Biomimetic bone scaffolds based on chitosan and calcium phosphates

This paper reports a novel biomimetic technique for obtaining chitosan-calcium phosphate composite scaffolds (Cs-CP) by calcium phosphate precipitation from its precursors,CaCl₂ and NaH₂PO₄ in the presence of ammonia and Cs, followed by product freeze-drying. The structural and morphological analysis showed the arrangement of CP onto Cs fibers, while Cs acting as glue to bind together the CP crystals. The presence of different forms of CP was evidenced, including hydroxyapatite, which is known to be involved in the formation of the new bone tissue in living organisms. Great mechanical properties of the scaffolds indicate their potential use in bone tissue engineering.     

Properties of scaffolds based on chitosan and collagen with bioglass 45S5

Scaffolds based on chitosan (CTS), collagen (Coll) and glycosaminoglycans (GAG) mixtures cross‐linked by tannic acid (TA) with bioglass 45S5 addition were obtained with the use of the freeze‐drying method. The prepared scaffolds were characterised for morphology, mechanical strength and degradation rate. Moreover, cell viability on the obtained scaffolds was measured with and without the presence of ascorbic acid and dexamethasone. The main purpose of the research was to compare the effectiveness of bioglass 45S5 influence on the physicochemical and biological properties of scaffolds. The results demonstrated that the scaffolds based on the blends of biopolymers cross‐linked by TA are stable in an aqueous environment. Scanning electron microscope images allowed the observation of a porous scaffold structure with interconnected pores. The addition of bioglass nanoparticles improved the mechanical properties and decreased the degradation rate of composite materials. The biological properties were improved for 20% tannic acid addition compared to 5%. However, the addition of bioglass 45S5 did not change to cells response significantly.Inspec keywords: biomedical materials, drying, porous materials, freezing, tissue engineering, proteins, nanofabrication, bone, scanning electron microscopy, polymers, molecular biophysics, cellular biophysics, nanoparticles, porosityOther keywords: chitosan, collagen, glycosaminoglycans, bioglass 45S5 addition, freeze‐drying method, degradation rate, ascorbic acid, dexamethasone, physicochemical properties, biological properties, porous scaffold structure, bioglass nanoparticles, mechanical properties, tannic acid addition, scanning electron microscopy     

Titanate nanotube coatings on biodegradable photopolymer scaffolds

Rigid, biodegradable photopolymer scaffolds were coated with titanate nanotubes (TNTs) by using a spin-coating method. TNTs were synthesized by a hydrothermal process at 150 °C under 4.7 bar ambient pressure. The biodegradable photopolymer scaffolds were produced by mask-assisted excimer laser photocuring at 308 nm. For scaffold coating, a stable ethanolic TNT sol was prepared by a simple colloid chemical route without the use of any binding compounds or additives. Scanning electron microscopy along with elemental analysis revealed that the scaffolds were homogenously coated by TNTs. The developed TNT coating can further improve the surface geometry of fabricated scaffolds, and therefore it can further increase the cell adhesion.     

Dextrine-modified chitosan marine polymer coatings

In trying to modify partially acetylated chitosan (CS) marine polymers ground from crab or shrimp shells for use as environmentally benign water-base coatings for aluminum (Al) substrates, CS was dissolved in a solution of HCL acid, and then mixed with corn starch-derived dextrine (DEX) containing Ce nitrate as oxidizing agent in aqueous medium. This blend of polysaccharides was deposited on Al surfaces by a simple dip-withdrawing method, and then heated at 200°C to transform the liquid layer into a solid film. The solution solid phase transition provided the changes in the molecular conformation of CS and DEX ; the former was transformed into deacetylated poly(D-glucosamine) and the latter referred to the formation of Ce-complexed carboxylate fragments. Furthermore, the chemical reactions between the NH₂ groups in deacetylated CS and the carboxylate fragments led to the creation of amide linkages that served in grafting DEX fragments onto the CS. Such fragment-grafted CS polymer coating films deduced from the proper proportions of CS to DEX offered great film-forming performance, low susceptibility to moisture, and low ionic conductivity, conferring a salt-spray resistance of 720 hours.     

Alginate-crosslinked chitosan scaffolds as pentoxifylline delivery carriers

To prevent fibrous encapsulation of implants, measures are taken to suppress inflammatory reactions around them. Sustained anti-inflammatory drug release from the scaffolds can potentially be a way to reduce inflammation around these implants. Alginate-crosslinked chitosan is often used to make biocompatible tissue engineered scaffolds. However, there is a lack of quantitative studies on the drug delivery properties of alginate-crosslinked chitosan scaffolds. For this study, chitosan, crosslinked with different concentrations of alginate, was made into porous scaffolds. Infrared and thermal gravimetric analyses showed polyelectrolyte complex formation between chitosan and alginate units. The alginate-crosslinked chitosan scaffolds were more hydrophilic, showed less swelling, had lower pentoxifylline (PTX) release efficacies, were more favorable for initial cell attachment, and were mechanically stronger and more resistant to enzymatic degradation when compared to non-crosslinked chitosan scaffolds. The differences became more significant as the concentrations of chitosan and alginate increased. Furthermore, in vitro tests showed that when PTX was slowly released from the scaffolds, it became more effective in suppressing the production of TNF-α and IL-6 by stimulated macrophage cells.     

Characterization of electrophoretic chitosan coatings on stainless steel

Electrophoretic chitosan deposits on stainless steel AISI 316 L were produced and characterized. The coating quality (thickness, defectiveness, corrosion protection ability) was seen to depend on the electric field used for EPD. Corrosion studies in concentrated simulated body fluid (SBF5) demonstrated that the surface characteristics of AISI 316 L can be positively influenced by the chitosan coating.     

Cell growth and function on calcium phosphate reinforced chitosan scaffolds

Macroporous chitosan scaffolds reinforced by calcium phosphate powders such as hydroxyapatite (HA) or calcium phosphate invert glass were fabricated using a thermally induced phase separation technique. Human osteoblast-like MG63 cells were cultured on the composite scaffolds for up to 11 days, and the cell growth and function were analyzed. The cell growth is much faster on the chitosan/HA scaffolds incorporated with the glass (CHG) than on the chitosan/HA scaffold without the glass (CH). The total protein content of cells were quantified and increased over time on both composites (CH, CHG) but was significantly higher on CHG after 7 days of culture. The cells on CHG also expressed significantly higher amount of alkaline phosphatase at days 7 and 11 and osteocalcin at day 7 than those on CH. The results suggested that the addition of glass in chitosan/hydroxyapatite composite scaffolds might enhance the proliferation and osteoblastic phenotype expression of MG63 cells. However, the chitosan-matrix scaffolds did not show higher phenotype expression of MG63 cells, in comparison with the TCPS plate, probably due to the degradation of chitosan and release of acidic byproducts. Larger amount of soluble calcium phosphate invert glasses should be added into the scaffolds to prevent chitosan from fast degradation that may affect the differentiation of osteoblast cells.     

Development of polycaprolactone/chitosan blend porous scaffolds

Polycaprolactone (PCL) and chitosan were blended to fabricate porous scaffolds for tissue-engineering applications by employing a concentrated acetic acid solution as solvent and salt particles as porogen. These scaffolds showed well-controlled and interconnected porous structures. The pore size and porosity of the scaffolds could be effectively modulated by selecting appropriate amounts and sizes of porogen. The results obtained from compressive mechanical measurements indicated that PCL/chitosan could basically retain their strength in their dry state compared to individual components. In a hydrated state, their compressive stress and modulus could be still well maintained even though the weight ratio of chitosan reached around 50 wt%.     

壳聚糖支架在组织工程中的应用

综述了壳聚糖作为细胞生长载体在软骨组织工程，骨组织工程和皮肤组织工程等方面的应用进展，表明壳聚糖有望成为优异的组织工程支架材料。     

Effect of collagen II coating on mesenchymal stem cell adhesion on chitosan and on reacetylated chitosan fibrous scaffolds

The biocompatibility and biomimetic properties of chitosan make it attractive for tissue engineering but its use is limited by its cell adhesion properties. Our objectives were to produce and characterize chitosan and reacetylated-chitosan fibrous scaffolds coated with type II collagen and to evaluate the effect of these chemical modifications on mesenchymal stem cell (MSC) adhesion. Chitosan and reacetylated-chitosan scaffolds obtained by a wet spinning method were coated with type II collagen. Scaffolds were characterized prior to seeding with MSCs. The constructs were analyzed for cell binding kinetics, numbers, distribution and viability. Cell attachment and distribution were improved on chitosan coated with type II collagen. MSCs adhered less to reacetylated-chitosan and collagen coating did not improve MSCs attachment on those scaffolds. These findings are promising and encourage the evaluation of the differentiation of MSCs in collagen-coated chitosan scaffolds. However, the decreased cell adhesion on reacetylated chitosan scaffold seems difficult to overcome and will limit its use for tissue engineering.     

Thermal-crosslinked porous chitosan scaffolds for soft tissue engineering applications

The aim of this study was to demonstrate the feasibility of using a steam autoclave process for sterilization and simultaneously thermal-crosslinking of lyophilized chitosan scaffolds. This process is of great interest in biomaterial development due to its simplicity and low toxicity. The steam autoclave process had no significant effect on the average pore diameter (~ 70 μm) and overall porosity (> 80%) of the resultant chitosan scaffolds, while the sterilized scaffolds possessed more homogenous pore size distribution. The sterilized chitosan scaffolds exhibited an enhanced compressive modulus (109.8 kPa) and comparable equilibrium swelling ratio (23.3). The resultant chitosan scaffolds could be used directly for in vitro cell culture without extra sterilization. The data of in vitro studies demonstrated that the scaffolds facilitated cell attachment and proliferation, indicating great potential for soft tissue engineering applications.     

木垛型壳聚糖多孔支架的磷酸化改性和仿生矿化

为提高壳聚糖支架材料的孔隙率及矿化程度,通过磷酸化表面改性和仿生矿化制备了磷酸化(PCSW)和生物矿化(BMCW)木垛型壳聚糖多孔支架.FTIR结果显示,壳聚糖分子中有磷酸根的引入.XRD结果表明,矿化24 h后支架上形成结晶度较高的磷酸钙盐晶体,矿化48 h后结晶度明显增加并形成单纯的羟基磷灰石(HA)结晶.SEM观察发现,在支架的内外表面均致密地沉积了HA晶体层.压缩强度测试结果表明,复合支架BMCW矿化48 h的压缩强度为(0.54±0.005) MPa,压缩模量为(5.47±0.65) MPa,BMCW可用作非承重骨组织修复材料.     

Preparation and cytocompatibility of silk fibroin/chitosan scaffolds

One challenge in soft tissue engineering is to find an applicable scaffold, not only having suitable mechanical properties, porous structures, and biodegradable properties, but also being abundant in active groups and having good biocompatibility. In this study, a three-dimensional silk fibroin/chitosan (SFCS) scaffold was successfully prepared with interconnected porous structure, excellent hydrophilicity, and proper mechanical properties. Compared with polylactic glycolic acid (PLGA) scaffold, the SFCS scaffold further facilitated the growth of HepG2 cells (human hepatoma cell line). Keeping the good cytocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold should be a prominent candidate for soft tissue engineering, for example, liver.     

木垛型壳聚糖多孔支架的磷酸化改性和仿生矿化

为提高壳聚糖支架材料的孔隙率及矿化程度, 通过磷酸化表面改性和仿生矿化制备了磷酸化(PCSW)和生物矿化(BMCW)木垛型壳聚糖多孔支架。FTIR结果显示, 壳聚糖分子中有磷酸根的引入。XRD结果表明, 矿化24 h后支架上形成结晶度较高的磷酸钙盐晶体, 矿化48 h后结晶度明显增加并形成单纯的羟基磷灰石(HA)结晶。SEM观察发现, 在支架的内外表面均致密地沉积了HA晶体层。压缩强度测试结果表明, 复合支架BMCW矿化48 h的压缩强度为(0.54±0.005) MPa, 压缩模量为(5.47±0.65) MPa, BMCW可用作非承重骨组织修复材料。     

Preparation and cytocompatibility of silk fibroin / chitosan scaffolds

One challenge in soft tissue engineering is to find an applicable scaffold, not only having suitable mechanical properties, porous structures, and biodegradable properties, but also being abundant in active groups and having good biocompatibility. In this study, a three-dimensional silk fibroin/chitosan (SFCS) scaffold was successfully prepared with interconnected porous structure, excellent hydrophilicity, and proper mechanical properties. Compared with polylactic glycolic acid (PLGA) scaffold, the SFCS scaffold further facilitated the growth of HepG2 cells (human hepatoma cell line). Keeping the good cytocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold should be a prominent candidate for soft tissue engineering, for example, liver.     

半乳糖修饰大孔壳聚糖支架的制备及表征

生物人工肝反应器中肝细胞的数量和功能是能否有效替代已衰竭肝脏功能的关键,与此紧密相关的是细胞支架材料结构和性能的优化.采用冷冻干燥法,制备了大孔壳聚糖支架,孔隙率在90%以上,平均孔径在100～200μm之间.以肝细胞表面去唾液酸糖蛋白受体(asialoglycoprotein receptor,ASGPR)的特异性配体-半乳糖,对材料表面进行糖基化修饰,制备了半乳糖基修饰的大孔壳聚糖支架,肝细胞在其上生长状况良好,细胞培养密度高,为高密度培养肝细胞提供了一种性能优良的支架材料.     

Electrode coatings for high temperature hydrogen electrolysis

A discussion of fabrication techniques and performance testing of solid oxide components for use in hydrogen steam electrolysis is presented. Novel plasma spray techniques are utilized to deposit the thin ceramic oxide electrode, electrolyte, and interconnect layers on a planar intermetallic bipolar plate. Optimal porosity is achieved within the electrode microstructure through mixed feed techniques that are a combination of dry powder feed and liquid solution injection. The perovskite anode coatings formed from liquid precursor feedstock require post-deposition annealing in an oxygen-rich atmosphere to form the desired perovskite structures. Electrical conductivity measurements were measured for all electrodes and interconnect materials as a function of temperature to 1000 °C. Supported by the U.S. Department of Energy, Office of Nuclear Energy Science and Technology, under the DOE Idaho Operations Contract No. DE-AC07-05ID14517     

Electrochemistry assisted reacting deposition of hydroxyapatite in porous chitosan scaffolds

Electrochemistry assisted reacting deposition method was employed to prepare porous chitosan/hydroxyapatite (CS/HA) composite scaffold with a new design device using ion exchange membranes to separate calcium salt and phosphate solutions. The results determined from XRD and SEM indicates hydroxyapatite can be electrochemically deposited in the chitosan scaffold using the device. After electrochemistry assisted reacting deposition, the surface of the chitosan scaffold was coated with low crystalline HA, particularly at the frame edge of the scaffold. The pores in the scaffold still kept interconnected well and the deposited hydroxyapatite has a cluster microsphere shape whose size is about 3-5 μm.     

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。     

Preparation,characterization and cytocompatibility of bioactive coatings on porous calcium-silicate-hydrate scaffolds

The major goal of this research was to investigate and characterize the deposition of a biomimetic apatite-like coating onto the surface of 3D porous calcium-silicate-hydrate scaffolds with suitable bioactivity for potential application in bone tissue engineering. Basically, Portland cement, water, sand and lime were mixed for preparing the slurry which was poured into molds, and fine aluminum powder was added as foaming agent resulting on the formation of porous 3D structures. After aging for 28 days, these porous inorganic scaffolds were immersed in calcium chloride supersaturated solution in PBS for 7 days at 37 °C for the biomimetic layer deposition. Scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier Transformed Infrared Spectroscopy (FTIR) techniques were used in order to characterize the porous scaffolds and the apatite-like biomimetic coating. The results have showed that 3D constructs were successfully produced with interconnected porosity, compressive strength and cytocompatibility appropriate for potential use as an alternative in trabecular bone repair.