Transplantation of Nogo-66 receptor gene-silenced cells in a poly(D,L-lactic-co-glycolic acid) scaffold for the treatment of spinal cord injury

Inhibition of neurite growth, which is in large part mediated by the Nogo-66 receptor, affects neural regeneration following bone marrow mesenchymal stem cell transplantation. The tissue engineering scaffold poly（D,L-lactide-co-glycolic acid） has good histocompatibility and can promote the growth of regenerating nerve fibers. The present study used small interfering RNA to silence Nogo-66 receptor gene expression in bone marrow mesenchymal stem cells and Schwann cells, which were subsequently transplanted with poly（D,L-lactide-co-glycolic acid） into the spinal cord lesion regions in rats. Simultaneously, rats treated with scaffold only were taken as the control group. Hematoxylin-eosin staining and immunohistochemistry revealed that at 4 weeks after transplantation, rats had good motor function of the hind limb after treatment with Nogo-66 receptor gene-silenced ceils prus the poly（O,L-lactide-co-glycolic acid） scaffold compared with rats treated with scaffold only, and the number of bone marrow mesenchymal stem cells and neuron-like cells was also increased. At 8 weeks after transplantation, horseradish peroxidase tracing and transmission electron microscopy showed a large number of unmyelinated and myelinated nerve fibers, as well as intact regenerating axonal myelin sheath following spinal cord hemisection injury. These experimental findings indicate that transplantation of Nogo-66 receptor gene-silenced bone marrow mesenchymal stem cells and Schwann cells plus a poly（D,L-lactide-co-glycolic acid） scaffold can significantly enhance axonal regeneration of spinal cord neurons and improve motor function of the extremities in rats following spinal cord injury.

Synthesis of the Biomimetic Polymer: Aliphatic Diamine and RGDS Modified Poly(d,l-lactic acid)

A novel poly（d, /-lactic acid） （PDLLA） based biomimetic polymer was synthesized by grafting maleic anhydride, butanediamine and arg-gly-asp-ser （RGDS） peptides onto the backbone of PDLLA, aiming to overcome the acidity and auto-accelerating degradation of PDLLA during degradation and to improve its biospecificity and biocompatibility. The synthetic copolymer was characterized by FTIR, ^13C NMR and amino acid analyzer （AAA）.

Synthesis and Characterization of Chitosan-g-poly- (D, L-lactic acid) Copolymer

Biodegradable chitosan-g-poly (D, L-lactic acid) copolymers were prepared via two methods. (1) The lactide was grafted onto hydroxyl groups of chitosan by using macromolecular initiator sodium of trimethylsilyl-chitosan, (2) poly (D,L-lactic acid)(PLA) with low molecular weight can be linked to the amino group by coupling activated PLA to trimethylsilyl-chitosan. Two graft copolymers had hydrophilic-hydrophobic character and can be applied as carriers for drug delivery.

Honeycomb-structured solid acid catalysts fabricated via the swelling-induced self-assembly of acidic poly(ionic liquid)s for highly efficient hydrolysis reactions

The development of heterogeneous acid catalysts with higher activity than homogeneous acid catalysts is critical and still challenging.In this study,acidic poly(ionic liquid)s with swelling ability(SAPILs)were designed and synthesized via the free radical copolymerization of ionic liquid monomers,sodium p-styrenesulfonate,and crosslinkers,followed by acidification.The 31P nuclear magnetic resonance chemical shifts of adsorbed trimethylphosphine oxide indicated that the synthesized SAPILs presented moderate and single acid strength.The thermogravimetric analysis results in the temperature range of 300–345°C revealed that the synthesized SAPILs were more stable than the commercial resin Amberlite IR-120(H)(245°C).Cryogenic scanning electron microscopy testing demonstrated that SAPILs presented unique three-dimensional(3D)honeycomb structure in water,which was ascribed to the swelling-induced self-assembly of the molecules.Moreover,we used SAPILs with micron-sized honeycomb structure in water as catalysts for the hydrolysis of cyclohexyl acetate to cyclohexanol,and determined that their catalytic activity was much higher than that of homogeneous acid catalysts.The equilibrium concentrations of all reaction components inside and outside the synthesized SAPILs were quantitatively analyzed using a series of simulated reaction mixtures.Depending on the reaction mixture,the concentration of cyclohexyl acetate inside SAPIL-1 was 7.5–23.3 times higher than that outside of it,which suggested the high enrichment ability of SAPILs for cyclohexyl acetate.The excellent catalytic performance of SAPILs was attributed to their 3D honeycomb structure in water and high enrichment ability for cyclohexyl acetate,which opened up new avenues for designing highly efficient heterogeneous acid catalysts that could eventually replace conventional homogeneous acid catalysts.

Microwave-assisted Polymerization of D,L-Lactide with Stannous Octanoate as Catalyst

Poly (lactic acid) (PLA) was synthesized by microwave-assisted ring-opening polymerization of D, L-lactide with stannous octanoate (SnOct(2)) as catalyst. Its weight-average molar mass (M-w) ranged from 39000 to 67000 and the polydispersity index from 1.3 to 1.7. The polymerization rate was much faster than that of the conventional thermal polymerization. A degradation of newly formed PLA in reaction mixture by microwave irradiation was observed.

Preparation and Degradation Characteristics of Poly(d,1-laetide) Composite Films

PDLLA/CHI/β-TCP/NGF composite films were prepared by a solvent evaporation method. The degradation characteristics of the poly （d, l-lactide） composite films were studied in vitro and in vivo. The acidity produced by poly （d, l-lactide） materials was not obvious. Adding chitosan and β-TCP can relieve the acidity problem and improve strength performance of films. The NGF has influences on the degradation characteristics of films. It is verified that PDLLA/CHI/β-TCP/NGF composite films prepared by solvent evaporation method have excellent degradation characteristics. It can be used as a perfect biomaterial for repairing nerve injuries.

Preparation of a novel biodegradable β-cyclodextrin-containing polymer

A novel cyclodextrin-containing polymer was prepared by graftingβ-cyclodextrin onto the backbone of poly(D,L-lactic acid)(PLA).First,mono(6-(2-aminoethyl)amino-6-deoxy)-β-cyclodextrin(β-CD-6-en)was prepared by sulfonylation and amination ofβ-cyclodextrin and modified poly(D,L-lactic acid)(MPLA)was prepared by free radical polymerization of maleic anhydride and PLA.Then,grafting ofβ-cyclodextrin derivative to MPLA backbone was carried out by N-acylation reaction of MPLA andβ-CD-6-en in dimethyl formamide.The...

Interaction of Immune System Protein with PEGylated and Un-PEGylated Polymeric Nanoparticles

Biodegradable Nanoparticles (NPs) are under intense investigation due to their potential application in targeted drug delivery. Upon their entry to the biological system, they encounter the immune system, which limits their availability at the intended site. Most importantly, the innate immune system is the one that acts as the first line of defense against foreign materials. It can be activated by collectin proteins which recognize the structural pattern of polysaccharide on the surface of microorganisms. NPs may interact with these proteins in a similar way, and the interaction may lead to beneficial outcomes in vaccine delivery. On the other hand, in targeted drug delivery, it is desirable for the NPs not to be recognized as foreign material as this may lead to their fast elimination from the system through mechanism such as opsonization. We investigated the interaction of PEGylated and un-PEGylated PLGA NPs with Recombinant Human Mannose-Binding Protein (HMBP) in an effort to understand the effect of surface modification on their binding to the protein. Results show that both PLGA-COOH and PLGA-PEG-NH2 bind to HMBP as studied using dynamic light scattering (DLS), fluoresce and UV-vis spectroscopy. However, their binding is shown to have different effect on the structure of the protein. Study done using fluorescence spectroscopy displayed a decrease in fluorescence emission of the protein upon binding to PLGA-COOH. On the other hand the fluorescence emission of the protein increased upon binding to the PLGA-PEG-NH2 indicating conformational changes in the protein structure.

线形聚(D,L-乳酸)和星形聚(D,L-乳酸)的合成研究

在辛酸亚锡Sn(Oct)_(2)催化下,分别用丙炔醇和季戊四醇引发D,L-丙交酯(DLLA)的开环聚合(ROP),合成了具有末端炔基的线形聚(D,L-乳酸)(PDLLA)和四臂星形聚(D,L-乳酸)(S-(PDLLA)4),通过核磁共振氢谱(^(1)H NMR)和凝胶渗透色谱(GPC)对聚合物的结构和相对分子质量进行了表征。合成产物可用于后续制备基于聚乳酸的两亲性嵌段共聚物,为生物可降解性高分子材料的开发应用提供实验依据。

Co-transplantation of neural stem cells and Schwann cells within poly （L-lactic-co-glycolic acid） scaffolds facilitates axonal regeneration in hemisected rat spinal cord

Background Various tissue engineering strategies have been developed to facilitate axonal regeneration after spinal cord injury. This study aimed to investigate whether neural stem cells （NSCs） could survive in poly（L-lactic-co-glycolic acid） （PLGA） scaffolds and, when cografted with Schwann cells （SCs）, could be induced to differentiate towards neurons which form synaptic connection and eventually facilitate axonal regeneration and myelination and motor function. Methods NSCs and SCs which were seeded within the directional PLGA scaffolds were implanted in hemisected adult rat spinal cord. Control rats were similarly injured and implanted of scaffolds with or without NSCs. Survival, migration, differentiation, synaptic formation of NSCs, axonal regeneration and myelination and motor function were analyzed. Student＇s t test was used to determine differences in surviving percentage of NSCs. One-way analysis of variance （ANOVA） was used to determine the differences in the number of axons myelinated in the scaffolds, the mean latency and amplitude of cortical motor evoked potentials （CMEPs） and Basso, Beattie ＆ Bresnahan locomotor rating scale （BBB） score. The X2 test was used to determine the differences in recovery percentage of CMEPs. Results NSCs survived, but the majority migrated into adjacent host cord and died mostly. Survival rate of NSCs with SCs was higher than that of NSCs without SCs （（1.7831±0.0402）% vs. （1.4911±0.0313）%, P 〈0.001）. Cografted with SCs, NSCs were induced to differentiate towards neurons and might form synaptic connection. The mean number of myelinated axons in PLGA＋NSCs＋SCs group was more than that in PLGA＋NSCs group and in PLGA group （（110.25±30.46） vs. （18.25±3.30） and （11.25±5.54）, P 〈0.01）. The percentage of CMEPs recovery in PLGA＋NSCs＋SCs group was higher than in the other groups （84.8% vs, 50.0% and 37.5%, P 〈0.05）. The amplitude of CMEPs in PLGA＋NSCs＋SCs group was higher than in the other groups （（1452.63±331.70） IJV vs. （428.84±193.01） IJV and （117.33±14.40） μV, P 〈0.05）. Ipsilateral retransection resulted in disappearance again and functional loss of CMEPs for a few days. But contralateral retransection completely damaged the bilateral motor function. Conclusions NSCs can survive in PLGA scaffolds, and SCs promote NSCs to survive and differentiate towards neurons in vivo which even might form synaptic connection. The scaffolds seeded with cells facilitate axonal regeneration and myelination and motor function recovery. But regenerating axons have limited contribution to motor function recovery.

Poly(D,L-lactic-co-glycolic acid)-based artesunate nanoparticles:formulation,antimalarial and toxicity assessments

Objective： The aim of this study was to formulate polymer-based artesunate nanoparticles for malaria treatment. Methods： Artesunate was loaded with poly（D,L-lactic-co-glycolic acid） （PLGA） by solvent evaporation from an oil-in-water single emulsion. Nanoparticles were characterized by X-ray diffraction and differential scanning calo- rimetry analyses. In vivo antimalarial studies at 4 mg/kg were performed on Swiss male albino mice infected with Plasmodium berghei. Hematological and hepatic toxicity assays were performed. In vitro cytotoxicity of free and en- capsulated artesunate （Art-PLGA） to cell line RAW 264.7 was determined at concentrations of 7.8-1000 pg/ml. Re- sults： The particle size of the formulated drug was （329.3±21.7） nm and the entrapment efficiency was （38.4±10.1）%. Art-PLGA nanoparticles showed higher parasite suppression （62.6%） compared to free artesunate （58.2%, P〈0.05）. Platelet counts were significantly higher in controls （305 000.00±148 492.40） than in mice treated with free artesunate （139 500.00±20 506.10） or Art-PLGA （163 500.00±3535.53） （P〈0.05）. There was no sign of hepatic toxicity following use of the tested drugs. The half maximal inhibitory concentration （IC50） of Art-PLGA （468.0 pg/ml） was significantly higher （P〈0.05） than that of free artesunate （7.3 pg/ml） in the in vitro cytotoxicity assay. Conclusions： A simple treatment of PLGA-entrapped artesunate nanoparticles with dual advantages of low toxicity and better antiplasmodial efficacy has been developed.

超高分子量聚D,L乳酸小夹板螺钉行颧弓骨折内固定的动物实验研究

目的：观察新型可吸收性骨夹板的骨内固定效果。方法：采用自身对照研究方法，用超高分子量生物降解材料聚Ｄ，Ｌ乳酸制作小夹板、螺钉内固定装置，对１０只狗进行颧弓骨折内固定，并与金属钛小夹板、螺钉内固定相比较。结果：对颧弓凹陷性骨折，分子量高达６０万ｋＤ以上的超高分子量聚Ｄ，Ｌ乳酸小夹板螺钉的机械性能足够维持骨段的稳定性并支撑其愈合。结论：聚Ｄ，Ｌ乳酸小夹板、螺钉同金属钛小夹板、螺钉内固定一样，对颧弓骨折具有良好的骨内固定效果，对骨折愈合过程无任何不良影响，其显著优点是无需二次手术取出。

聚乙二醇单甲醚一聚(D,L-乳酸)嵌段共聚物的研究

采用熔融缩聚反应合成一系列聚(D,L-乳酸)(PDLLA)/聚乙二醇单甲醚(mPEG)两亲性二嵌段共聚物(PEDLLA),采用IR、1H-NMR、DSC、WAXD和TEM等手段分析和研究PEDLLA的结构与性能。实验结果表明,PEDLLA的结构和组成与设计相一致,结晶度和熔点均低于均聚物,且随着PEDLLA中PDLLA含量的增加,mPEG嵌段熔点降低,随着PDLLA嵌段相对分子质量的增大,PEDLLA降解速率增大。载药纳米粒呈核壳结构,载药量达30%。

聚D,L-乳酸中间体——D,L-丙交酯的合成

聚D,L-乳酸(PDLLA)的合成方法主要有直接法和间接法两种。间接法合成PDLLA的关键在于中间体D,L-丙交酯(DLLA)的制备。文中以D,L-乳酸为原料,在催化剂辛酸亚锡(ZnO ct2)的作用下,合成DLLA。研究温度、压力、时间等因素对DLLA产率的影响,利用DSC、XRD、IR等分析测试技术对DLLA的性能和结构进行相关的测试和表征。研究结果表明,在反应体系中加入乙二醇作为稀释剂,能降低DLLA的馏出温度,避免了DLLA在蒸馏过程中产生炭化现象,并使DLLA的平均产率由36.8%提高至51.3%。

聚(D,L-乳酸)微球的制备与释药性能研究

以溶液缩聚法制得的聚(D,L-乳酸)(PDLLA)为载体材料,聚乙烯醇为乳化剂,二氯甲烷为溶剂,采用乳化-溶剂挥发法制备了PDLLA微球和PDLLA/吡虫啉、PDLLA/啶虫脒载药缓释微球。考察了油相PDLLA用量、水相聚乙烯醇用量以及水油相体积比对微球粒径的影响,并采用紫外分光光度法探讨了PDLLA/吡虫啉、PDLLA/啶虫脒缓释微球的释药性能。结果表明,工艺条件对微球粒径有显著影响,当PDLLA用量为8%(质量分数),聚乙烯醇用量1%(质量分数),水油相体积比2∶1时,可得光滑完整的微球。

胆固醇-聚(D,L-乳酸)的制备及软骨细胞相容性研究

依据胆固醇对细胞膜良好的热力学亲和力及可以调节细胞膜磷脂双分子层的结构和动态行为的生理学的特点,采用本体聚合的方法制备了胆固醇-聚(D,L-乳酸),经GPC,1H-NMR分析证明产物分子量与理论分子量有较好的一致性,证明了聚合物的分子量可以通过调节D,L-丙交酯和胆固醇的量来控制.软骨细胞粘附率、增殖率以及3-(4,5-二甲基噻唑)-2,5-二苯基四氮唑溴盐(MTT)细胞活性的观测结果表明相比纯聚乳酸,胆固醇-聚乳酸明显促进了细胞的粘附与生长.通过激光共聚焦显微镜观察细胞形貌结果显示相比胆固醇-聚乳酸表面,纯聚乳酸表面的细胞数量较少且大多团聚没有正常铺展,而胆固醇-聚乳酸表面的软骨细胞则分布均匀,铺展充分.以上结果证明了胆固醇的存在有利于软骨细胞的生长和粘附,预示着胆固醇-聚乳酸在软骨组织工程以及其他相关的生物医疗领域有着广阔的应用前景.

聚乙二醇甲醚聚(D,L-乳酸)嵌段共聚物纳米胶束的制备

采用熔融缩聚反应合成了一系列聚 (D ,L 乳酸 ) (PDLLA) /聚乙二醇单甲醚 (mPEG)两亲性二嵌段共聚物 (PEDLLA) ,采用纳米沉淀技术制备PEDLLA纳米胶束 ,运用动态光散射法、氢核磁光谱法、芘荧光探针技术、紫外光谱法研究了PEDLLA纳米胶束。结果表明 ,胶束呈核壳结构 ,粒径为纳米级 ,受PDLLA链段相对分子质量和有机相中共聚物浓度的影响 ,PEDLLA的临界胶束浓度 (CMC)小于 2 4 3× 10 -6mol/L ,且随着PDLLA链段相对分子质量的增大而减小。

外消旋乳酸直接熔融聚合-二异氰酸酯TDI熔融扩链的研究

用2,4-甲苯二异氰酸酯(TDI)对以外消旋乳酸(D,L-LA)直接熔融聚合合成的低分子量聚外消旋乳酸(PDLLA)进行扩链,用粘均分子量、红外光谱(IR)、核磁共振氢谱(1H NMR)、DSC、X-射线衍射等对扩链产物进行了表征。在SnCl2催化下,NCO/OH=1,160℃下反应60min,可得到Mη为扩链前3.42倍的可溶性聚乳酸类生物降解材料。IR1、H NMR表明熔融聚合-TDI熔融扩链产物引入了NH和苯环,DSC、XRD等表征发现扩链使Tg、Tm和结晶度等相应提高。但是,温度过高、时间过长、NCO/OH过大时,易生成可溶性差的交联副产物,较多副产物的混杂会导致Tg和结晶性的降低。

胆固醇-聚(D,L-乳酸)的直接熔融聚合法制备与表征

取代传统的丙交酯开环共聚法,直接以外消旋乳酸(D,L-LA)为原料,以廉价的氯化亚锡为催化剂,采用熔融聚合法合成胆固醇-聚(D,L-乳酸)共聚物。分别考察不同投料比、预聚时间和熔融共聚时间的影响,并用FT-IR1、HNMR、GPC、DSC等进行了系统表征。当投料比n(胆固醇)∶n(乳酸)=1∶66、预聚4 h、熔融共聚10 h、催化剂用量0.3%时,以85%的产率生成了重均相对分子质量(Mw)达3 600(分散度Mw/Mn=1.24)的共聚物。新方法步骤少、路线简捷,相对分子质量可以达到丙交酯开环共聚法的水平,且成本更加低廉。