Gelatin Methacryloyl (GelMA) - 45S5 Bioactive Glass (BG) Composites for Bone Tissue Engineering: 3D Extrusion Printability and Cytocompatibility Assessment Using Human Osteoblasts.

3D extrusion printing has been widely investigated for low-volume production of complex-shaped scaffolds for tissue regeneration. Gelatin methacryloyl (GelMA) is used as a baseline material for the synthesis of biomaterial inks, often with organic/inorganic fillers, to obtain a balance between good printability and biophysical properties. The present study demonstrates how 45S5 bioactive glass (BG) addition and GelMA concentrations can be tailored to develop GelMA composite scaffolds with good printability and buildability. The experimental results suggest that 45S5 BG addition consistently decreases the compression stiffness, irrespective of GelMA concentration, albeit within 20% of the baseline scaffold (without 45S5 BG). The optimal addition of 2 wt % 45S5 BG in 7.5 wt % GelMA was demonstrated to provide the best combination of printability and buildability in the 3D extrusion printing route. The degradation decreases and the swelling kinetics increases with 45S5 BG addition, irrespective of GelMA concentration. Importantly, the dissolution in simulated body fluid over 3 weeks clearly promoted the nucleation and growth of crystalline calcium phosphate particles, indicating the potential of GelMA-45S5 BG to promote biomineralization. The cytocompatibility assessment using human osteoblasts could demonstrate uncompromised cell proliferation or osteogenic marker expression over 21 days in culture for 3D printable 7.5 wt % GelMA -2 wt % 45S5 BG scaffolds when compared to 7.5 wt % GelMA. The results thus encourage further investigations of the GelMA/45S5 BG composite system for bone tissue engineering applications.

Potential magnetic drug targeting with magnetite nanoparticles in cancer treatment by enhancer-modifier natural herb and loaded drug.

In the present study, we prepared magnetite nanoparticles (MNPs) loaded with natural Moringa oleifera (M. olf) herb and Epilim (Ep) drug to evaluate the anti-cancerous activity against brain cancer cells. All the samples were prepared via co-precipitation approach modified with different concentrations of M. olf and Ep drug at room temperature. The MNPs loaded with drug and natural herb were studied in terms of crystal structure, morphology, colloidal stability, size distribution, and magnetic properties. Field emission scanning electron microscopy (FESEM) images exhibited the morphologies of samples with spherical shape as well as the particles size of 9 nm for MNPs and up to 23 nm for its composites. The results of vibrating sample magnetometer (VSM) indicated the magnetization saturation (Ms) of 42.510 emu/g for MNPs. This value reduced to 16-35 emu/g upon loading MNPs with different concentrations of M. olf and Ep. Fourier transform infrared spectroscopy (FTIR) indicated the chemical interaction between the Ep, M.olf and MNPs. Brunauer-Emmett-Teller (BET) analysis confirmed the largest surface area for MNPs (422.61 m2/g) which gradually reduced on addition of M. olf and Ep indicating the successful loading. The zeta potential measurements indicated that the MNPs and MNPs loaded with M. olf and Ep are negatively charged and can be dispersed in the suspension. Furthermore, U87 human glioblastoma cell line was used for the in vitro cellular studies to determine the efficacy of synthesized MNPs against cancer cells. The results confirmed the anti-proliferative activity of the MNPs loaded with M. olf and Ep.

3D-printed polyether-ether ketone/carboxymethyl cellulose scaffolds coated with Zn-Mn doped mesoporous bioactive glass nanoparticles.

Patient-specific fabrication of scaffold/implant requires an engineering approach to manufacture the ideal scaffold. Herein, we design and 3D print scaffolds comprised of polyether-ether-ketone (PEEK) and sodium-carboxymethyl cellulose (Na-CMC). The fabricated scaffold was dip coated with Zn and Mn doped bioactive glass nanoparticles (Zn-Mn MBGNs). The synthesized ink exhibit suitable shear-thinning behavior for direct ink write (DIW) 3D printing. The scaffolds were crafted with precision, featuring 85% porosity, 0.3 mm layer height, and 1.5 mm/s printing speed at room temperature. Scanning electron microscopy images reveal a well-defined scaffold with an average pore size of 600 ± 30 µm. The energy dispersive X-ray spectroscopy analysis confirmed a well dispersed/uniform coating of Zn-Mn MBGNs on the PEEK/Na-CMC scaffold. Fourier transform infrared spectroscopy approved the presence of PEEK, CMC, and Zn-Mn MBGNs. The tensile test revealed a Young's modulus of 2.05 GPa. Antibacterial assays demonstrate inhibition zone against Staphylococcus aureus and Escherichia Coli strains. Chick Chorioallantoic Membrane assays also present significant angiogenesis potential, owing to the antigenic nature of Zn-Mn MBGNs. WST-8 cell viability assays depicted cell proliferation, with a 103% viability after 7 days of culture. This study suggests that the PEEK/Na-CMC scaffolds coated with Zn-Mn MBGNs are an excellent candidate for osteoporotic fracture treatment. Thus, the fabricated scaffold can offer multifaceted properties for enhanced patient outcomes in the bone tissue regeneration.

Oxidized alginate-gelatin (ADA-GEL)/silk fibroin/Cu-Ag doped mesoporous bioactive glass nanoparticle-based hydrogels for potential wound care treatments.

The present work focuses on developing 5% w/v oxidized alginate (alginate di aldehyde, ADA)-7.5% w/v gelatin (GEL) hydrogels incorporating 0.25% w/v silk fibroin (SF) and loaded with 0.3% w/v Cu-Ag doped mesoporous bioactive glass nanoparticles (Cu-Ag MBGNs). The microstructural, mechanical, and biological properties of the composite hydrogels were characterized in detail. The porous microstructure of the developed ADA-GEL based hydrogels was confirmed by scanning electron microscopy, while the presence of Cu-Ag MBGNs in the synthesized hydrogels was determined using energy dispersive x-ray spectroscopy. The incorporation of 0.3% w/v Cu-Ag MBGNs reduced the mechanical properties of the synthesized hydrogels, as investigated using micro-tensile testing. The synthesized ADA-GEL loaded with 0.25% w/v SF and 0.3% w/v Cu-Ag MBGNs showed a potent antibacterial effect againstEscherichia coliandStaphylococcus aureus. Cellular studies using the NIH3T3-E1 fibroblast cell line confirmed that ADA-GEL films incorporated with 0.3% w/v Cu-Ag MBGNs exhibited promising cellular viability as compared to pure ADA-GEL (determined by WST-8 assay). The addition of SF improved the biocompatibility, degradation rate, moisturizing effects, and stretchability of the developed hydrogels, as determinedin vitro. Such multimaterial hydrogels can stimulate angiogenesis and exhibit desirable antibacterial properties. Therefore further (in vivo) tests are justified to assess the hydrogels' potential for wound dressing and skin tissue healing applications.

Synthesis and characterization of mesoporous bioactive glass nanoparticles loaded with peganum harmala for bone tissue engineering.

Globally, there is an increase in a number of bone disorders including osteoarthritis (OA), osteomyelitis, bone cancer, and etc., which has led to a demand for bone tissue regeneration. In order to take use of the osteogenic potential of natural herbs, mesoporous bioactive glass nanoparticles (MBGNs) have the ability to deliver therapeutically active chemicals locally. MBGNs influence bioactivity and osteointegration of materials making them suitable for bone tissue engineering (BTE). In the present study, we developed Peganum Harmala (P. harmala) loaded MBGNs (PH-MBGNs) synthesized via modified Stöber process. The MBGNs were analyzed in terms of surface morphology, chemical make-up, amorphous nature, chemical interaction, pore size, and surface area before and after loading with P. harmala. A burst release of drug from PH-MBGNs was observed within 8 h immersion in phosphate buffer saline (PBS). PH-MBGNs effectively prevented Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) from spreading. Furthermore, PH-MBGNs developed a hydroxyapatite (HA) layer in the presence of simulated body fluid (SBF) after 21 days, which confirmed the in-vitro bioactivity of MBGNs. In conclusion, PH-MBGNs synthesized in this work are potential candidate for scaffolding or a constituent in the coatings for BTE applications.

Biodegradable Packaging Based on Poly(vinyl Alcohol) and Carboxymethyl Cellulose Films Incorporated with Ascorbic Acid for Food Packaging Applications.

Petroleum-based plastics are used as packaging materials because of their low cost and high availability; however, continuous use of these nondegradable materials especially in the food industry has led to environmental pollution. The present study aimed to synthesize antibacterial and biodegradable films based on natural biopolymers carboxymethyl cellulose (CMC), poly(vinyl alcohol) (PVA), and ascorbic acid (AA) cross-linked in the presence of glutaraldehyde (GA). The films were synthesized in two different concentrations, 60PVA:40CMC:AA and 70PVA:30CMC:AA with a fixed amount of AA. Films with smooth texture and overall uniform thickness were obtained. Fourier transform infrared spectroscopy (FTIR) confirmed the cross-linking between the aldehyde group of GA and hydroxyl of PVA through detection of acetal and ether bridges. The synthesized films were thermally stable in the temperature range of 180-300 °C; however, 70PVA:30CMC:AA showed higher weight loss in this range as compared to the 60PVA:40CMC:AA film. Soil burial test demonstrated that the 60PVA:40CMC:AA film was more degradable (71% at day 15) as compared to the 70PVA:30CMC:AA film (65% at day 15). The films exhibited excellent antimicrobial activity against Gram-positive staphylococcus aureus(inhibition zone of 21 mm) and Gram-negative Escherichia coli (inhibition zone of 15 mm). In comparison, the 60PVA:40CMC:AA film showed better results in terms of high mechanical strength, uniform morphology, higher soil burial degradation, and lower water vapor transmission rate. Therefore, the prepared film could be used as a promising candidate in the food packaging industry.

Corrosion, surface, and tribological behavior of electrophoretically deposited polyether ether ketone coatings on 316L stainless steel for orthopedic applications.

Electrophoretic deposition (EPD) of polyether ether ketone (PEEK) coatings on metallic implants has recently attracted a great deal of interest; however, further investigation into their corrosion, surface, and tribological properties is required for their clinical application. Using Potentiodynamic polarization and Mott-Schottky analysis of PEEK coatings, we analyzed the electrochemical corrosion behavior of electrophoretically deposited PEEK coatings on 316L stainless steel (SS) substrates. In addition, the tribological behavior of the coatings was determined through pin-on-disc and scratch testing. Initially, the EPD parameters were optimized using a Taguchi Design of Experiment (DoE) approach. The coatings exhibited irregular shaped grains along with ∼66 µm of thickness. Fourier transform infrared spectroscopy confirmed the presence of functional groups ascribed with PEEK. The coatings were moderately hydrophobic and had an average roughness of ∼2 µm. The corrosion studies demonstrated promising features of current density and corrosion potential, indicating that corrosion resistance significantly improves with PEEK coating. Electrochemical impedance spectroscopy also confirmed the corrosion resistance of PEEK coating. The coatings exhibited a slightly lower wear resistance than SS samples, but still possessed adequate wear and scratch resistance for biomedical applications. The current study confirmed that the PEEK coatings on metallic implants is effective for orthopedic applications where corrosion and tribology are major concerns.

A study on the effect of bioactive glass and hydroxyapatite-loaded Xanthan dialdehyde-based composite coatings for potential orthopedic applications.

The most important challenge faced in designing orthopedic devices is to control the leaching of ions from the substrate material, and to prevent biofilm formation. Accordingly, the surgical grade stainless steel (316L SS) was electrophoretically deposited with functional composition of biopolymers and bioceramics. The composite coating consisted of: Bioglass (BG), hydroxyapatite (HA), and lawsone, that were loaded into a polymeric matrix of Xanthan Dialdehyde/Chondroitin Sulfate (XDA/CS). The parameters and final composition for electrophoretic deposition were optimized through trial-and-error approach. The composite coating exhibited significant adhesion strength of "4B" (ASTM D3359) with the substrate, suitable wettability of contact angle 48°, and an optimum average surface roughness of 0.32 µm. Thus, promoting proliferation and attachment of bone-forming cells, transcription factors, and proteins. Fourier transformed infrared spectroscopic analysis revealed a strong polymeric network formation between XDA and CS. scanning electron microscopy and energy dispersive X-ray spectroscopy analysis displayed a homogenous surface with invariable dispersion of HA and BG particles. The adhesion, hydrant behavior, and topography of said coatings was optimal to design orthopedic implant devices. The said coatings exhibited a clear inhibition zone of 21.65 mm and 21.04 mm with no bacterial growth against Staphylococcus aureus (S. Aureus) and Escherichia coli (E. Coli) respectively, confirming the antibacterial potential. Furthermore, the crystals related to calcium (Ca) and HA were seen after 28 days of submersion in simulated body fluid. The corrosion current density, of the above-mentioned coating was minimal as compared to the bare 316L SS substrate. The results infer that XDA/CS/BG/HA/lawsone based composite coating can be a candidate to design coatings for orthopedic implant devices.

Development and Characterization of Natural Chromite Coating on Metal Substrate Using the Plasma Spray Process.

Natural materials are gaining interest as coating feedstock because their "quality to cost" ratio is better and they are more environmentally friendly than most of the synthetic ceramics. They give sufficient protection to metal surfaces against harsh conditions such as corrosion, wear, and high temperature. In the current study, chromite mineral was beneficiated and reduced to two different sizes to be used as feedstock material for thermal spray coating. Powders were upgraded by gravity and magnetic separation, respectively, and thermally sprayed onto mild steel samples by using atmospheric plasma spray (APS) equipment. Morphology, structure, phases, elemental distribution of chromite powder, and coatings were studied using field emission scanning electron microscopy, X-ray diffraction, X-ray fluorescence spectroscopy, and energy-dispersive X-ray spectroscopy. Tribological properties of APS chromite coatings were investigated by using a ball-on-disk tribometer, and corrosion resistance properties were evaluated by carrying out potentiodynamic polarization testing in 3.5% NaCl solution. It is observed that the coating has better wear and corrosion resistance and is worn by abrasive wear that includes scratching and particles pull out. Coating efficiency, surface morphology, and microhardness of the coating developed by fine powder were better than those of coarse powder coating.

Electrophoretic Deposition, Microstructure, and Selected Properties of Poly(lactic-co-glycolic) Acid-Based Antibacterial Coatings on Mg Substrate.

There is an urgent need to develop biodegradable implants that can degrade once they have fulfilled their function. Commercially pure magnesium (Mg) and its alloys have the potential to surpass traditional orthopedic implants due to their good biocompatibility and mechanical properties, and most critically, biodegradability. The present work focuses on the synthesis and characterization (microstructural, antibacterial, surface, and biological properties) of poly(lactic-co-glycolic) acid (PLGA)/henna (Lawsonia inermis)/Cu-doped mesoporous bioactive glass nanoparticles (Cu-MBGNs) composite coatings deposited via electrophoretic deposition (EPD) on Mg substrates. PLGA/henna/Cu-MBGNs composite coatings were robustly deposited on Mg substrates using EPD, and their adhesive strength, bioactivity, antibacterial activity, corrosion resistance, and biodegradability were thoroughly investigated. Scanning electron microscopy and Fourier transform infrared spectroscopy studies confirmed the uniformity of the coatings' morphology and the presence of functional groups that were attributable to PLGA, henna, and Cu-MBGNs, respectively. The composites exhibited good hydrophilicity with an average roughness of 2.6 µm, indicating desirable properties for bone forming cell attachment, proliferation, and growth. Crosshatch and bend tests confirmed that the adhesion of the coatings to Mg substrates and their deformability were adequate. Electrochemical Tafel polarization tests revealed that the composite coating adjusted the degradation rate of Mg substrate in a human physiological environment. Incorporating henna into PLGA/Cu-MBGNs composite coatings resulted in antibacterial activity against Escherichia coli and Staphylococcus aureus. The coatings stimulated the proliferation and growth of osteosarcoma MG-63 cells during the initial incubation period of 48 h (determined by the WST-8 assay).

Shape Memory Hybrid Composites.

Smart structures can help to resolve many issues related to conventional materials that are being used in different industries. Shape memory alloys (SMAs) are smart materials with better actuation response, vibration damping characteristics, and large strain recovery, making them good candidates due to their high strength and corrosion resistance for various engineering applications. The performance of fiber-reinforced polymer (FRP) composite materials that are replacing many conventional materials due to their good strength, stiffness, and lightweight potential especially in fuel-consuming industries such as aerospace and automotive, can further be improved by impregnation with SMAs. This review discusses the SMA-reinforced FRP composites, leading to shape memory hybrid composite materials, the issues and limitations in composite manufacturing, and their uses in different research arenas including impact and damping applications, seismic protection applications, crack closure applications, shape morphing applications, and self-deployable structures.

Zn-Mn-Doped Mesoporous Bioactive Glass Nanoparticle-Loaded Zein Coatings for Bioactive and Antibacterial Orthopedic Implants.

In recent years, natural polymers have replaced synthetic polymers for antibacterial orthopedic applications owing to their excellent biocompatibility and biodegradability. Zein is a biopolymer found in corn. The lacking mechanical stability of zein is overcome by incorporating bioceramics, e.g., mesoporous bioactive glass nanoparticles (MBGNs). In the present study, pure zein and zein/Zn-Mn MBGN composite coatings were deposited via electrophoretic deposition (EPD) on 316L stainless steel (SS). Zn and Mn were co-doped in MBGNs in order to make use of their antibacterial and osteogenic potential, respectively. A Taguchi design of experiment (DoE) study was established to evaluate the effect of various working parameters on the morphology of the coatings. It was observed that coatings deposited at 20 V for 5 min with 4 g/L concentration (conc.) of Zn-Mn MBGNs showed the highest deposition yield. Uniform coatings with highly dispersed MBGNs were obtained adopting these optimized parameters. Scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) were employed to investigate the morphology and elemental composition of zein/Zn-Mn MBGN composite coatings. Surface properties, i.e., coating roughness and wettability analysis, concluded that composite coatings were appropriate for cell attachment and proliferation. For adhesion strength, various techniques, including a tape test, bend test, pencil hardness test, and tensile test, were performed. Wear and corrosion analysis highlighted the mechanical and chemical stability of the coatings. The colony forming unit (CFU) test showed that the zein/Zn-Mn MBGN composite coating was highly effective against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) due to the presence of Zn. The formation of a hydroxyapatite (HA)-like structure upon immersion in the simulated body fluid (SBF) validated the in vitro bioactivity of the coating. Moreover, a WST-8 assay depicted that the MG-63 cells proliferate on the composite coating. It was concluded that the zein/Zn-Mn MBGN coating synthesized in this work can be used for bioactive and antibacterial orthopedic applications.

Synthesis and Characterization of Silver-Strontium (Ag-Sr)-Doped Mesoporous Bioactive Glass Nanoparticles.

Biomedical implants are the need of this era due to the increase in number of accidents and follow-up surgeries. Different types of bone diseases such as osteoarthritis, osteomalacia, bone cancer, etc., are increasing globally. Mesoporous bioactive glass nanoparticles (MBGNs) are used in biomedical devices due to their osteointegration and bioactive properties. In this study, silver (Ag)- and strontium (Sr)-doped mesoporous bioactive glass nanoparticles (Ag-Sr MBGNs) were prepared by a modified Stöber process. In this method, Ag+ and Sr2+ were co-substituted in pure MBGNs to harvest the antibacterial properties of Ag ions, as well as pro-osteogenic potential of Sr2 ions. The effect of the two-ion concentration on morphology, surface charge, composition, antibacterial ability, and in-vitro bioactivity was studied. Scanning electron microscopy (SEM), X-Ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) confirmed the doping of Sr and Ag in MBGNs. SEM and EDX analysis confirmed the spherical morphology and typical composition of MBGNs, respectively. The Ag-Sr MBGNs showed a strong antibacterial effect against Staphylococcus carnosus and Escherichia coli bacteria determined via turbidity and disc diffusion method. Moreover, the synthesized Ag-Sr MBGNs develop apatite-like crystals upon immersion in simulated body fluid (SBF), which suggested that the addition of Sr improved in vitro bioactivity. The Ag-Sr MBGNs synthesized in this study can be used for the preparation of scaffolds or as a filler material in the composite coatings for bone tissue engineering.

Fabrication and Characterization of Ag-Sr-Substituted Hydroxyapatite/Chitosan Coatings Deposited via Electrophoretic Deposition: A Design of Experiment Study.

In this study, silver-strontium-doped hydroxyapatite (AgSr-HA)/chitosan composite coatings were deposited on a 316L stainless steel (SS) substrate via electrophoretic deposition (EPD). The Taguchi design of experiment (DoE) approach was used to optimize the EPD parameters such as the applied voltage, interelectrode spacing, and deposition time. Furthermore, the concentration of AgSr-HA particles in the suspension was also optimized via the DoE approach. DoE results demonstrated that the "homogeneous" coatings were obtained at the deposition time of 7 min, deposition voltage of 20 V, and at a concentration of 5 g/L AgSr-HA particles in the suspension. Scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), antibacterial studies, contact angle, and roughness measurements were performed to characterize the optimized coatings. SEM images confirmed the deposition of chitosan/AgSr-HA on the SS substrate. The wettability studies indicated the hydrophilic nature of the chitosan/AgSr-HA composite coatings, which confirmed that the developed coatings are suitable for biomedical applications, e.g., orthopedics. The average surface roughness of the chitosan/AgSr-HA composite coatings was in a suitable range used to attach the bone marrow stromal cells. Chitosan/AgSr-HA composite coatings showed an effective antibacterial effect against Gram-positive and Gram-negative bacteria. Moreover, the coatings developed apatite crystals on their surface upon immersion in simulated body fluid.

Electrophoretic deposition of lawsone loaded bioactive glass (BG)/chitosan composite on polyetheretherketone (PEEK)/BG layers as antibacterial and bioactive coating.

In this study, chitosan/bioactive glass (BG)/lawsone coatings were deposited by electrophoretic deposition (EPD) on polyetheretherketone (PEEK)/BG layers (previously deposited by EPD on 316-L stainless steel) to produce bioactive and antibacterial coatings. First, the EPD of chitosan/BG/lawsone was optimized on stainless steel in terms of suspension stability, homogeneity and thickness of coatings. Subsequently, the optimized EPD parameters were used to produce bioresorbable chitosan/bioactive glass (BG)/lawsone coatings on PEEK/BG layers. The produced layered coatings were characterized in terms of composition, microstructure, corrosion resistance, in vitro bioactivity, drug release kinetics and antibacterial activity. Ultraviolet/Visible (UV/VIS) spectroscopic analyses confirmed the release of lawsone from the coatings. Moreover, the deposition of chitosan/BG coatings was confirmed by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared spectroscopy (FTIR). The coated specimens presented higher corrosion resistance (10 times) in comparison to that of bare 316-L stainless steel and showed convenient wettability for initial protein attachment. The presence of lawsone in the top layer provided antibacterial effects against Staphylococcus carnosus. Moreover, the developed coatings formed apatite-like crystals upon immersion in simulated body fluid, indicating the possibility of achieving close interaction between the coating surface and bone. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 3111-3122, 2018.

Antibacterial and Bioactive Coatings Based on Radio Frequency Co-Sputtering of Silver Nanocluster-Silica Coatings on PEEK/Bioactive Glass Layers Obtained by Electrophoretic Deposition.

Bioactive and antibacterial coatings on stainless steel substrates were developed and characterized in this study. Silver nanocluster-silica composite coatings of 60-150 nm thickness were deposited using radio frequency (RF) co-sputtering on PEEK/bioactive glass (BG) layers (of 80-90 µm thickness) which had been electrophoretically deposited onto stainless steel. Two sputtering conditions were used by varying the deposition time (15 and 40 min); the resulting microstructure, composition, adhesion strength, in vitro bioactivity, and antibacterial activity were investigated. Scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and energy dispersive spectroscopy (EDX) confirmed the presence of silver nanoclusters, which were homogeneously embedded in the silica matrix. The isoelectric point of the coatings and their charge at physiological pH were determined by zeta potential measurements. The presence of BG particles in the PEEK/BG layer allows the coatings to form apatite-like crystals upon immersion in simulated body fluid (SBF). Moreover, silver nanoclusters embedded in the silica matrix as a top layer provided an antibacterial effect against Escherichia coli and Staphylococcus carnosus.