透明质酸及其衍生物作药物载体

透明质酸(HA)具有良好的生物相容性和生物降解性，是优良的药物载体。但其稳定性差，对强酸、强碱、热、自由基及透明质酸酶敏感，容易发生降解而限制了其应用。本文简要介绍HA的基本特性及应用，重点阐述了HA经不同的化学改性方法如酯化、交联、接枝所得衍生物作药物载体的最新研究进展。化学改性赋予了HA一系列的优良特性,如适当的机械强度、特殊的流变学特性、良好的稳定性、靶向性等,可提高、扩大HA作药物载体的性能和应用范围。最后展望了透明质酸及其衍生物作药物载体的应用前景。     

Enhanced Transdermal Absorption of Hyaluronic Acid via Fusion with Pep-1 and a Hyaluronic Acid Binding Peptide

It is always big challenges for hyaluronic acid (HA) in transmembrane absorbing and efficient delivering to the skin. Pep-1, as one of the cell-penetrating peptides, has been documented to permeate various substances across cellular membranes without covalent binding. Here, a novel hyaluronic acid binding peptide (named HaBP) is designed, and then combined with Pep-1 to enhance the cell-penetrating efficiency of HA. The results of ELISA and immunofluorescence assay show that HaBP could bind with HA very well, and a combination of Pep-1 and HaBP could efficiently improve the transmembrane ability of HA. Furthermore, HA gradually enters the dermis from the surface of the skin in mice when it is administrated with both HaBP and Pep-1, while there are no obvious allergies or other adverse reactions during this process. This study finds a new method to promote the efficient transmembrane and transdermal absorption of HA, and throws some light on further research on the development of hyaluronic acid and its related cosmetics or drugs.     

Characterization of Composite Networks Made of Type I Collagen,Hyaluronic Acid and Decorin

Summary: To ultimately assess the ability of macromolecular medicines e.g. liposomes, non viral gene delivery systems, to penetrate one of the least studied physiological barriers, the extracellular matrix (ECM), composite networks made of different ECM components i.e. type I collagen, hyaluronic acid and a proteoglycan, decorin, were prepared. These composite networks were characterized by rheology, Confocal Reflection Microscopy, Fluorescence Recovery After Photobleaching and Transmission Electron Microscopy. While being at the low end of its physiological concentration, collagen appears to be the backbone of the composite networks as it provides the elastic modulus. On the other hand, 15 kDa and 1.1 MDa hyaluronic acid, when present at physiological levels interpenetrate the collagen network. When approaching their overlap concentration, hyaluronic acid chains lead to an increase of the population of collagen fibrils. Finally, while decorin increased the population of fibrils in pure collagen networks, its role in presence of hyaluronic acid remains unclear as it does not alter the diameter of fibrils nor their population.     

TEMPO/TCC as a Chemo Selective Alternative for the Oxidation of Hyaluronic Acid

Hyaluronic acid (HA)-based hydrogels are very commonly applied as cell carriers for different approaches in regenerative medicine. HA itself is a well-studied biomolecule that originates from the physiological extracellular matrix (ECM) of mammalians and, due to its acidic polysaccharide structure, offers many different possibilities for suitable chemical modifications which are necessary to control, for example, network formation. Most of these chemical modifications are performed using the free acid function of the polymer and, additionally, lead to an undesirable breakdown of the biopolymer’s backbone. An alternative modification of the vicinal diol of the glucuronic acid is oxidation with sodium periodate to generate dialdehydes via a ring opening mechanism that can subsequently be further modified or crosslinked via Schiff base chemistry. Since this oxidation causes a structural destruction of the polysaccharide backbone, it was our intention to study a novel synthesis protocol frequently applied to selectively oxidize the C6 hydroxyl group of saccharides. On the basis of this TEMPO/TCC oxidation, we studied an alternative hydrogel platform based on oxidized HA crosslinked using adipic acid dihydrazide as the crosslinker.     

基于透明质酸的缺氧响应型胶束的制备及性能研究

以透明质酸（HA）及硝基咪唑（NI）衍生物为原料,通过酰胺反应合成了一种两亲性接枝聚合物（HA-NI）.该聚合物具有缺氧响应性和肿瘤靶向性.利用傅里叶红外光谱（FT-IR）和核磁共振（NMR）对接枝聚合物的结构进行了表征,同时基于¹H NMR计算出接枝聚合物中NI的取代度.HA-NI在水中自组装形成胶束,利用动态光散射仪（DLS）表征胶束大小并对胶束的稳定性进行了研究,利用原子力显微镜（AFM）和透射电镜（TEM）观察胶束形貌.载药胶束的载药率（DL）和包封率（EE）通过紫外-可见吸收光谱（UV）测试并计算得到.胶束的缺氧响应性利用DLS、AFM、TEM、UV等探究.实验证明,该聚合物胶束具有缺氧响应性并对药物表现出良好的控制释放能力,在常氧环境下,胶束稳定存在;而在缺氧环境下,胶束散开,药物快速且完全地释放.     

Construction of Hyaluronic Acid-Covered Hierarchically Porous MIL-nanoMOF for Loading and Controlled Release of Doxorubicin

The porous nano-sized metal–organic framework (nanoMOF) and its proper surface modification could greatly promote the drug loading capability and introduce biocompatibility, biodegradability, and targeting functions into nano-drug delivery systems. Herein, the HACD@ADA-PA/MIL-101_NH₂ (Fe)-P nanoparticle was successfully fabricated through supramolecular and coordination interactions from three building blocks, including hierarchically porous MIL-101_NH₂ (Fe)-P nanoMOF, phosphite-modified adamantane (ADA-PA), and β-cyclodextrin (β-CD)-modified hyaluronic acid (HACD). The obtained HACD@ADA-PA/MIL-101_NH₂ (Fe)-P nanoparticle was nano-sized and highly stable in physiological fluids. The porous structure of HACD@ADA-PA/MIL-101_NH₂ (Fe)-P nanoparticle could effectively load the commercial chemotherapeutic drug doxorubicin (DOX) with an encapsulation rate of 41.20 % and a loading rate of 48.84 %. The obtained drug-loaded HACD@ADA-PA/MIL-101_NH₂ (Fe)-P@DOX nanoparticle was pH-sensitive and relatively stable at neutral condition (pH 7.2) but could release DOX in a controlled way in subacid solution at pH 5.7. The simulated in vitro DOX release experiment signified that the HACD@ADA-PA/MIL-101_NH₂ (Fe)-P@DOX nanoparticle could realize the controlled release of DOX in tumor issues.     

Gel‐Like Structure of a Hexadecyl Derivative of Hyaluronic Acid for the Treatment of Osteoarthritis

Hyaluronic acid is a polysaccharide with viscoelastic and mechanical properties that are crucial for the normal functioning of osteoarticular junctions. It is demonstrated that introduction of a hexadecyl side chain into HA yields an injectable polysaccharide capable of forming physical hydrogels, which are stable at very low polymer concentrations, whereas native hyaluronic acid forms viscous solutions at concentrations that are ten times higher. Characterization of this system showed that the driving force for its gel‐like behavior is the occurrence of hydrophobic interactions involving aliphatic side chains, despite the low degree of substitution, as confirmed by molecular dynamics simulations of HYADD4 and HA hydrogels.

     

谷胱甘肽过氧化物酶模拟物碲化透明质酸的合成及催化动力学研究

 以透明质酸(HA)作为谷胱甘肽过氧化物酶(GPX)的酶模型,将碲(Te)引入HA中,合成了一种新型的高活力的GPX模拟物碲化透明质酸(TeHA). 用红外光谱和核磁共振技术对TeHA的结构进行了研究,证明Te的修饰位点位于HA的N-乙酰氨基葡萄糖的羟甲基(-CH2OH)上. 采用Wilson辅酶偶联法测定得到TeHA催化还原型谷胱甘肽(GSH)还原H2O2的GPX活力为163.6 U/μmol, 高于文献报道的其它模拟酶. TeHA还能够催化GSH还原异丙苯基过氧化物(CuOOH)和叔丁基过氧化物(t-BuOOH)的反应,并且CuOOH为该模拟酶的最适底物. 通过研究TeHA催化GSH还原三种不同过氧化物的反应动力学发现, TeHA的催化遵循乒乓机制.     

Fabrication and Characterization of an Electro-Compacted Collagen/Elastin/Hyaluronic Acid Sheet as a Potential Skin Scaffold

The development of biomimetic structures with integrated extracellular matrix (ECM) components represents a promising approach to biomaterial fabrication. Here, an artificial ECM, comprising the structural protein collagen I and elastin (ELN), as well as the glycosaminoglycan hyaluronan (HA), is reported. Specifically, collagen and ELN are electrochemically aligned to mimic the compositional characteristics of the dermal matrix. HA is incorporated into the electro-compacted collagen-ELN matrices via adsorption and chemical immobilization, to give a final composition of collagen/ELN/HA of 7:2:1. This produces a final collagen/ELN/hyaluronic acid scaffold (CEH) that recapitulates the compositional feature of the native skin ECM. This study analyzes the effect of CEH composition on the cultivation of human dermal fibroblast cells (HDFs) and immortalized human keratinocytes (HaCaTs). It is shown that the CEH scaffold supports dermal regeneration by promoting HDFs proliferation, ECM deposition, and differentiation into myofibroblasts. The CEH scaffolds are also shown to support epidermis growth by supporting HaCaTs proliferation, differentiation, and stratification. A double-layered epidermal-dermal structure is constructed on the CEH scaffold, further demonstrating its ability in supporting skin cell function and skin regeneration.     

Hybrid Pericardium with VEGF‐Loaded Hyaluronic Acid Hydrogel Coating to Improve the Biological Properties of Bioprosthetic Heart Valves

Bioprosthetic heart valves (BHVs) used in the clinic are mostly fixed by glutaraldehyde and the lack of endothelialization is a major problem for glutaraldehyde‐fixed pericardia. Hyaluronic acid is a major glycosaminoglycan that exists in native heart valves. Coupled with its inherent biocompatibility, it may enhance endothelial adhesion and proliferation when associated with vascular endothelial growth factor (VEGF). In this study, an optimized system is developed to improve the endothelialization of glutaraldehyde‐fixed pericardium. A hybrid pericardium with VEGF‐loaded hyaluronic acid hydrogel coating is developed by the crosslinking of 1,4‐butanediol diglycidyl ether. The adhesion and growth potential of human umbilical vein endothelial cells (HUVECs) on pericardia, platelet adhesion, and calcification by an in vivo rat subdermal implantation model are investigated. The results show improved HUVEC adhesion and proliferation, less platelet adhesion, and less calcification for hybrid pericardium by introducing the coating of VEGF‐loaded hyaluronic acid hydrogel. Thus, the coating of VEGF‐loaded hyaluronic acid hydrogel on pericardium is a promising approach to obtain bioprosthetic valves for clinical applications with increased endothelialization and antithrombotic and anticalcification properties.     

Fabrication and Properties of Porous Collagen/Hyaluronic Composite Scaffolds Containing Nano-Bioactive Glass for Tissue Engineering

In this work, nano-structured scaffolds were designed for tissue engineering using collagen, hyaluronic acid (HA) and nano-bioactive glass (NBAG) as their main components. The scaffold was prepared via freeze-drying method and the properties including morphology, porosity, compressive strength, swelling ratio and cytotoxicity in-vitro, were also evaluated. The composite scaffolds showed well interconnected macropores with the pore size of ranging from 100 to 500 μm. The porosity percent and swelling ability were decreased with the introduction of NBAG into the collagen/HA hydrogel; however, the compressive strength was enhanced. The cytotoxicity in-vitro study shows that the collagen-HA/NBAG scaffolds have good biocompatibility with improving effect on fibroblastic cells growth. It could be concluded that this scaffold fulfills the main requirements to be considered as a bone substitute.     

Crosslinked Hyaluronic Acid with Liposomes and Crocin Confers Cytoprotection in an Experimental Model of Dry Eye

Dry eye disease (DED) is a multifactorial condition caused by tear deficiency and accompanied by ocular surface damage. Recent data support a key role of oxidative and inflammatory processes in the pathogenesis of DED. Hyaluronic acid (HA) is widely used in artificial tears to treat DED by improving ocular hydration and reducing surface friction. Crocin (Cr), the main constituent of saffron, is a renowned compound that exhibits potent antioxidant and anti-inflammatory effects. The present study was undertaken to assess the viscosity and muco-adhesiveness of a photoactivated formulation with crosslinked HA (cHA), Cr, and liposomes (cHA-Cr-L). Our aim was also to evaluate whether cHA-Cr-L may exert cytoprotective effects against oxidative and inflammatory processes in human corneal epithelial cells (HCECs). Viscosity was measured using a rotational rheometer, and then the muco-adhesiveness was evaluated. Under hyperosmolarity (450 mOsm), the HCECs were treated with cHA-Cr-L. Interleukin-1β (IL-1β) and tumor necrosis factor α (TNFα) were quantified by quantitative real-time polymerase chain reaction (RT-qPCR). The levels of reactive oxygen species (ROS) were measured using the DCF assay. The combined action of cHA-Cr-L produced a higher viscosity and muco-adhesiveness compared to the control. The anti-inflammatory effect of cHA-Cr-L was achieved through a significant reduction of IL-1β and TNFα (p < 0.001). The results also showed that cHA-Cr-L reduces ROS production under conditions of hyperosmolarity (p < 0.001). We conclude that cHA-Cr-L has potential as a therapeutic agent in DED, which should be further investigated.     

Preparation and properties of redox responsive modified hyaluronic acid hydrogels for drug release

A series of oxidized hyaluronic acid (oxi‐HA)/3,3′‐dithiobis (propionohydrazide) (DTP) redox responsive hydrogels by Schiff base reaction under physiological conditions were designed and prepared. The influence of the concentration of oxi‐HA and DTP on rheological properties, equilibrium swelling ratio, and degradation rate were investigated. All oxi‐HA/DTP hydrogels exhibited good rheological properties, high equilibrium swelling ratio, low degradation rate, and sustainable drug release properties, and the comprehensive performance of oxi‐HA5/DTP6 hydrogel was better than that of others. The redox responsiveness was evaluated by means of degradation and in vitro bovine serum albumin release behavior investigation with the stimulus of different concentration of dithiothreitol as reducing agent. The intelligent hydrogels could be potentially applied in the fields of drug delivery system, tissue engineering, or cell scaffold materials. Copyright © 2017 John Wiley & Sons, Ltd.     

Gelatin—Hyaluronic Acid Hydrogels with Tuned Stiffness to Counterbalance Cellular Forces and Promote Cell Differentiation

Cells interact mechanically with their environment, exerting mechanical forces that probe the extracellular matrix (ECM). The mechanical properties of the ECM determine cell behavior and control cell differentiation both in 2D and 3D environments. Gelatin (Gel) is a soft hydrogel into which cells can be embedded. This study shows significant 3D Gel shrinking due to the high traction cellular forces exerted by the cells on the matrix, which prevents cell differentiation. To modulate this process, Gel with hyaluronic acid (HA) has been combined in an injectable crosslinked hydrogel with controlled Gel–HA ratio. HA increases matrix stiffness. The addition of small amounts of HA leads to a significant reduction in hydrogel shrinking after cell encapsulation (C2C12 myoblasts). We show that hydrogel stiffness counterbalanced traction forces of cells and this was decisive in promoting cell differentiation and myotube formation of C2C12 encapsulated in the hybrid hydrogels.

     

软骨素酶ABC酶解高效液相法测定鱼翅中的透明质酸

 建立了一种测定鲨鱼翅中透明质酸的酶解 高效液相方法。在37℃,0 2mol/LTris HCl缓冲溶液中,鱼翅中的透明质酸被硫酸软骨素酶ABC酶解成透明质酸二糖。条件为:ZORBAX糖分析柱(4 6mmi d ×250mm,5μm)柱;紫外检测波长226nm;流动相为乙腈 0 5%磷酸(体积比为2∶98),流速1mL/min;进样量10μL。透明质酸二糖的线性范围为25g/L～600g/L。将此法应用到鱼翅的实际样品检测中,取得了良好的结果,透明质酸在鱼翅样品中的质量分数为0 86%～1 96%。     

基于透明质酸及硫堇层层自组装的前列腺特异性抗原免疫传感器的研究

将带负电荷的透明质酸(HA)和带正电荷的电子媒介体硫堇(Thi)层层自组装到壳聚糖修饰的玻碳电极表面,利用硫堇上的氨基固定纳米金(nano-Au)并固载前列腺特异性抗体(anti-PSA),制得新型前列腺特异性抗原电流型免疫传感器。通过循环伏安法(CV)、微分脉冲伏安法(DPV)考察了该免疫传感器的电化学特性,并对影响该免疫传感器性能的各种因素进行详细研究。在优化实验条件下,此免疫传感器的线性范围为0.5~4μg/L和4~25μg/L,检出限为0.2μg/L。该免疫传感器具有制备简单、灵敏度高、稳定性好等优点。     

Validation of an Analytical Method for the Simultaneous Determination of Hyaluronic Acid Concentration and Molecular Weight by Size-Exclusion Chromatography

The hyaluronic acid (HA) global market growth can be attributed to its use in medical, cosmetic, and pharmaceutical applications; thus, it is important to have validated, analytical methods to ensure confidence and security of its use (and to save time and resources). In this work, a size-exclusion chromatography method (HPLC-SEC) was validated to determine the concentration and molecular distribution of HA simultaneously. Analytical curves were developed for concentration and molecular weight in the ranges of 100–1000 mg/L and 0.011–2.200 MDa, respectively. The HPLC-SEC method showed repeatability and reproducibility greater than 98% and limits of detection and quantification of 12 and 42 mg/L, respectively, and was successfully applied to the analysis of HA from a bacterial culture, as well as cosmetic, and pharmaceutical products.