新型可降解聚合物聚乙二醇对苯二甲酸酯/聚对苯二甲酸丁二醇酯的细胞相容性研究

目的：研究新型可降解聚合物聚乙二醇对苯二甲酸酯／聚对苯二甲酸丁二醇酯(PEGT／PBT)的人脐静脉内皮细胞(HUVEC)相容性，对其在组织工程血管中的应用进行探讨。方法：对PEGT／PBT进行细胞毒性评价。观察并检测HUVEC在PEGB／PBT、Ⅰ型胶原改性(Col-)的PEGT／PBT、纤维连接蛋白改性(Fn-)的PEGT／PBT上的粘附和增殖，对细胞在粘附过程中的粘着斑蛋白进行免疫荧光染色观察。结果：PEGT／PBT细胞毒性不大于1级，能支持脐静脉内皮细胞的粘附和增殖。纤维连接蛋白和Ⅰ型胶原处理可促进HLIVEC在PEGT／PBT膜上的增殖，而且纤维连接蛋白可增加HUVEC在PEGT／PBT上20min、2h的粘附率，并促进细胞形成局部粘附结构。结论：新型可降解聚合物PEGT／PBT具有良好的血管细胞相容性，对其在组织工程血管中的应用值得进一步开发研究。     

Cytocompatibility of Three Corneal Cell Types with Amniotic Membrane

Rabbit limbal corneal epithelial cells, corneal endothelial cells and keratocytes were cultured on amniotic membrane. Phase contrast microscope examination was performed daily. Histological and scan electron microscopic examinations were carried out to observe the growth, arrangement and adhesion of cultivated cells. Results showed that three corneal cell types seeded on amniotic membrane grew well and had normal cell morphology. Cultured cells attached firmly on the surface of amniotic membrane. Corneal epithelial cells showed singular layer or stratification. Cell boundaries were formed and tightly opposed. Corneal endothelial cells showed cobblestone or polygonal morphologic characteristics that appeared uniform in size. The cellular arrangement was compact. Keratocytes elongated and showed triangle or dendritic morphology with many intercellular joints which could form networks. In conclusion, amniotic membrane has good scaffold property, diffusion effect and compatibility with corneal cells. The basement membrane side of amniotic membrane facilitated the growth of corneal epithelial cells and endothelial cells and cell junctions were tightly developed. The spongy layer of amniotic membrane facilitated the growth of keratocytes and intercellular joints were rich. Amniotic membrane is an ideal biomaterial for layering tissue engineered cornea.     

γ射线低温辐照对胶原膜体外稳定性和细胞相容性的影响

低温下,胶原膜经γ射线辐照改性,采用剂量率为22 KG y/h,辐照剂量分别为15、25、35 KG y。测定辐照前后胶原膜抗胶原酶酶解能力,对胶原膜的体外稳定性进行了初步评价,结合红外光谱分析对辐照改性机理进行了探讨。结果表明,在实验所设计的条件下,辐照改性后胶原膜的交联度及稳定性均增加。采用M TT法结合扫描电镜观察,对辐照后胶原膜的细胞相容性进行了研究,表明在一定的辐照剂量范围内(<25 KG y),辐照对胶原膜的细胞相容性没有明显的影响。当超过一定剂量后,辐照改性将在一定程度上影响胶原膜的细胞相容性。     

Cytocompatibility study of NHLBI primary reference materials using human endothelial cells

Four different materials, low density polyethylene (LDPE), polydimethylsiloxane (PDMS), polyvinylcholoride (PVC) and cellulose, were selected by the Devices and Technology Branch of the National Heart Lung and Blood Institute (NHLBI) as primary reference materials for blood contacting. Among the wide variety of tests proposed to assess hemocompatibility of short-term blood contacting catheters, it was desirable to rule out whether these materials could release toxics for vascular cells of the physiological environment. Thus, the cytocompatibility of these materials have been checked towards human umbilical vein endothelial cells: the method used avoids direct contact between cells and materials but evaluates the effect of possible toxic substances leached from materials. These substances were obtained under defined conditions according to a standard. The results show that the extracts of cellulose and LDPE provoke an important cytotoxic effect on the endothelial cell cultures, while the extracts of PDMS and PVC allow the obtention of endothelial cell lining of the reference surface, with a correct global metabolic activity and the intracellular presence of von Willebrand factor.     

肝素缓释膜材料的制备与评价*

目的 为建立一种长效的肝素给药途径,制备肝素缓释膜材料,并评价其性质。方法 利用明胶与肝素交联成膜,测定其肝素缓释性质、细胞相容性及动物组织相容性。结果 该材料具有良好的肝素缓释效果, L929成纤维细胞在材料表面的生长受到一定抑制作用,载药膜片皮下植入和肌肉植入能逐步降解,但在降解过程中有一定的炎症反应,组织相容性还需要进一步完善。结论 该材料肝素缓释能力持久,为肝素缓释给药提供了实验依据。     

两种壳聚糖修饰材料细胞相容性及蛋白吸附行为的评价

目的对甘油明胶/壳聚糖(CGG)和玉米醇溶蛋白/壳聚糖(ZC)两种壳聚糖修饰生物材料的细胞相容性和蛋白吸附作用进行初步评价。方法应用M TT比色法,评价两种材料对L 929小鼠细胞的细胞毒性,测定他们对大鼠原代表皮细胞的细胞粘附率和细胞增殖率;同时应用考马斯亮兰G-250染色法测定材料对牛血清蛋白的吸附作用。结果本实验结果显示,两种壳聚糖修饰材料对L 929细胞生长抑制反应级别为1级或0级,属于无明显毒性作用。大鼠皮肤成纤维细胞在CGG上的粘附率和增殖率分别达到88.8%和96.0%;在ZC上的粘附率和增殖率分别达到80.7%和94.7%。两种材料不同时间对蛋白的吸附量存在有差异。结论两种修饰材料具有良好的细胞相容性,其中CGG在细胞体外增殖作用和蛋白吸附作用方面要优于单纯壳聚糖和ZC。     

Mechanical and Biological Properties of a Biodegradable Mg‐Zn‐Ca Porous Alloy

Objectives

As promising alternative to current metallic biomaterials, the porous Mg scaffold with a 3‐D open‐pore framework has drawn much attention in recent years due to its suitable biodegradation, biocompatibility, and mechanical properties for human bones. This experiment's aim is to study the mechanical properties, biosafety, and osteogenesis of porous Mg‐Zn alloy.

Methods

A porous Mg‐2Zn‐0.3Ca (wt%) alloy was successfully prepared by infiltration casting, and the size of NaCl particles was detected by a laser particle size analyzer. The microstructure of the Mg‐2Zn‐0.3Ca alloy was characterized by the stereoscopic microscope and Sirion Field emission scanning electron microscope. X‐ray computerized tomography scanning (x‐CT) was used to create the 3‐D image. The degradation rate was measured using the mass loss method and the pH values were determined together. The engineering stress–strain curve, compressive modulus, and yield strength were tested next. The bone marrow stromal cells (BMSC) were cultured in vitro. The CCK‐8 method was used to detect the proliferation of the BMSC. Alkaline phosphatase (ALP) and alizarin red staining were used to reflect the differentiation effects. After co‐culturing, cell growth on the material's surface was observed by scanning electron microscope (SEM). The cell adhesion was tested by confocal microscopy.

Results

The obtained results showed that by using near‐spherical NaCl filling particles, the porous Mg alloy formed complete open‐cell foam with a very uniform size of pores in the range of 500–600 μm. Benefitting from the small size and uniform distribution of pores, the present porous alloy exhibited a very high porosity, up to 80%, and compressive yield strength up to 6.5 MPa. The degradation test showed that both the pH and the mass loss rate had similar change tendency, with a rapid rise in the early stage for 1–2 day's immersion and subsequently remaining smooth after 3 days. In vitro cytocompatibility trials demonstrated that in comparison with Ti, the porous alloy accelerated proliferation in 1, 3, 5, and 7 days (P < 0.001), and the osteogenic differentiation test showed that the ALP activity in the experimental group was significantly higher (P = 0.017) and has more osteogenesis nodules. Cell adhesion testing showed good osteoconductivity by more BMSC adhesion around the holes. The confocal microscopy results showed that cells in porous Mg‐based alloy had better cytoskeletal morphology and were larger in number than in titanium.

Conclusions

These results indicated that this porous Mg‐based alloy fabricated by infiltration casting shows great mechanical properties and biocompatibilities, and it has potential as an ideal bone tissue engineering scaffold material for bone regeneration.

     

Assessment of cytocompatibility of surface-modified CdSe/ZnSe quantum dots for BALB/3T3 fibroblast cells

With the widespread use of quantum dots (QDs), the likelihood of exposure to QDs has been assumed to have increased substantially. Recently, QDs have been employed in numerous biological and medical applications. However, there is a lack of toxicological data pertaining to QDs. In this study, we aimed to investigate the cytocompatibility of surface-modified CdSe/ZnSe QDs for BALB/3T3 fibroblast cells. The ligands used for surface modification are mercaptopropionic acid (MPA) and Gum arabic (GA)/tri-n-octylphosphine oxide (TOPO). Cells were exposed to different concentrations of QDs followed by illustrative cytotoxicity analyses. Furthermore, we used a confocal microscope to assess intracellular uptake of QDs. Confocal images showed that MPA-coated QDs were distributed inside the cytoplasmic region of cells. In contrast, GA/TOPO-coated QDs were not found inside cells. MPA-coated QDs were highly cytocompatible, whereas GA/TOPO-coated QDs were toxic to the cells. Cells treated with GA/TOPO-coated QDs showed altered morphology, decreased viability, significant concentrations of intracellular free cadmium, detectable reactive oxygen species (ROS) formation, depolymerized cytoskeleton, and irregular-shaped nuclei. This study suggests that surface modification by ligands plays a significant role in the prevention of cytotoxicity of QDs.     

Hemocompatibility and biocompatibility of antibacterial biomimetic hybrid films

In previous work, we developed novel antibacterial hybrid coatings based on dextran containing dispersed Ag NPs (~ 5 nm, DEX-Ag) aimed to offer dual protection against two of the most common complications associated with implant surgery, infections and rejection of the implant. However, their blood-material interactions are unknown. In this study, we assess the hemocompatibility and biocompatibility of DEX-Ag using fresh blood and two cell lines of the immune system, monocytes (THP-1 cells) and macrophages (PMA-stimulated THP-1 cells). Glass, polyurethane (PU) and bare dextran (DEX) were used as reference surfaces. PU, DEX and DEX-Ag exhibited non-hemolytic properties. Relative to glass (100%), platelet attachment on PU, DEX and DEX-Ag was 15%, 10% and 34%, respectively. Further, we assessed cell morphology and viability, pro-inflammatory cytokines expression (TNF-α and IL-1β), pro-inflammatory eicosanoid expression (Prostaglandin E₂, PGE₂) and release of reactive oxygen species (ROS, superoxide and H₂O₂) following incubation of the cells with the surfaces. The morphology and cell viability of THP-1 cells were not affected by DEX-Ag whereas DEX-Ag minimized spreading of PMA-stimulated THP-1 cells and caused a reduction in cell viability (16% relative to other surfaces). Although DEX-Ag slightly enhanced release of ROS, the expression of pro-inflammatory cytokines remained minimal with similar levels of PGE₂, as compared to the other surfaces studied. These results highlight low toxicity of DEX-Ag and hold promise for future applications in vivo.     

Design of new titanium alloys for orthopaedic applications

Parallel to the biofunctionalisation of existing materials, innovation in biomaterials engineering has led to the specific design of titanium alloys for medical applications. Studies of the biological behaviour of metallic elements have shown that the composition and structure of the material should be carefully tailored to minimise adverse body reactions and to enhance implant longevity, respectively. Consequently, interest has focused on a new family of titanium alloys: Ti−6Mo−3Fe−5Ta, Ti−4Mo−2Fe−5Ta and Ti−6Mo−3Fe−5Zr−5Hf alloys. The non-toxicity of the specially designed titanium alloys compared with osteoblastic cells has been ascertained using MTT and RN tests. In addition, phase transformations upon thermal processing have been investigated, with comparison with a well-defined β titanium alloy. Optimum thermal processing windows (above 550°C) have been designed to generate a stable and nanostructured α phase from the isothermal ω phase that precipitates in a low temperature range (150–350°C). The generation of such nano-structured microstructures should provide a promising opportunity to investigate tissue-biomaterial interactions at the scale of biomolecules such as proteins.