Formation of nematic ordered cellulose and chitin

We proposed in a previous paper a unique form of β-glucan association, nematic ordered cellulose (NOC) that is molecularly ordered, yet non-crystalline. NOC has unique characteristics; in particular, its surface properties provide with a function of tracks or scaffolds for regulated movements and fiber-production of Acetobacter xylinum [Kondo et al. 2002. Proc. Natl. Acad. Sci. USA 99: 14008–14013]. In order to extend the usage of this NOC film as a functional template, the present article attempts to clarify how β-glucan association is initiated and established by uniaxial stretching of water swollen cellulose gel films. Wide angle X-ray diffraction, high-resolution transmission electron microscopy and atomic force microscopy were employed to exhibit molecular behavior of the ordering at various scales. Then, the preparative method for NOC was applied to the other carbohydrate polymers such as α-chitin and cellulose/α-chitin blends, leading to nematic ordered states as well as cellulose. However, the method did not necessarily provide the typical structure like NOC at the molecular scale. Instead, it yielded a variety of hierarchical nematic ordered states at various scales, which allows development of new artificial ordered sheet structures.     

Molecular orientation in the Nematic Ordered Cellulose film using polarized FTIR accompanied with a vapor-phase deuteration method

Previously, the authors reported “Nematic Ordered Cellulose (NOC)” that is a well-ordered state of β-1,4-glucan chains without exhibiting typical X-ray diffraction patterns of any cellulose polymorphs (Togawa and Kondo 1999; Kondo et al. 2001; Kondo 2007). The NOC was prepared by stretching water-swollen gel-like films at the draw ratio of 2.0 to provide highly oriented β-1,4-glucan molecular chains of cellulose, which was proved by the high resolution TEM observation. In this paper, a detailed study of the unique ordered state of the NOC was attempted to characterize orientation of the main chains as well as the OH groups of molecules using polarized FTIR accompanied with a vapor-phase deuteration method. The dichroic analysis suggested that the main chains were fairly oriented in the stretching direction whereas the OH groups remained unoriented. The disordered state of the OH groups regardless of the oriented state for the main chain may hinder the oriented crystallization during the preparation of NOC films.     

Solvothermal Transformation of a Calcium Oleate Precursor into Large‐Sized Highly Ordered Arrays of Ultralong Hydroxyapatite Microtubes

Hydroxyapatite (HAP), a well‐known member of the calcium phosphate family, is the major inorganic component of bones and teeth in vertebrates. The highly ordered arrays of HAP structures are of great significance for hard tissue repair and for understanding the formation mechanisms of bones and teeth. However, the synthesis of highly ordered HAP structure arrays remains a great challenge. In this work, inspired by the ordered structure of tooth enamel, we have successfully synthesized three‐dimensional bulk materials with large sizes (millimeter scale) that are made of highly ordered arrays of ultralong HAP microtubes (HOAUHMs) by solvothermal transformation of calcium oleate precursor. The core–shell‐structured oblate sphere consists of a core that is composed of HAP nanorods and a shell that consists of highly ordered HAP microtube arrays. The prepared HOAUHMs are large: 6.0 mm in diameter and up to 1.4 mm in thickness. With increasing solvothermal reaction time, the HOAUHMs grow larger; the microtubes become more uniform and more ordered. This work provides a new synthetic method for synthesizing highly ordered arrays of uniform HAP ultralong microtubes that are promising for biomedical applications.     

Quartz Crystal Microbalance Analysis of DNA-Templated Calcium Phosphate Mineralization

A quartz crystal microbalance (QCM) sensor was developed for the quantitation of calcium phosphate mineralization and the assessment of DNA as a template molecule. Inherent advantages of QCM, such as nanogram sensitivity, temporal resolution, surface-based measurements, and flow capabilities, were leveraged in the design of this sensor, and in-line fluidic mixing was used to control precursor reaction. This research shows that DNA, a highly programmable anionic polymer, is able to template and control mineralization of calcium phosphate, with nucleation occurring in less than 15 min and initial rates ranging from 4 to 8 ng/min. FT-IR measurements show mineralized material to be calcium phosphate resembling hydroxyapatite (HAP) when a DNA template is used. DNA is a promising mineralization template, and the QCM proves to be a dynamic technique for a broad range of heterogeneous mineralization experiments in complement to classic, diffusion-limited, end-point analysis techniques.     

Calcium phosphate mineralization controlled by carboxymethyl cellulose-g-polymethacrylic acid

Carboxymethyl cellulose-grafted polymethacrylic acid (CMC-g-PMAA) was synthesized by graft copolymerization process onto carboxymethyl cellulose backbone using methacrylic acid as a monomer and ammonium persulfate as a free radical initiator. CMC-g-PMAA was employed as dispersed template for controlling calcium phosphate mineralization from aqueous solutions at different copolymer contents and pHs. Hybrids with different morphologies and particles diameter were investigated by adjusting of preparation conditions. Synthesized hybrids were characterized by FT-IR, SEM, XRD, and particle size analyzer. Such functionalized hybrids with complex morphologies can be manipulated as a novel reinforcing fillers, ceramic precursors, or biomedical implants.     

仿生矿化与硬组织修复

生物矿化(Biomineralization)是生物硬组织(软体动物的外壳,脊椎动物的骨和牙等)形成的重要环节,是生物体调控矿物沉积,并利用矿物增强硬组织机能的重要生物策略。生物矿化所形成的生物矿物具有多级有序的结构、优异的机械性能和重要的生理功能,启发了有机-无机复合生物材料的设计和仿生矿化制备,为体内外硬组织修复提供研究思路和奠定材料基础。本文主要综述了生物矿化的基本原理和主要生物矿物,矿物结晶原理和新认识,与硬组织修复密切相关的胶原矿化机制和最新进展,硬组织材料的多级结构特征,以及仿生矿化在硬组织修复中的前沿进展。     

The effect of PVP-<Emphasis Type="Italic">b</Emphasis>-PMAA block copolymer on morphologies control of calcium carbonate

A novel double-hydrophilic block copolymer (DHBC) poly(vinyl pyrrolidone)–block–poly(methacrylic acid) (PVP-b-PMAA) was synthesized via reversible addition–fragmentation chain transfer polymerization. The structure of the resulting copolymer was characterized by ¹H nuclear magnetic resonance, and the molecular weight of the block copolymer was determined by gel permeation chromatography. The study of morphological control of calcium carbonate (CaCO₃) has been performed in the presence of the PVP-b-PMAA block copolymer. Various morphologies of CaCO₃ particles such as rhombohedral, multilayered, and aggregated with cavities can be produced by varying the copolymer concentrations. The all-obtained CaCO₃ particles were calcite, which was confirmed by either X-ray diffraction or Fourier transform infrared spectra. Such calcium carbonate/polymer hybrids with complex morphologies may find valuable applications in biomimic mineralization.     

Interaction between nanosized crystalline components of a composite based on <Emphasis Type="Italic">Acetobacter xylinum</Emphasis> cellulose and calcium phosphates

A composite consisting of two nanosized biocompatible components, Acetobacter xylinum cellulose and calcium phosphate, is prepared through aggregation in an aqueous suspension. The structures of initial components and composite are investigated by the methods of X-ray and electron diffraction and electron microscopy. The mineral component consists of two crystalline phases, hydroxyapatite and whitlockite (magnesium-containing tricalcium phosphate), which are nanosized platelike crystals. The composite preserves the crystalline structures of initial calcium phosphates and cellulose. In the course of composite formation, hydroxyapatite and whitlockite crystallites are adsorbed on the surfaces of nanofibrillar cellulose ribbons. Whitlockite nanocrystals are predominantly deposited on the surface of cellulose ribbons. The mutual orientation of the surfaces of crystalline structures of cellulose and two types of calcium phosphates, hydroxyapatite and whitlockite, is analyzed by means of computer simulation, and the variants of mutual arrangement of their surfaces during formation of the interfacial boundary are suggested.     

仿生矿化的机理和应用研究进展

碳酸钙、磷酸钙为代表的生物矿物广泛分布于自然界中,经过不同的矿化过程,在生物体内呈现出多样的结构、形貌和功能,构成生物体多种组织和器官.在人工材料合成领域,仿生矿化通过调控碳酸钙、磷酸钙等矿物的成核与生长,获得具有复杂高级结构和特殊生物功能的无机或无机/有机复合材料.本文重点介绍仿生矿化机理和应用的最近研究进展,包括仿...     

The influence of isothermal aging,surfactant and inorganic precursors concentrations on pore size and structural order of mesoporous bioactive glass

Ordered hexagonal mesoporous bioactive glasses (MBGs) are synthesized by an evaporation-induced self-assembly (EISA) method using tetraethylorthosilicate (TEOS), calcium nitrate tetrahydrate (Ca(NO₃)₂.4H₂O) and triethyl phosphate (TEP) as the silicon, calcium, phosphorous sources, respectively, and pluronic P123 as the structure directing agent. The influences of P123, TEOS, TEP, and Ca(NO₃)₂.4H₂O on the structural order and pore size of MBGs at different environmental conditions are studied. The prepared MBGs are characterized by the small angle X-ray diffraction (SXRD), N₂ adsorption-desorption isotherms, and transmission electron microscopy (TEM). The study revealed that polymerization degree of inorganic precursors should be low enough at the initial assembling stage of inorganic species with organic surfactants to form highly ordered mesoporous structure. Concentration of Ca(NO₃)₂ governs the formation of ordered mesostructure in MBGs by complexation of Ca²⁺ with the hydrophilic group of surfactant P123. Isothermal aging also plays an important role in the formation of highly ordered MBGs as it permits formation of rigid inorganic framework.     

Multistep growth mechanism of calcium phosphate in the earliest stage of morphology-controlled biomineralization

We studied the effect of surface-functional-group position on precipitate morphology in the earliest stage of calcium phosphate biomineralization and determined the detailed mechanism of precipitation starting from nucleation to precipitate growth. The biomineralization template was a β-sheet peptide scaffold prepared by adsorption with carboxyl groups arranged at strict 7 ? intervals. Phosphate was then introduced. Within 10 s, highly ordered embryos of calcium phosphate were formed and confined by a peptide nanofiber pattern. They repeatedly nucleated and dissolved, with the larger embryos absorbing the smaller ones in a clear demonstration of an Ostwald-ripening-like phenomenon, then aggregated in a line pattern, and finally formed highly ordered nanofibers of amorphous calcium phosphate. This multistep growth process constitutes the earliest stage of biomineralization.     

Organic acid-assisted soft-templating synthesis of ordered mesoporous carbons

A series of soft-templated ordered mesoporous carbons (OMCs) was synthesized by using resorcinol and formaldehyde as carbon precursors, triblock copolymer Pluronic F127 as a soft-template, and an organic acid (acetic, benzoic, citric, oxalic, or succinic) as a polymerization reaction catalyst. The aforementioned organic acids were strong enough to facilitate the formation of ordered mesophases by the block copolymer template used and to catalyze the polymerization reaction of resorcinol and formaldehyde in this template. The use of weak organic acids instead of strong inorganic acids such as HCl eliminated inorganic anions from the reaction environment and resulted in high surface area OMCs. Basically, the resulting carbons showed the surface areas and pore volumes comparable to those reported for the carbons prepared under similar conditions but in the presence of strong inorganic acids. Electron microscopy analysis proved the presence of ordered mesopores, whereas thermogravimetric analysis showed a good thermal stability of these carbons.     

Layer-by-layer self-assembled polyelectrolyte multilayers with embedded liposomes: immobilized submicronic reactors for mineralization

The development of chemical reactions in nanospaces is of paramount importance for the development of active nanodevices, particularly in nanofluidics. It has been shown in a previous paper that phospholipid vesicles can be incorporated without spontaneous bilayer rupture into poly-L-glutamic acid/poly(allylamine) (PGA/PAH) multilayered polyelectrolyte films. The aim of the present study was to use such a system as an "embedded submicronic reactor" able to trigger precipitation of calcium phosphates within closed spaces through an enzymatic reaction, the enzyme also being encapsulated in the vesicle interior. To this aim, large unilamellar vesicles (LUVs) were produced containing calcium ions as active ions in the mineralization process, spermine as an activator of crystal growth, and alkaline phosphatase as a catalyst to convert phosphate esters into phosphates. After stabilization by adding a layer of poly-(D-lysine), these vesicles were embedded in a (PGA-PAH)n film. A paranitrophenyl phosphate containing solution was then put in contact with this film. It is shown by means of infrared spectroscopy in the attenuated total reflection mode that, consecutively to this contact, calcium phosphates are growing inside the embedded vesicles. By using scanning near-field fluorescence microscopy, it is demonstrated that the alkaline phosphatase enzymes are most probably located inside the vesicles after their embedding. In addition, atomic force microscopy was used to show, after chemical removal of the organic top layer of the film, that the inorganic platelets produced after the precipitation reaction are localized in volumes of similar size and shape as that of the vesicles into which the phosphate ester hydrolysis and subsequent precipitation reaction did occur.     

Unique structural characteristics of nematic ordered cellulose—Stability in water and its facile transformation

Nematic Ordered Cellulose (NOC) film that exhibits a noncrystalline yet highly ordered form was prepared by stretching a water‐swollen cellulose gel obtained in a unique manner with coagulation of cellulose molecules dissolved in the N,N‐dimethylacetamide/LiCl solvent system. In this article, structural characteristics of this unique film were investigated. Orientation of the molecular chains in the noncrystalline regions across the entire film were stable after immersing in water at room temperature, though conventional amorphous cellulose regions are in any forms believed fairly to be recrystallized under a humid atmosphere. Even 30 days after immersing in water at 50 °C, neither crystallization nor disordering of the chains occurred in the NOC film. On the contrary, the film was capable of being transformed into films composed of cellulose polymorphs domains where the molecular orientation was still maintained as the initial film under various mild conditions that both cotton and cellophane did not show any changes on their structure. These contradictory properties of the NOC film proved to be dependent on its unique supermolecular structure. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 2850–2859, 2007     

Calcium phosphate mineralization with linear poly(ethylene imine): a time-resolved study

We have earlier shown that linear poly(ethylene imine) (LPEI) is an efficient growth modifier for calcium phosphate mineralization from aqueous solution (Shkilnyy et al., Langmuir, 2008, 24 (5), 2102). The current study addresses the growth process and the reason why LPEI is such an effective additive. To that end, the solution pH and the calcium and phosphate concentrations were monitored vs. reaction time using potentiometric, complexometric, and photometric methods. The phase transformations in the precipitates and particle morphogenesis were analyzed by X-ray diffraction and transmission electron microscopy, respectively. All measurements reveal steep decreases of the pH, calcium, and phosphate concentrations along with a rapid precipitation of brushite nanoparticles early on in the reaction. Brushite transforms into hydroxyapatite (HAP) within the first 2 h, which is much faster than what is reported, for example, for calcium phosphate precipitated with poly(acrylic acid). We propose that poly(ethylene imine) acts as a proton acceptor (weak buffer), which accelerates the transformation from brushite to HAP by taking up the protons that are released from the calcium phosphate precipitate during the phase transformation.     

L-半胱氨酸自组装膜诱导草酸钙结晶研究

在CaCl2.H2O和Na2C2O4配制的过饱和溶液中,利用L-半胱氨酸(L-Cys)在金片上形成的自组装膜为模板,研究了草酸钙(CaOxa)在自组装膜上的结晶行为,并探讨了溶液pH对CaOxa晶体组成、晶型及其形貌的影响。采用X射线衍射(XRD)和扫描电子显微镜(SEM)等技术对CaOxa晶体的结构和形貌进行了表征。实验结果表明:当溶液pH=3.0时,溶液中可以形成一水草酸钙(CaC2O4.H2O,COM)和二水草酸钙(CaC2O4.2H2O,COD)晶体,而在同样pH条件下,在L-Cys自组装膜上只形成COD晶体,表明自组装单层对CaOxa晶体的成核和生长有重要影响。通过改变溶液的pH,在自组装单层上可以得到不同晶型和不同形状的CaOxa晶体。当pH=3.0时得到四方块状的COD晶体,而pH=5.0和pH=7.0时分别得到六边形和拉长六边形的COM晶体。     

Calcium Phosphate‐Reinforced Photosensitizer‐Loaded Polymer Nanoparticles for Photodynamic Therapy

Calcium phosphate‐reinforced photosensitizer‐loaded polymer nanoparticles have been developed for photodynamic therapy. Chlorin e6 (Ce6)‐loaded core–shell–corona polymer micelles of poly(ethylene glycol)‐b‐poly(L ‐aspartic acid)‐b‐poly(L ‐phenylalanine) ( PEG-PAsp-PPhe ) were employed as template nanoparticles for mineralization with calcium phosphate (CaP). CaP deposition was performed by the electrostatic localization of calcium ions at the anionic PAsp middle shells and the subsequent addition of phosphate anions. CaP‐reinforced nanoparticles exhibited enhanced stability. The CaP mineral layer effectively inhibited Ce6 release from the Ce6‐loaded mineralized nanoparticles (Ce6‐NP‐CaP) at physiological pH value. At an acidic endosomal pH value of 5.0, Ce6 release was enhanced, owing to rapid dissolution of the CaP minerals. Upon irradiation of Ce6‐NP‐CaP‐treated MCF‐7 breast‐tumor cells, the cell viability dramatically decreased with increasing irradiation time. The phototoxicity of Ce6‐NP‐CaP was much higher than that of free Ce6. Non‐invasive optical‐imaging results indicated that Ce6‐NP‐CaP exhibited enhanced tumor specificity compared with free Ce6 and Ce6‐loaded non‐mineralized polymer nanoparticles (Ce6‐NP).     

Biomimetic gelatin-octacalcium phosphate core-shell microspheres

Calcium phosphate/polymeric microparticles synthesized through a biomimetic approach are regarded with increasing interest for their various potential applications, including tissue engineering and regenerative medicine. Herein we report the synthesis and characterization of gelatin/octacalcium phosphate core/shell microspheres. Deposition of the calcium phosphate shell on the polymeric microspheres was obtained through bio-inspired mineralization on the surface of functionalized gelatin microparticles. Gelatin microspheres stabilized by alginate dialdehyde were prepared using an inverse microemulsion. Functionalization was achieved by enriching the microspheres composition with calcium ions or, alternatively, with alendronate, a bisphosphonate widely employed for the treatment of bone diseases. Functionalization and synthesis of the inorganic phase in the microemulsion environment were key factors for the achievement of a complete coating of the microspheres with calcium phosphate. The inorganic shell is constituted of small crystals of octacalcium phosphate, which control gelatin and alendronate release.     

Divalent cation-induced variations in polyelectrolyte conformation and controlling calcite morphologies: direct observation of the phase transition by atomic force microscopy

In the biomineralization process, the changes in conformation of organic matrix may be a widespread phenomenon. Investigation of the structural relationship between organic and inorganic materials is the main subject. The approach taken was to extract quantitative information of the variations in polyelectrolyte conformation during the mineralization process using atomic force microscopy. The results infer the evidence of the role of polyelectrolyte conformation in mineralization of calcium carbonate and the methods for understanding the principle that govern biomineralization.     

Thermal, UV, FTIR, and XRD studies of urinary stones

Various crystals are seen in human urine. Oxalate, Phosphate, Uric acid, and Urate crystals are generally seen in urinary calculi. Calcium stones are most common, comprising 75 % of all urinary calculi. They may be pure calcium oxalate or calcium phosphate or a mixture of both. Many stones are not homogeneous. Low calcium intake increases the intestinal absorption of calcium, thus decreasing the amount of calcium available in the intestinal tract to form insoluble complexes with Oxalate. Consequently, a higher amount of oxalate is available for intestinal absorption and as a result, urinary oxalate excretion increases. Mineral water consumption did not reduce urinary oxalate excretion. High urinary excretion and concentration of magnesium decrease both the nucleation and growth rates of calcium oxalate crystals in urine, because of the higher solubility of magnesium oxalate compared with calcium oxalate. Analytical results show calcium oxalate to be one of the major inorganic components of renal stones and found to be present in almost all kidney and bladder stones. About 39.5 % of the total composition of the calculi is found to contain purely calcium oxalate and also hydroxyl apatite. The ten samples are a mixture of calcium oxalate and phosphate stones. Four samples are calcium oxalate as major composition and the remaining are calcium phosphate as major composition. These kidney stones are taken photographically and size of the stone are measured using optical microscopy. These qualitative analyses are also confirmed by UV, FTIR, DSC, and XRD analysis.