Fabrication and characterization of novel hydroxyapatite/porous carbon composite scaffolds

Novel hydroxyapatite (HA)/porous carbon composite scaffolds were prepared by applying sonoelectrodeposition and a subsequent hydrothermal treatment to previous carbonized phenolic resin coated polyurethane sponges. The interconnected pore network and morphology of HA/porous carbon composite scaffolds were determined by scanning electron microscope (SEM), and the whole surface of porous carbons were evenly coated with the deposited HA layer which was confirmed by EDS and XRD. The porosity (83.5 ± 0.3%) and the bulk density (0.297 ± 0.009 g·cm⁻³) of HA/porous carbon scaffolds were detected by the Archimedes method. The compressive and flexural strength of the scaffolds is 1.187 ± 0.064 MPa and 0.607 ± 0.268 MPa, respectively. Compared with the polymeric surface of 24-well cell culture plates, these novel scaffolds significantly promote the proliferation of human osteoblast-like MG-63 cells, indicating that this novel HA/porous carbon composite scaffold could be used for in vitro 3D culture of osteoblasts.     

Improving the strength and biocompatibility of porous titanium scaffolds by creating elongated pores coated with a bioactive, nanoporous TiO2 layer

This paper reports a novel way of improving the mechanical properties and biocompatibility of porous Ti scaffolds using a combination of the modified sponge replication method and anodization process. The use of a stretched polymeric sponge as a novel template allowed the creation of elongated pores in a porous Ti scaffold, which, accordingly, led to a high compressive strength of 24.2 ± 2.08 MPa at a porosity of approximately 70 vol%. Furthermore, the surfaces of the Ti walls were coated successfully with a bioactive nanoporous TiO₂ layer using the anodization process, which enhanced the biocompatibility remarkably, as assessed by the attachment of MC3T3-E1 cells.     

壳聚糖/羟基磷灰石表面修饰聚己内酯多孔骨支架的制备及性能

采用选择性激光烧结技术构建多孔聚己内酯(PCL)骨支架,用原位合成的方法制得壳聚糖/羟基磷灰石(CS/HA)悬浮液,并采用真空浸泡、低速离心和冷冻凝胶的方法使CS/HA黏附在PCL支架的表面,以改善骨支架的生物相容性和细胞增殖活性。通过X射线衍射(XRD)和扫描电子显微镜(SEM)观测复合支架的物相和形貌,测量支架的压缩强度和杨氏模量,测量支架表面的水接触角,并通过体外细胞实验研究复合支架的生物学性能。实验结果表明,原位合成的方法制得了羟基磷灰石(HA);CS/HA凝胶与PCL骨支架表面黏附良好;CS/HA改善了PCL支架表面的亲水性,提升了骨支架的生物相容性和细胞增殖活性。     

Highly porous polycaprolactone-45S5 Bioglass® scaffolds for bone tissue engineering

Highly porous biocompatible composites made of polycaprolactone (PCL) and 45S5 Bioglass® (BG) were prepared by a solid–liquid phase separation method (SLPS). The composites were obtained with BG weight contents varying in the range 0–50%, using either dimethylcarbonate (DMC) or dioxane (DIOX) as solvent, and ethanol as extracting medium. The porosity of the scaffolds was estimated to be about 88–92%. Mechanical properties showed a dependence on the amount of BG in the composites, but also on the kind of solvent used for preparation, composites prepared with DIOX showing enhanced stress at deformation with respect to composites prepared with DMC (stress at 60% of deformation being as high as 214 ± 17 kPa for DIOX-prepared composites and 98 ± 24 kPa for DMC-prepared ones, with 50 wt/wt_PCL% of glass), as well as higher elastic modulus (whose value was 251 ± 32 kPa for DIOX-prepared scaffolds and 156 ± 36 kPa for DMC-prepared ones, always with 50 wt/wt_PCL% of glass). The ability of the composites to induce precipitation of hydroxyapatite was positively evaluated by means of immersion in simulated body fluid and the best results were achieved with high glass amounts (50 wt/wt_PCL%). In vitro tests of cytotoxicity and osteoblast proliferation showed that, even if the scaffolds are to be considered non-cytotoxic, cells suffer from the scarce wettability of the composites.     

Quality Monitoring of Porous Zein Scaffolds: A Novel Biomaterial

Our previous studies have shown that zein has good biocompatibility and good mechanical properties. The first product from a porous scaffold of zein, a resorbable bone substitute, has passed the biological evaluation of medical devices (ISO 10993) by the China Food and Drug Administration. However, Class III medical devices need quality monitoring before being placed on the market, and such monitoring includes quality control of raw materials, choice of sterilization method, and evaluation of biocompatibility. In this paper, we investigated four sources of zein through amino acid analysis (AAA) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in order to monitor the composition and purity, and control the quality of raw materials. We studied the effect of three kinds of sterilization method on a porous zein scaffold by SDS-PAGE. We also compared the changes in SDS-PAGE patterns when irradiated with different doses of gamma radiation. We found that polymerization or breakage did not occur on peptide chains of zein during gamma-ray (γ-ray) sterilization in the range of 20–30 kGy, which suggested that γ-ray sterilization is suitable for porous zein scaffolds. Regarding cell compatibility, we found a difference between using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and a cell-counting kit-8 (CCK-8) assay to assess cell proliferation on zein film, and concluded that the CCK-8 assay is more suitable, due to its low background optical density.     

Novel method to produce β-TCP scaffolds

The production of ceramic scaffolds by a novel method was reported in this work. The method comprised the mixture of colloidal silica and β-tricalcium phosphate (β-TCP) powder, where paraffin microspheres were further added to provide the porosity in the scaffold after firing. Thermo-gravimetric analysis showed that the paraffin was completely degraded before 550 °C, where the heat treatment at 1100 °C was enough to remove the paraffin and provide porosity and mechanical strength. The scaffold had an open porosity, but with poor pore interconnection. The density of the scaffold was 1.11 ± 0.03 g/cm³ with a porosity of 64.98 ± 1.1%, and the compression strength was 5.02 ± 1.2 MPa.     

Nanostructured titanium-45S5 Bioglass scaffold composites for medical applications

Titanium–10 wt.% 45S5 Bioglass scaffold nanocomposites were synthesized by the combination of mechanical alloying and by a “space-holder” sintering process. The porous structure and corrosion properties were investigated. In vitro biocompatibility of these materials was evaluated and compared with a conventional microcrystalline titanium, where normal human osteoblast (NHOst) cells from Cambrex (CC-2538) were cultured on the disks of the materials and cell growth was examined. The morphology of the cell cultures obtained on Ti–10 wt.% 45S5 Bioglass nanocomposite was similar to those obtained on the microcrystalline titanium. On the other hand, on porous scaffold, the cells adhered with their whole surface to the insert penetrating the porous structure, while on the polished surface, more spherical cells were observed with a smaller surface of adhesion. The present study has demonstrated that titanium–10 wt.% 45S5 Bioglass scaffold nanocomposite is a promising biomaterial for bone tissue engineering.     

Physicochemical properties and enhanced cellullar responses of biocompatible polymeric scaffolds treated with atmospheric pressure plasma using O2 gas

Biocompatible polymeric scaffolds were fabricated by mixing 5 wt.% poly(ε-caprolactone) (P) with 4 wt.% gelatin (G) and 1.6 wt.% Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (D). These PGD scaffolds were also treated with atmospheric pressure (AP) plasma using O₂ reactive gas (to create O-PGD scaffolds). The physicochemical and mechanical properties of the PGD scaffolds were characterized by in vitro biodegradability tests, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, contact angle measurements, and tensile strength measurements. The wettability and hydrophilic properties of the scaffold surface were improved remarkably by adding G and D to P, and by subsequent oxygen-assisted AP plasma treatment. An MTT assay, a cell attachment efficiency assay, scanning electron microscopy, and confocal microscopy revealed that Chinese Hamster Ovary (CHO)-K1 cells exhibited higher cell attachment and viability on the PGD and O-PGD scaffolds than on the P and PG scaffolds. Furthermore, the long-term viability of the CHO cells on the PGD and O-PGD scaffolds without exchanging the cell culture media was significantly improved compared to their viability on the P and PG scaffolds. Overall, the PGD and O-PGD scaffolds are expected to be useful as cell growth supporting biomaterials in tissue engineering.     

CuInSe2 monograin growth in the liquid phase of potassium iodide

The results of monograin CuInSe₂ synthesis from Cu-In alloy and Se in liquid KI are presented. The amounts of CuInSe₂ and KI were nearly equal to fulfil the criterion for the monograin growth (all free volume between the particles has to be filled with liquid). All the grown powder materials with narrow-disperse granularity were chalcopyrite CuInSe₂. The grown crystallites had tetrahedral shapes and homogeneous composition. Particle size distribution was used to describe the growth process. The activation energy of linear growth of crystals was E_d = 0.25 ± 0.05 eV, and the power of time dependence of the crystal growth was l/n = 0.26 ± 0.06. The solubility of CuInSe₂ in KI at 990 K was 0.17 ± 0.05 wt. %. The solubility of potassium and iodine in CuInSe₂ at 990 K was 0.094 wt. %, and 0.0086 wt. %, respectively. As a result, homogeneous p-type CuInSe₂ monograin materials were synthesised in KI solvent.     

Highly porous titanium (Ti) scaffolds with bioactive microporous hydroxyapatite/TiO2 hybrid coating layer

Highly porous Ti scaffolds with a bioactive microporous hydroxyapatite (HA)/TiO₂ hybrid coating layer were fabricated using the sponge replication process and micro-arc oxidation (MAO) treatment to produce the porous Ti scaffold and hybrid coating layer, respectively. In particular, the morphology and chemical composition of the hybrid coating layer were controlled by carrying out the MAO treatment in electrolyte solutions containing various concentrations of HA, ranging from 0 to 30 wt.%. The fabricated sample showed high porosity of approximately 70 vol.% with interconnected pores and reasonably high compressive strength of 18 ± 0.3 MPa. Furthermore, the surfaces could be coated successfully with a bioactive microporous HA/TiO₂ hybrid layer. The amount of HA particles in the hybrid coating layer increased with increasing HA content in the electrolyte solution, while preserving the microporous morphology. This hybrid coating improved the osteoblastic activity of the porous Ti scaffolds significantly.     

Biological response to pre-mineralized starch based scaffolds for bone tissue engineering

It is known that calcium-phosphate (Ca-P) coatings are able not only to improve the bone bonding behaviour of polymeric materials, but at the same time play a positive role on enhancing cell adhesion and inducing the differentiation of osteoprogenitor cells. Recently an innovative biomimetic methodology, in which a sodium silicate gel was used as a nucleative agent, was proposed as an alternative to the currently available biomimetic coating methodologies. This methodology is especially adequate for coating biodegradable porous scaffolds. In the present work we evaluated the influence of the referred to treatment on the mechanical properties of 50/50 (wt%) blend of corn starch/ethylene-vinyl alcohol (SEVA-C) based scaffolds. These Ca-P coated scaffolds presented a compressive modulus of 224.6 ± 20.6 and a compressive strength of 24.2 ± 2.20. Cytotoxicity evaluation was performed according ISO/EN 10993 part 5 guidelines and showed that the biomimetic treatment did not have any deleterious effect on L929 cells and did not inhibit cell growth. Direct contact assays were done by using a cell line of human osteoblast like cells (SaOS-2). 3 × 10⁵ cells were seeded per scaffold and allowed to grow for two weeks at 37C in a humidified atmosphere containing 5% CO₂. Total protein quantification and scanning electron microscopy (SEM) observation showed that cells were able to grow in the pre-mineralized scaffolds. Furthermore cell viability assays (MTS test) also show that cells remain viable after two weeks in culture. Finally, protein expression studies showed that after two weeks osteopontin and collagen type I were being expressed by SaOS-2 cells seeded on the pre-mineralized scaffolds. Moreover, alkaline phosphatase (ALP) activity was higher in the supernatants collected from the pre-mineralized samples, when compared to the control samples (non Ca-P coated). This may indicate that a faster mineralization of the ECM produced on the pre-mineralized samples was occurring. Consequently, biomimetic pre-mineralization of starch based scaffolds can be a useful route for applying these materials on bone tissue engineering.     

Characterisation of blends between poly(ε-caprolactone) and polysaccharides for tissue engineering applications

In this work, bioartificial binary blends between poly(ε-caprolactone) (PCL) and a polysaccharide (chitosan (CS) or starch (S)) with different contents of the natural polymer (5–30 wt.%) were produced. Melt-mixing and double-precipitation were the methods used for the obtainment of PCL/S and PCL/CS blends, respectively. Tubular scaffolds were produced from bioartificial blends by melt-extrusion. Physico-chemical characterisation was performed by differential scanning calorimetry analysis (DSC), thermogravimetry (TGA), scanning electron microscopy (SEM), infrared analysis (FTIR-ATR and micro-ATR mapping), atomic force microscopy (AFM) and stress–strain tests. Blends were not miscible, phase-separated systems, showing a homogeneous composition and morphology only at low polysaccharide content (≤ 10 wt.%). The biocompatibility of bioartificial guides was investigated by culturing NIH-3T3 mouse fibroblasts. Cells response showed the following order: PCL/S > PCL > PCL/CS. For each blend type, biocompatibility increased with decreasing the polysaccharide content. In vitro cell tests using S5Y5 neuroblastoma cells, carried out on the most biocompatible blends, assessed their absence of cytotoxicity towards these model cells of the nervous tissue. Results showed that blends with a low chitosan or starch content (≤ 10 wt.%) are promising for the regeneration of tissues requiring tubular scaffolds, such as the peripheral nerves.     

Construction of collagen II/hyaluronate/chondroitin-6-sulfate tri-copolymer scaffold for nucleus pulposus tissue engineering and preliminary analysis of its physico-chemical properties and biocompatibility

To construct a novel scaffold for nucleus pulposus (NP) tissue engineering, The porous type II collagen (CII)/hyaluronate (HyA)–chondroitin-6-sulfate (6-CS) scaffold was prepared using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) cross-linking system. The physico-chemical properties and biocompatibility of CII/HyA–CS scaffolds were evaluated. The results suggested CII/HyA–CS scaffolds have a highly porous structure (porosity: 94.8 ± 1.5%), high water-binding capacity (79.2 ± 2.8%) and significantly improved mechanical stability by EDC/NHS crosslinking (denaturation temperature: 74.6 ± 1.8 and 58.1 ± 2.6°C, respectively, for the crosslinked scaffolds and the non-crosslinked; collagenase degradation rate: 39.5 ± 3.4 and 63.5 ± 2.0%, respectively, for the crosslinked scaffolds and the non-crosslinked). The CII/HyA–CS scaffolds also showed satisfactory cytocompatibility and histocompatibility as well as low immunogenicity. These results indicate CII/HyA–CS scaffolds may be an alternative material for NP tissue engineering due to the similarity of its composition and physico-chemical properties to those of the extracellular matrices (ECM) of native NP.     

Carbon nanotube-coated silicone as a flexible and electrically conductive biomedical material

Artificial cell scaffolds that support cell adhesion, growth, and organization need to be fabricated for various purposes. Recently, there have been increasing reports of cell patterning using electrical fields. We fabricated scaffolds consisting of silicone sheets coated with single-walled (SW) or multi-walled (MW) carbon nanotubes (CNTs) and evaluated their electrical properties and biocompatibility. We also performed cell alignment with dielectrophoresis using CNT-coated sheets as electrodes. Silicone coated with 10 μg/cm² SWCNTs exhibited the least sheet resistance (0.8 kΩ/sq); its conductivity was maintained even after 100 stretching cycles. CNT coating also improved cell adhesion and proliferation. When an electric field was applied to the cell suspension introduced on the CNT-coated scaffold, the cells became aligned in a pearl-chain pattern. These results indicate that CNT coating not only provides electro-conductivity but also promotes cell adhesion to the silicone scaffold; cells seeded on the scaffold can be organized using electricity. These findings demonstrate that CNT-coated silicone can be useful as a biocompatible scaffold.     

Preparation and characterization of bimodal porous poly(γ-benzyl-L-glutamate) scaffolds for bone tissue engineering

An ideal scaffold in bone tissue-engineering strategy should provide biomimetic extracellular matrix-like architecture and biological properties. Poly(γ-benzyl-L-glutamate) (PBLG) has been a popular model polypeptide for various potential biomedical applications due to its good biocompatibility and biodegradability. This study developed novel bimodal porous PBLG polypeptide scaffolds via a combination of biotemplating method and in situ ring-opening polymerization of γ-benzyl-L-gIutamate N-carboxyanhydride (BLG-NCA). The PBLG scaffolds were characterized by proton nuclear magnetic resonance spectroscopy, X-ray diffraction, differential scanning calorimetry, scanning electron microscope (SEM) and mechanical test. The results showed that the semi-crystalline PBLG scaffolds exhibited an anisotropic porous structure composed of honeycomb-like channels (100–200 μm in diameter) and micropores (5–20 μm), with a very high porosity of 97.4 ± 1.6%. The compressive modulus and glass transition temperature were 402.8 ± 20.6 kPa and 20.2 °C, respectively. The in vitro biocompatibility evaluation with MC3T3-E1 cells using SEM, fluorescent staining and MTT assay revealed that the PBLG scaffolds had good biocompatibility and favored cell attachment, spread and proliferation. Therefore, the bimodal porous polypeptide scaffolds are promising for bone tissue engineering.     

In vitro biocompatibility study of calcium phosphate glass ceramic scaffolds with different trace element doping

Porous calcium phosphate based glass ceramics (CaO-P₂O₅-Na₂O) containing different trace elements (2.0 mol% Mg, Sr and Zn respectively) were prepared by coating polyurethane foams with sol-gel derived glass slurry. After heat treatment at suitable temperatures, main phase catena hexaphosphate (Ca₄P₆O₁₉) and minor phase calcium pyrophosphate (β-Ca₂P₂O₇) crystallized from the glass matrix. These scaffolds were soaked in simulated body fluid (SBF) to determine the solubility and apatite formation, and mouse MC3T3-E1 cells were used to investigate the bioactivity and biocompatibility. The Sr doped scaffold showed a higher degradability than those samples containing Zn or Mg, inducing the formation of an apatite layer with a high (Sr + Ca)/P molar ratio of 1.64, whereas only some discontinuous CaP layers and spare apatite agglomerates were found on the scaffolds doped with Mg ((Mg + Ca)/P = 1.12) and Zn ((Zn + Ca)/P = 1.55) respectively. In vitro cell culture, a high degree of cell adhesion and spreading was achieved on the samples containing Sr or Zn, while only a few cells adhered to the Mg doped sample. These results implied that the bioactivity and biocompatibility of the scaffolds were not only strongly associated with the apatite forming ability, but also related with the Ca/P molar ratios of the deposits.     

A novel composite membrane of chitosan-carboxymethyl cellulose polyelectrolyte complex membrane filled with nano-hydroxyapatite I. Preparation and properties

A novel tri-component composite membranes of chitosan/carboxymethyl cellulose (CS/CMC) polyelectrolyte complex membranes filled with different weight ratios of nano-hydroxyapatite (n-HA)(0, 20, 40 and 60 wt%), namely, n-HA/CS/CMC composite membrane, were prepared by self-assembly of static electricity. The structure and the properties of the composite membranes were investigated by Fourier transformed infrared spectroscopy(IR), X-ray diffraction(XRD), Scanning electron microscopy(SEM), mechanical performance measurement, swelling behavior test, and soaking behavior study in phosphate buffered saline (PBS) and simulate body fluid (SBF). The results showed that the n-HA/CS/CMC composite membrane was formed though superficial static electricity interaction among n-HA, CS and CMC. For the n-HA/CS/CMC composite membrane, the microstructure compatibility, mechanical property, swelling behavior, the degradation and bioactivity in vitro of the composite membrane were improved by the addition of n-HA, compared with CS/CMC polyelectrolyte complex membrane. Moreover, the n-HA/CS/CMC composite membrane with 40 wt% n-HA had the most highest mechanical property, which suggested that the novel n-HA/CS/CMC composite membrane with 40 wt% n-HA was more suitable to be used as guided bone tissue regeneration membrane than CS/CMC polyelectrolyte complex membrane.     

Radiation synthesis of gelatin/CM-chitosan/β-tricalcium phosphate composite scaffold for bone tissue engineering

A series of biodegradable composite scaffolds was fabricated from an aqueous solution of gelatin, carboxymethyl chitosan (CM-chitosan) and β-tricalcium phosphate (β-TCP) by radiation-induced crosslinking at ambient temperature. Ultrasonic treatment on the polymer solutions significantly influenced the distribution of β-TCP particles. An ultrasonic time of 20 min, followed by 30 kGy irradiation induced a crosslinked scaffold with homogeneous distribution of β-TCP particles, interconnected porous structure, sound swelling capacity and mechanical strength. Fourier Transform Infrared Spectroscopy and X-ray Diffraction analysis indicated that β-TCP successfully incorporated with the network of gelatin and CM-chitosan. In vivo implantation of the scaffold into the mandible of beagle dog revealed that the scaffolds had excellent biocompatibility and the presence of β-TCP can accelerate bone regeneration. The comprehensive results of this study paved way for the application of gelatin/CM-chitosan/β-TCP composite scaffolds as candidate of bone tissue engineering material.     

原位水化法制备羟基磷灰石/壳聚糖复合支架材料

以含Ca2+和PO34-的溶液为无机相,壳聚糖(chitosan,CS)溶液为高分子相,采用原位水化法制备羟基磷灰石(hydroxyapatite,HAP)/CS复合多孔支架材料。XRD和IR的表征和分析表明水化24h后,复合支架中的钙磷盐从磷酸氢钙(dicalciumphos phate dehydrate,DCPD)转化为HAP。SEM和EDS显示15μm左右的棒状HAP颗粒均匀地分散在多孔支架的孔壁上,压缩强度的测试结果表明这种结构显著提高复合支架的力学性能。     

Fabrication of highly porous titanium (Ti) scaffolds with two interlaced periodic pores

This paper reports a novel type of porous titanium (Ti) scaffolds with two interlaced periodic pores that were produced by coating the surfaces of a dual-channeled hydroxyapatite (HA) scaffold, as a supporting framework, with a titanium hydride (TiH₂) slurry followed by heat-treatment at 1200 °C for 3 h in a vacuum to convert TiH₂ to Ti metal. This method allowed the porous Ti scaffolds to mimic the original pore structure of the dual-channeled HA scaffold in a tightly controlled manner. It was observed that the Ti layer was strongly adhered to the HA layer, owing to the diffusion of P ions into the Ti layer. The fabricated sample showed a high compressive strength of 6.0 ± 0.77 MPa and a porosity of 78 vol.% due to its unique pore structure, as well as perfect interconnections between the pores.