胶原-磺化羧甲基壳聚糖/硅橡胶皮肤再生材料的制备及其对小型猪烫伤创面全层皮肤缺损的修复研究

制备了一种胶原-磺化羧甲基壳聚糖/硅橡胶皮肤再生材料,并以小型猪为模型,考察了其对烫伤全层皮肤缺损的修复性能.首先合成了磺化羧甲基壳聚糖,并对其结构进行了表征.制备了胶原-磺化羧甲基壳聚糖多孔支架,采用扫描电子显微镜(SEM)研究了磺化羧甲基壳聚糖含量对支架微结构的影响.随着磺化羧甲基壳聚糖含量的增大,胶原-磺化羧甲基壳聚糖支架从纤维结构向片状结构转化,且支架的孔径相对变大.采用体外成纤维细胞培养实验证明胶原-磺化羧甲基壳聚糖支架无明显细胞毒性.进一步将胶原-磺化羧甲基壳聚糖支架与硅橡胶膜复合,构建具有双层结构的皮肤再生材料.以小型猪为模型,评价了其对深度烫伤创面的修复性能.大体观察和组织学分析结果显示,胶原-磺化羧甲基壳聚糖/硅橡胶皮肤再生材料具有更快的血管化性能,且经该材料处理的创面能有效支持薄自体皮片的移植成活,实现深度烫伤创面的全层修复.     

乙酰化对壳聚糖-明胶海绵结构和性能的影响

用乙酸酐对壳聚糖与明胶共混物进行乙酰化,然后冷冻干燥制备乙酰化壳聚糖-明胶海绵,并研究乙酰化对海绵结构与性能的影响。用盐酸环丙沙星作模型药物,探讨载药海绵的抑菌效果。结果表明:制得的海绵具有多孔结构,随壳聚糖在混合物中含量的增加,海绵的吸水率和保水率先增后减;随着乙酰化程度的提高,海绵的吸水率先减后增,而保水率与吸水率的变化规律相反。酶对海绵的降解率不受壳聚糖和明胶混合比例的影响,但随海绵乙酰化程度的增加而增加。载药海绵的抑菌效果与海绵中壳聚糖含量有关。     

皮肤组织工程支架材料的研究进展

皮肤组织工程支架为种子细胞提供粘附、生长、增殖和代谢的环境,并起支撑和模板作用,引导组织再生和控制组织结构,是人工皮肤的重要组成部分。探索理想的支架材料是当前皮肤组织工程领域的热点,本文综述了近年来皮肤组织工程支架材料的国内外研究进展,包括天然支架材料、合成支架材料和复合支架材料三大类,全面探讨了皮肤组织工程支架材料的制备方法及应用情况,本文还分析了皮肤组织工程支架材料存在的一些问题,并对其未来的发展进行了展望。     

温度对牛胶原聚集态的影响

本研究采用酶解法制备了牛胶原。分别用粘度法、紫外分光光度计、红外光谱和原子力显微镜分析方法,考察了不同温度下胶原的物理化学特性。实验结果表明:粘度法和紫外分光光度计测定的胶原溶液的变性温度约为38℃;红外光谱测定表明,随着温度的提高,胶原膜在1650cm-1的吸收峰减弱;原子力显微镜实验表明了在40℃时牛胶原分子由螺旋结构的长链断裂为小链段和一些小碎片,说明此方法制备的胶原在40℃以上其三股螺旋结构将被破坏。     

磺化壳聚糖的制备及其对生长因子活性的保护作用

以壳聚糖为基材,分别以氯磺酸、三甲胺.三氧化硫和丙磺酸内酯为磺化试剂,制备了3,6-O-磺化壳聚糖(OCS)、2-N-磺化壳聚糖(NCS)和2-N-磺丙基壳聚糖(PCS)3种磺化壳聚糖.采用红外光谱、核磁共振谱证明了磺化壳聚糖的结构,元素分析测定了磺化壳聚糖的磺化率.以组织修复过程中一种重要的活性因子——碱性成纤维细胞生长因子(bFGF)为目标因子,通过体外成纤维细胞培养实验,评价了不同磺化壳聚糖对bFGF活性的保护能力.结果显示,相对于未改性壳聚糖(CS),磺化壳聚糖对成纤维细胞的形态无显著影响.细胞活性检测结果显示,OCS和PCS对bFGF活性无明显的保护能力;NCS能有效提高bFGF促进成纤维细胞活性的能力,其对bFGF活性的保护能力可达肝素70%左右.磺化壳聚糖的bFGF活性保护能力不随磺化率的改变而变化.     

仿生材料电活性聚合物“人工肌肉”的研究进展

自古以来,自然界就是人类各种技术思想、工程原理及重大发明的源泉.20世纪中期,人们越来越深刻认识到大自然的启发对于开发新材料和新技术的重要性,从而提出仿生学概念并建立仿生学这一学科.随着研究的发展,仿生学已成为自然科学的一个前沿和焦点.进入21世纪以来,随着机器人开发的不断深入以及人们对智能机械系统的强烈需求,作为机器人和智能机械系统驱动关键的人工肌肉已成为仿生领域的研究重点.电活性聚合物驱动器具有应变高、柔软性好、质轻、无噪声等特点,与肌肉有着极为相似的特性,甚至在一些方面的性能已经超过了肌肉,被公认为是最合适的仿肌肉材料,称之为"人工肌肉".近二十年来,在电活性聚合物驱动材料方面取得的研究进展使得仿生的"人工肌肉"研究得以飞速发展.     

紫外光照射对皮胶原热降解活化能的影响

将皮胶原经不同时间的紫外光照射,采用热重法(TG)和微分热重分析(DTG)研究了照射前后皮胶原的热降解行为,用Horowitz-Metzger法和Coats-Redfern法计算了其热降解活化能,得到了未照射皮胶原和经过不同紫外线照射时间处理后的皮胶原的热降解活化能。研究发现,在较短时间(0~4h)的照射后,皮胶原的热降解活化能略有增大;在较长时间(8~64h)的照射后,皮胶原的热降解活化能大幅降低。这可能是由于在紫外光照射的过程中,皮胶原分子链发生了以交联为主和以断链为主的复杂光化学反应。     

胶原液晶膜的制备及对细胞生长的影响

从仿生角度出发, 制备了一系列液晶态胶原薄膜. 采用偏光显微镜、扫描电镜和原子力显微镜分别研究了各种膜的液晶织构和拓扑结构, 并通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法及细胞骨架荧光染色观察研究了各种膜对鼠成纤维细胞3T3的黏附、增殖以及形貌的影响. 结果表明, 胶原液晶薄膜呈现典型的胆甾型液晶织构, 与非液晶薄膜相比, 它不仅能更好地促进细胞的黏附、生长及增殖, 而且其拓扑结构对细胞生长具有接触引导效应.     

硫酸对壳聚糖膜分离性能影响的本质

研究了Ｈ２ＳＯ４对壳聚糖（ＣＳ）膜醇水分离性能的影响．结果表明，Ｈ２ＳＯ４使ＣＳ上的—ＮＨ２质子化为—ＮＨ＋３，降低了极性较差的乙醇分子进入膜内的能力，提高了膜对水分子的吸附选择性．又因为ＳＯ２－４可以与２个—ＮＨ＋３作用，使部分ＣＳ链产生交联，缩小了分子透过的通道．由于乙醇分子比水分子大，故大大提高了膜对水的渗透选择性．所以，Ｈ２ＳＯ４能提高ＣＳ膜的醇水分离性能是以上两种因素综合作用的结果．     

A New Kind of Biomaterials——Bullfrog Skin Collagen

Pepsin-soluble collagen was prepared from bullfrog skin and partially characterized. This study revealed interesting differences, such as molecular weight, amino acid composition,denaturation temperature (Ta), in the frog skin collagen when compared to the known vertebrate collagens. This study gives hints that bullfrog skin can be a potential, safe alternative source of collagen from cattle for use in various fields.     

Mineralization of Phosphorylated Fish Skin Collagen/Mangosteen Scaffolds as Potential Materials for Bone Tissue Regeneration

In this study, a potential hard tissue substitute was mimicked using collagen/mangosteen porous scaffolds. Collagen was extracted from Tilapia fish skin and mangosteen from the waste peel of the respective fruit. Sodium trimetaphosphate was used for the phosphorylation of these scaffolds to improve the nucleation sites for the mineralization process. Phosphate groups were incorporated in the collagen structure as confirmed by their attenuated total reflection Fourier transform infrared (ATR-FTIR) bands. The phosphorylation and mangosteen addition increased the thermal stability of the collagen triple helix structure, as demonstrated by differential scanning calorimetry (DSC) and thermogravimetry (TGA) characterizations. Mineralization was successfully achieved, and the presence of calcium phosphate was visualized by scanning electron microscopy (SEM). Nevertheless, the porous structure was maintained, which is an essential characteristic for the desired application. The deposited mineral was amorphous calcium phosphate, as confirmed by energy dispersive X-ray spectroscopy (EDX) results.     

Crosslinked Networks Based on Polysaccharides and Collagen for Pilocarpine Sustained Release

The paper presents the experimental studies regarding synthesis and characterization of hydrogels based on gellan (Gel)/chitosan (CS) and collagen (Col), obtained by crosslinking with glutaraldehyde (GLA). The influence of the polysaccharide content and GLA ratio on the final composition and swelling characteristics was evaluated. Hydrogels swelling analysis, in distilled water and phosphate buffer (PBS, pH 7.2) has shown higher swelling degrees at increased concentration of polysaccharide into hydrogels. In vitro release of pilocarpine has demonstrated the possibility to use gellan-collagen and chitosan-collagen hydrogels as ophthalmic drug delivery matrix.     

Thermal and Rheological Behavior of Collagen. Chitosan blends

Collagen:chitosan blends in 1:1 ratio were prepared and characterized by Fourier transform infrared spectroscopy, thermal (DSC, TG) and rheological studies. Apparently each material maintains its behavior and addition of chitosan does not denature collagen fibers. The rheological behavior showed that adding chitosan to collagen causes a decrease of storage modulus (G’),viscous loss modulus (G”) and apparent viscosity when measured as a function of frequency. Both anionic and native collagen presented more solid-like behavior than fluid-like viscoelastic behavior. Collagen:chitosan blends exhibits a more fluid-like viscoelastic behavior. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Characterization of Collagen from Three Genetic Lines (Gray,Red and F1) of Oreochromis niloticus (Tilapia) Skin in Young and Old Adults

From tilapia (Oreochromis niloticus) farming, the by-products have been identified as a source of collagen that could be used for the development of dermocosmetics or pharmaceutical products. However, the characteristics of collagen related to a specific strain or culture must be well defined prior to its application. Collagen was extracted from the skin of three strains of tilapia: red YY males (YY: two Y-type sex chromosomes), XX gray females, and the F1: offspring of crossing red YY males with XX gray females; at different ages in the adult phase, using acetic acid and pepsin enzyme. The characteristics of acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were shown by SDS-PAGE band profiles to be similar to bovine collagen type I (SIGMA), the PSC of gray tilapia being more fragile to temperature changes, consistent with the results of fractional viscosity. The characteristics of the F1 progeny were prioritized for being a commercially productive and sustainable source for the extraction of collagen, and the ASC form, being the one with the greatest stability and advantage over PSC, of importance to our investigations, leads to a controlled digestion as in the case of peptide induction, and also in the development of natural products in the pharmaceutical and/or dermocosmetic industry. Evaluations of the triple helix structure by FT-IR, X-ray diffraction and UV–visible spectroscopy give similar results between the strains: red, gray, and F1, and between ages in the adult form F1 (15, 24, and 36 months of age). Consequently, the skin of tilapia in adult form is recommended sustainably for up to 24 months of age where the collagen is obtained with the use of acetic acid without enzymatic treatment.     

污染肝素中多硫酸化硫酸软骨素的分步醋酸纤维素薄膜电泳分析

采用离子交换色谱法从污染肝素原料中分离出多硫酸化硫酸软骨素(OSCS),建立了分步醋酸纤维素薄膜电泳法分析污染肝素中OSCS含量的方法.结果表明,先以0.05 mol/L醋酸钡缓冲液(pH 5.0)电泳,再以0.15 mol/L醋酸锌缓冲液(pH 6.3)电泳,可以将肝素和OSCS完全分开,检出限为0.1 g/L; 通过灰度积分建立定量校准曲线,相关系数为0.9934,平均回收率为102.1%～106.1%; RSD为4.1%～6.0%.     

胶原-壳聚糖共混膜的微观结构

胶原-壳聚糖共混膜的微观结构;胶原;壳聚糖;共混膜;原子力显微镜(AFM)     

Kinetic and Structural Aspects of Glycosaminoglycan–Monkeypox Virus Protein A29 Interactions Using Surface Plasmon Resonance

Monkeypox virus (MPXV), a member of the Orthopoxvirus genus, has begun to spread into many countries worldwide. While the prevalence of monkeypox in Central and Western Africa is well-known, the recent rise in the number of cases spread through intimate personal contact, particularly in the United States, poses a grave international threat. Previous studies have shown that cell-surface heparan sulfate (HS) is important for vaccinia virus (VACV) infection, particularly the binding of VACV A27, which appears to mediate the binding of virus to cellular HS. Some other glycosaminoglycans (GAGs) also bind to proteins on Orthopoxviruses. In this study, by using surface plasmon resonance, we demonstrated that MPXV A29 protein (a homolog of VACV A27) binds to GAGs including heparin and chondroitin sulfate/dermatan sulfate. The negative charges on GAGs are important for GAG–MPXV A29 interaction. GAG analogs, pentosan polysulfate and mucopolysaccharide polysulfate, show strong inhibition of MPXV A29–heparin interaction. A detailed understanding on the molecular interactions involved in this disease should accelerate the development of therapeutics and drugs for the treatment of MPXV.