Biomimetic Mineralization of Protein Nanogels for Enzyme Protection

Protein nanogels have found a wide variety of applications, ranging from biocatalysis to drug/protein delivery. However, in practical applications, proteins in nanogels may suffer from enzymic hydrolysis and denaturation. Inspired by the structure and functionalities of the fowl eggshells, biomimetic mineralization of protein nanogels was studied in this research. Protein nanogels with embedded porcine pancreas lipase (PPL) in the cross-linked nanostructures were synthesized through the thiol–disulfide reaction between thiol-functionalized PPL and poly(N-isopropylacrylamide) with pendant pyridyl disulfide groups. The nanogels were further reacted with reduced bovine serum albumin (BSA) and BSA molecules were coated on the nanogels. Mineralization of BSA leads to the synthesis of biomineralized shells on the nanogels. With the growth of CaCO₃ on the shells, the nanogels aggregate into suprastructures. Thermogravimetric analysis, XRD, dynamic light scattering, and TEM were employed to study the mechanism of the biomineralization process and analyze the structures of the mineralized nanogels. The biomineralized shells can effectively protect the PPL molecules from hydrolysis by trypsin; meanwhile, the nanosized channels on the mineralized shells allow the transport of small-molecule substrates across the shells. Bioactivity measurements indicate that PPL in the nanogels maintains more than 80 % bioactivity after biomineralization.     

Protein‐Assisted Synthesis of Double‐Shelled CaCO3 Microcapsules and Their Mineralization with Heavy Metal Ions

Calcium carbonate (CaCO₃) is one of the most abundant and important biominerals in nature. Due to its biocompatibility, biodegradability and nontoxicity, CaCO₃ has been investigated extensively in recent years for various fundamental properties and technological applications. Inspired by basic wall structures of cells, we report a protein‐assisted approach to synthesize CaCO₃ into a double‐shelled structural configuration. Due to varying reactivities of outer and inner shells, the CaCO₃ microcapsules exhibit different sorption capacities and various resultant structures toward different kinds of heavy metal ions, analogical to biologically controlled mineralization (BCM) processes. Surprisingly, three mineralization modes resembling those found in BCM were found with these bacterium‐like “CaCO₃ cells”. Our investigation of the cytotoxicity (MTT assay protocol) also indicates that the CaCO₃ microcapsules have almost no cytotoxicity against HepG2 cells, and they might be useful for future application of detoxifying heavy metal ions after further study.     

单分子膜诱导下的矿物晶体生长

介绍了近几年单分子膜诱导下矿物晶体生长的最新进展。讨论了单分子膜诱导下的矿物晶体生长与溶溶中的异同,成膜材料,单分子膜性质和聚集态,矿物晶体的选择性及晶体的表征方法。     

An Effective Osteogenesis Porous CaP/Collagen Interface Compatible with Various Substrates Fabricated by Controlled Mineralization in a Delicately Adjustable Organic Matrix

Increasing bone formation on the surfaces of implants such as screws, plates, or shims holds great significance for clinical medicine. However, osteogenesis implant coatings that mimic natural bone in terms of both their components and structural features are still lacking. Here we report the biomimetic interface of calcium phosphate (CaP) in a collagen matrix fabricated by controlled mineralization that presents biomimetic porous features. The porous CaP/collagen interface, with a thickness of about 1 μm, significantly enhances osteogenesis, as verified at both the gene and protein levels as well as by in vivo experiments. Taking advantage of the generality of the method, the biomimetic interface was prepared on a variety of substrates, including conductive substrates, 3D metal meshes, plastic or elastic substrates, and even on filter papers. The adjustability and generality of the method have enabled new characterization tests to be developed during experiments on cells and thus should greatly facilitate clinical medicine and tissue engineering.     

Pigmentation Chemistry and Radical‐Based Collagen Degradation in Alkaptonuria and Osteoarthritic Cartilage

Alkaptonuria (AKU) is a rare disease characterized by high levels of homogentisic acid (HGA); patients suffer from tissue ochronosis: dark brown pigmentation, especially of joint cartilage, leading to severe early osteoarthropathy. No molecular mechanism links elevated HGA to ochronosis; the pigment's chemical identity is still not known, nor how it induces joint cartilage degradation. Here we give key insight on HGA‐derived pigment composition and collagen disruption in AKU cartilage. Synthetic pigment and pigmented human cartilage tissue both showed hydroquinone‐resembling NMR signals. EPR spectroscopy showed that the synthetic pigment contains radicals. Moreover, we observed intrastrand disruption of collagen triple helix in pigmented AKU human cartilage, and in cartilage from patients with osteoarthritis. We propose that collagen degradation can occur via transient glycyl radicals, the formation of which is enhanced in AKU due to the redox environment generated by pigmentation.     

胶原矿化与仿生修复

脊椎动物硬组织（牙和骨）是通过生命系统的矿化过程形成的,其中矿化胶原是这些生物材料的基本结构单元。矿化胶原是由胶原分子与纳米磷酸钙矿物形成的有机-无机复合材料,其所特有的纳米有序多级结构赋予了生物硬组织材料优异的机械性能（如硬度和韧性）。该结构特性和矿化过程可为新型硬组织修复材料制备提供有益的启示。其中,胶原纤维内有序矿化是仿生重构的难点,也是开展硬组织修复的关键。本文综述了骨的分层结构特征、胶原分子的组装和矿化胶原的多级结构特点,胶原分子和非胶原蛋白与磷酸钙材料的相互作用,功能调控分子对胶原和矿物的界面修饰,以及胶原矿化技术在硬组织修复中的应用;指出了目前胶原矿化亟须解决的一些关键问题,如调控无定形矿物进入胶原纤维、胶原矿化速度和程度,大规模有序胶原纤维制备等。     

生物矿物及其矿化过程

生物体内有机基质指导矿物晶体的成核、生长和聚集,使得生物矿物具有特定的形貌、取向和组装方式,从而产生特殊的功能.本文从有机基质与矿物的晶格匹配、立体化学互补和空间定位、静电作用和电荷匹配,以及分子间弱相互作用力等方面综述了生物矿化过程中所涉及的机理,并讨论了生物矿物的分布和特性、生物矿化过程中的有机基质、生物矿化的过程和类型.     

Amino Acid Assisted Incorporation of Dye Molecules within Calcite Crystals

Biomineralisation processes invariably occur in the presence of multiple organic additives, which act in combination to give exceptional control over structures and properties. However, few synthetic studies have investigated the cooperative effects of soluble additives. This work addresses this challenge and focuses on the combined effects of amino acids and coloured dye molecules. The experiments demonstrate that strongly coloured calcite crystals only form in the presence of Brilliant Blue R (BBR) and four of the seventeen soluble amino acids, as compared with almost colourless crystals using the dye alone. The active amino acids are identified as those which themselves effectively occlude in calcite, suggesting a mechanism where they can act as chaperones for individual molecules or even aggregates of dyes molecules. These results provide new insight into crystal–additive interactions and suggest a novel strategy for generating materials with target properties.     

In vitro study of magnesium-calcite biomineralization in the skeletal materials of the seastar Pisaster giganteus

The mechanisms of formation of biogenic magnesium-rich calcite remain an enigma. Here we present ultrastructural and compositional details of ossicles from the seastar Pisaster giganteus (Echinodermata, Asteroidea). Powder X-ray diffraction, infrared spectroscopy and elemental analyses confirm that the ossicles are composed of magnesium-rich calcite, whilst also containing about 0.01 % (w/w) of soluble organic matrix (SOM) as an intracrystalline component. Amino acid analysis and N-terminal sequencing revealed that this mixture of intracrystalline macromolecules consists predominantly of glycine-rich polypeptides. In vitro calcium carbonate precipitation experiments indicate that the SOM accelerates the conversion of amorphous calcium carbonate (ACC) into its final crystalline product. From this observation and from the discovery of ACC in other closely related taxa, it is suggested that substitution of magnesium into the calcite lattice through a transient precursor phase may be a universal phenomenon prevalent across the phylum echinodermata.     

Mollusk shell formation: a source of new concepts for understanding biomineralization processes

The biological approach to forming crystals is proving to be most surprising. Mollusks build their shells by using a hydrophobic silk gel, very acidic aspartic acid rich proteins, and apparently also an amorphous precursor phase from which the crystals form. All this takes place in a highly structured chitinous framework. Here we present ideas on how these disparate components work together to produce the highly structured pearly nacreous layer of the mollusk shell.     

Morphogenetic control of calcite crystal growth in sulfonic acid based hydrogels

In this paper the mineralization of CaCO(3) in various hydrogel matrices is presented. Sulfonic acid based hydrogels were prepared by introduction of sulfonate-containing monomers into a polyacrylamide network. The sulfonate content of polyacrylamide-co-vinylsulfonate and polyacrylamide-co-allylsulfonate decreases during elution of the copolymers in demineralized water, indicating insufficient linking of the sulfonate-bearing monomers within the hydrogel. In contrast to this, acrylamidomethylpropanesulfonate (AMPS) effectively copolymerizes with acrylamide (AAm) monomers. To study the influence of spatial arrangement of ionic functional groups within hydrogel networks on the mineralization of CaCO(3), AMPS copolymers with different degrees of AMPS cross-linking were synthesized. For the mineralization experiments the copolymers were placed into a double-diffusion arrangement. Calcite as the thermodynamically stable modification of CaCO(3) was obtained with a particular morphology. The pseudocubic habitus resembles aggregates obtained by mineralization in pure polyacrylamide. However, closer examination of the aggregates by scanning electron microscopy (SEM) shows that the crystal growth in the AMPS copolymers is different from that observed in polyacrylamide. Whereas the morphology of the calcite aggregates could be fine-tuned by using copolymers with different sulfonate content, the spatial distribution of the ionic functional groups alters the course of crystallization. Calcium ions are locally accumulated due to the heterogeneous distribution of functional sulfonate groups within the copolymer network. Thereby the nucleation of calcite is triggered, resulting in enhanced mineralization.     

单分子膜诱导生物矿物晶体生长中的晶格匹配和电荷匹配

有机基质与无机晶体的晶格几何匹配和静电相互作用是导致生物体内矿物有序生长并具有特殊理化性质的重要因素,但有机基质的作用机理至今没有完全弄清.作为模拟生物矿化的重要模板之一,Langmuir单分子膜具有独特的优势.本文综述了单分子膜诱导下生物矿物碳酸钙(文石、方解石和球霰石)、羟磷灰石、硫酸钡和纤铁矿等生长过程中的晶格匹配和电荷匹配,讨论了单分子膜亲水头基、膜的电荷性质、膜聚集态等因素对膜控晶体生长过程中晶格匹配和电荷匹配的影响,指出了该领域所面临的问题和将来的发展方向.