Healing of Skin Wounds in Rats Using Creams Based on Symphytum Officinale Extract.

Rosmarinic acid is a well-known natural antioxidant and anti-inflammatory compound, and it is one of the polyphenolic compounds found in comfrey plants. Comfrey root also contains allantoin, which helps with new skin regeneration. This study aimed to investigate the healing and skin regeneration process of skin wounds in Wistar rats using creams based on comfrey extract and to correlate the results with active compounds in the extract. The obtained results showed that comfrey root is rich in bioactive compounds, including allantoin, salvianolic acid, and rosmarinic acid, which are known for their great free radical scavenging activity, and the high antioxidant activity of the extract may be mainly due to these compounds. The obtained extract has an antimicrobial effect on Staphylococcus aureus (1530.76/382.69), Escherichia coli (6123.01/6123.01), and Pseudomonas aeruginosa (6123.01/6123.01). The macroscopic evaluation and the histological analysis of the skin defects 14 days after the intervention showed faster healing and complete healing in the skin excisions treated with oil-in-water cream with 20% extract of comfrey as the active ingredient.

Gelatin-assisted fabrication of reduced nanographene oxide for dual-modal imaging of melanoma cells.

In this work we report a gelatin-based, simple two-steps approach for fabrication of reduced graphene oxide (rGO-GEL) possessing high stability and biocompatibility, as novel label-free intracellular contrast agents. Gelatin, a biopolymer that is known for its versatility, was employed not only to biocompatibilize the rGO, but also to prevent the aggregation of the GO nanosheets during the reduction process. To confirm the successful reduction process and the attachment of the gelatin to the rGO nanosheets, we employed multiple spectroscopic analyses such as FT-IR, Raman, UV-VIS and photoluminescence, while the morphology and the lateral dimensions of the resulting hybrid rGO-GEL were investigated by Scanning-Transmission Electron Microscopy (STEM). Cellular toxicity test proved that the rGO-GEL nanoflakes are nontoxic for melanoma B16-F10 cells, even at high concentrations. Finally, the intracellular tracking after 24 h of treatment was performed by non-invasive Super-resolution re-scan confocal microscopy as well as Confocal Raman imaging, thus implementing our nanoflakes as a suitable contrast agent candidate for cellular imaging of interest.

The evaluation of the osteopromoting capabilities of composites based on biopolymers and gold/silver nanoparticles doped bioactive glasses on an experimental rat bone defect.

The most important concept behind using bone scaffolds is the biocompatibility of the material to avoid a local inflammatory response and must have the following properties: osteoinduction, osteoconductivity, angiogenesis, and mechanical support for cell growth. Gold nanoparticles/gold and silver nanoparticles -containing bioactive glasses in biopolymer composites have been used to enhance bone regeneration. These composites were testedin vitroon fibroblast and osteoblast cell lines using MTT tests, immunofluorescence, scanning electron microscopy analysis, andin vivoin an experimental bone defect in Sprague-Dawley rats. Both composites promoted adequate biological effects on human fibroblastic BJ (CRL 2522TM) cell lines and human osteoblastic cells isolated from the human patella in terms of cell proliferation, morphology, migration, and attachment. Most importantly, they did not cause cellular apoptosis and necrosis. According to the histological and immunohistochemical results, both composites were osteoinductive and promoted new bone formation at 60 d. Evidence from this study suggests that the small amount of silver content does not influence negatively thein vitroorin vivoresults. In addition, we obtained accurate results proving that the existence of apatite layer and proteins on the surface of the recovered composite, supports the validity ofin vitrobioactivity research.

Anionic Polysaccharide Cryogels: Interaction and In Vitro Behavior of Alginate-Gum Arabic Composites.

In the present study, polysaccharide-based cryogels demonstrate their potential to mimic a synthetic extracellular matrix. Alginate-based cryogel composites with different gum arabic ratios were synthesized by an external ionic cross-linking protocol, and the interaction between the anionic polysaccharides was investigated. The structural features provided by FT-IR, Raman, and MAS NMR spectra analysis indicated that a chelation mechanism is the main process linking the two biopolymers. In addition, SEM investigations revealed a porous, interconnected, and well-defined structure suitable as a scaffold in tissue engineering. The in vitro tests confirmed the bioactive character of the cryogels through the development of the apatite layer on the surface of the samples after immersion in simulated body fluid, identifying the formation of a stable phase of calcium phosphate and a small amount of calcium oxalate. Cytotoxicity tests performed on fibroblast cells demonstrated the non-toxic effect of alginate-gum arabic cryogel composites. In addition, an increase in flexibility was noted for samples with a high gum arabic content, which determines an appropriate environment to promote tissue regeneration. The newly obtained biomaterials that exhibit all these properties can be successfully involved in the regeneration of soft tissues, wound management, or controlled drug release systems.

Insights into the Stability of Graphene Oxide Aqueous Dispersions.

Understanding graphene oxide's stability (or lack thereof) in liquid solvents is critical for fine-tuning the material's characteristics and its potential involvement in future applications. In this work, through the use of structural and surface investigations, the alteration of the structural and edge-surface properties of 2D graphene oxide nanosheets was monitored over a period of eight weeks by involving DLS, zeta potential, XRD, XPS, Raman and FT-IR spectroscopy techniques. The samples were synthesized as an aqueous suspension by an original modified Marcano-Tour method centred on the sono-chemical exfoliation of graphite. Based on the acquired experimental results and the available literature, a phenomenological explanation of the two underlying mechanisms responsible for the meta-stability of graphene oxide aqueous dispersions is proposed. It is based on the cleavage of the carbon bonds in the first 3-4 weeks, while the bonding of oxygen functional groups on the carbon lattice occurs, and the transformation of epoxide and hydroxyl groups into adsorbed water molecules in a process driven by the availability of hydrogen in graphene oxide nanosheets.

Bioactive Properties of Composites Based on Silicate Glasses and Different Silver and Gold Structures.

Using an ideal biomaterial to treat injured bones can accelerate the healing process and simultaneously exhibit antibacterial properties; thus protecting the patient from bacterial infections. Therefore, the aim of this work was to synthesize composites containing silicate-based bioactive glasses and different types of noble metal structures (i.e., AgI pyramids, AgIAu composites, Au nanocages, Au nanocages with added AgI). Bioactive glass was used as an osteoconductive bone substitute and Ag was used for its antibacterial character, while Au was included to accelerate the formation of new bone. To investigate the synergistic effects in these composites, two syntheses were carried out in two ways: AgIAu composites were added in either one step or AgI pyramids and Au nanocages were added separately. All composites showed good in vitro bioactivity. Transformation of AgI in bioactive glasses into Ag nanoparticles and other silver species resulted in good antibacterial behavior. It was observed that the Ag nanoparticles remained in the Au nanocages, which was also beneficial in terms of antibacterial properties. The presence of Au nanoparticles contributed to the composites achieving high cell viability. The most outstanding result was obtained by the consecutive addition of noble metals into the bioactive glasses, resulting in both a high antibacterial effect and good cell viability.

Co-Crystals of Etravirine by Mechanochemical Activation.

The co-crystals formation of etravirine with three carboxylic acids was investigated. New co-crystals of etravirine with adipic acid, benzoic acid, and 4-hydroxybenzoic acid have been synthesized by wet milling of ingredients for 120 min. The novelty of these solid forms was first evidenced by powder X-ray diffraction. Their different morphology was evidenced by SEM microscopy. Spectroscopic analyses (FT-IR, MAS-NMR, and XPS) highlighted the hydrogen bonds between etravirine and co-formers, as a result of the solid-state reaction of the ingredients by wet milling. Thermal analyses pointed out that the milling process caused in co-crystals a reduction in the fusion enthalpy and the melting temperature, compared to the values obtained for etravirine. These co-crystals are stable up to four months on storage under extreme conditions, excepting the co-crystal with benzoic acid which begins to transform into a polymorph of etravirine after 30 days. The UV absorption spectra of the samples tested in three simulated physiological media with pH values of 6, 6.3, and 7 have evidenced the conformation change of etravirine due to hydrogen bonds between etravirine and carboxylic acids.

Rapid Synthesis Method of Ag3PO4 as Reusable Photocatalytically Active Semiconductor.

The widespread use of Ag3PO4 is not surprising when considering its higher photostability compared to other silver-based materials. The present work deals with the facile precipitation method of silver phosphate. The effects of four different phosphate sources (H3PO4, NaH2PO4, Na2HPO4, Na3PO4·12 H2O) and two different initial concentrations (0.1 M and 0.2 M) were investigated. As the basicity of different phosphate sources influences the purity of Ag3PO4, different products were obtained. Using H3PO4 did not lead to the formation of Ag3PO4, while applying NaH2PO4 resulted in Ag3PO4 and a low amount of pyrophosphate. The morphological and structural properties of the obtained samples were studied by X-ray diffractometry, diffuse reflectance spectroscopy, scanning electron microscopy, infrared spectroscopy, and X-ray photoelectron spectroscopy. The photocatalytic activity of the materials and the corresponding reaction kinetics were evaluated by the degradation of methyl orange (MO) under visible light. Their stability was investigated by reusability tests, photoluminescence measurements, and the recharacterization after degradation. The effect of as-deposited Ag nanoparticles was also highlighted on the photostability and the reusability of Ag3PO4. Although the deposited Ag nanoparticles suppressed the formation of holes and reduced the degradation of methyl orange, they did not reduce the performance of the photocatalyst.

Bioactive glass-biopolymers­gold nanoparticle based composites for tissue engineering applications.

Biomaterials based on bioactive glass with gold nanoparticle composites have many applications in tissue engineering due to their tissue regeneration and angiogenesis capacities. The objectives of the study were to develop new composites using bioactive glass with gold nanospheres (BGAuSP) and gold nanocages (BGAuIND), individually introduced in alginate-pullulan (Alg-Pll) polymer, to evaluate their biocompatibility potential, and to compare the obtained results with those achieved when ß-tricalcium phosphate-hydroxyapatite (ßTCP/HA) replaced the BG. The novel composites underwent structural and morphological characterization followed by in vitro viability testing on fibroblast and osteoblast cell lines. Additionally, the biomaterials were subcutaneously implanted in Sprague Dawley rats, for in vivo biocompatibility assessment during 3 separate time frames (14, 30 and 60 days). The biological effects were evaluated by histopathology and immunohistochemistry. The physical characterization revealed the cross-linking between polymers and glasses/ceramics and demonstrated a suitable thermal stability for sterilization processes. The in vitro assays demonstrated adequate form, pore size of composites ranging from few micrometers up to 100 µm, while the self-assembled apatite layer formed after simulated body fluid immersion confirmed the composites' bioactivity. Viability assays have highlighted optimal cellular proliferation and in vitro biocompatibility for all tested composites. Furthermore, based on the in vivo subcutaneous analyses the polymer composites with BGAuNP have shown excellent biocompatibility at 14, 30 and 60 days, exhibiting marked angiogenesis while, tissue proliferation was confirmed by high number of Vimentin positive cells, in comparison with the polymer composite that contains ßTCP/HA, which induced an inflammatory response represented by a foreign body reaction. The obtained results suggest promising, innovative, and biocompatible composites with bioactive properties for future soft tissue and bone engineering endeavours.

The Impact of Composites with Silicate-Based Glasses and Gold Nanoparticles on Skin Wound Regeneration.

The silver content of the skin regeneration ointments can influence its regeneration process but in the meantime, it can take the benefit of the antibacterial properties of silver by avoiding the bacterial infection of an open wound. In the current study, the skin healing and regeneration capacity of bioactive glass with spherical gold nanocages (BGAuIND) in the Vaseline ointments were evaluated in vivo comparing the bioactive glass (BG)-Vaseline and bioactive glass with spherical gold (BGAuSP)-Vaseline ointments. Spherical gold nanocages are stabilized with silver and as a consequence the BGAuIND exhibits great antibacterial activity. Histological examination of the cutaneous tissue performed on day 8 indicates a more advanced regeneration process in rats treated with BGAuSP-Vaseline. The histopathological examination also confirms the results obtained after 11 days post-intervention, when the skin is completely regenerated at rats treated with BGAuSP-Vaseline compared with the others groups where the healing was incomplete. This result is also confirmed by the macroscopic images of the evolution of wounds healing. As expected, the silver content influences the wound healing process but after two weeks, for all of the post-interventional trials from the groups of rats, the skin healing was completely.

Shape tailoring of AgBr microstructures: effect of the cations of different bromide sources and applied surfactants.

Investigations regarding AgBr-based photocatalysts came to the center of attention due to their high photosensitivity. The present research focuses on the systematic investigation regarding the effect of different alkali metal cation radii and surfactants/capping agents applied during the synthesis of silver-halides. Their morpho-structural and optical properties were determined via X-ray diffractometry, diffuse reflectance spectroscopy, scanning electron microscopy, infrared spectroscopy, and contact angle measurements. The semiconductors' photocatalytic activities were investigated using methyl orange as the model contaminant under visible light irradiation. The correlation between the photocatalytic activity and the obtained optical and morpho-structural properties was analyzed using generalized linear models. Moreover, since the (photo)stability of Ag-based photoactive materials is a crucial issue, the stability of catalysts was also investigated after the degradation process. It was concluded that (i) the photoactivity of the samples could be fine-tuned using different precursors and surfactants, (ii) the as-obtained AgBr microcrystals were transformed into other Ag-containing composites during/after the degradation, and (iii) elemental bromide did not form during the degradation process. Thus, the proposed mechanisms in the literature (for the degradation of MO using AgBr) must be reconsidered.

Innovative and Cost-Efficient BiOI Immobilization Technique on Ceramic Paper-Total Coverage and High Photocatalytic Activity.

In the present work, visible light active bismuth oxyiodide (BiOI) was immobilized on a commercial, non-conductive support (an Al2O3 based ceramic paper) using a novel two-step spray coating technique and investigated with different characterization methods (e.g., SEM, Raman, XPS). Our main goal was to eliminate the separation costs after the photocatalytic measurement and investigate the chemical relevance and opportunity to use this technique in the industry. Our as-prepared uniform BiOI layer had similar properties to the well-known reference BiOI powder. The Raman and XPS measurements confirmed that the enriched amount of the surface iodine defined the color and as well the band gap of the BiOI layer. The durable BiOI layers have prominent photocatalytic activity under UV and visible light irradiation as well. The scale-up procedure proved that the designed BiOI coated paper was reusable and potentially applicable in the industry by straightforward scale-up, which is due to the elaborated non-conventional BiOI coverage estimation method. This immobilization technique could open several opportunities for immobilizing many other visible light active photocatalysts with simple materials and low cost.

Multi-analyses of gallstones and correlation between their properties with the laboratory results.

This study explores the morpho-structure of gallstones (GSs) removed from 36 patients in NW Romania and correlate it with the laboratory results of the patients. GSs were analyzed by SEM-EDS, X-ray diffraction and IR, UV-Vis and X-ray photoelectron spectroscopy. The laboratory studies consisted in hematological, coagulation, biochemistry, immunological and tumor markers tests. The morphological and structural investigations allowed to classify the GS in five different types and to establish their mechanism of formation. Only macroscopic evaluation, SEM microscopy, FTIR and UV-Vis spectroscopy give different easily noticeable information for each GS type. EDS, XPS and XRD diffraction are recommended to distinguish pigment and carbonate stones from the other GS types and a carefully examination is needed to establish the differences between the pure cholesterol, the mixed cholesterol and the composite cholesterol stones, due to the high similarities. The variation of specific markers cannot differentiate the patients with pure cholesterol GS from those with mixed cholesterol and pigment GS and those with mixed cholesterol from those with composite cholesterol stones. Seven laboratory parameters (RDW-CV, MPV, PCT, GLUC-HK, WBC, PT, GPT) are the key indicators for the GS disease and trend to present generally higher values than normal.

Bone regeneration response in an experimental long bone defect orthotopically implanted with alginate-pullulan-glass-ceramic composite scaffolds.

In the present study, scaffolds based on alginate-pullulan-bioactive glass-ceramic with 0.5 and 1.5 mol % copper oxide were orthotopically implanted in experimental rat models to assess their ability to heal an induced bone defect. By implying magnetic resonance and imaging scans together with histological evaluation of the processed samples, a progressive healing of bone was observed within 5 weeks. Furthermore, as the regenerative process continued, new bone tissue was formed, enhancing the growth of irregular bone spicules around the scaffolds. A significantly higher amount of new bone was formed (37%) in the defect that received the composite with 1.5 mol % CuO (in glass-ceramic matrix) content implant. Nevertheless, the bone regeneration obtained by scaffold with 0.5 mol % CuO implanted is comparable with the alginate-pullulan-ß-tricalcium phosphate/hydroxiapatite composite implant. The assessed amount of new bone formed was found to be between 29.75 and 37.15% for all the composition involved in the present study. During this process a regeneration process was shown when the alginate-pullulan composite materials were involved, fact that indicate the great potential of these materials to be used in tissue engineering.

Utilization of Carbon Nanospheres in Photocatalyst Production: From Composites to Highly Active Hollow Structures.

Titanium dioxide-carbon sphere (TiO2-CS) composites were constructed via using prefabricated carbon spheres as templates. By the removal of template from the TiO2-CS, TiO2 hollow structures (HS) were synthesized. The CS templates were prepared by the hydrothermal treatment of ordinary table sugar (sucrose). TiO2-HSs were obtained by removing CSs with calcination. Our own sensitized TiO2 was used for coating the CSs. The structure of the CSs, TiO2-CS composites, and TiO2-HSs were characterized by scanning electron microscopy (SEM), infrared spectroscopy (IR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and diffuse reflectance spectroscopy (DRS). The effect of various synthesis parameters (purification method of CSs, precursor quantity, and applied furnace) on the morphology was investigated. The photocatalytic activity was investigated by phenol model pollutant degradation under visible light irradiation (λ > 400 nm). It was established that the composite samples possess lower crystallinity and photocatalytic activity compared to TiO2 hollow structures. Based on XPS measurements, the carbon content on the surface of the TiO2-HS exerts an adverse effect on the photocatalytic performance. The synthesis parameters were optimized and the TiO2-HS specimen having the best absolute and surface normalized photocatalytic efficiency was identified. The superior properties were explained in terms of its unique morphology and surface properties. The stability of this TiO2-HS was investigated via XRD and SEM measurements after three consecutive phenol degradation tests, and it was found to be highly stable as it entirely retained its crystal phase composition, morphology and photocatalytic activity.

Intracellular Dynamic Disentangling of Doxorubicin Release from Luminescent Nanogold Carriers by Fluorescence Lifetime Imaging Microscopy (FLIM) under Two-Photon Excitation.

There is still a lack of available techniques to follow noninvasively the intracellular processes as well to track or disentangle various signals from the therapeutic agents at the site of action in the target cells. We present here the assessment of the intracellular kinetics of doxorubicin (DOX) and gold nanoparticle (AuNP) carriers by mapping simultaneously fluorescence and photoluminescence signals by fluorescence lifetime imaging microscopy under two-photon excitation (TPE-FLIM). The new nano-chemotherapeutic system AuNPs@gelatin-hyd-DOX has been fabricated by DOX loading onto the surface of gelatin-biosynthesized AuNPs (AuNPs@gelatin) through a pH-sensitive hydrazone bond. The successful loading of DOX onto the AuNPs was studied by spectroscopic methods and steady-state fluorescence, and the nanosystem pH-responsive character was validated under simulated biological conditions at different pH values (i.e., pH 4.6, 5.3, and 7.4). Considering that the fluorescence lifetime of DOX molecules at a specific point in the cell is a reliable indicator of the discrimination of the different states of the drug in the internalization path, i.e., released versus loaded, the kinetics of AuNPs@gelatin-hyd-DOX cellular uptake and DOX release was compared to that of free DOX, resulting in two different drug internalization pathways. Finally, cell viability tests were conducted against NIH:OVCAR-3 cell line to prove the efficiency of our chemotherapeutic nanosystem. TPE-FLIM technique could be considered promising for noninvasive, high-resolution imaging of cells with improved capabilities over current one-photon-excited FLIM.

New alginate-pullulan-bioactive glass composites with copper oxide for bone tissue regeneration trials.

Composites based on sodium alginate, pullulan, and bioactive SiO2 -CaO-P2 O5 glass-ceramics with copper oxide were prepared as capsules. The obtained samples were structurally characterized by Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM), and their bioactivity and biocompatibility properties were also tested both in vitro and in vivo by XRD, FT-IR, SEM, and high-resolution transmission electron microscopy. The fibroblast and osteoblast cell viability assays have shown good proliferation rates for all investigated samples, whereas all composites exhibited a good in vivo tolerance. The recovered composites after 5 weeks' in vivo and in vitro trials evidenced clear macroscopic alterations; particularly, after soaking in simulated body fluid, they have a corn flake aspect, and after their in vivo inoculation, a globular shape is retained. Different crystalline shapes of hydroxyapatite were formed after in vitro and in vivo trials for the glass-ceramic-polymer composites, the in vitro precipitated apatite was found to be nodular, and the in vivo experiment led to needlelike crystallites formation. Histopathological results showed a good biocompatibility with no significant signs of rejection by the host tissue. These assessments performed on the composites indicate that the studied materials can be considered without any doubt suitable candidates for future bone regeneration applications.

Surface Plasmon Resonance or Biocompatibility-Key Properties for Determining the Applicability of Noble Metal Nanoparticles.

Metal and in particular noble metal nanoparticles represent a very special class of materials which can be applied as prepared or as composite materials. In most of the cases, two main properties are exploited in a vast number of publications: biocompatibility and surface plasmon resonance (SPR). For instance, these two important properties are exploitable in plasmonic diagnostics, bioactive glasses/glass ceramics and catalysis. The most frequently applied noble metal nanoparticle that is universally applicable in all the previously mentioned research areas is gold, although in the case of bioactive glasses/glass ceramics, silver and copper nanoparticles are more frequently applied. The composite partners/supports/matrix/scaffolds for these nanoparticles can vary depending on the chosen application (biopolymers, semiconductor-based composites: TiO2, WO3, Bi2WO6, biomaterials: SiO2 or P2O5-based glasses and glass ceramics, polymers: polyvinyl alcohol (PVA), Gelatin, polyethylene glycol (PEG), polylactic acid (PLA), etc.). The scientific works on these materials' applicability and the development of new approaches will be targeted in the present review, focusing in several cases on the functioning mechanism and on the role of the noble metal.

Bioactivity evolution of the surface functionalized bioactive glasses.

The formation of a calcium phosphate layer on the surface of the SiO2 -CaO-P2 O5 glasses after immersion in simulated body fluid (SBF) generally demonstrates the bioactivity of these materials. Grafting of the surface by chemical bonding can minimize the structural changes in protein adsorbed on the surface. Therefore, in this study our interest was to evaluate the bioactivity and blood biocompatibility of the SiO2 -CaO-P2 O5 glasses after their surface modification by functionalization with aminopropyl-triethoxysilane and/or by fibrinogen. It is shown that the fibrinogen adsorbed on the glass surfaces induces a growing of the apatite-like layer. It is also evidenced that the protein content from SBF influences the growth of the apatite-like layer. Furthermore, the good blood compatibility of the materials after fibrinogen and bovine serum albumin adsorption is proved from the assessment of the ß-sheet-ß-turn ratio.

Addressing the optimal silver content in bioactive glass systems in terms of BSA adsorption.

Bioactive glasses doped with silver are aimed to minimize the risk of microbial contamination; therefore, the influence of silver on the bioactive properties is intensely investigated. However, information related to the role played by silver, when added to the bioactive glass composition, on biocompatibility properties is scarce. This aspect is essential as long as the silver content can influence blood protein adsorption onto the surface of the glass, thus affecting the material's biocompatibility. Therefore, from the perspective of the biocompatibility standpoint, the finding of an optimal silver content in a bioactive glass is an extremely important issue. In this study, silver-doped bioactive glasses were prepared by a melt-derived technique, which eliminates the pores' influence in the protein adsorption process. The obtained glasses were characterized by X-ray diffraction, UV-vis, X-ray photoelectron (XPS) and Fourier transform infrared (FT-IR) spectroscopy; afterwards, they were investigated in terms of protein adsorption. Both UV-vis and XPS spectroscopy revealed the presence of Ag+ ions in all silver containing samples. By increasing the silver content, metallic Ag0 appears, the highest amount being observed for the sample with 1 mol% AgO2. Electron paramagnetic resonance measurements evidenced that the amount of spin-labeled serum albumin attached to the surface increases with the silver content. The results obtained by analyzing the information derived from atomic force microscopy and FT-IR measurements indicate that the occurrence of metallic Ag0 in the samples' structure influences the secondary structure of the adsorbed protein. Based on the results derived from the protein response upon interaction with the investigated glass calcium-phosphate based system, the optimal silver oxide concentration was determined for which the secondary structure of the adsorbed protein is similar with that of the free one. This concentration was found to be 0.5 mol%.