinforced with molten PCL was prepared. PCL was used to improve the mechanical properties of a porous apatite by a simple polymer infiltration method, in which the molten PCL was deposited through the interconnected channel of porous Si‐CO3Ap. The PCL covered and penetrated into the pores of the Si‐CO3Ap to form an excellent physical interaction with Si‐CO3Ap leading to a significant increase in diametral tensile strength from 0.23 MPa to a maximum of 2.04 MPa. The Si‐CO3Ap/PCL composite has a porosity of about 50–60% and an interconnected porous structure, with pore sizes of 50–150 μm which are necessary for bone tissue formation. These results could pave the way for producing a porous, structured biocomposite which could be used for bone replacement. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

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     

Nano‐Hydroxyapatite Improves the Properties of β‐tricalcium Phosphate Bone Scaffolds

β‐Tricalcium phosphates have been widely used as biomaterials for bone substitutes; however, the poor mechanical properties limit the application in bearing loading bones. In this study, nano‐hydroxyapatite has been introduced to improve the mechanical properties for porous bioceramic scaffolds. Nanocomposites containing 0–10 wt% needle‐like nano‐hydroxyapatite were prepared for investigation. It has been found that needle‐like nano‐hydroxyapatite improves the toughness, hardness, and compressive strength of the porous β‐tricalcium phosphates scaffolds, as well as the microstructure properties. The study provides a reference for the development of safe, excellent bone scaffolds for bone tissue engineering.     

The effect of ethyl-lactate and ethyl-acetate plasticizers on PCL and PCL–HA composites foamed with supercritical CO2

The present study reports foaming of polycaprolactone (PCL) and PCL nano- and micro-composites with dispersed hydroxyapatite (HA) particles by means of binary mixtures of supercritical CO₂ (scCO₂) and either ethyl lactate (EL) or ethyl acetate (EA) as plasticizer. The effect of the size and concentration of HA particles, as well as the effects of the plasticizer type and the incorporation route were investigated aiming to fabricate porous scaffolds with uniform morphology and controlled pore size distribution. For this purpose, foaming experiments were carried out by selecting two operating temperatures, 40 and 45 °C, and two soaking times, 1 and 17 h. Furthermore, a double step of depressurization was used to promote the development of a double-scale pore size structure in porous scaffolds useful for tissue engineering.The results of this study indicated that supercritical foaming of PCL and PCL–HA composites is enhanced when the selected operating temperature and time are 45 °C and 17 h, respectively. Furthermore, although both EL and EA plasticizers enhanced the low temperature foaming of the materials, we demonstrated that the route of incorporation of the plasticizer is a critical aspect for enhancing composite foaming and scaffold fabrication. From this point of view, the best results were achieved when EA was pre-mixed with the polymeric powder for preparing a dough for the foaming process.     

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.     

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     

聚醚砜超临界CO2发泡行为研究

以超临界CO₂为发泡剂,采用间歇发泡法制备了聚醚砜（PES）泡沫。采用旋转流变仪和扫描电子显微镜分析表征了PES发泡的均相成核行为,继而分别以滑石粉和二氧化硅（SiO₂）作为异相成核剂,探究了PES发泡过程中的异相成核行为。结果表明,间歇发泡法制备PES泡沫的发泡区间为200~230 ℃;最佳浸泡压力为20 MPa;最佳浸泡时间为3 h;未改性PES的泡孔直径均可在10 μm以下,泡孔密度在10⁹~10¹⁰ 个/cm³之间,但泡孔壁较厚;SiO₂相对于Talc,表现出更显著的异相成核作用;在210 ℃、3 h、20 MPa的发泡条件下,添加0.9 %（质量分数,下同）的SiO₂,可得到泡孔直径为0.77 μm,泡孔密度为7.14×10¹¹ 个/cm³的PES微孔泡沫。     

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.     

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     

壳聚糖/生物活性玻璃多孔复合支架的制备及界面增强作用

采用卵磷脂作为表面活性剂对溶胶-凝胶生物玻璃进行表面改性,并采用冷冻干燥法制备用于骨和软骨组织工程的壳聚糖/生物活性玻璃复合多孔支架(chitosan/bioglass porous composite scaffolds,CS/BGS),观察CS/BGS的显微形貌并测定抗压强度,探讨生物玻璃的表面改性对CS/BGS显微结构及力学强度的影响。研究表明:采用冷冻干燥法可以制备具有一定强度的三维连通的CS/BGS,且孔隙率达到90%以上。通过对生物玻璃表面改性可以在一定程度上提高CS/BGS的抗压强度。