Long-Term Effect of Gamma Irradiation on the Functional Properties and Cytocompatibility of Multiblock Co-Polymer Films

Abstract

The purpose of this work was to investigate the long-term effect of gamma-irradiation treatment on the functional properties of PEG-PDLLA and PEG-PLGA films and to evaluate the cytocompatibility of sterilized samples. Chemical and thermal properties, and cytocompatibility of sterilized films were detected for samples at time zero and after storage at 5 ± 3°C for 60 days. An in vitro degradation study was carried out on polymer samples to examine the effect of sterilization on the degradation performances of co-polymer films. Incubated samples were characterized in terms of film surface structure (SEM), chemical (GPC) and thermal (DSC) properties. The study performed on films upon gamma sterilization showed no significant changes of the PEG-PDLLA and PEG-PLGA film structure, while GPC analysis highlighted that the effect of gamma irradiation was dependent on the M_w and composition of polymers. DSC traces suggested more pronounced gamma-ray effects on the PEG-PLGA multiblock co-polymer. During the stability study important changes in terms of structure surface, thermal properties and cytocompatibility were observed and investigated. Data collected during the in vitro degradation study emphasized the need to know and investigate the degradation performances and behaviour of polymer or polymer systems (as DDS, scaffolds and bandage) treated with gamma rays.     

微沟槽图形结构对硅橡胶表面性能和细胞相容性的影响

目的 制作具有微沟槽图形的硅橡胶(silicone rubber,SR)表面改性材料,检测改性前后材料表面理化性质,初步评价细胞相容性.方法 利用具有微沟槽图形的方形母版,采用浇灌的方法成功制作表面具有微沟槽图形的硅橡胶改性材料,同时进行碳离子注入处理,并采用扫描电镜(scanning electron microscope,SEM)、原子力显微镜(atomic force microscope,AFM)、水接触角等技术测量改性前后硅橡胶的表面理化性质.通过细胞黏附实验、CCK-8细胞增殖实验以及Western blot等技术观察人真皮成纤维细胞的生长情况以及Talin、Zyxin蛋白的表达.结果 AFM观察到各组材料表面形貌出现明显的差异,微沟槽硅橡胶(patterned SR,P-SR)以及微沟槽-碳硅橡胶(patterned carbon ion implanted SR,PC-SR)均可在表面见微沟槽图形,而碳硅橡胶(carbon ion implanted SR,C-SR)出现不规则凸凹结构.水接触角显示材料进行微沟槽图形化处理后,可增加材料的水接触角.细胞黏附和增殖实验提示对材料进行微沟槽图形化处理后,细胞的黏附和增殖能力较C-SR降低,但两者差异无统计学意义.免疫荧光观察到人真皮成纤维细胞在P-SR和PC-SR组排列有序.Western blot检测细胞黏附相关蛋白Talin、Zyxin的表达在C-SR和PC-SR组高于SR组(P＜0.05).结论 微沟槽图形可影响硅橡胶的表面性能和细胞的增殖、排列等功能,对减轻包膜挛缩发生率起到积极作用.     

Cytocompatibility of novel extracellular matrix protein analogs of biodegradable polyester polymers derived from α-hydroxy amino acids

One of the challenges in regenerative medicine is the development of novel biodegradable materials to build scaffolds that will support multiple cell types for tissue engineering. Here we describe the preparation, characterization, and cytocompatibility of homo- and hetero-polyesters of α-hydroxy amino acid derivatives with or without lactic acid conjugation. The polymers were prepared by a direct condensation method and characterized using gel permeation chromatography, ¹H-nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, optical activity, and solubility. The surface charge of the polymers was evaluated using zeta potential measurements. The polymers were coated onto glass cover slips followed by characterization using nano-surface profiler, thin film reflectometry, and atomic force microscopy (AFM). Their interaction with endothelial and neuronal cells was assessed using adhesion, proliferation, and differentiation assays. Of the characterized polymers, Poly-HOVal-LA, but not Poly-(D)HOPhe, significantly augmented nerve growth factor (NGF)-induced neuronal differentiation of the PC12 pheochromcytoma cells. In contrast, Poly-HOLeu increased by 20% the adhesion of endothelial cells, but did not affect PC12 cell differentiation. NGF-induced Erk1/2 phosphorylation in PC12 cells grown on the different polymers was similar to the effect observed for cells cultured on collagen type I. While no significant association could be established between charge and the differentiative/proliferative properties of the polymers, AFM analysis indicated augmentation of NGF-induced neuronal differentiation on smooth polymer surfaces. We conclude that overall selective cytocompatibility and bioactivity might render α-hydroxy amino acid polymers useful as extracellular matrix-mimicking materials for tissue engineering.     

Synthesis,biocompatible, and self-assembly properties of poly (ethylene glycol)/lactobionic acid-grafted chitosan

Polymers with targeted ligands are widely used as the anti-cancer drug delivery materials. For applications of chitosan as an anti-liver cancer drug delivery, poly (ethylene glycol)/lactobionic acid-grafted chitosan (PEG/LA-CS) was prepared and investigated since lactobionic acid can be specifically recognized by the hepatocytes. The structure of the PEG/LA-CS was characterized by Fourier transform infrared spectrometry and elemental analysis. The self-assembly behaviors of the PEG/LA-CS were monitored by steady-state fluorescence spectroscopy and electronic transmission microscope. The protein adsorption of the PEG/LA-CS was detected with bovine serum albumin (BSA) by electrochemical impedance spectroscopy. The results showed that the PEG/LA-CS almost did not adsorb protein. To study the effects of PEG/LA-CS on the structure of BSA, the interactions between the PEG/LA-CS and BSA were detected by ultraviolet spectrum, fluorescence spectrum, and circular dichroism. All the data gave one result that BSA maintained its original folded confirmation in PEG/LA-CS solution. The hemocompatibility of PEG/LA-CS was investigated by observing the effects of PEG/LA-CS on the hemolysis rate and the plasma recalcification time (PRT). The results showed that the PRT was prolonged greatly and the hemolysis rate was less than 5%. Furthermore, PEG/LA-CS also showed good cytocompatibility with K562, Hep G2, and LO2 cells. Therefore, the PEG/LA-CS is believed to have great potential for producing injectable anti-liver cancer drug delivery.     

Nanolayered Metal Phosphates as Biocompatible Reservoirs for Antimicrobial Silver Nanoparticles

There is an increasing demand on synthesizing pharmaceuticals and biomaterials that possess antimicrobial and/or antiviral activities. In this respective silver nanoparticles are known for their excellent antimicrobial activity. Nevertheless, their uncontrolled release in a biological medium can induce a cytotoxic effect. For this, we explored the use of nanolayered metal phosphates based on titanium and zirconium as materials that can be enriched with silver nanoparticles. Employing the hydrothermal route, crystalline α-phases of zirconium and titanium phosphates (α-ZrP, α-TiP) were synthesized and there after surface-enriched with silver nanoparticles. The structural assessment confirmed the stability of the structures and their sizes are in the nanoscale at least in one dimension. The cytocompatibility assays confirmed the biocompatibility of the pristine phases and the antimicrobial assay confirmed that both silver-enriched nanolayered structures maintain an antibacterial effect at reasonably low concentrations.     

Phospholipid‐induced silk fibroin hydrogels and their potential as cell carriers for tissue regeneration

Silk fibroin (SF) hydrogels can be obtained via self‐assembly, but this process takes several days or weeks, being unfeasible to produce cell carrier hydrogels. In this work, a phospholipid, namely, 1,2‐dimyristoyl‐sn‐glycero‐3‐phospho‐(1′‐rac‐glycerol) sodium salt (DMPG), was used to induce and accelerate the gelation process of SF solutions. Due to the amphipathic nature and negative charge of DMPG, electrostatic and hydrophobic interactions between the phospholipids and SF chains will occur, inducing the structural transition of SF chains to the beta sheet and consequently a rapid gel formation is observed (less than 50 min). Moreover, the gelation time can be controlled by varying the lipid concentration. To assess the potential of the hydrogels as cell carriers, several mammalian cell lines, including L929, NIH/3T3, SaOS‐2, and CaSki, were encapsulated into the hydrogel. The silk‐based hydrogels supported the normal growth of fibroblasts, corroborating their cytocompatibility. Interestingly, an inhibition in the growth of cancer‐derived cell lines was observed. Therefore, DMPG‐induced SF hydrogels can be successfully used as a 3D platform for in situ cell encapsulation, opening promising opportunities in biomedical applications, such as in cell therapies and tissue regeneration.     

In Vitro Direct and Indirect Cytotoxicity Comparative Analysis of One Pre-Hydrated versus One Dried Acellular Porcine Dermal Matrix

Aim: The aim of the present study was to compare the direct and indirect cytotoxicity of a porcine dried acellular dermal matrix (PDADM) versus a porcine hydrated acellular dermal matrix (PHADM) in vitro. Both are used for periodontal and peri-implant soft tissue regeneration. Materials and methods: Two standard direct cytotoxicity tests—namely, the Trypan exclusion method (TEM) and the reagent WST-1 test (4-3-[4-iodophenyl]-2-[4-nitrophenyl]-2H-[5-tetrazolio]-1,3-benzol-desulphonated)—were performed using human primary mesenchymal stem cells (HPMSCs) seeded directly onto a PDADM and PHADM after seven days. Two standard indirect cytotoxicity tests—namely, lactate dehydrogenase (LTT) and MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazoliumbromide)—were performed using HPMSCs cultivated in eluates from the matrices incubated for 0.16 h (10 min), 1 h, and 24 h in a serum-free cell culture medium. Results: The WST and the TEM tests revealed significantly lower direct cytotoxicity values of HPMSCs on the PHADM compared with the PDADM. The indirect cytotoxicity levels were low for both the PHADM and PDADM, peaking in short-term eluates and decreasing with longer incubation times. However, they were lower for the PHADM with a statistically significant difference (p < 0.005). Conclusions: The results of the current study demonstrated a different biologic behavior between the PHADM and the PDADM, with the hydrated form showing a lower direct and indirect cytotoxicity.     

Manufacture and Cytotoxicity of a Lead-free Piezoelectric Ceramic as a Bone Substitutc Consolidation of Porous Lithium Sodium Potassium Niobate by Cold Isostatic Pressing

Aim The piezoelectric properties and cytotoxicity of a porous lead-free piezoelectric ceramic for use as a direct bone substitute were investigated. Methodology Cold isostatic pressing （CIP） was applied to fabricate porous lithium sodium potassium niobate （Li0.06Na0.5K0.44） NbO3 specimens using a pore-forming method. The morphologies of the CIP-processed specimens were characterized and compared to those of specimens made by from conventional pressing procedures. The effects of the ceramic on the attachment and proliferation of osteoblasts isolated from the cranium of 1-day-old Sprague- Dawley rats were examined by a scanning electron microscopy （SEM） and metbylthiazol tetrazolium （MTT） assay. Results The results showed that CIP enhanced piezoelectricity and biological performance of the niobate specimen, and also promoted an extracellular matrix-like topography of it. In vitro studies showed that the CIP-enhanced material had positive effects on the attachment and proliferation of osteoblasts. Conclusion Niobate ceramic generated by CIP shows a promise for being a piezoelectric composite bone substitute.