Characterization of stimuli-sensitive polymers for biomedical applications

Stimuli-sensitive polymers were synthesized by copolymerizing varying ratios of N-isopropyl acrylamide(NIPAAm) and acrylic acid(AAc). The influence of polyelectrolytes on the lower critical solution temperatures(LCSTs) of these temperature/pH sensitive polymers was investigated in the pH range of 2-12. Polyelectrolyte complexes were prepared by mixing poly(NIPAAm-co-AAc) as anionic polyelectrolyte with poly(allyl amine)(PAA) or poly(L-lysine)(PLL) as cationic polyelectrolytes, respectively. Back titration was performed to determine the pK_a values of PAAc in poly(NIPAAm-co-AAc) and to study the effect of comonomer ionization on the cloud point temperature. The effect of polyelectrolyte complex formation on the conformation of PLL was studied as a function of temperature by means of circular dichroism(CD). The swelling ratio of poly(NIPAAm-co-AAc) hydrogels as a function of pH at various temperature was obtained by measuring the weight of the hydrogels in buffer solutions. The LCSTs of the poly(NIPAAm-co-AAc) were strongly affected by pH, polyelectrolyte solutes, AAc content, and charge density. The influence of more hydrophobic PLL as a polyelectrolyte on the cloud point of PNIPAAm/water in the copolymer was stronger than that of poly(allyl amine)(PAA). Indomethacin was loaded into these hydrogels, and controlled release of this molecule from the hydrogel was determined under various temperature and pH conditions using UV/Vis spectrophotometry.     

Synthesis and Properties of Poly(N-isopropylacrylamide-co-acrylamide) Hydrogels

Poly (N-isopropylacrylamide-co-acrylamide) [Poly(NIPAAm-co-AAm)] with different feed ratios were obtained by radiation polymerization using Co⁶⁰γ-rays. Swelling equilibrium data in various media: deionized water, aqueous NaCl solutions and different pH buffer solutions, were determined. It appeared that the lower critical transition temperature (LCST) of the hydrogels increased with an increasing acrylamide content and decreased with increasing ionic strength. Moreover, LCST was affected by pH.     

Cell-Encapsulating Hydrogel Puzzle: Polyrotaxane-Based Self-Healing Hydrogels

Slide-ring hydrogels using polyrotaxanes have been developed as highly tough soft materials. However, they have never been used as biomaterials because of the lack of biocompatibility. Meanwhile, self-healing hydrogels are expected to improve fatigue resistance and extend the period of use. However, owing to the lack of high mechanical strength, they are limited in their use as biomaterials. Here we first developed a biocompatible self-healing/slide-ring hydrogel using glycol chitosan and a water-soluble polyrotaxane. We obtained excellent mechanical toughness and biocompatibility to promote the proliferation of human umbilical vein endothelial cells (HUVECs) encapsulated in the hydrogel. Owing to the rapid self-healing property, the cell-encapsulating gels adjusted arbitrarily, maintaining good cell proliferation function. Therefore, slide-ring hydrogels enable the use of biomaterials for soft-tissue engineering.     

新型阴离子型温敏水凝胶

新型阴离子型温敏水凝胶王昌华,曹维孝（北京大学化学与分子工程学院,北京,１００８７１）关键词温敏水凝胶,溶胀比,Ｎ－异丙基丙烯酰胺１９８４年Ｔａｎａｋａ等人［１］发现Ｎ－异丙基丙烯酰胺（ＮＩＰＡ）与Ｎ,Ｎ－亚甲基双丙烯酰胺（ＭＢＡ）的微交联共聚体,在...     

Synthesis and Swelling Properties of Thermosensitive Hydrogels based on Terpolymerization

Novel thermosensitive hydrogels based on polymerization of N-isopropyl acrylamide, Sodium acrylate, and diacetone acrylamide were synthesized, The swelling ratio and dynamic swelling were investigated. The results indicated that the hydrogels exhibited high water uptake and themosensitivity. The swelling properties and volume phase transition temperature could be adjusted by contents of the comonomers in the gels.     

Human Albumin-Based Hydrogels for Their Potential Xeno-Free Microneedle Applications

Nowadays, hydrogels-based microneedles (MNs) have attracted a great interest owing to their outstanding qualities for biomedical applications. For the fabrication of hydrogels-based microneedles as tissue engineering scaffolds and drug delivery carriers, various biomaterials have been tested. They are required to feature tunable physiochemical properties, biodegradability, biocompatibility, nonimmunogenicity, high drug loading capacity, and sustained drug release. Among biomaterials, human proteins are the most ideal biomaterials for fabrication of hydrogels-based MNs; however, they are mechanically weak and poorly processible. To the best of the knowledge, there are no reports of xeno-free human protein-based MNs so far. Here, human albumin-based hydrogels and microneedles for tissue engineering and drug delivery by using relatively new processible human serum albumin methacryloyl (HSAMA) are engineered. The resultant HSAMA hydrogels display tunable mechanical properties, biodegradability, and good biocompatibility. Moreover, the xeno-free HSAMA microneedles display a sustained drug release profile and significant mechanical strength to penetrate the model skin. In vitro, they also show good biocompatibility and anticancer efficacy. Sustainable processible human albumin-based biomaterials may be employed as a xeno-free platform in vivo for tissue engineering and drug delivery in clinical trials in the future.     

The effect of external stimuli on the Bovine Serum Albumin adsorption capacity of poly(acrylamide/maleic acid) hydrogels prepared by gamma rays

The effects of external stimuli such as pH of the buffer solution, ionic strength, temperature and the amount of poly-electrolyte monomer in the hydrogel system on the Bovine Serum Albumin (BSA) adsorption capacity of poly(acrylamide/maleic acid) [P(AAm/MA)] hydrogels were investigated. Poly-electrolyte P(AAm/MA) hydrogels with varying compositions were prepared by irradiating acrylamide/maleic acid/water mixtures with γ rays at ambient temperature. Langmuir type adsorption isotherms were observed for all prepared hydrogels. Increase of ionic strength of the buffer solution from 0.01 to 0.1 mol dm⁻³ decreased the adsorption capacity of hydrogels and zero adsorption was observed in the presence of 0.1 mol dm⁻³ Na⁺ and Ca²⁺ ion in the adsorption medium. The adsorption capacity of hydrogels was found to increase from 0 to 120 mg BSA/g dry gel, by changing external stimuli and hydrogel composition.     

Synthesis and network structure of ionic poly(N,N-dimethylacrylamide-co-acrylamide) hydrogels: Comparison of swelling degree with theory

At four different charge densities, ionic hydrogels based on N,N-dimethylacrylamide (DMAAm), acrylamide (AAm), and itaconic acid (IA) were synthesized by free-radical cross-linking copolymerization in water with N,N-methylenebis(acrylamide) (BAAm) as the cross-linker, ammonium persulfate (APS) as the initiator, and N,N,N′,N′-tetramethylenediamine (TEMED) as the activator. The swelling behaviors of these hydrogels were analyzed in buffer solutions at various pH. It was observed that the swelling behavior of cross-linked ionic poly(N,N-dimethylacrylamide-co-acrylamide) [P(DMAAm-co-AAm)] hydrogels at different pHs agreed with the modified Flory-Rehner equation based both on the phantom network and affine network models and the ideal Donnan theory. In addition, the kinetics of swelling of the hydrogels was studied in pH 2, 5 and 9 buffer solutions. The swelling curves exhibited the characteristic features of transport process, apparently the Fickian diffusion of fast rates.     

离子键交联的聚两性电解质凝胶在不同pH和不同电解质溶液中溶胀行为研究

以丙烯酸(AA)和甲基丙烯酸二乙氨基乙酯(DEAM)形成的离子复合物与丙烯酰胺(AAm)共聚,合成了一种新型的离子键交联的聚两性电解质凝胶(PADA).由于分子之间的氢键作用,PADA凝胶并不是在A/C(负正离子单体摩尔比)为1,而是在A/C为1.55处有最大消溶胀.与共价键交联的聚两性电解质凝胶相比,PADA凝胶的溶胀行为具有更强的pH敏感性.PADA凝胶在不同pH缓冲溶液中的溶胀行为表明,在pH 3~4之间消溶胀程度最大.在偏离该pH区域时凝胶均发生溶胀.但凝胶的溶胀程度在pH<3的酸性溶液中随A/C的增加而降低;而在pH>4的偏碱性溶液中随其增加而增加.在不同价数的离子溶液中,离子浓度对于PADA凝胶的平衡溶胀有着不同的影响.对于一价的NaCl溶液,PADA凝胶有典型的反聚电解质效应.但对于高价的CaCl2和柠檬酸三钠溶液,只在较低的浓度下,才表现出反聚电解质效应.而在较高盐浓度时,随盐浓度的增加其溶胀比反而降低.这可能与高价离子形成的离子键交联有关.与pH对PADA凝胶溶胀程度的影响相似,在CaCl2溶液中,PADA凝胶的溶胀程度随A/C的增加而降低;而在柠檬酸三钠溶液中则刚好相反.这种独特的溶胀行为似乎与高价离子电荷的正负性有关.     

Enhanced selectivity of hydrogel-based molecularly imprinted polymers (HydroMIPs) following buffer conditioning

We have investigated the effect of buffer solution composition and pH during the preparation, washing and re-loading phases within a family of acrylamide-based molecularly imprinted polymers (MIPs) for bovine haemoglobin (BHb), equine myoglobin (EMb) and bovine catalyse (BCat). We investigated water, phosphate buffer saline (PBS), tris(hydroxymethyl)aminomethane (Tris) buffer and succinate buffer. Throughout the study MIP selectivity was highest for acrylamide, followed by N-hydroxymethylacrylamide, and then N-iso-propylacrylamide MIPs. The selectivity of the MIPs when compared with the NIPs decreased depending on the buffer conditions and pH in the order of Tris > PBS > succinate. The Tris buffer provided optimum imprinting conditions at 50 mM and pH 7.4, and MIP selectivities for the imprinting of BHb in polyacrylamide increased from an initial 8:1 to a 128:1 ratio. It was noted that the buffer conditions for the re-loading stage was important for determining MIP selectivity and the buffer conditions for the preparation stage was found to be less critical. We demonstrated that once MIPs are conditioned using Tris or PBS buffers (pH7.4) protein reloading in water should be avoided as negative effects on the MIP's imprinting capability results in low selectivities of 0.8:1. Furthermore, acidifying the pH of the buffer solution below pH 5.9 also has a negative impact on MIP selectivity especially for proteins with high isoelectric points. These buffer conditioning effects have also been successfully demonstrated in terms of MIP efficiency in real biological samples, namely plasma and serum.     

Network structure and swelling–shrinking behaviors of pH‐sensitive poly(acrylamide‐co‐itaconic acid) hydrogels

pH‐sensitive poly(acrylamide‐co‐itaconic acid) [P(AAm/IA)] hydrogels were prepared by radiation induced copolymerization of acrylamide (AAm) and itaconic acid (IA) at various ratios. Swelling and shrinking behaviors of these hydrogels were found greatly dependent on the composition of the hydrogel and pH of the buffer solution. The basic structural parameters of the P(AAm/IA) networks such as the molecular weight between crosslinks (M _c) and polymer–solvent interaction parameter (χ) were also determined using the modified Flory‐Rehner equations. © 2004 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 42: 2586–2594, 2004     

Synthesis of hydrogels based on poly(NIPAM) inserted into collagen sponge

The study presents the preparation of a semi-synthetic hydrogel based on poly(N-isopropyl acrylamide-co-diethylene glycol diacrylate) inserted onto the collagen porous membrane. The synthesis of the hydrogels was performed through radical copolymerization of N-isopropyl acrylamide (NIPAM) with diethylene glycol diacrylate (DEGDA) also as crosslinking agent, using ammonium persulfate as initiator and N,N,N',N'-tetramethylethylene diamine as activator, and it was achieved in the presence of the collagen matrix. The prepared hydrogels were characterized by Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy and Scanning Electron Microscopy. The swelling behaviour of the semi-interpenetrated polymer network related on the hydrogel composition, it was also evaluated. The pore sizes of the synthesized hydrogels, much larger than the typical mesh size of a conventional hydrogel, allow to consider the hybrid hydrogel based on the inserted poly(NIPAM-co-DEGDA) onto collagen fibrils as a super-porous hydrogel.     

SWELLING BEHAVIOR OF ACRYLAMIDE HYDROGEL IN DIFFERENT SOLVENTS AND pHs

Swelling property of acrylamide hydrogels, prepared from aqueous solutions of acrylamide monomer having concentrations in the range of 10-60 wt% by γ-ray irradiation method using a Co-60 gamma radiation source at doses ranging 1-30.0 kGy, has been investigated under various swelling media. These swelling media were basically solvents (solutions), produced by dissolving methanol, ethanol, glucose, sucrose, sodium chloride and sodium persulfate individually with distilled water, and solutions prepared with pHs = 3, 7 and 10. The investigation was performed in order to observe the effect of these solvents and pHs as well as the influence of monomer concentrations, radiation doses and times on swelling behavior of hydrogels. Swelling values were found higher for hydrogels prepared with lower monomer concentrations (ca.20 wt%) and radiation doses (ca. 5 kGy) and showed a leveling off tendency within 24 h. The glucose solvent and the buffer solution of pH = 10 revealed significant increase of swelling of hydrogels as compared to other solutions. Results are explained based on crosslinking density in hydrogel, polymer-solvent/polymer-polymer interactions in solutions,permeability of molecules in solutions and ionization capacity of hydrogel in pH.     

Single Amino-Acid Based Self-Assembled Biomaterials with Potent Antimicrobial Activity

The design and development of soft biomaterials based on amino acid and short-peptide have gained much attention due to their potent biomedical applications. A slight alteration in the side-chain of single amino acid in a peptide or protein sequence has a huge impact on the structure and function. Phenylalanine is one of the most studied amino acids, which contains an aromatic phenyl group connected through a flexible −CH₂− unit. In this work, we have examined whether flexibility and aromatic functionality of phenylalanine (Phe) are important in gel formation of model gelator Fmoc-Phe-OH or not. To examine this hypothesis, we synthesized Fmoc-derivatives of three analogues unnatural amino acids including cyclohexylalanine, phenylglycine, and homophenylalanine; which are slightly varied from Phe. Interestingly, all these three new analogues formed hydrogels in phosphate buffer at pH 7.0 having different gelation efficacy and kinetics. This study suggests that the presence of aromatic side-chain and flexibility are not mandatory for the gelation of this model gelator. Newly synthesized unnatural amino acid derivatives have also exhibited promising antimicrobial activity towards gram-positive bacteria by inhibiting cellular oxygen consumption. We further determined the biocompatibility of these amino acid derivatives by using a hemolysis assay on human blood cells. Overall studies described the development of single amino acid-based new injectable biomaterials with improved antimicrobial activity by the slight alteration in the side-chain of amino acid.     

羧甲基纤维素与丙烯酸接枝共聚凝胶的溶胀动力学及过溶胀平衡特性的研究

采用接枝共聚合成了羧甲基纤维素钠、丙烯酸与N,N′-亚甲基双丙烯酰胺的交联凝胶, 研究了这类凝胶在不同pH值的缓冲溶液中的溶胀行为, 发现在酸性介质中凝胶的溶胀动力学行为表现出过溶胀平衡特性(overshooting effect), 即凝胶先发生溶胀到最大值, 然后再逐渐消溶胀到平衡. 这种现象可归因于凝胶溶胀过程中羧基之间通过氢键所产生的协同物理交联. 较之凝胶的组成, 缓冲溶液的pH值对过溶胀平衡现象的影响更为显著. 前者是因为凝胶羧基的总摩尔分数并不随两组分结构单元摩尔数的改变而改变, 羧基之间通过氢键形成的物理交联程度在交联剂摩尔分数接近的条件下变化不大; 后者是由于溶液的pH值显著影响凝胶羧基的质子化程度, 进而影响羧基之间通过氢键形成的物理交联程度. 凝胶在酸性介质中的溶胀过程符合E. Díez-Peña等提出的溶胀动力学定量模型, 理论曲线与实验数据有较好的相关性. 凝胶在pH≥5.0的缓冲溶液中的溶胀不产生过溶胀平衡现象, 这一现象归因于完全离子化的羧基之间不能形成物理交联. 凝胶的溶胀过程遵循Schott二级溶胀动力学.     

光控水凝胶的构筑及在生物医药领域的应用

水凝胶由于具有优越的保水性、良好的生物相容性和可降解性,被认为是最接近人体组织的生物医用材料。通过构建环境敏感水凝胶可以高度拟合生物组织的微环境,实现其在组织工程与再生医学领域的应用。由于光具有非物理接触和时空分辨等优势,利用光调控技术可实现水凝胶微环境的精确构筑与调控。本文重点介绍了近年来光控水凝胶的构筑,以及在生物医学和材料领域的应用进展。     

Swelling behavior of ionically cross-linked polyampholytic hydrogels in varied salt solutions

Ionically cross-linked polyampholytic hydrogels were synthesized by redox copolymerization of acrylamide and an ionic complex of (N,N-diethylamino)ethyl methacrylate and acrylic acid (designated as PADA hydrogel). The swelling behavior of the hydrogels in water indicated that a minimal equilibrium swelling ratio is found when the molar ratio of anionic/cationic monomers was 1.55. In NaCl solution, the hydrogels exhibited the typical swelling behavior of conventional polyampholytic gels. Their equilibrium swelling ratios increased with an increase in the NaCl concentration. In solutions of multivalent ions (CaCl₂ and trisodium citrate solutions), the equilibrium swelling ratios of the hydrogels increased first and were then followed by a decrease with an increase in salt concentration. Interestingly, an unexpected abrupt swelling phenomenon was observed when the fully swollen hydrogels in salt solution were transmitted to pure water. The unique swelling behavior of PADA hydrogels depends not only on the molar ratio of the anionic/cationic monomers but also on the valency of the ions.     

pH‐responsive ionic poly(N,N‐diethylaminoethyl methacrylate‐co‐N‐vinyl‐2‐pyrrolidone) hydrogels: Synthesis and swelling properties

A series of an ionic hydrogels composed of N,N‐diethylaminoethyl methacrylamide (DEAEMA), N‐vinyl‐2‐pyrrolidone (VP), and itaconic acid were synthesized by free‐radical cross‐linking copolymerization in water–ethanol mixture by using N,N‐methylenebis(acrylamide) as the cross‐linker, ammonium persulfate as the initiator, and N,N,N′,N′‐tetramethylenediamine as the activator. The swelling behaviors of these hydrogels were analyzed in buffer solutions at various pH. It was observed that the swelling behavior of cross‐linked ionic poly(N,N‐diethylaminoethyl methacrylamide‐co‐N‐vinyl‐2‐pyrrolidone) [P(DEAEMA/VP)] hydrogels at different pH agreed with the modified Flory–Rehner equation based on the affine network model and the ideal Donnan theory. The swelling process in buffer solutions at various pH was found to be Fickian‐type diffusion. The pH‐reversibility and on–off switching properties of the P(DEAEMA/VP) hydrogels may be considered as good candidate to design novel drug‐delivery system. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 2819–2828, 2005     

纤维素纳米纤维接枝聚(N-异丙基丙烯酰胺)水凝胶的制备与表征

将具有温度响应的聚N-异丙基丙烯酰胺(PNIPAm)接枝到电纺纤维素纳米纤维膜上,制备温度响应型纤维素接枝聚N-异丙基丙烯酰胺(PNIPAm-g-Cell)纳米纤维水凝胶。 研究接枝单体(N)与纤维素(c)的质量比、反应温度、反应时间和引发剂浓度对产物接枝率、溶胀性和形貌的影响。 结果表明,最佳聚合反应条件为m(N):m(c)=15:1、反应温度40 ℃、反应时间3 h、引发剂浓度为10 mmol/L,得到PNIPAm-g-Cell接枝率和溶胀率分别为35%和31%。 与PNIPAm相比,PNIPAm-g-Cell水凝胶的低临界相转变温度(LCST)显著升高,说明亲水性纤维素的引入改变了体系的亲疏水平衡。 去溶胀动力学测试表明,0.5 min内接枝率为25%和35%的水凝胶保水率分别降低至93%和61%。 说明接枝率越高PNIPAm-g-Cell水凝胶对温度的响应速度越快,对温度越敏感。     

Antigen Responsive Hydrogels Based on Polymerizable Antibody Fab′ Fragment

Novel antigen responsive hydrogels were prepared by using polymerizable antibody Fab′ fragment from monoclonal anti‐fluorescein BDC1 antibody (IgG2a). To form Fab′ containing hydrogels, the polymerizable Fab′ fragment was copolymerized with N‐isopropylacrylamide (NIPAAm) and N,N′‐methylenebis(acrylamide) (MBAAm; crosslinker) using redox initiators. The thermosensitivity of the hydrogels decreased with increasing Fab′ fragment content. The antigen responsiveness of the hydrogels depended on the Fab′ content, pH, and temperature. When the hydrogels were alternately exposed to antigens fluorescein (FL) and polyamidoamine dendrimer (PAMAM)‐fluorescein (FD), significant reversible volume changes were observed for the hydrogel containing 50% (w/w) Fab′ fragment at 33.7 and 36.8 °C in acetate buffer (10 mM , pH 5.0), respectively, but not at 27.7 °C or in PBS buffer (10 mM , pH 7.4). No noticeable reversible volume changes were observed with pure PNIPAAm hydrogel and the gel containing 10% (w/w) Fab′ fragment.

Structure of Fab′ containing hydrogels.