Simulated Biological Fluids Research Articles

Development and Validation of RP-HPLC Method for Estimation of Empagliflozin and its Stability Studies In vitro Simulated Gastric and Intestinal Fluids

A simple and precise high performance liquid chromatography method for determination of Empagliflozin and its stability studies invitro simulated gastric and intestinal fluids. The separation was carried out on Cosmosil C18 (250mm X 4.6, 5μ) RP-HPLC column using a 0.8ml/min as flow rate with methanol: water (80: 20 v/v) mobile phase and RT was 5.017min. Quantification was performed with a UV detector at 224nm, solubility and stability was determined individually in simulated biological fluids at various pH was investigated. The calibration curves of EMPA were linear in the range of 10–50 μg/ml(R²=0.999). The developed method was applied to pharmaceutical formulation successfully with no interfering peaks. The percentage recovery was 99.71–100.29%. It was observed that solubility of EMPA was increased in all in vitro interaction medium, freely soluble in pH 6.8 and slightly in 1.2 and 7.4 pH and the stability of EMPA was stable or less degraded in pH 6.8 for 24 hours.

Study of behavior alloy Ti and 316L in to simulated body fluid by electrochemical techniques

In this work, Ti-316L stainless steel was produced, and its electrochemical behavior was characterized. Tisteel alloys were produced using an induction furnace, by mixing commercial Ti and 316L stainless steel. Xray diffraction analyses showed the presence of both the materials, Ti and 316L, in the samples produced. The elemental composition of the materials was determined by optical emission spectroscopy and energydispersive spectroscopy, which showed similar quantities of both the elements. Both commercial 316L stainless steel and Ti–steel were studied in simulated biological fluid for imitate a like composition of body blood plasma. Electrochemical experiments conducted at 37°C indicated the stable passive polarization behavior of the Ti-steel alloy. Furthermore, the electrochemical behavior of the 316L stainless steel was also analyzed for comparison purposes. Corrosion velocities were determined using the Tafel method and corrosion resistances were obtained using the electrochemical impedance spectroscopy. The Ti-steel exhibited better protection capacity, compared with the commercial 316L steel. The corrosion velocity and the passive current density of the Ti–steel alloy were lower than those exhibited by the 316L stainless steel. Keywords: XRD, elemental composition, Tafel curves, corrosion velocity.

Biopolymers – Calcium phosphates composites with inclusions of magnetic nanoparticles for bone tissue engineering

Composites based on combination of biopolymers (chitosan, hyaluronic acid and bovine serum albumin or gelatin), calcium phosphates (CP) and magnetic nanoparticles have been prepared by a biomimetic co-precipitation method. The biomimetic strategy is inspired by natural mineralization processes, where the synthesized minerals are usually combined with proteins, polysaccharides or other mineral forms to form composite, in physiological conditions of temperature and pH. The morphology of the magnetic composites, studied using scanning electron microscopy (SEM) indicated a macroporous structure, which influenced the retention of simulated biological fluids. Fourier transformed infrared spectroscopy and X-ray diffraction and Energy-dispersive X-ray spectroscopy (EDX) confirmed the composition of the scaffolds and the formation of various types of calcium phosphates with amorphous nature. The in vitro degradation studies showed a slow degradation process for magnetic composites that confirmed the tightly connection of the polymeric matrix with calcium phosphates, which limits the enzyme access to the degradable components and material disintegration. The magnetic scaffolds exhibited no negative effect on osteoblasts cell, emphasizing a good biocompatibility. Considering the scaffolds properties, some compositions based on calcium phosphates, chitosan, Hya/Bsa and more than 3% of MNPs are recommended for further optimization and in vivo tests.

Comparative study between natural and synthetic Hydroxyapatite: structural, morphological and bioactivity properties

In this work, a study of hydroxyapatite (HAp) powders obtained using both, porcine bones and chemical precursors was carried put. In the case of HAp obtained by means of porcine bones, physical processes as cooking, washing and milling were developed, for removing the organic material from the bones; after that, the powders were submitted to a thermal treatment at 800 °C, during 12 h. This procedure was carried out without adding chemical alkalines that are harmful for the environment and the human health. On the other hand, HAp powders were also synthetized using the chemical precipitation method widely reported, showing successful results. Moreover, both kind of powders were characterized using x ray diffraction, Fourier transformer infrared spectroscopy, scanning electron microscopy and energy dispersive spectroscopy. Furthermore, the bioactivity of the materials was determined using the simulated biological fluid (SBF) method. Results showed that the natural HAp exhibited better crystallographic properties. Moreover, according to these results, HAp obtained from porcine bones contains traces of elements as Na and Mg that are favorable for the bioactivity, according to the materials behavior when they are immersed in SBF. Keywords: Hydroxyapatite, porcine bones, chemical precipitation, simulated biological fluid.

Synthesis and characterization of amoxicillin loaded poly (vinyl alcohol)-g-poly (acrylamide) (PVA-g-PAM) hydrogels and study of swelling triggered release of antibiotic drug

In the present work, amoxicillin drug-loaded films of poly (vinyl alcohol)-g-poly (acrylamide) of varying compositions were prepared by graft copolymerization method. The as-prepared drug-loaded grafted hydrogels were characterized by various analytical techniques such as Fourier transform infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy, and X-ray diffraction analysis, respectively. The grafted hydrogels were investigated for their water intake behavior under varying experimental conditions of their chemical composition, pH and temperature of the swelling bath, and simulated biological fluids. The water sorption data of hydrogels were used to calculate network parameters of the grafted hydrogel such as average molecular weight between crosslinks (Mc), crosslink density (Ve), number of elastically effective chains (qe), swelling exponent (n), and diffusion constant (D), respectively. The swelling controlled drug release behavior of amoxicillin loaded hydrogel was investigated under in vitro conditions, and the influence of various factors such as chemical composition of grafted hydrogel, percent loading of drug (amoxicillin), pH and temperature of the release media was studied on the release profiles of the drug. The amoxicillin loaded grafted hydrogels were also examined for their antibacterial activity against gram-negative bacteria.

Postsynthetic Metalation Metal-Organic Framework as a Fluorescent Probe for the Ultrasensitive and Reversible Detection of PO43- Ions.

The self-assembly of a zinc salt with the novel ligand 4,4',4″-[(1,3,5-triazine-2,4,6-triyl)tris(sulfanediyl)]tribenzoic acid generated 3D microporous metal-organic frameworks (MOFs), namely, {[Zn4(L3-)2(O2-)(H2O)2]·4EtOH} n. The frameworks with multiple Lewis basic sites exhibit easily sensitized properties. After encapsulation of the Tb3+ cation in this Zn-MOF, the as-obtained fluorescent-functionalized Tb@Zn-MOFs not only maintain distinguished chemical stabilities but also exhibit strong characteristic emissions of trivalent terbium ions. Interestingly, Tb@Zn-MOF has been chosen to be a potential highly selective and sensitive luminescent platform for the reversible detection of PO43- ions in aqueous and living cell buffer solutions with a fast response time of 10 s and a low detection limit (0.1 ppm). Strikingly, this work presents the first example of a fluorescent sensor that can quantitate PO43- in simulated biological fluids and monitor these ions in the water system in a wide range of concentrations from 10-6 to 10-3 M.

Development and characterization of xyloglucan-poly(vinyl alcohol) hydrogel membrane for Wireless Smart wound dressings

Hydrogel-based smart wound dressings that combine the traditional favourable properties of hydrogels as skin care materials with sensing functions of relevant biological parameters for the remote monitoring of wound healing are under development. In particular, lightweight, ultra-high frequency radiofrequency identification (UHF RFID) sensor are adjoined to xyloglucan-poly(vinyl alcohol) hydrogel films to battery-less monitor moisture level of the bandage in contact with the skin, as well as wireless transmit the measured data to an off-body reader. This study investigates the swelling behavior of the hydrogels in contact with simulated biological fluids, and the modification of their morphology, mechanical properties, and dielectric properties in a wide range of frequencies (100–106 Hz and 108–1011 Hz).The films absorb simulated body fluids up to approximately four times their initial weight, without losing their integrity but undergoing significant microstructural changes. We observed relevant linear increases of electric conductivity and permittivity with the swelling degree, with an abrupt change of slope that is related to the network rearrangements occurring upon swelling.

Magnetically responsive release of 5-FU from superparamagnetic egg albumin coated iron oxide core-shell nanoparticles

The present study aimed at preparing egg albumin coated iron oxide (EAIO) core shell nanoparticles and evaluating their ability to release 5-FU drug under applied external magnetic field. EAIO nanoparticles were prepared by simultaneous co-precipitation and emulsion crosslinking technique and characterized by FTIR, DSC, TEM, XRD, particle size analysis and surface potential measurements. The influence of chemical composition of EAIO nanoparticles, pH and temperature of release media, presence of simulated biological fluids as the release media, and applied magnetic field was investigated on the release profiles of 5-FU drug. The drug release was analyzed kinetically using Fickan power law, zero order, first order and Korsmeyer-Peppas models and the amount of released drug was correlated and the relation was drawn between the quantity of released drug and water holding capacity of nanoparticles. The nanoparticles were judged for in vitro cytotoxicity and the chemical integrity of the drug was also accessed.

Characterization of tantalum and tantalum nitride films on Ti6Al4V substrate prepared by filtered cathodic vacuum arc deposition for biomedical applications

This paper explores tantalum and tantalum nitride thin films produced by filtered cathodic vacuum arc deposition method as bio-stable surface treatment for Ti6Al4V titanium alloy. Effect of nitrogen to argon gas ratio on microstructure of the deposited film has been investigated. Corrosion behaviour of the films in simulated biological fluid solution was evaluated by electrochemical impedance spectroscopy. It was found that both the Ta and TaN films enhanced corrosion resistance of the Ti6Al4V substrate with the best protective characteristics achieved by the TaN film deposited at 0.25 N2/Ar gas ratio. The protective characteristic was attributed to the formation of tantalum oxide and oxynitride compound at the surface, as verified by X-ray photoelectron spectroscopy. Increasing N2/Ar gas ratio increased susceptibility to localized corrosion. The corrosion resistance decreased as the thickness of the film increased to 1 μm.

Enhanced uptake in 2D- and 3D- lung cancer cell models of redox responsive PEGylated nanoparticles with sensitivity to reducing extra- and intracellular environments

In the treatment of lung cancer, there is an urgent need of innovative medicines to optimize pharmacological responses of conventional chemotherapeutics while attenuating side effects. Here, we have exploited some relatively unexplored subtle differences in reduction potential, associated with cancer cell microenvironments in addition to the well-known changes in intracellular redox environment. We report the synthesis and application of novel redox-responsive PLGA (poly(lactic-co-glycolic acid)) -PEG (polyethylene glycol) nanoparticles (RR-NPs) programmed to change surface properties when entering tumor microenvironments, thus enhancing cell internalization of the particles and their drug cargo.The new co-polymers, in which PEG and PLGA were linked by ‘anchiomeric effector’ dithiylethanoate esters, were synthesized by a combination of ring-opening polymerization and Michael addition reactions and employed to prepare NPs. Non redox-responsive nanoparticles (nRR-NPs) based on related PLGA-PEG copolymers were also prepared as comparators. Spherical NPs of around 120 nm diameter with a low polydispersity index and negative zeta potential as well as good drug loading of docetaxel were obtained. The NPs showed prolonged stability in relevant simulated biological fluids and a high ability to penetrate an artificial mucus layer due to the presence of the external PEG coating. Stability, FRET and drug release studies in conditions simulating intracellular reductive environments demonstrated a fast disassembly of the external shell of the NPs, thus triggering on-demand drug release.FACS measurements and confocal microscopy showed increased and faster uptake of RR-NPs in both 2D- and 3D- cell culture models of lung cancer compared to nRR-NPs. In particular, the ‘designed-in’ reductive instability of RR-NPs in conditioned cell media, the fast PEG release in the extracellular compartment, as well as a diminution of uptake rate in control experiments where extracellular thiols were neutralized, suggested a partial extracellular release of the PEG fringe that promoted rapid internalization of the residual NPs into cells.Taken together, these results provide further evidence of the effectiveness of PEGylated reducible nanocarriers to permeate mucus layer barriers, and establish a new means to enhance cancer cell uptake of drug carriers by extra-and intra-cellular cleavage of protein- and cell-shielding hydrophilic blocks.

Scaffolds Based on Collagen, Hyaluronan and Sericin with Potential Applications as Controlled Drug Delivery System.

Natural proteins have been extensively studied as matrices for tissue engineering, due to their excellent biocompatibility and biological properties associated with increasing cell proliferation. By generating complex materials, cell and tissue functions can be tailored to obtain a specific direction, according to the medical needs. The aim of this paper was to obtain scaffolds based on collagen, hyaluronan and sericin, with morphology and physical-chemical properties adequate for controlled drug delivery systems. In this aim various tests were performed: in vitro swelling and degradation studies, Fourier Transform Infrared spectroscopy (FT-IR), Scanning Electronic Microscopy (SEM) and thermogravimetric analysis. Loading and releasing of ibuprofen is also discussed. The results indicate that scaffolds based on collagen, hyaluronan and sericin have a porous structure, strength and stability adequate for skin tissue engineering. The obtained scaffolds swell, degrade and have controlled drug release properties in simulated biological fluids.

Easy fabrication and characterization of gelatin nanocarriers and in vitro investigation of swelling controlled release dynamics of paclitaxel

The purpose of the present investigation was to prepare gelatin nanoparticles, capable of releasing the paclitaxel drug in a controllable fashion by regulating the extent of swelling of the nanoparticles. Gelatin nanoparticles were prepared by a single water in oil (W/O) emulsion crosslinking technique. The nanoparticles were characterized by FTIR, SEM, TEM, XRD, particle size analysis, and surface potential measurements. The influence of experimental conditions such as chemical composition of gelatin nanoparticles, pH, temperature, and presence of simulated biological fluids as the release media were investigated on the release profiles of paclitaxel. The drug release processes were analyzed kinetically using Ficks power law, zero-order, first-order, and Korsmeyer–Peppas models, and a correlation was drawn between the quantity of released drug and swelling of the nanoparticles. The nanoparticles were judged for in vitro cytotoxicity and the chemical stability of the drug was also accessed.

Effect of concentration and surface roughness on corrosion behavior of Co–Cr–Mo alloy in hyaluronic acid

The work presented in this paper is based on the study of electrochemical corrosion behavior of Co–Cr–Mo alloy as a function of the concentration of simulated biological fluid (hyaluronic acid), the influence of Cl- on hyaluronic acid (HA) and machining configurations. For the electrochemical tests, electrochemical impedance spectroscopy and potentiodynamic polarization test were undertaken. With the increase in HA concentration, high metal dissolution was observed. Whereas with the addition of NaCl in the solution, the corrosion resistance of the sample was enhanced. Also, an increase in surface roughness of sample surface HA caused a decrease in icorr. This is due to the adhesion property of the HA on the sample surface. X-ray photoelectron spectroscopy was executed to analyze the passive film formed due to HA and NaCl on Co–Cr–Mo. Scanning electron microscope were carried out to analyze the surface morphology. Energy dispersive spectroscopy shows lower Cr at the pit. The study reveals the adsorption of HA on the surface. This adsorbed HA on the surface caused the metal dissolution and hence initiation sites for pitting. When compared with the sample in HA + NaCl solution, better passive film property was observed to cause better corrosion resistance.

Glutaraldehyde crosslinked and alkaline denaturation induced self association of haemoglobin to design nanocarriers for In vitro release of insulin in simulated gastrointestinal fluids (SGFs)

Adopting a novel synthetic route the haemoglobin nanoparticles were prepared by its simultaneous microemulsion mediated crosslinking with glutaraldehyde and alkaline induced denaturation. The as prepared nanoparticles were characterized by techniques like Fourier transform infrared spectroscopy (FTIR), Transmission electron microscopy (TEM), Scanning electron microcopy (SEM), Zeta potential and Dynamic Light Scattering measurements. The particle size analysis revealed that size of the nanoparticles lays in the range 60–150 nm with surface charge of −27.1 mV. The TEM images suggested for aggregated structures which confirm denaturation of haemoglobin macromolecules under alkaline conditions. The nanoparticles were assessed for water intake capacity and the effect of various factors like chemical composition of nanoparticles, pH and temperature of the swelling bath, and simulated biological fluids on water sorption capacity was investigated. The insulin was loaded on to the prepared nanoparticles and investigated for swelling controlled release of insulin in simulated gastrointestinal fluid (SGIFs). The influence of chemical composition of nanoparticles, pH and temperature of the release media, and simulated physiological fluids was studied on the released amount of insulin and an optimized formulation was achieved. The prepared haemoglobin nanoparticles were also investigated for their in-vitro cytotoxicity.

Solubility of nano-sized metal oxides evaluated by using in vitro simulated lung and gastrointestinal fluids: implication for health risks

The solubility of nano-sized metal oxides (nZnO, nCuO, nTiO2, nCeO2, and nFe3O4, 17–42 nm) and some non-nano-mineral powders (ZnO, ZnSiO3, ZnS, and CuO) were evaluated by using gastrointestinal solubility bioavailability research consortium (SBRC), in vitro gastrointestinal (IVG) method, pulmonary artificial lysosomal fluid (ALF), and Gamble solution method, respectively. It is found that these nano-sized metal oxides aggregated more or less when suspending in the simulated biological fluids analyzed by dynamic light scattering (2 mg L−1) and UV-Vis spectrometry (100 mg L−1). The aggregation and sedimentation of nano-metal oxides in a simulated biofluid are influenced by its surface property and the ingredient of the liquid. The dissolution in fluids may decrease the aggregating radius of a nano-metal oxide. In return, the aggregative effect can influence the solubility of metal elements and result in their weakened bioaccessibility. The suspending stability was consistent in the order of nFe3O4 < nCuO < nTiO2 < nCeO2 < nZnO in all the simulated biological fluids. Nano-ZnO and nCuO showed higher gastrointestinal and pulmonary bioaccessibility than nFe3O4, nTiO2, and nCeO2. The further comparisons on the bioaccessibility for nCuO and nZnO with non-nano-powder CuO and ZnO indicated that the aggregating size in suspension could play more important role in influencing the bioaccessibility than single particle size does. The present study reveals that aggregation of all studied nano-sized metal oxides occurred in body physiologic fluids and that nZnO and nCuO were easily dissolved in simulated physiologic fluids, suggesting more potential health risks from nZnO and nCuO’s exposure.

Effect of concentration of hyaluronic acid and NaCl on corrosion behavior of 316L austenitic stainless steel

Due to low cost and easily available material, 316L stainless steel (SS) is used for biomedical implants. The electrochemical corrosion behavior of 316L (SS) was studied as a function of the concentration of simulated biological fluid (hyaluronic acid), the influence of Cl− and the combined effect of NaCl and hyaluronic acid (HA). For the electrochemical tests, potentiodynamic polarization test and electrochemical impedance spectroscopy (EIS) were undertaken. With the increase in HA concentration, corrosion rate increases. Whereas, with the addition of NaCl to HA the solution, the corrosion resistance of the sample was enhanced. Also, in pure NaCl solution, the corrosion current density (icorr) increased as compared to bare HA and HA + NaCl. This is due to the adhesion property of the HA on the sample surface. EIS result agrees with the findings of potentiodynamic polarization tests. X-ray photoelectron spectroscopy (XPS) was executed to analyze the passive film formed in the solution of HA and NaCl on 316L SS. XPS spectra confirms the formation of the passive film containing chromium oxide and hydroxides. Also, the formation of MoO2 helps in improving better corrosion resistance. The peak of nitrogen was observed in the sample immersed in HA solution. Scanning electron microscope (SEM) was carried out to analyze the surface morphology.

Preparation and characterization of amorphous ciprofloxacin-amino acid salts

The amorphization of the poorly soluble drug ciprofloxacin (CIP) may be facilitated by the use of a suitable stabilizer. In this study seven amino acids, with various side chain properties, were evaluated in this regard. Solid dispersions were prepared by ball milling 1:1 molar ratios of CIP with the amino acids, and their solid-state and pharmaceutical properties were then examined. Fully X-ray amorphous solid dispersions were obtained with aspartic acid, glutamic acid, cysteine and arginine. In each case, evidence of salt formation between the drug and amino acids was found via Fourier transform infrared spectroscopy and solid-state nuclear magnetic resonance. In contrast, semi-crystalline solid dispersions were obtained with serine, alanine and glycine. The glass transition temperatures of the amorphous salts were significantly higher than those of the starting materials, and they remained fully X-ray amorphous during long-term stability studies. Significant improvements in the solubility of CIP were also observed with the amorphous salts in water and simulated biological fluids, over and above that of the corresponding physical mixtures. In permeability studies on the other hand, the amorphous aspartate and glutamate salts were found to be less permeable than the pure drug, whereas formulation as an amorphous salt containing cysteine or arginine increased the permeability of CIP. Therefore, while amorphous salt formation with amino acids appears to be a suitable means of improving the thermal stability and solubility of CIP, in some cases this is associated with a decrease in permeability.

Excited State Dynamics of a Photobiologically Active Ru(II) Dyad Are Altered in Biologically Relevant Environments.

In this study femtosecond and nanosecond time-resolved transient absorption spectroscopy was used to investigate the influence of ionic strength and complexity on the excited state dynamics of a Ru(II)-based metal-organic dyad. The bis-heteroleptic complex [Ru(bpy)2(ippy)]2+ (1), where bpy = 2,2'-bipyridine and ippy = 2-(1-pyrenyl-1H-imidazo[4,5-f][1,10]phenanthroline, is a potent photosensitizer for in vitro photodynamic therapy (PDT) and photodynamic inactivation (PDI) of microorganisms owing to a long-lived triplet excited state derived from a metal-to-ligand charge-transfer (3MLCT) state that is equilibrium with an intraligand (3IL) state. The prolonged lifetime provides ample opportunity for bimolecular quenching of this state by oxygen; thus singlet oxygen (1O2) sensitization is very efficient. In simple aqueous solution, fast cooling within the 3MLCT manifold is followed by energy transfer to an 3IL state, which is facilitated by rotation of a pyrenyl unit about the imidazo-pyrenyl (ip) coannular bond. For solutions of 1 in high ionic strength simulated biological fluid (SBF), a more physiologically relevant solvent that contains a complex mixture of ions at pH 7.4, femtosecond studies revealed an additional excited state, possibly based on an ion-ligand interaction. This new state appearing in high ionic strength SBF was not observable in water, simple buffers, or low ionic strength SBF. These photoinduced dynamics were also affected by the presence of biomolecules such as DNA in simple buffer, whereby relaxation on the picosecond time scale was accelerated from 39 to 18 ps with DNA intercalation by 1. The increased rate of coplanarization of the pyrene and the imidazole units was attributed to DNA-induced conformational restriction of the pyrenyl unit relative to the ip bond. Quantitative changes to excited state decay rates of 1 in solutions of high ionic strength were also observed when probed on the microsecond time scale. Notably, the thermalized excited state decay pathways were altered substantially with DNA intercalation, with access to some states being completely blocked. Experimentally, this manifested in the absence of the slowest microsecond decay channel, which is normally observed for 1 in solution. The quantitative and qualitative observations from this study highlight the importance of employing biologically relevant solvents and potential biomolecule targets when the excited state dynamics and photophysical properties (under cell-free conditions) responsible for the potent photobiological effects are assessed in the context of photodynamic therapy and photodynamic inactivation.

Magnetic Nanoparticles Inclusion into Scaffolds Based on Calcium Phosphates and Biopolymers for Bone Regeneration

Considering its functions (support, protection, assisting in movement and storage of minerals), the bone is an essential organ for the human body and the bone trauma/damages have a great impact on the human body functionality. For that reason a variety of biomaterials are studied for potential applications in bone regeneration or substitution. Bone substitution materials, with similar chemical composition to that of natural bone, and specifically those obtained by processes which mimic the natural bone formation in vivo, has been shown to be among the best. In this study, using a process of co-precipitation of calcium phosphate precursors on a mixture of biopolymers (chitosan, collagen, hialuronic acid) and magnetic nanoparticles (magnetite functionalized with chitosan), biodegradable biomimetic scaffolds have been obtained. In order to study their chemical structure, the biodegradable scaffolds have been characterized by Fourier Transform Infrared Spectroscopy (FTIR). The morphology of the biodegradable scaffolds, studied using scanning electron microscopy (SEM) indicated a macroporous morphology, which influenced the retention of simulated biological fluids. A direct relationship between the scaffolds’ degradation rate and the concentration of the polymeric phase has been observed. The in vitro cytocompatibility tests indicate that the prepared scaffolds are biocompatible and assure and adequate mediums for osteoblasts.

Structural, morphological and thermal characterization of poly (2-hydroxyethyl methacrylate-co-acrylonitrile) (P (HEMA-co-AN)) Cryogels: evaluation of water sorption potential and cytotoxicity

Cryogels are macroporous hydrogels which are synthesized through cryogelation method. In the present study cryogels of poly (2-hydroxyethyl methacrylate-co-acrylonitrile) (P (HEMA-co-AN)) were synthesized by copolymerization of 2-hydroxyethyl methacrylate (HEMA) and acrylonitrile (AN) monomers by redox polymerization using cryogelation technique. The synthesized cryogels were characterized by FTIR, SEM, XRD, DSC and TGA techniques. Different compositions of the cryogels were prepared by varying concentrations of the monomers and redox initiators in the feed mixture. These cryogels were then subjected to swelling studies and porosity determination. The swelling behavior was studied as function of concentration of the monomers, redox initiators, temperature, pH, and simulated biological fluids. The prepared cryogels were also characterized for their network parameters using water sorption data. The biocompatibility of P (HEMA-co-AN) cryogel was evaluated by in vitro cytotoxicity test. The results indicated that the P (HEMA-co-AN) cryogel had macroporous morphology and exhibited good water absorption capacity. Moreover, the cryogel was thermally stable and biocompatible in nature.