Surface modification of titanium thin film with chitosan via electrostatic self-assembly technique and its influence on osteoblast growth behavior

Chitosan (Chi) and poly (styrene sulfonate) (PSS) were employed to surface modify titanium thin film via electrostatic self-assembly (ESA) technique in order to improve its biocompatibility. The surface chemistry, wettability and surface topography of the coated films with different number of deposited layers were investigated by using X-ray photoelectron spectroscopy (XPS), water contact angle measurement and atomic force microscopy (AFM), respectively. The results indicated that a full surface coverage for the outmost layer was achieved at least after deposition of five layers, i.e., PEI/(PSS/Chi)₂ on the titanium films. The formed multi-layered structure of PEI(PSS/Chi) _x (x ≥ 2) on the titanium film was stable in air at room temperature and in phosphate buffered solution (PBS) for at least 3 weeks. Cell proliferation, cell viability, DNA synthesis as well as differentiation function (alkaline phosphatase) of osteoblasts on chitosan-modified titanium film (PEI/(PSS/Chi)₆) and control sample were investigated, respectively. Osteoblasts cultured on chitosan-modified titanium film displayed a higher proliferation tendency than that of control (p < 0.01). Cell viability, alkaline phosphatase as well as DNA synthesis measurements indicated that osteoblasts on chitosan-modified titanium films were greater (p < 0.01) than those for the control, respectively. These results suggest that surface modification of titanium film was successfully achieved via deposition of PEI/(PSS/Chi) _x layers, which is useful to enhance the biocompatibility of the titanium film.     

Effects of baicalin-modified poly(D,L-lactic acid) surface on the behavior of osteoblasts

In the present study, the functions of rat calvaria osteoblasts on baicalin-modified poly(D,L-lactic acid) (PDLLA) films were investigated in vitro. The surface characteristics of surfaces (both modified and control) were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). Cell morphologies on these surfaces were examined by scanning electron microscopy (SEM). Cell adhesion and proliferation were used to assess cell growth on the modified and control surfaces. The MTT assay was used to determine cell viability and alkaline phosphatase (ALP) activity was performed to evaluate differentiated cell function. Compared to control films, cell attachment of osteoblasts on baicalin-modified PDLLA film was significantly higher (P<0.05 and P<0.01) after 6 h and 8 h culture, and cell proliferation was also significantly greater (P<0.05 and P<0.01) at the end of 4th and 7th day, respectively. The MTT assay suggested that the cell viability of osteoblasts cultured on baicalin-modified PDLLA film was significantly higher (P<0.05) than that seeded on the control. Meanwhile, the ALP activity of osteoblasts cultured on modified films was also considerably enhanced (P<0.01) compared to that found on control. These results revealed that the biocompatibility PDLLA could be improved by surface modification with baicalin.     

聚(D,L-乳酸)基仿生聚合物材料的合成与表征

探索一种新型聚乳酸基仿生聚合物材料的制备新方法.具体实验步骤是:利用聚乳酸上叔碳原子的自由基反应活性,在过氧化二苯甲酰的催化作用下,将马来酸酐引入聚乳酸侧链上,以此提供高反应活性的酸酐键;然后利用酸酐基团与-NH2发生N-酰化反应这一特点,将脂肪族二胺引入聚乳酸侧链上,从而克服聚乳酸降解产物的体液环境呈酸性的缺陷;再用碳二亚胺作缩合剂,在二胺改性聚乳酸材料中共价引入一种细胞粘附肽段Arg-Gly-Asp-Ser(RGDS),赋予材料生物活性和生物特异性,这样就制备了一种新型聚乳酸基仿生材料.采用MALLS、FTIR和XPS对仿生材料进行结构表征;采用罗丹明比色法、茚三酮显色法和氨基酸分析仪检测法对仿生材料中的马来酸酐、二胺和粘附肽RGDS进行定量测定.结果表明,按文中所述之制备技术,在不改变聚乳酸材料主链结构的前提下,该仿生材料中粘附肽RGDS的含量是5.12μmol/g.这就形成了一种具有类似细胞外基质的新型仿生材料.     

新型聚乙二醇接枝聚乳酸及其降解特性研究

通过直接酰胺化反应,以马来酸酐本体改性聚乳酸(MPLA)和氨基封端聚乙二醇(H2N-PEG-NH2)为原料,合成了聚乙二醇本体改性聚乳酸(PPLA).通过红外、核磁共振技术对改性聚合物的结构进行了表征;采用FITC标记牛血清白蛋白(FITC-BSA)为模型蛋白质,测试了聚合物对蛋白质的非特异性吸附;利用吸水率表征了聚合物的亲水性;利用聚合物在12周降解过程中pH值、失重率的变化评价了它们的降解特性.结果表明:H2N-PEG-NH2已成功接枝到MPLA上;与聚乳酸(PLA)、MPLA相比,PPLA明显降低了对FITC-BSA的吸附;其亲水性和降解性增加.预计该材料将是一种更适合组织工程和药物缓释应用的聚乳酸类可降解材料.     

聚L-乳酸的固态缩聚

采用熔融 /固态缩聚方法制备了聚L 乳酸。考察了不同催化剂、反应温度、反应时间和预聚物颗粒大小对固态缩聚反应速度的影响。结果表明 ,利用真空干燥箱 ,在适宜的反应条件下反应 ,有利于最终聚合物分子量的控制。     

高分子量聚乳酸用于SLS快速成型的研究

通过选区激光烧结技术(selective laser sintering,SLS),对高分子量聚(D,L-乳酸)(PDLLA)快速成型加工,研究该技术制造PDLLA组织工程支架的可行性。重均分子量80000,最大粒径120μm的PDLLA粉末,采用SLS技术快速成型,优化得到最佳控制参数,即预热温度为40℃、激光功率为12W、扫描速度1200mm/s。按照该参数加工测试件及人耳状支架,通过GPC-十八角激光光散射联用技术分析材料分子量的变化,力学测试仪分析成型材料的力学性能,扫描电镜分析支架材料的微观结构。结果显示高分子量的PDLLA可用于SLS加工成型,加工过程中高温会引起分子量部分降低,这种变化主要是激光接触面分子量降低所致,烧结作用不会改变材料的主要化学基团,力学测试显示烧结成型的材料能保留加工前聚合物20%的压缩模量,扫描电镜观察成型材料具有多孔结构,显示PDLLA用于SLS加工组织工程支架的潜能。     

Synthesis and characterization of cholesterol-poly(ethylene glycol)-poly(D,L-lactic acid) copolymers for promoting osteoblast attachment and proliferation

A novel cholesterol-poly(ethylene glycol)-poly(D,L-lactic acid) copolymer (CPEG-PLA) has been synthesized as a potential surface additive for promoting osteoblast attachment and proliferation. The gel permeation chromatography (GPC) and nuclear magnetic resonance spectroscopy (NMR) results indicated the product had expected structure with low polydispersities in the range of 1.1–1.5. By blending the poly(D,L-lactic acid) (PLA) with CPEG-PLA, the surface of modified PLA membrane was investigated by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and contact angle. The results revealed the enrichment of PEG chain on the surface. Osteoblast cell line (MC3T3) was chosen to test the cell behavior on modified PLA membranes. The osteoblast test about cell attachment, proliferation, cell viability and cell morphology investigation on CPEG-PLA modified PLA substrates showed the CPEG-PLA with 15 and 5 ethylene glycol units promoted osteoblast attachment and growth, while the CPEG-PLA with 30 ethylene glycol units prevent osteoblast adhesion and proliferation. This simple surface treatment method may have potentials for tissue engineering and other biomedical applications.     

Preparation and characterization of bioactive and biodegradable Wollastonite/poly(D,L-lactic acid) composite scaffolds

Composite scaffolds of poly(D,L-lactic acid) (PDLLA) with bioactive wollastonite were fabricated by the conventional solvent casting-particulate leaching method. The pore structures and morphology of the scaffolds were determined by scanning electron microscopy (SEM). The bioactivity of the composites was evaluated by soaking in a simulated body fluid (SBF), and the formation of the hydroxyapatite (HAp) layer was determined by SEM and energy-dispersive spectrometer. The results showed that the wollastonite/PDLLA composites were bioactive as it induced the formation of HAp on the surface of the composite scaffolds after soaking in SBF for seven days. In addition, pH and ion concentration changes of SBF solutions with composite scaffolds were examined. The results showed that the composites could release Ca and Si ions, which could neutralize the acidic degradation by-products of the PDLLA, and stabilize the pH of the SBF solutions between 6.7 and 7.2 within a three-week soaking period. Furthermore, the measurements of the water contact angles suggested that incorporation of wollastonite into PDLLA could improve the hydrophilicity of the composites and the enhancement was dependent on the wollastonite content. All these results suggest that incorporation of wollastonite into PDLLA might be a useful approach for the preparation of composite scaffolds for tissue repair and tissue-engineering applications.     

Change in cell adhesion property on cytocompatible interface using phospholipid polymer grafted with poly(D,L-lactic acid)segment for tissue engineering

Tissue engineering is a multi-disciplinary science that utilizes basic principles from materials engineering and molecular biology to reconstruct tissues from polymer matrices and cellular components. Artificial skins were well known as one of the concrete examples. Technological innovation of the tissue engineering must be contributed to improve quality of life. From the viewpoint, design of cytocompatible materials for tissue engineering would be the most important candidate to reconstruct tissue. 2-Methacryloyloxyethyl phosphorylcholine (MPC), n-butyl methacrylate, and polylactic acid (PLA) macromonomer were polymerized for the preparation of cytocompatible interface. The polymer may involve following novel properties: (i) cytocompatibility by phospholipid groups, and (ii) enhancement of cell adhesion by PLA segment. The results of X-ray photoelectron spectroscopy showed the MPC unit and PLA segment on the membrane, which was prepared by dip coating. The surface mobility by contacting water was estimated with static contact angle measurement. The contact angle by water decreased after contact with water due to the chain rearrangement of hydrophilic MPC unit. Fibroblast cells adhesion and protein adsorption on the membranes were studied. The number of cell adhesion and cell proliferation on the membrane was well correlated with each other. Furthermore, the number of cell adhesion was proportional to the PLA macromonomer (MaPLA) composition. The adherent cell morphology showed round shape, because of the existence of MPC unit. However, the cell morphology would be spread after the cell proliferation. These findings suggest that the change in the polymer composition by combination of MPC and MaPLA could regulate the number of cell adhesion and the morphology.     

立构选择性催化剂对D,L-乳酸聚合物微结构的影响

以L-和D,L-乳酸为原料熔融缩聚制备聚乳酸,采用旋光度法、1H核磁共振法和0.7C核磁共振法和x-射线衍射等方法研究催化剂对聚乳酸微结构的影响.D,L-乳酸采用非立构选择性催化剂萘磺酸缩聚时,所得的聚乳酸为消旋的非晶态聚合物.而采用SnCl2/对甲苯磺酸和SnCl2/蔡磺酸等立构选择性催化体系时,D,L-乳酸缩聚所得的聚乳酸是L_构型为主的结晶聚合物,且SnCl2/蔡磺酸体系的聚乳酸具有更高的光学活性.采用不同催化剂,D,L-乳酸可缩聚制备不同结构和性能的聚乳酸.     

两亲性嵌段共聚物聚乳酸-聚亮氨酸-聚乙二醇的合成及性能研究

具有温度敏感的生物水凝胶因可根据温度变化发生相变而受到人们重视。采用直接熔融聚合法,先以外消旋乳酸(D,L-LA)和L-亮氨酸(Leu)为原料,辛酸亚锡为催化剂,合成了聚乳酸-聚亮氨酸;再以一定比例将上述两组分共聚物与一定分子量的聚乙二醇(PEG)混合,在辛酸亚锡为催化剂的情况下熔融聚合制备聚乳酸-聚亮氨酸-聚乙二醇三组分嵌段共聚物。采用傅里叶红外光谱(FTIR)、核磁共振波谱仪(1 H-NMR)和凝胶渗透色谱仪(GPC)对两组分和三组分共聚物进行了系统表征,所得三组分共聚物的一定浓度的水溶液在20～35℃时具有溶胶-凝胶转变特性,能够满足该生物友好材料在药物缓释领域中的应用。     

有机玻璃表面的静电自组装超亲水防雾改性

研究利用聚烯丙基胺盐酸盐与SiO2纳米粒子在有机玻璃片表面的静电自组装来进行超亲水防雾改性。水接触角分析显示随着双电层数目的增加,有机玻璃的亲水性明显改善,并在20双电层时呈现超亲水性。原子力显微镜观察表明,有机玻璃片的超亲水性与其表面形成规整排列的纳米微孔结构,有利于水汽在有机玻璃片表面形成水膜有关。防雾测试结果进一步表明,表面自组装改性能明显改善有机玻璃片的防雾性能。此外,透光率分析结果还得出有机玻璃片改性后并不影响其透光性,反而还会因为反射光强减弱而略微提高。     

Synthesis and characterization of poly(L-lactic acid-co-ethylene oxide-co-aspartic acid) and its interaction with cells

Multiblock terpolymer of poly(L-lactic acid)/ poly(ethylene oxide)/poly(L-aspartic acid), (PLLA/PEO/ PAsp) was synthesized by ring opening polymerization of β -benzyl L-aspartate N-carboxyanhydride, Asp(OBzl)-NCA with α-ω -hydroxy terminated triblock PLLA/PEO/PLLA copolymer. The resulting multiblock terpolymer was characterized by several techniques including Fourier transform infrared spectroscopy and differential scanning calorimetry.¹H nuclear magnetic resonance spectra indicated the molar ratio of PLLA/PEO/PAsp (OBzl) to be 86/10/4. Thermal gravimetric analysis and environmental scanning electron microscopy data showed that PLLA/PEO/PAsp had crystalline and brittle structure. In order to improve its mechanical and physical properties, the terpolymer was blended with high molecular weight poly(L-lactic-co-glycolic acid) copolymer, PLGA(85/15) (M_w: 95000 gmol^{− 1}) in 25/75 and 50/50 mole ratios. The hydrolytical degradation properties of these polymers were studied. Degradation experiments were performed during a 48-day period in pH:7.4 phosphate-buffered saline (PBS) at 37^∘C. The observed molecular weight losses were 91% and 67% for the 25/75 and 50/50 mixtures, respectively. In vitro attachment and growth of L929 mouse fibroblasts on these biopolymers were also investigated. Cell growth experiments indicated that the copolymer blend allowed the attachment and growth of cells.     

Biodegradation of porous versus non-porous poly(L-lactic acid) films

The influence of porosity on the degradation rate of poly(L-lactic acid) (PLLA) films was investigated in vitro and in vivo. Non-porous, porous and combi (porous with a non-porous layer) PLLA films were used. Changes in Mw, Mn, polydispersity (Mw/Mn) ratio, melting temperature (T _m), heat of fusion, tensile strength, E-modulus, mass and the remaining surface area of cross-sections of the PLLA films were measured. In general, during the degradation process, the porous film has the highest Mw, Mn, Mw/Mn ratio and T _m, while the non-porous film has the lowest. In contrast, the highest heat of fusion values were observed for the non-porous film, indicating the presence of relatively smaller molecules forming crystalline domains more easily. The tensile strength and E-modulus of the non-porous film decrease faster than those of the porous and the combi film. None of the three types of films showed massive mass loss in vitro nor a significant decrease in remaining polymer surface area in light microscopical sections in vitro and in vivo. Heavy surface erosion of the non-porous layer of the combi film was observed after 180 days, turning the combi film into a porous film. This is also indicated by the changes in tensile strength, Mw, Mw/Mn, T _m and heat of fusion as a function of time. It is concluded that non-porous PLLA degrades faster than porous PLLA. Thus, in our model, porosity is an important determinant of the degradation rate of PLLA films.     

熔融缩聚法直接合成聚L-乳酸的研究

以L-乳酸为原料,熔融缩聚法直接合成聚L-乳酸.考察了预聚工艺、催化剂配比及用量、反应温度、反应时间及真空度等条件对乳酸聚合反应的影响.通过红外光谱(FTIR)和核磁共振波谱(1H-NMR)分析和表征聚乳酸的结构,凝胶渗透色谱(GPC)测试其分子量及分子量分布.研究表明,较为适宜的工艺条件为:以二水合氯化亚锡(SnCl2·2H2O)和对甲基苯磺酸(TSA)为催化剂,且SrCl2·2H2O相对于乳酸预聚体的质量比为0.5%,真空度-0.98MPa,温度180℃,反应时间10h.得到的聚L-乳酸的粘均分子量为6.17×104,分子量分布1.37,且产率较高,色泽较浅.     

MGF-Ct24E改性聚乳酸的合成、表征及其对成骨细胞增殖的评价

力生长因子(MGF)是骨修复重建的一种重要生长因子。以二环己基碳二亚胺为缩合剂,将MGF羧基端E结构域24肽(MGF-Ct24E)共价接枝到丁二胺改性的聚乳酸上(DPLA),制得了新型MGF-Ct24E改性聚乳酸仿生材料(MGF-Ct24E-DPLA)。采用氨基酸分析和高效液相色谱对MGF-Ct24E含量进行了定性定量表征,静态水接触角和吸水率测定了MGF-Ct24E-DPLA材料的亲水性,MTT法评价了其对成骨细胞的增殖作用。结果表明,MGF-Ct24E成功引入到DPLA中,接枝效率为24.7%,并且和DPLA相比,MGF-Ct24E-DPLA材料具有更好的亲水性和促进成骨细胞增殖的能力。因此这种新型MGF-Ct24E改性聚乳酸仿生材料有望成为骨组织工程领域中一种卓越的生物材料。     

生物基聚乳酸微孔膜的制备及透析性能

通过浸没沉淀相转化法制备生物基聚乳酸(PLLA)微孔膜,并考察溶剂种类和致孔剂种类及致孔剂分子量对微孔膜结构和性能的影响.分别考察N-甲基吡咯烷酮(NMP)、N,N-二甲基乙酰胺(DMAc)和1,4-二氧六环(DO)作为溶剂对PLLA膜微孔结构的影响,研究用聚乙二醇(PEG6000)和聚乙烯基吡咯烷酮(PVP-K30)作为致孔剂对PLLA膜微孔结构、平均孔径、水通量及接触角的影响;不同分子量的致孔剂PEG(600、6000、10000)对PLLA膜微孔结构、平均孔径及水通量的影响.通过扫描电镜照片发现,NMP和DMAc作为溶剂可得到指状孔,而DO作为溶剂得到胞状孔,PEG和PVP-K30作为致孔剂均可促进PLLA膜大孔的形成,PVP-K30可使PLLA膜表面更为多孔,不同分子量的PEG作为致孔剂会同时影响PLLA膜的孔径和皮层厚度.通过对制备的PLLA微孔膜的水通量及尿素、溶菌酶和牛血清蛋白的透过性能分析表明聚乳酸膜具有较好的溶质选择透过性,有望成为具有良好生物相容性和透析性能的新一代血液透析膜材料.     

Porous and dense poly(L-lactic acid) and poly(D,L-lactic acid-co-glycolic acid) scaffolds: <Emphasis Type="Italic">In vitro</Emphasis> degradation in culture medium and osteoblasts culture

The use of bioresorbable polymers as a support for culturing cells has received special attention as an alternative for the treatment of lesions and the loss of tissue. The aim of this work was to evaluate the degradation in cell culture medium of dense and porous scaffolds of poly(L-lactic acid) (PLLA) and poly(D,L-lactic acid-co-glycolic acid) (50:50) (PLGA₅₀) prepared by casting. The adhesion and morphology of osteoblast cells on the surface of these polymers was evaluated. Thermal analyses were done by differential scanning calorimetry and thermogravimetric analysis and cell morphology was assessed by scanning electron microscopy. Autocatalysis was observed in PLGA₅₀ samples because of the concentration of acid constituents in this material. Samples of PLLA showed no autocatalysis and hence no changes in their morphology, indicating that this polymer can be used as a structural support. Osteoblasts showed low adhesion to PLLA compared to PLGA₅₀. The cell morphology on the surface of these materials was highly dispersed, which indicated a good interaction of the cells with the polymer substrate.     

有机苯磷酸钙对聚L-乳酸结晶性能和熔融行为的影响

为拓宽聚乳酸结晶成核剂范畴,以苯磷酸和氯化钙为原料采用溶液法制备得到层间距为1.544 nm的有机苯磷酸钙(CP),再将其添加到聚L-乳酸(PLLA)基体进行非等温结晶性能和熔融行为研究。非等温结晶性能测试显示CP对PLLA具有异相成核作用,能显著提高其结晶性能;0.7%CP对PLLA的结晶成核效应最佳,在1℃/min的降温速率下,可使PLLA的结晶起始温度从105.88℃增加到115.47℃,结晶焓从1.379 J/g提高到33.90 J/g。熔融行为测试表明PLLA/CP材料的熔融双峰遵循熔融―再结晶机理。     

Formation of nano-patterned protein layers on poly (lactic acid) surface: Induced by dewetting process and self-assembly

A simple method was developed to fabricate nano-patterned protein layers on poly (lactic acid) (PLA) surface by dewetting method. It was found that nano-patterned protein layers were formed on PLA surface by slow drying in high humidity. The influence of wettability and viscoelastic properties of substrates were also investigated. As a result, typical spinodal dewetting patterns formed for BSA. However, network structures were obtained for collagen at acidic condition and well-oriented fibrils at neutral pH. It was attributed to dewetting on hydrophobic substrate as well as the plasticity and variety of collagen self-assemblies.