Whey Protein Isolate/Calcium Silicate Hydrogels for Bone Tissue Engineering Applications-Preliminary In Vitro Evaluation.

Whey protein isolate (WPI) hydrogels are attractive biomaterials for application in bone repair and regeneration. However, their main limitation is low mechanical strength. Therefore, to improve these properties, the incorporation of ceramic phases into hydrogel matrices is currently being performed. In this study, novel whey protein isolate/calcium silicate (WPI/CaSiO3) hydrogel biomaterials were prepared with varying concentrations of a ceramic phase (CaSiO3). The aim of this study was to investigate the effect of the introduction of CaSiO3 to a WPI hydrogel matrix on its physicochemical, mechanical, and biological properties. Our Fourier Transform Infrared Spectroscopy results showed that CaSiO3 was successfully incorporated into the WPI hydrogel matrix to create composite biomaterials. Swelling tests indicated that the addition of 5% (w/v) CaSiO3 caused greater swelling compared to biomaterials without CaSiO3 and ultimate compressive strength and strain at break. Cell culture experiments demonstrated that WPI hydrogel biomaterials enriched with CaSiO3 demonstrated superior cytocompatibility in vitro compared to the control hydrogel biomaterials without CaSiO3. Thus, this study revealed that the addition of CaSiO3 to WPI-based hydrogel biomaterials renders them more promising for bone tissue engineering applications.

Hydroxyapatite or Fluorapatite-Which Bioceramic Is Better as a Base for the Production of Bone Scaffold?-A Comprehensive Comparative Study.

Hydroxyapatite (HAP) is the most common calcium phosphate ceramic that is used in biomedical applications, e.g., as an inorganic component of bone scaffolds. Nevertheless, fluorapatite (FAP) has gained great attention in the area of bone tissue engineering in recent times. The aim of this study was a comprehensive comparative evaluation of the biomedical potential of fabricated HAP- and FAP-based bone scaffolds, to assess which bioceramic is better for regenerative medicine applications. It was demonstrated that both biomaterials had a macroporous microstructure, with interconnected porosity, and were prone to slow and gradual degradation in a physiological environment and in acidified conditions mimicking the osteoclast-mediated bone resorption process. Surprisingly, FAP-based biomaterial revealed a significantly higher degree of biodegradation than biomaterial containing HAP, which indicated its higher bioabsorbability. Importantly, the biomaterials showed a similar level of biocompatibility and osteoconductivity regardless of the bioceramic type. Both scaffolds had the ability to induce apatite formation on their surfaces, proving their bioactive property, that is crucial for good implant osseointegration. In turn, performed biological experiments showed that tested bone scaffolds were non-toxic and their surfaces promoted cell proliferation and osteogenic differentiation. Moreover, the biomaterials did not exert a stimulatory effect on immune cells, since they did not generate excessive amounts of reactive oxygen species (ROS) and reactive nitrogen species (RNS), indicating a low risk of inflammatory response after implantation. In conclusion, based on the obtained results, both FAP- and HAP-based scaffolds have an appropriate microstructure and high biocompatibility, being promising biomaterials for bone regeneration applications. However, FAP-based biomaterial has higher bioabsorbability than the HAP-based scaffold, which is a very important property from the clinical point of view, because it enables a progressive replacement of the bone scaffold with newly formed bone tissue.

Could Curdlan/Whey Protein Isolate/Hydroxyapatite Biomaterials Be Considered as Promising Bone Scaffolds?-Fabrication, Characterization, and Evaluation of Cytocompatibility towards Osteoblast Cells In Vitro.

The number of bone fractures and cracks requiring surgical interventions increases every year; hence, there is a huge need to develop new potential bone scaffolds for bone regeneration. The goal of this study was to gain knowledge about the basic properties of novel curdlan/whey protein isolate/hydroxyapatite biomaterials in the context of their use in bone tissue engineering. The purpose of this research was also to determine whether the concentration of whey protein isolate in scaffolds has an influence on their properties. Thus, two biomaterials differing in the concentration of whey protein isolate (i.e., 25 wt.% and 35 wt.%; hereafter called Cur_WPI25_HAp and Cur_WPI35_HAp, respectively) were fabricated and subjected to evaluation of porosity, mechanical properties, swelling ability, protein release capacity, enzymatic biodegradability, bioactivity, and cytocompatibility towards osteoblasts in vitro. It was found that both biomaterials fulfilled a number of requirements for bone scaffolds, as they demonstrated limited swelling and the ability to undergo controllable enzymatic biodegradation, to form apatite layers on their surfaces and to support the viability, growth, proliferation, and differentiation of osteoblasts. On the other hand, the biomaterials were characterized by low open porosity, which may hinder the penetration of cells though their structure. Moreover, they had low mechanical properties compared to natural bone, which limits their use to filling of bone defects in non-load bearing implantation areas, e.g., in the craniofacial area, but then they will be additionally supported by application of mechanically strong materials such as titanium plates. Thus, this preliminary in vitro research indicates that biomaterials composed of curdlan, whey protein isolate, and hydroxyapatite seem promising for bone tissue engineering applications, but their porosity and mechanical properties should be improved. This will be the subject of our further work.

Gypsum-related compensation of ions uptake by highly porous hydroxyapatite ceramics - Consequences for osteoblasts growth and proliferation.

Hydroxyapatites of high calcium and phosphate ions adsorption capacity are highly bioactive. However, they cause the removal of these ions from tissue liquids and cell culture media, thus reducing viability and proliferation potential of osteoblasts. Addition of small amount of gypsum (calcium sulfate dihydrate) to such hydroxyapatite-based composites may help to compensate the ions removal and stimulate the osteoblasts growth and proliferation. Therefore, the aim of this work was to enrich the highly porous hydroxyapatite-based composite with gypsum and verify its effect on ions adsorption as well as osteoblasts viability and proliferation. The results showed that addition of 1.5-1.75% gypsum caused short-term calcium ions compensation in media incubated with the composite and time-shifted increase of osteoblasts proliferation. Moreover, presence of gypsum in the composite increased the content of large pores in SBF-incubated biomaterials with no effect on their microstructure or mechanical parameters. Overall, gypsum addition improves the compatibility of hydroxyapatite-based materials with no critical disadvantages for other properties.

Miscibility and Optimization of the Liquid Rubber Content in the Resins of Light-Cured Dental Composites.

Fracture toughness is one of the main factors influencing the durability of light-cured composites used for dental restorations and fillings. One of the methods of increasing the fracture toughness is the modification of the matrix with liquid acrylonitrile-free liquid rubber. This study aimed to assess the miscibility of acrylonitrile-free liquid rubber with a blend of resins and their stability over time, and to determine the optimal amount of liquid rubber (LR) in the blend due to mechanical properties. Two blends of dimethacrylate resins were used: resin "F" composed of BisGMA (60 wt.%), TEGDMA (20 wt.%), BisEMA (10 wt.%) and UDMA (10 wt.%), and "C" resin containing BisGMA (40 wt.%), TEGDMA (40 wt.%), BisEMA (10 wt.%) and UDMA (10 wt.%). The modifier Hypro® 2000X168LC VTB liquid rubber was used in at 1%, 2%, 3%, 4%, 5%, 10%, 15% and 20% by weight in the resin blend. The miscibility was assessed by microscopy. The fracture toughness, flexural strength and Young's modulus were determined in the bending test. The results showed that the solubility of the liquid rubber depends on the ratio of BisGMA/TEGDMA in the resins. In resins with 40 wt.% TEGDMA, the LR solubility was as high as 5%, while resins with 20 wt.% TEGDMA, the liquid rubber did not dissolve. The LR-resin mixtures showed good time stability, and no changes in the size or morphology of the rubber domains were found after 24 h of mixing. The maximum fracture toughness (2.46 MPa m1/2) was obtained for 5 wt.% LR in resin F and for 15 wt.% LR in resin C (2.53 MPa m1/2). The modification with liquid rubber resulted in an exponential reduction in both flexural strength and Young's modulus. The analysis of the results of the mechanical tests allowed us to determine the optimal amount of LR for both resins. For resin F it was 5.4 wt.%, and for resin C it was 8.3 wt.%. It can be stated that the optimal amount of liquid rubber increases with its solubility in the resin.

Highly Porous Fluorapatite/ß-1,3-Glucan Composite for Bone Tissue Regeneration: Characterization and In-Vitro Assessment of Biomedical Potential.

A novel fluorapatite/glucan composite ("FAP/glucan") was developed for the treatment of bone defects. Due to the presence of polysaccharide polymer (ß-1,3-glucan), the composite is highly flexible and thus very convenient for surgery. Its physicochemical and microstructural properties were evaluated using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), mercury intrusion, mechanical testing and compared with the reference material, which was a hydroxyapatite/glucan composite ("HAP/glucan") with hydroxyapatite granules (HAP) instead of FAP. It was found that FAP/glucan has a higher density and lower porosity than the reference material. The correlation between the Young's modulus and the compressive strength between the materials is different in a dry and wet state. Bioactivity assessment showed a lower ability to form apatite and lower uptake of apatite-forming ions from the simulated body fluid by FAP/glucan material in comparison to the reference material. Moreover, FAP/glucan was determined to be of optimal fluoride release capacity for osteoblasts growth requirements. The results of cell culture experiments showed that fluoride-containing biomaterial was non-toxic, enhanced the synthesis of osteocalcin and stimulated the adhesion of osteogenic cells.

Curdlan-Based Hydrogels for Potential Application as Dressings for Promotion of Skin Wound Healing-Preliminary In Vitro Studies.

The aim of this work was to establish whether novel curdlan-based hydrogels enriched with Ca2+ ions may be considered as potential candidates for dressings, for the acceleration of skin wound healing. Firstly, biomaterials were allocated for evaluation of structural and mechanical properties. Subsequently, the ability of hydrogels to absorb simulated wound fluid and water vapor permeability, as well their capacity to release calcium ions, was evaluated. The biocompatibility of biomaterials was assessed using normal human skin fibroblasts. Importantly, the main features of the obtained curdlan-based hydrogels were compared with those of KALTOSTAT® (a commercial calcium sodium alginate wound dressing). The obtained results showed that curdlan-based biomaterials possessed a mesoporous structure (pore diameter ranged from 14-48 nm) and exhibited a good ability to absorb simulated wound fluid (swelling ratio close to 974-1229%). Moreover, in a wet state, they enabled proper water vapor transmission rate (>2000 g/m2/day), thanks to their hydrogel structure. Finally, it was found that biomaterial composed of 11 wt.% of curdlan (Cur_11%) possessed the most desirable biological properties in vitro. It released a beneficial amount of calcium ions to the aqueous environment (approximately 6.12 mM), which significantly enhanced fibroblast viability and proliferation. Taking into account the beneficial properties of Cur_11% biomaterial, it seems justified to subject it to more advanced cell culture experiments in vitro and to in vivo studies in order to determine its precise influence on skin wound healing.

Superabsorbent curdlan-based foam dressings with typical hydrocolloids properties for highly exuding wound management.

Effective management of chronic wounds with excessive exudate may be challenging for medical doctors. Over the years, there has been an increasing interest in the engineering of biomaterials, focusing on the development of polymer-based wound dressings to accelerate the healing of exuding wounds. The aim of this study was to use curdlan, which is known to support wound healing, as a base for the production of superabsorbent hybrid biomaterials (curdlan/agarose and curdlan/chitosan) with the intended use as wound dressings for highly exuding wound management. To evaluate the biomedical potential of the fabricated curdlan-based biomaterials, they were subjected to a comprehensive assessment of their microstructural, physicochemical, and biological properties. The obtained results showed that foam-like biomaterials with highly porous structure (66-77%) transform into soft gel after contact with the wound fluid, acting as typical hydrocolloid dressings. Novel biomaterials have the superabsorbent ability (1 g of the biomaterial absorbs approx. 15 ml of exudate) with horizontal wicking direction while keeping dry edges, and show water vapor transmission rate of approx. 1700-1800 g/m2/day which is recommended for optimal wound healing. Moreover, they are stable in the presence of collagenases, but prone to biodegradation in lysozyme solution (simulated infected wound environment). Importantly, the developed biomaterials are non-toxic and their surface hinders fibroblast attachment, which is essential during dressing changes to avoid damage to newly formed tissues in the wound bed. All mentioned features make the developed biomaterials promising candidates to be used as the wound dressings for the management of chronic wounds with moderate to high exudate.

Highly Porous and Superabsorbent Biomaterial Made of Marine-Derived Polysaccharides and Ascorbic Acid as an Optimal Dressing for Exuding Wound Management.

There are many modern wound dressings that have promising properties for repairing skin damage. However, due to various types of wounds and the problems they cause, there is still a great demand for new, effective healing strategies. The aim of this study was to create superabsorbent wound dressing made of marine-derived polysaccharides (agarose and chitosan) using the freeze-drying method. The secondary goal was its comprehensive evaluation for potential use as an external superabsorbent bandage for wounds with high exudation. Due to the well-known positive effect of ascorbic acid (vitamin C) on the healing process, biomaterial enriched with vitamin C was prepared and compared to the variant without the addition of ascorbic acid. It was shown that the produced foam-like wound dressing had a very porous structure, which was characterized by hydrophilicity, allowing a large amount of human fluids to be absorbed. According to in vitro tests on human fibroblasts, biomaterial was nontoxic and supportive to cell proliferation. Vitamin C-enriched dressing also had the ability to significantly reduce matrix metalloproteinase-2 production and to promote platelet-derived growth factor-BB synthesis by fibroblasts, which is desired during chronic wound treatment. The material has features of the eco-friendly wound care product since it was made of naturally-derived polysaccharides and was proved to be biodegradable. Importantly, despite degradable character, it was stable in the chronic and infected wound microenvironment, maintaining high integrity after 8-week incubation in the enzymatic solutions containing lysozyme and collagenases. The obtained results clearly showed that developed biomaterial possesses all necessary features of the external dressing for the management of exudate from both acute and chronic non-healing wounds.

The Effect of Liquid Rubber Addition on the Physicochemical Properties, Cytotoxicity, and Ability to Inhibit Biofilm Formation of Dental Composites.

The aim of this study was to evaluate the effect of modification with liquid rubber on the adhesion to tooth tissues (enamel, dentin), wettability and ability to inhibit bacterial biofilm formation of resin-based dental composites. Two commercial composites (Flow-Art-flow type with 60% ceramic filler and Boston-packable type with 78% ceramic filler; both from Arkona Laboratorium Farmakologii Stomatologicznej, Nasutów, Poland) were modified by addition of 5% by weight (of resin) of a liquid methacrylate-terminated polybutadiene. Results showed that modification of the flow type composite significantly (p < 0.05) increased the shear bond strength values by 17% for enamel and by 33% for dentine. Addition of liquid rubber significantly (p < 0.05) reduced also hydrophilicity of the dental materials since the water contact angle was increased from 81-83° to 87-89°. Interestingly, modified packable type material showed improved antibiofilm activity against Steptococcus mutans and Streptococcus sanguinis (quantitative assay with crystal violet), but also cytotoxicity against eukaryotic cells since cell viability was reduced to 37% as proven in a direct-contact WST-8 test. Introduction of the same modification to the flow type material significantly improved its antibiofilm properties (biofilm reduction by approximately 6% compared to the unmodified material, p < 0.05) without cytotoxic effects against human fibroblasts (cell viability near 100%). Thus, modified flow type composite may be considered as a candidate to be used as restorative material since it exhibits both nontoxicity and antibiofilm properties.

Polydopamine-coated curdlan hydrogel as a potential carrier of free amino group-containing molecules.

Curdlan hydrogel obtained after thermal gelling exhibits elasticity and high water-absorbing capacity. However, its modifications leading to the increase of biofunctionality usually alter its solubility and reduce mechanical parameters. Therefore, curdlan hydrogel was modified by deposition of polydopamine to improve its capacity to bind biologically active molecules with free amino groups. It exhibited the unchanged structure, mechanical properties and increased soaking capacity. Aminoglycoside antibiotic (gentamicin) as a model molecule was effectively immobilized to such modified curdlan via quinone moiety (but not amino groups) of polydopamine. Approximately 50 % of the immobilized drug was released following Fickian diffusion and inhibited the bacterial growth in matrix-surrounding medium in prolonged manner. The remaining drug amount was stably attached and prevented the hydrogel against bacterial adhesion even when all the mobile drug has been released. Therefore, polydopamine-modified curdlan hydrogel shows the potential for fabrication of functional materials for different purposes, including drug-loaded biomaterials.

Efficient non-contact heat generation on flexible, ternary hydroxyapatite/curdlan/nanomagnetite hybrids for temperature controlled processes.

The ternary HAp/curdlan/nanomagnetite hybrids with ceramic and polymer phase incorporation of magnetite nanoparticles (MNPs) were fabricated to study their heating ability under action of the alternating magnetic field (AMF), 808 nm near infrared laser radiation (NIR) and their synergic stimulation. The energy conversion was evaluated in terms of the specific absorption rate (SAR) as a function of the MNPs concentration in composites and to estimate their potential in temperature-controlled regenerative processes and hyperthermia. Measurements were carried out on dry and Ringer's solution soaked composite materials in order to mimic in situ conditions. It was found that the MNPs release during prolonged experiment is limited and has no significant effect on energy conversion emphasizing stability of the hybrids. Incorporation of the MNPs in polymer phase of the hybrid can additionally limit particle leaking as well as plays a role as insulating layer for the heat dissipation lowering the risk of sample overheating. In general, it was shown that maximum temperature of hybrid can be achieved in a relatively short time of exposure to stimulating factors whereas its control can be done through optimization of experiment conditions. MNPs incorporation into the curdlan (polymer phase) lead to strengthening of the mechanical properties of the whole network.

Effect of artificial saliva on the mechanical properties of a polymer material reinforced with fiber, used in esthetic tooth restorations.

BACKGROUND: The oral environment can negatively affect the physical properties of fiber-reinforced composite (FRC) materials, which can lead to the deterioration of mechanical stability and reduce the span of their clinical usefulness. OBJECTIVES: The aim of this study was to determine the influence of artificial saliva on the selected mechanical properties of FRC. MATERIAL AND METHODS: The core of the polymer material selected for the study was a bundle of ultrahighmolecular-weight polyethylene (UHMWPE) fibers. Fourteen samples were stored in an incubator at 37°C, in 20 mL of artificial saliva solution, and weighed on days 1 and 28. At the same time, mechanical tests were performed, including the measurements of Young's modulus, tensile stress, maximum tensile force, and tensile deformation. RESULTS: The analysis of basic statistics together with the results of the Shapiro-Wilk test and the distribution of Spearman's rho coefficient showed a strong negative relationship between the pair of variables - tensile deformation and the sorption of synthetic saliva. The results related to Young's modulus of elasticity and tensile stress were not statistically significant. CONCLUSIONS: Water penetration into the space between the fibers does not adversely affect the mechanical properties of the material tested. In the static tensile test, high and desired mechanical strength was observed, which may justify the effective use of this type of material in clinical practice and may be a good alternative to prosthetic restorations, whose retention is obtained only through a mechanical connection with the abutment tooth.

Fluorapatite ceramics for bone tissue regeneration: Synthesis, characterization and assessment of biomedical potential.

Calcium phosphates, due to their similarity to the inorganic fraction of mineralized tissues, are of great importance in treatment of bone defects. In order to improve the biological activity of hydroxyapatite (HAP), its fluoride-substituted modification (FAP) was synthesized using the sol-gel method and calcined at three different temperatures in the range of 800-1200 °C. Physicochemical and biological properties were evaluated to indicate which material would support bone regeneration the best. X-ray diffraction (XRD), scanning electron microscopy (SEM) with energy dispersive X-ray spectrometry (EDS), and Fourier transform infrared spectroscopy (FTIR) revealed that fluoride ions were incorporated into the apatite lattice structure. In studies it was found that fluorapatite sintered at the highest temperature had the lowest porosity, no internal pores and the highest density. In vitro ion reactivity assessments showed that during the 28-day immersion of the samples in the simulated body fluid, the uptake of calcium and phosphorus ions was inversely correlated to the calcination temperature. All tested materials were non-toxic since the cytotoxicity MTT assay demonstrated that the viability of preosteoblast cells incubated with sample extracts was high. Fluorapatite sintered at 800 °C was determined to be of optimal porosity and fluoride release capacity and then used in cell proliferation studies. The results showed that it significantly shortened the doubling time and thus enhanced the proliferation of osteogenic cells, as compared to the fluoride solutions and control group. Therefore, this material is proposed for the use in orthopedic applications and bone tissue engineering.

Porous Curdlan-Based Hydrogels Modified with Copper Ions as Potential Dressings for Prevention and Management of Bacterial Wound Infection-An In Vitro Assessment.

Bacterial infections at the wound site still remain a huge problem for current medicine, as they may lead to development of chronic wounds. In order to prevent such infections, there is a need to use wound dressings that possess ability to inhibit bacterial colonization. In this study, three new curdlan-based biomaterials modified with copper ions were fabricated via simple and inexpensive procedure, and their structural, physicochemical, and biological properties in vitro were evaluated. Received biomaterials possessed porous structure, had ability to absorb high amount of simulated wound fluid, and importantly, they exhibited satisfactory antibacterial properties. Nevertheless, taking into account all evaluated properties of new curdlan-based biomaterials, it seems that Cur_Cu_8% is the most promising biomaterial for management of wounds accompanied with bacterial infections. This biomaterial exhibited the best ability to reduce Escherichia coli and Staphylococcus aureus growth and moreover, it absorbed the highest amount of simulated wound fluid as well as enabled optimal water vapor transmission. Furthermore, Cur_Cu_8% biomaterial possessed the best values of selective indexes, which determine its potential safety in vitro. Thus, Cur_Cu_8% hydrogel may be considered as a promising candidate for management of infected wounds as well as it may constitute a good platform for further modifications.

Elastic and biodegradable chitosan/agarose film revealing slightly acidic pH for potential applications in regenerative medicine as artificial skin graft.

The aim of this work was to develop new method for the production of chitosan/agarose (CHN/A) film that could serve as a potential artificial skin substitute for regenerative medicine applications. Within this new production method, the concentrations of biomaterial components (chitosan in CH3COOH and agarose in NaOH) were optimized to provide neutralization of the resultant mixture up to pH approx. 6.0 without uncontrolled precipitation (gelation) of chitosan constituent. Produced thin film was subjected to comprehensive evaluation of its biomedical potential by determination of mechanical, physicochemical, and biological properties. It was demonstrated that the developed CHN/A film reveals slightly acidic pH of 5.98 known to support skin regeneration, high exudate absorption capacity, and is prone to biodegradation in simulated enzymatic wound environment. Performed tensile test proved high elastic deformations of the film (Young's modulus = 0.02 MPa, elongation at break = 23%) in a wet state, which would allow suitable stretching after application at the wound site. Moreover, novel biomaterial is non-toxic and supports fibroblast growth on its surface. Based on the obtained results it may be concluded that the produced CHN/A film possesses all necessary features of the artificial skin substitute.

Improved Fracture Toughness and Conversion Degree of Resin-Based Dental Composites after Modification with Liquid Rubber.

There are many methods widely applied in the engineering of biomaterials to improve the mechanical properties of the dental composites. The aim of this study was to assess the effect of modification of dental composites with liquid rubber on their mechanical properties, degree of conversion, viscosity, and cytotoxicity. Both flow and packable composite consisted of a mixture of Bis-GMA, TEGDMA, UDMA, and EBADMA resins reinforced with 60 and 78 wt.% ceramic filler, respectively. It was demonstrated that liquid rubber addition significantly increased the fracture toughness by 9% for flow type and 8% for condensable composite. The influence of liquid rubber on flexural strength was not statistically significant. The addition of the toughening agent significantly reduced Young's modulus by 7% and 9%, respectively, while increasing deformation at breakage. Scanning electron microscopy (SEM) observations allowed to determine the mechanisms of toughening the composites reinforced with ceramic particles. These mechanisms included bridging the crack edges, blocking the crack tip by particles and dissipation of fracture energy by deflection of the cracks on larger particles. The degree of conversion increased after modification, mainly due to a decrease in the matrix resin viscosity. It was also shown that all dental materials were nontoxic according to ISO 10993-5, indicating that modified materials have great potential for commercialization and clinical applications.

Ion-exchanging dialysis as an effective method for protein entrapment in curdlan hydrogel.

The purpose of this study was to determine whether dialysis method allows for efficient protein entrapment in curdlan-based hydrogel. Thus, bovine serum albumin, a model of bioactive protein, was incorporated into curdlan matrix using ion-exchanging dialysis method against two concentrations of CaCl2 solution - 2% and 10%, respectively. Then, physicochemical, mechanical, and biological properties of the bovine serum albumin-loaded curdlan hydrogels were evaluated. Received results show that neither the polymer nor the entrapment procedure change the bovine serum albumin conformation (as proven by Fourier transform infrared spectroscopy and circular dichroism spectroscopy) and the process guarantees high protein entrapment efficiency (above 95%). The curdlan-based carrier obtained against 2% of CaCl2 solution was found to possess higher swelling ability, release greater amounts of bovine serum albumin (up to 4 weeks), and exhibit superior biocompatibility compared to curdlan-based carrier obtained against 10% of CaCl2 solution. Thus, dialysis method enables efficient protein entrapment in curdlan hydrogel and obtained protein carrier via dialysis method into 2% of CaCl2 solution may be considered as a promising protein delivery system especially for tissue engineering applications. It should be noted that we are the first who presented effective method for protein entrapment in curdlan hydrogel.

Development and Optimization of the Novel Fabrication Method of Highly Macroporous Chitosan/Agarose/Nanohydroxyapatite Bone Scaffold for Potential Regenerative Medicine Applications.

Bone scaffolds mimicking the three-dimensional bone structure are of essential importance for bone regeneration. The aim of this study was to develop and optimize the production method of highly macroporous bone scaffold composed of polysaccharide matrix (chitosan-agarose) reinforced with nanohydroxyapatite. The highly macroporous structure was obtained by the simultaneous application of a gas-foaming agent and freeze-drying technique. Fabricated variants of biomaterials (produced using different gas-foaming agent and solvent concentrations) were subjected to porosity evaluation and compression test in order to select the scaffold with the best properties. Then, bioactivity, cytotoxicity, and cell growth on the surface of the selected biomaterial were assessed. The obtained results showed that the simultaneous application of gas-foaming and freeze-drying methods allows for the production of biomaterials characterized by high total and open porosity. It was proved that the best porosity is obtained when solvent (CH3COOH) and foaming agent (NaHCO3) are applied at ratio 1:1. Nevertheless, the high porosity of novel biomaterial decreases its mechanical strength as determined by compression test. Importantly, novel scaffold is non-toxic to osteoblasts and favors cell attachment and growth on its surface. All mentioned properties make the novel biomaterial a promising candidate to be used in regenerative medicine in non-load bearing implantation sites.

Behavior of new hydroxyapatite/glucan composite in human serum.

Biomaterials for bone tissue regeneration, including polymer-based composites, are typically evaluated in vitro prior to the clinical trials. However, such composites tested in vivo may behave different due to the specific body conditions. For example, some composites implanted into the tissue acidified due to transient postoperative inflammation may unexpectedly swell which delays the wound healing. Such massive swelling in acidic medium was previously observed for new elastic hydroxyapatite (HAp)/ß-glucan biomaterial. However, in further clinical cases concerning the composite implantation in patients without significant inflammation indicators, no side effects were observed. Therefore, it was reasonable to test the effect of human serum of neutral pH (typical for noninflamed tissues) on the composite parameters, in particular volume changes. Thus, this article shows the characterization of physicochemical parameters of the composite after incubation (5 days) in human serum of neutral pH by means of weight and volume measurement, scanning electron microscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, microcomputed tomography, mercury intrusion, and biochemical techniques. Results showed that human serum collected from healthy people caused no uncontrolled changes in weight and volume, porosity and mechanical properties of the composite. Therefore, this suggests the lack of volume change-related side effects of HAp/glucan composite in bone defects treatment if postoperative inflammation is prevented. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2653-2664, 2018.