The Mechanical and Biological Performance of Photopolymerized Gelatin-Based Hydrogels as a Function of the Reaction Media

From the first experiments with biomaterials to mimic tissue properties, the mechanical and biochemical characterization has evolved extensively. Several properties can be described, however, what should be essential is to conduct a proper and physiologically relevant characterization. Herein, the influence of the reaction media (RM) and swelling media (SM)–phosphate buffered saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) with two different glucose concentrations–is described in gelatin methacrylamide (GelMA) hydrogel mechanics and in the biological behavior of two tumoral cell lines (Caco-2 and HCT-116). All scaffolds are UV-photocrosslinked under identical conditions and evaluated for mass swelling ratio and stiffness. The results indicate that stiffness is highly susceptible to the RM, but not to the SM. Additionally, PBS-prepared hydrogels exhibited a higher photopolymerization degree according to high resolution magic-angle spinning (HR-MAS) NMR. These findings correlate with the biological response of Caco-2 and HCT-116 cells seeded on the substrates, which demonstrated flatter morphologies on stiffer hydrogels. Overall, cell viability and proliferation are excellent for both cell lines, and Caco-2 cells displayed a characteristic apical-basal polarization based on F-actin/Nuclei fluorescence images. These characterization experiments highlight the importance of conducting mechanical testing of biomaterials in the same medium as cell culture.     

Poly(acrylamidoglycolic acid) nanocomposite hydrogels reinforced with cellulose nanocrystals for pH-sensitive controlled release of diclofenac sodium

In this study, a non-cytotoxic and pH-sensitive poly(acrylamidoglycolic acid) based nanocomposite (PAGA-NC) hydrogels reinforced with cellulose nanocrystals (CNCs) was synthesized using redox free radical polymerization. The successful formation and crystalline behaviour of PAGA-NC hydrogels was verified by fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analyses. The results showed that morphological, rheological and mechanical properties of the PAGA-NC hydrogels were strongly influenced by the CNCs content. Moreover, swelling properties were investigated, and the results suggested that they behaved as pH sensitive manner. The in vitro MTT assay showed that the PAGA-NC hydrogels are cytocompatibile to NIH-3T3 fibroblast cells. In addition, diclofenac sodium (DCF) model drug was successfully encapsulated into these PAGA-NC hydrogels via equilibrium swelling method. The in vitro release of DCF from PAGA-NC hydrogels was retained at pH 1.2 and maximum release was observed at 7.4, revealing as potential candidates for controlled release carriers for oral drug delivery applications.     

（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶的制备及其释药作用

将壳聚糖改性为（2-羟基-3-丁氧基）丙基 羟丙基壳聚糖（2-H-3-B-P-HPCS）,并以（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖和聚乙二醇（PEG）为原料制备（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶,研究了（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖浓度、聚乙二醇的用量、交联剂戊二醛用量、反应温度对该凝胶溶胀性能的影响。 通过红外光谱分析和扫描电子显微镜的方法比较了壳聚糖、（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖和（2-羟基-3-丁氧基）丙基-羟丙基壳聚糖/聚乙二醇互穿网络凝胶结构和形态上的不同。 以阿昔洛韦为模型药物研究了其释药性能。 结果表明,该凝胶均具有良好的溶胀性、pH敏感性和药物缓释作用,有望用作新型的药物载体。     

Studies on Intestinal Transport of Ginsenoside Compatibility with Veratrum nigrum via Caco-2 Cell Monolayer Model Coupled with UPLC-ESI-MS Method

The Caco-2 cells have been recognized as effective tools to be applied to imitating the drug absorption in human intestine for the transport of drug. Herein, Caco-2 cell monolayer model was used to study the transport of the ginsenoside compatibility with Veratrum nigrum in different proportions. A specific high performance liquid chromatography-electrospray ionization-mass spectrometry(HPLC-ESI-MS) method was developed for the semiquantitative determination of ginsenoside in intestinal transport with Dioscin as an internal standard. For the Caco-2 model constructed, two influencing factors were investigated, including time and concentration. The results suggest that the absorption of ginsenoside Re, Rg1, Rb1, Rc, Rb2 and Rd are time- and concentration-dependent and the excretions of Rb1, Rc, Rb2 and Rd have a relatronship with some transport proteins. The bioavailability of the ginsenosides has reduced compared to the single Panax ginseng extract when compatibility with a certain amount of Veratrum nigrum.     

Physicochemical characterization and drug release properties of PDMAEMA/OSA Semi‐IPN hydrogels with microporous structure

A new poly(2‐(dimethylamino) ethyl methacrylate)/oxidized sodium alginate (PDMAEMA) semi‐interpenetrating network (Semi‐IPN) hydrogel with microporous structure was prepared by using PDMAEMA microgels as an additive during the polymerization/crosslinking process. The interior morphology characterized by scanning electron microscopy showed the Semi‐IPN hydrogels have different pore sizes by changing the amount of microgels. The hydrogels were also characterized by using Fourier transform infrared and DSC. The swelling behaviors of hydrogels indicated that the hydrogels have excellent pH and temperature sensitivity. Bovine serum albumin was entrapped in the hydrogels and the in vitro drug release profiles were established in different buffer solutions at various temperatures. The release behaviors of the model drug were dependent on the pore size of the hydrogels and environmental temperature/pH, which suggested that these materials have potential application as intelligent drug carriers. Copyright © 2011 John Wiley & Sons, Ltd.     

医用高分子水凝胶的设计与合成

作为一类重要的医用功能材料,高分子水凝胶可望在药物控释、软骨支架构建、活性细胞封装等方面获得广泛应用。综述了基于化学交联和物理交联的有关水凝胶的设计与合成方法,重点介绍了通过自由基共聚反应、结构互补基团间化学反应形成的化学交联水凝胶以及通过荷电相反离子问相互作用、两亲性嵌段或接枝共聚物疏水缔合、结晶与氢键相互作用形成的物理交联水凝胶。     

Hydrophilic silicone hydrogels with interpenetrating network structure for extended delivery of ophthalmic drugs

In the current work, hydrophilic silicone hydrogels were prepared for extended drug delivery applications. The preparation method was based on sequential interpenetrating network synthesis. A hydrophilic network was prepared by radical copolymerization of hydrophilic monomers 2‐hydroxyethyl methacrylate and poly(ethylene glycol) diacrylate. A hydrophobic silicone network was obtained by crosslinking polymerization of bifunctional methacrylated polydimethylsiloxanes macromonomer. The morphology of the silicone hydrogels was characterized by transmission electron microscopy. The result showed that the silicone hydrogels exhibited heterogeneous morphology. The properties of the silicone hydrogels such as equilibrium swelling ratio (ESR), mechanical property, oxygen permeability, contact angle, and protein repelling ability were investigated. Finally, the silicone hydrogels were loaded with timolol by pre‐soaking in drug solution to evaluate drug‐loading capacity and in vitro release behavior. The results showed that mechanical strength and oxygen permeability increased, and the ESR decreased with the increase of silicone component in the silicone hydrogels. The result of the contact angle measurement indicated that the silicone hydrogels possessed hydrophilic surfaces. The drug loading and in vitro releases were dependent on the composition of hydrophilic/hydrophobic phase of silicone hydrogels. Copyright © 2011 John Wiley & Sons, Ltd.     

Drug‐Loaded Elastin‐Like Polypeptide–Collagen Hydrogels with High Modulus for Bone Tissue Engineering

Emphasizing the role of hydrogel stiffness and cellular differentiation, this study develops collagen and elastin‐like polypeptide (ELP)–based bone regenerative hydrogels loaded with recombinant human bone morphogenetic protein‐2 (rhBMP‐2) and doxycycline with mechanical properties suitable for osteogenesis. The drug‐incorporated collagen–ELP hydrogels has significantly higher modulus of 35 ± 5 kPa compared to collagen‐only hydrogels. Doxycycline shows a bi‐phasic release with an initial burst release followed by a gradual release, while rhBMP‐2 exhibits a nearly linear release profile for all hydrogels. The released doxycycline shows anti‐microbial activity against Pseudomonas aeruginosa, Streptococcus sanguinis, and Escherichia coli. Microscopic observation of the hydrogels reveals their interconnected, macroporous, 3D open architecture with pore diameters between 160 and 400 µm. This architecture supports human adipose–derived stem cell attachment and proliferation from initial days of cell seeding, forming a thick cellular sheath by day 21. Interestingly, in collagen and collagen–ELP hydrogels, cell morphology is elongated with stretched slender lamellipodial formation, while cells assemble as spheroidal aggregates in crosslinked as well as drug‐loaded hydrogels. Osteogenic markers, alkaline phosphatase and osteocalcin, are expressed maximally for drug‐loaded hydrogels compared to those without drugs. The drug‐loaded collagen–ELP hydrogels are thus promising for combating bacterial infection and promoting guided bone regeneration.     

Swelling and drug release behavior of poly(2-hydroxyethyl methacrylate/itaconic acid) copolymeric hydrogels obtained by gamma irradiation

The new copolymeric hydrogels based on 2-hydroxyethyl methacrylate (HEMA) and itaconic acid (IA) were prepared by gamma irradiation, in order to examine the potential use of these hydrogels in controlled drug release systems. The influence of IA content in the gel on the swelling characteristics and the releasing behavior of hydrogels, and the effect of different drugs, theophylline (TPH) and fenethylline hydrochloride (FE), on the releasing behavior of P(HEMA/IA) matrix were investigated in vitro. The diffusion exponents for swelling and drug release indicate that the mechanisms of buffer uptake and drug release are governed by Fickian diffusion. The swelling kinetics and, therefore, the release rate depends on the matrix swelling degree. The drug release was faster for copolymeric hydrogels with a higher content of itaconic acid. Furthermore, the drug release for TPH as model drug was faster due to a smaller molecular size and a weaker interaction of the TPH molecules with(in) the P(HEMA/IA) copolymeric networks.     

Synthesis and Evaluation of Biocompatible pH‐Sensitive Hydrogels as Colon‐Specific Drug Delivery Systems

Because of the growing importance of pH‐sensitive hydrogels as drug delivery systems, biocompatible copolymeric hydrogels based N‐vinyl‐2‐pyrrolidinone (NVP) and methacrylic acid (MAA) were designed and synthesized. These hydrogels were investigated for oral drug delivery. Radical copolymerizations of N‐vinyl‐2‐pyrrolidinone (NVP) and methacrylic acid (MAA) with the various ratios of cross‐linking agent were carried out at 70 °C. Azabisisobutyronitrile (AIBN) was the free‐radical initiator employed and Cubane‐1,4‐dicarboxylic acid (CDA) linked to two 2‐hydroxyethyl methacrylate (HEMA) group was the crosslinking agent (CA) used for hydrogel preparations. The hydrogels were characterized by differential scanning calorimetry and FT‐IR. Equilibrium swelling studies were carried out in enzyme‐free simulated gastric and intestinal fluids (SGF and SIF, respectively). A model drug, olsalazine [3,3′‐azobis (6‐hydroxy benzoic acid)] (OSZ) as an azo derivative of 5‐aminosalicylic acid (5‐ASA), was entrapped in these gels and the in‐vitro release profiles were established separately in both enzyme‐free SGF and SIF. The drug‐release profiles indicated that the amount of drug released depended on the degree of swelling. The swelling was modulated by the amount of crosslinking of the polymer bonded drug (PBDs) prepared. Based on the great difference in hydrolysis rates at pH 1 and 7.4, these pH‐sensitive hydrogels appear to be good candidates for colon‐specific drug delivery.     

Cellulose–halloysite nanotube composite hydrogels for curcumin delivery

Halloysite nanotubes (HNTs) were added to cellulose NaOH/urea solution to prepare composite hydrogels using epichlorhydrine crosslinking at an elevated temperature. The shear viscosity, mechanical properties, microstructure, swelling properties, cytocompatibility, and drug delivery behavior of the cellulose/HNT composite hydrogels were investigated. The viscosity of the composite solution increases with the addition of HNT. The compressive mechanical properties of composite hydrogels are significantly improved compared with pure cellulose hydrogel. The compressive strength of the composite hydrogels with 66.7% HNTs is 128 kPa, while that of pure cellulose hydrogel is only 29.8 kPa in compressive strength. Rheological measurement suggests the resistance to deformation is improved for composite hydrogels. X-ray diffraction and Fourier transform infrared spectroscopy show that the crystal structure and chemical structure of HNT are not changed in the composite hydrogels. Hydrogen bonding interactions between HNT and cellulose exist in the composites. A porous structure of the composite hydrogels with pore size of 200–400 μm was found by scanning electron microscopy. The addition of HNT leads to decreased swelling ratios in NaCl solution and pure water for the composite hydrogels. Cytotoxicity assays show that the cellulose/HNT composite hydrogels have a good biocompatibility with MC3T3-E1 cells and MCF-7 cells. Curcumin is further loaded into the composite hydrogel via physical adsorption. The curcumin-loaded composite hydrogels show a strong inhibition effect on the cancer cells. All the results illustrate that the cellulose/HNT composite hydrogels have promising applications such as anticancer drug delivery systems and anti-inflammatory wound dressings.     

Designing of pH-responsive ketorolac tromethamine loaded hydrogels of alginic acid: Characterization,in-vitro and in-vivo evaluation

pH-responsive hydrogels based on alginic acid grafted with acrylic acid and ethylene glycol dimethylacrylate in the presence of ammonium persulfate were developed for controlled delivery of Ketorolac tromethamine. The alginic acid based hydrogels were prepared by free radical polymerization technique. Increase in gel fraction was observed with the increase in alginic acid, acrylic acid, and ethylene glycol dimethylacrylate content. The dynamic swelling and drug release studies were conducted at two different pH values (pH 1.2 and 7.4). Maximum swelling and drug release were observed at pH 7.4. The characterization of prepared hydrogels was carried out by using Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, powder x-ray diffraction, and scanning electron microscopy. Similarly, in-vivo study was performed on rabbits and greater plasma drug concentration was achieved by fabricated hydrogels as compared to drug solution and commercial product Keten. Conclusively, the fabricated hydrogels can be considered as a potential candidate for controlled delivery of Ketorolac tromethamine.     

Synthesis, characterisation and evaluation of poly[lactose acrylate-N-vinyl-2-pyrrolidinone] hydrogels for drug delivery

New biodegradable polymeric hydrogels based on biocompatible materials, lactose acrylate (LA) and N-vinyl-2-pyrrolidinone were designed and synthesized. LA was synthesized and characterized by Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy. Hydrogel synthesis was carried out by free-radical polymerization of the co-monomers using azobisisobutyronitrile as initiator and N,N^′-methylenebisacrylamide as crosslinker. These hydrogels were also characterized. Equilibrium swelling of the hydrogels was studied in phosphate buffer of physiological pH, 7.4 and at 37 °C. Propranolol hydrochloride was entrapped into these hydrogels and the in vitro release profile of this drug was established in phosphate buffer. The drug release followed a near zero-order fashion in the first 6 h and thereafter slowed down releasing more than 90% of the entrapped drug at the end of 48 h.     

Carnosic Acid Encapsulated in Albumin Nanoparticles Induces Apoptosis in Breast and Colorectal Cancer Cells

Carnosic acid (CA) is a natural phenolic compound with several biomedical actions. This work was performed to study the use of CA-loaded polymeric nanoparticles to improve the antitumor activity of breast cancer cells (MCF-7) and colon cancer cells (Caco-2). CA was encapsulated in bovine serum albumin (BSA), chitosan (CH), and cellulose (CL) nanoparticles. The CA-loaded BSA nanoparticles (CA-BSA-NPs) revealed the most promising formula as it showed good loading capacity and the best release rate profile as the drug reached 80% after 10 h. The physicochemical characterization of the CA-BSA-NPs and empty carrier (BSA-NPs) was performed by the particle size distribution analysis, transmission electron microscopy (TEM), and zeta potential. The antitumor activity of the CA-BSA-NPs was evaluated by measuring cell viability, apoptosis rate, and gene expression of GCLC, COX-2, and BCL-2 in MCF-7 and Caco-2. The cytotoxicity assay (MTT) showed elevated antitumor activity of CA-BSA-NPs against MCF-7 and Caco-2 compared to free CA and BSA-NPs. Moreover, apoptosis test data showed an arrest of the Caco-2 cells at G₂/M (10.84%) and the MCF-7 cells at G2/M (4.73%) in the CA-BSA-NPs treatment. RT-PCR-based gene expression analysis showed an upregulation of the GCLC gene and downregulation of the BCL-2 and COX-2 genes in cells treated with CA-BSA-NPs compared to untreated cells. In conclusion, CA-BSA-NPs has been introduced as a promising formula for treating breast and colorectal cancer.     

Microscale 3-D hydrogel scaffold for biomimetic gastrointestinal (GI) tract model

Here we describe a simple and efficient method for fabricating natural and synthetic hydrogels into 3-D geometries with high aspect ratio and curvature. Fabricating soft hydrogels into such shapes using conventional techniques has been extremely difficult. Combination of laser ablation and sacrificial molding technique using calcium alginate minimizes the stress associated with separating the mold from the hydrogel structure, and therefore allows fabrication of complex structures without damaging them. As a demonstration of this technique, we have fabricated a microscale collagen structure mimicking the actual density and size of human intestinal villi. Colon carcinoma cell line, Caco-2 cells, was seeded onto the structure and cultured for 3 weeks until the whole structure was covered, forming finger-like structures mimicking the intestinal villi covered with epithelial cells. This method will enable construction of in vitro tissue models with physiologically realistic geometries at microscale resolutions.     

pH and glucose dually responsive injectable hydrogel prepared by in situ crosslinking of phenylboronic modified chitosan and oxidized dextran

Injectable hydrogels with pH and glucose triggered drug release capability were synthesized based on biocompatible phenylboronic modified chitosan and oxidized dextran through the formation of covalent imine bond and phenylboronate ester. Rheological characterization demonstrated that the gelation rate was rapid, and the moduli of the hydrogels were able to be tuned with chemical composition as well as pH and glucose concentration of the polymer solution. Anticancer drugs could be incorporated inside the hydrogel through the in situ gel forming process and undergo a controlled release by altering pH or glucose concentration. The hydrogels had good biocompatibility with viable and proliferated cells cultured in the three dimensional matrix, and the cell proliferation was suppressed when a small amount of DOX was added, which is benefit for the application of the hydrogels as smart anticancer drug delivery system. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2015 , 53, 1235–1244     

Chondroitin/polypyrrole nanocomposite hydrogels for the accurate release of 5-fluorouracil by electrical stimulation

The development of electro-stimulated drug release devices is an innovative approach to attain the drug delivery in accurate doses at target sites in a programmed manner. In this work, novel electroactive nanocomposite hydrogels were prepared by encapsulating green-synthesized polypyrrole (PPy) colloids within chondroitin sulfate (CS) networks during the self-crosslinking of CS via N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide chemistry. The structural and morphological properties of CS/PPy hydrogels were studied by Fourier-transformed infrared spectroscopy, scanning electron microscopy, and swelling kinetic measurements. The chemotherapeutic agent 5-fluorouracil (5-FU) was loaded into CS/PPy samples by hydrogel swelling method, or alternatively, by pre-incubating the drug in polymer mixture before crosslinking. Different electrical stimulations can be used to switch ON and accurately tune the 5-FU delivery from GG/PPy hydrogels. A single pulse potential of 5 V switched on the drug delivery up to 90% from nanocomposite hydrogel, in contrast to the low 5-FU amount released in a passive form (< 20%). PPy electroactive behavior played a determining role as the main driving force in 5-FU release activation. Cytotoxicity of hydrogels with and without 5-FU was examined in normal and cancer cells. Considering the high cytotoxicity of 5-FU, the ON/OFF 5-FU release patterns evidenced the potential of CS/PPy hydrogels for electrically controlled drug delivery in implantable or transdermal drug release devices.     

pH敏感型聚谷氨酸/透明质酸基互穿网络医用水凝胶的制备及表征

生物材料是推动生物医学领域日新月异变化的基石,医用水凝胶作为重要成员,近年来表现出蓬勃发展的态势。文章介绍了一种新型可注射的、以生物相容性方法交联的聚谷氨酸(Poly (γ-glutamic acid), PGA)/透明质酸(Hyaluronic acid, HA)复合水凝胶。研究首先采用EDC/NHS方法合成了酪胺(Tyramine,Ty)接枝聚谷氨酸的PGA-Ty前体大分子及半胱胺(Cysteamine, CA)修饰透明质酸的HA-CA前体大分子。两种前体大分子的结构分别使用核磁和红外进行了确证。得到的两种前体大分子在低浓度双氧水和辣根过氧化物酶(Horseradish Peroxidase, HRP)的共同作用下,于水相中交联得到互穿网络(Interpenetrating Network, IPN)水凝胶。实验对IPN水凝胶样品的系列性能,如平衡含水量、内部形貌、酶降解速率以及力学性能等进行了测试,并选取了盐酸四环素为药物模型对凝胶的体外药物释放行为、体外抗菌效果进行了测评。凝胶材料的细胞毒性及凝胶支架对细胞3D培养的效果证明其生物相容性优异,体外包埋的细胞经72h培养,未表现出明显细胞毒性。系列数据证明,该种水凝胶可以设计成为pH敏感型的药物控释载体材料,并因其良好的生物相容性,也有作为细胞支架、创伤辅料等其它生物医用材料的潜力。     

Fabrication of supramolecular hydrogels for drug delivery and stem cell encapsulation

Supramolecular hydrogels self-assembled by alpha-cyclodextrin and methoxypolyethylene glycol-poly(caprolactone)-(dodecanedioic acid)-poly(caprolactone)-methoxypolyethylene glycol (MPEG-PCL-MPEG) triblock polymers were prepared and characterized in vitro and in vivo. The sustained release of dextran-fluorescein isothiocyanate (FITC) from the hydrogels lasted for more than 1 month, which indicated that the hydrogels were promising for controlled drug delivery. ECV304 cells and marrow mesenchymal stem cells (MSC) were encapsulated and cultured in the hydrogels, during which the morphologies of the cells could be kept. The in vitro cell viability studies and the in vivo histological studies demonstrated that the hydrogels were non-cytotoxic and biocompatible, which indicated that the hydrogels prepared were promising candidates as injectable scaffolds for tissue engineering applications.     

Predicting Caco-2 cell permeation coefficients of organic molecules using membrane-interaction QSAR analysis

A methodology termed membrane-interaction QSAR (MI-QSAR) analysis has been developed in order to predict the behavior of organic compounds interacting with the phospholipid-rich regions of biological membranes. One important application of MI-QSAR analysis is to estimate ADME properties including the transport of organic solutes through biological membranes as a computational approach to forecasting drug intestinal absorption. A training set of 30 structurally diverse drugs, whose permeability coefficients across the cellular membranes of Caco-2 cells were measured, was used to construct significant MI-QSAR models of Caco-2 cell permeation. Cellular permeation is found to depend primarily upon aqueous solvation free energy (solubility) of the drug, the extent of drug interaction with a model phospholipid (DMPC) monolayer, and the conformational flexibility of the solute within the model membrane. A test set of eight drugs was used to evaluate the predictivity of the MI-QSAR models. The permeation coefficients of the test set compounds were predicted with the same accuracy as the compounds of the training set.