Fibroblast Cell Cultures Research Articles

All Trans-Retinoic Acids Facilitate the Remodeling of 2D and 3D Cultured Human Conjunctival Fibroblasts.

Vitamin A derivative, all-trans-retinoic acid (ATRA), is known to be a potent regulator of the growth and differentiation of various types of cells. In the present study, the unidentified effects of ATRA on superficial and vertical spreading conjunctival scarring were examined. The study involved the use of two-dimensional (2D) and three-dimensional (3D) cultures of human conjunctival fibroblast (HconF) cells in the presence or absence of TGF-β2. The effects of ATRA (1 μM) on superficial or vertical spreading conjunctival scarring were evaluated by the barrier function by trans-endothelial electrical resistance (TEER) and FITC dextran permeability measurements and real-time metabolic analysis, as well as the physical properties, namely, the size and stiffness, of 3D spheroids, respectively. In addition, the expressions of several related molecules, including extracellular matrix (ECM) molecules, ECM modulators including a tissue inhibitor of metalloproteinases (TIMPs), matrix metalloproteinases (MMPs), and ER stress-related factors, were examined. ATRA significantly induced (1) an increase in TEER values and a decrease in FITC dextran permeability, respectively, in the 2D monolayers, and (2) relatively and substantially increased the size and stiffness, respectively, of the 3D spheroids. These ATRA-induced effects were further enhanced in the TGF-β2-treated cells, whereas the TGF-β2-induced enhancement in glycolytic capacity was canceled by the presence of ATRA. Consistent with these physical and morphological effects, the mRNA expressions of several molecules were significantly but differently induced between 2D and 3D cultures by ATRA, although the presence of TGF-β2 did not substantially affect these gene expression levels. The findings reported in this study indicate that ATRA may exacerbate both superficial and vertical conjunctival fibrosis spreading independently of TGF-β2-induced changes.

Curcumin encapsulated polylactic acid nanoparticles embedded in alginate/gelatin bioinks for in situ immunoregulation: Characterization and biological assessment

Curcumin is a known naturally occurring anti-inflammatory agent derived from turmeric, and it is commonly used as a herbal food supplement. Here, in order to overcome the inherent hydrophobicity of curcumin (Cur), polylactic acid (PLA) nanoparticles (NPs) were synthesised using a solvent evaporation, and an oil-in-water emulsion method used to encapsulate curcumin. Polymeric NPs also offer the ability to control rate of drug release. The newly synthesised NPs were analysed using a scanning electron microscope (SEM), where results show the NPs range from 50 to 250 nm. NPs containing graded amounts of curcumin (0 %, 0.5 %, and 2 %) were added to cultures of NIH3T3 fibroblast cells for cytotoxicity evaluation using the Alamar Blue assay. Then, the curcumin NPs were incorporated into an alginate/gelatin solution, prior to crosslinking using a calcium chloride solution (200 nM). These hydrogels were then characterised with respect to their chemical, mechanical and rheological properties. Following hydrogel optimization, hydrogels loaded with NP containing 2 % curcumin were selected as a candidate as a bioink for three-dimensional (3D) printing. The biological assessment for these bioinks/hydrogels were conducted using THP-1 cells, a human monocytic cell line. Cell viability and immunomodulation were evaluated using lactate dehydrogenase (LHD) and a tumour necrosis factor alpha (TNF-α) enzyme-linked immunosorbent (ELISA) assay, respectively. Results show that the hydrogels were cytocompatible and supressed the production of TNF-α. These bioactive hydrogels are printable, supress immune cell activation and inflammation showing immense potential for the fabrication of tissue engineering constructs.

3D Printing of Cellulase-Laden Cellulose Nanofiber/Chitosan Hydrogel Composites: Towards Tissue Engineering Functional Biomaterials with Enzyme-Mediated Biodegradation.

The 3D printing of a multifunctional hydrogel biomaterial with bioactivity for tissue engineering, good mechanical properties and a biodegradability mediated by free and encapsulated cellulase was proposed. Bioinks of cellulase-laden and cellulose nanofiber filled chitosan viscous suspensions were used to 3D print enzymatic biodegradable and biocompatible cellulose nanofiber (CNF) reinforced chitosan (CHI) hydrogels. The study of the kinetics of CNF enzymatic degradation was studied in situ in fibroblast cell culture. To preserve enzyme stability as well as to guarantee its sustained release, the cellulase was preliminarily encapsulated in chitosan–caseinate nanoparticles, which were further incorporated in the CNF/CHI viscous suspension before the 3D printing of the ink. The incorporation of the enzyme within the CHI/CNF hydrogel contributed to control the decrease of the CNF mechanical reinforcement in the long term while keeping the cell growth-promoting property of chitosan. The hydrolysis kinetics of cellulose in the 3D printed scaffolds showed a slow but sustained degradation of the CNFs with enzyme, with approximately 65% and 55% relative activities still obtained after 14 days of incubation for the encapsulated and free enzyme, respectively. The 3D printed composite hydrogels showed excellent cytocompatibility supporting fibroblast cell attachment, proliferation and growth. Ultimately, the concomitant cell growth and biodegradation of CNFs within the 3D printed CHI/CNF scaffolds highlights the remarkable potential of CHI/CNF composites in the design of tissue models for the development of 3D constructs with tailored in vitro/in vivo degradability for biomedical applications.

Bi-allelic loss-of-function variants in TMEM147 cause moderate to profound intellectual disability with facial dysmorphism and pseudo-Pelger-Huët anomaly

SummaryThe transmembrane protein TMEM147 has a dual function: first at the nuclear envelope, where it anchors lamin B receptor (LBR) to the inner membrane, and second at the endoplasmic reticulum (ER), where it facilitates the translation of nascent polypeptides within the ribosome-bound TMCO1 translocon complex. Through international data sharing, we identified 23 individuals from 15 unrelated families with bi-allelic TMEM147 loss-of-function variants, including splice-site, nonsense, frameshift, and missense variants. These affected children displayed congruent clinical features including coarse facies, developmental delay, intellectual disability, and behavioral problems. In silico structural analyses predicted disruptive consequences of the identified amino acid substitutions on translocon complex assembly and/or function, and in vitro analyses documented accelerated protein degradation via the autophagy-lysosomal-mediated pathway. Furthermore, TMEM147-deficient cells showed CKAP4 (CLIMP-63) and RTN4 (NOGO) upregulation with a concomitant reorientation of the ER, which was also witnessed in primary fibroblast cell culture. LBR mislocalization and nuclear segmentation was observed in primary fibroblast cells. Abnormal nuclear segmentation and chromatin compaction were also observed in approximately 20% of neutrophils, indicating the presence of a pseudo-Pelger-Huët anomaly. Finally, co-expression analysis revealed significant correlation with neurodevelopmental genes in the brain, further supporting a role of TMEM147 in neurodevelopment. Our findings provide clinical, genetic, and functional evidence that bi-allelic loss-of-function variants in TMEM147 cause syndromic intellectual disability due to ER-translocon and nuclear organization dysfunction.

Biophotonic Effects of Low-Level Laser Therapy at Different Wavelengths for Potential Wound Healing

Our objective was to assess the effect of low-level laser therapy (LLLT) administered using a diode laser on the growth processes of human fibroblast cells involved in wound healing. Initially, studies were conducted using a diode laser at wavelengths of 633, 520, and 450 nm with an irradiance of 3 mW/cm2. The distance between the light source and culture plate was 3 cm. The mechanism(s) of action of the diode laser illumination on human fibroblast cells were studied by examining different wavelengths to determine the relevant light parameters for optimal treatment. In addition, the percentages of fibroblast-mediated procollagen and matrix metallopeptidase (MMP)-1, -2, and -9 production were compared. In the clinical study, the changes in basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and fibroblast collagen production were assessed in 60 patients with complicated wounds who received LLLT (633 nm). No statistically significant difference was observed between red light versus green and blue light in the viability analysis. In addition, the effects of LLLT on the cell cultures of fibroblast cells in vitro demonstrated a decrease in the relative expression of MMP-1, -2, and -9 while using light with a wavelength of 633 nm. In the clinical study, 633 nm diode laser LLLT at 2–8 J/cm2 was administered to 60 patients with complicated wounds; all patients showed increased levels of bFGF and VEGF and the occurrence of collagen synthesis. Our studies demonstrated that LLLT might affect fibroblast cell growth processes involved in wound healing.

Abstract P2025: BET Bromodomain Proteins Are Necessary For TGFβ2-Mediated Myofibroblast Activation

Fibrosis is a key feature of dilated cardiomyopathy (DCM). Stress signaling triggers the activation of quiescent fibroblasts to myofibroblasts. Bromodomain and extraterminal (BET) epigenetic reader proteins integrate stress-responsive signaling. We explored the role of BETs on distinct inflammatory signaling pathways in cardiac fibroblasts. Fibroblasts were isolated from wild type hearts using the Langendorff procedure. Cell cultures were treated with vehicle, the pan-BET inhibitor JQ1, or siRNA targeting Smad3 or Rela (to inhibit TGFβ and NFκB pathways, respectively) followed by TGFβ2 stimulation. Gene expression was assessed by QPCR, α-smooth muscle actin (αSMA) was quantified by western blot, and NFκB transcription was evaluated using a dual luciferase reporter assay. The interaction between BRD4 and SMAD3 was assessed by coimmunoprecipitation (coIP) in immortalized adult mouse cardiac fibroblast cell cultures treated with TGFβ2, JQ1, or vehicle. TGFβ2-induced fibroblast activation was ablated by JQ1 and Smad3 siRNA (reduced expression of Acta2, Col3a1, Postn and αSMA). CoIP revealed a direct interaction between BRD4 and SMAD3 in fibroblasts, which increased 2.2-fold (p<0.05) after TGFβ2 stimulation. By contrast, Rela siRNA alone induced myofibroblast gene expression ( Acta2 , 6.9-fold; Col3a1 , 7.0-fold; Postn , 11.2-fold; p<0.05 vs. vehicle for all), and increased αSMA levels (2.2-fold; p<0.05). This effect of Rela siRNA was similar to that observed with TGFβ2 stimulation. Further, TGFβ2 inhibited NFκB-driven gene transcription (-2.9-fold; p<0.05 vs. vehicle). These data demonstrate that BRD4 is a critical co-activator of TGFβ2-SMAD3 signaling and cardiac fibroblast activation. We also found that NFκB signaling is downregulated by TGFβ2, suggesting that NFκB may modulate inflammatory gene network activation.

Polyhedral Oligomeric Silsesquioxane as a Polarity Mediator and Reinforced Nanofiller for Fabricating Robust and Hierarchical Porous Film for Cell Bioengineering

A hierarchical porous polymer film that can provide a balance of mechanical properties and specific surface areas and sustain cell functions has become a major target for material scientists and bioengineers in the past decade. However, the limited availability of materials that exhibit high mechanical performance and easily form hierarchical porous structures hinder the progress in practical application. Here, we show the design of a polyhedral oligomeric silsesquioxane (POSS) hybrid polymer by incorporating a bi-functional glycidyl-propyl POSS (G-POSS) into the epoxy polymer backbone, in which POSS functions as both a nanofiller to reinforce the mechanical performance and as the polarity mediator to induce self-assembly of the polymer chains into a three-dimensional (3D) connected porous structure. Nanoindentation shows that the incorporation of POSS leads to a 25 and 36% increase in modulus and hardness, respectively. Scanning electron microscopy results show that POSS helps tune the hydrophobicity of the polymer and assists in the interface assembly, allowing easy formation of a densely packed two-dimensional or 3D hierarchical porous structure. Culture of mouse embryonic fibroblast cells demonstrates that the hybrid polymer scaffold provides prominent advantages ranging from enhanced cell adhesion to proliferation. Particularly, the 3D hierarchical porous film allows good nutrient supply and cell communication, which better mimics the in vivo microenvironment with less cell morphology change during cell proliferation. Hence, the successful integration of high material strength and easy formation of an interconnected porous structure shows that POSS hybrid polymers have strong potential for applications in the fabrication of cell culture platforms and future translation in tissue repair and regeneration.

THE CYTOTOXIC AND PROLIFERATIVE ACTIVITY OF COCOA POD HUSK EXTRACT (THEOBROMA CACAO L.) ON PERIODONTAL LIGAMENT FIBROBLASTS

Background: Theobroma cacao was considered agricultural wastes. Cocoa pod husk contain various bioactive compounds as natural antioxidants,antibacterial and anti-inflammatory. A good biomaterial must be non-toxic and have no detrimental effect on the biological environment. The aim of research was to determine the cytotoxicity and proliferative activity of cocoa pod husk extract on primary cells of periodontal ligament fibroblasts. Method: This experimental laboratory used primary cell culture of periodontal ligament fibroblasts. The cytotoxicity and proliferative activity were analyzed by MTT assay [2-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide]. Extraction of cacao pods were obtained using the ultrasonic homogenizer method with ethanol 70%. The cytotoxicity test was observed for 24 hours and the doubling time proliferative test was observed for 24, 48, and 72 hours. Result: Cocoa pod husk extract showed there were no cytotoxic effects with viability of primary cells of periodontal ligament fibroblasts at the concentration of 1.56 pg/ml, 3.125 pg/ml, 6.25 pg/ml, 12.5 pg/ml, 25 pg/ml, 50 pg/ml, and 100 pg/ml. The proliferation test showed increasing of fibroblasts at 24 and 48 hours. Kruskal Wallis analysis obtained a significance value (p<0.05) Conclusion: Cytotoxicity and proliferation assays of Cocoa pod husk extract increased cell viability and cell proliferation of periodontal ligament fibroblast.

Presence of Chlamydia abortus in colostrum, milk and vaginal discharge samples of sheep

Background: The main transmission route of Chlamydia abortus is by ingesting the microorganism that has been eliminated in vaginal secretions, placental membranes or abortions that contaminate the environment and, possibly, through milk and colostrum. Elimination through vaginal secretions is well documented. However, there are no reports about isolation and identification of C. abortus in the colostrum or milk of infected sheep, so it is important to determine whether or not C. abortus may be present in these secretions, which are the only food of lambs. Objective: To detect C. abortus in colostrum, milk, and vaginal secretions of sheep with a history of reproductive disorders. Methods: Colostrum, milk, and vaginal exudates were collected from 66 sheep. The samples were inoculated in mouse fibroblast cell cultures and the presence of C. abortus determined by direct immunofluorescence. Results: 19 out of 66 colostrum samples (28.7%), 14 out of 66 milk samples (21.2%) and 17 out of 66 vaginal swabs (25.7%) were positive for C. abortus. The 50 samples positive for isolation and detected by immunofluorescence, together with 42 negative samples were subjected to qPCR to amplify a fragment of the ompA gene from C. abortus. Thirty-eight of the 92 samples processed by this technique were positive for C. abortus. Conclusion: The results demonstrated the presence of C. abortus in a high proportion in colostrum, milk and vaginal secretions of infected sheep. To the best of our knowledge, this is the first field study confirming the presence of C. abortus in colostrum, which shows that excretion of Chlamydia by lactogenesis could occur in the first hours after birth.

USE OF KNOTTIN AS A PSMA-TROPIC PEPTIDE CARRIER

Prostate cancer is the most common type of cancer in males. Approximately 1.3 million cases of prostate cancer and over 400,000 deaths from the disease are diagnosed annually. The number of deaths is expected to double by 2040. Common methods of prostate cancer treatment have many disadvantages; one of them is the relapse risk. The shortcomings of traditional therapy have led to peptide-receptor radionuclide therapy.
 The aim of the study is to examine binding efficiency of Lu177 labeled knottins containing PSMA-tropic peptide in different domains in vitro and their biodistribution in vivo.
 Materials and Methods. We used prostate cancer cell (LNCaP, PC3) and ovarian fibroblast cell (CHO-K1) cultures. The peptides were synthesized using a peptide synthesizer (ResPepSL, Intavis).
 We studied peptide stability, their toxicity, binding to cell cultures, and biodistribution on the example of breast adenocarcinoma-bearing BALB/c mice. Chromatographic methods and radiometric techniques 
 were used.
 Results. The synthesized peptides with GTIQPYPFSWGY sequence inserted into U5-cytotoxin-Sth1a node are more stable in blood plasma and saline than 177Lu-PSMA-617a, but have a similar degree of binding. Biodistribution studies in BALB/c mice show a higher binding index of the synthesized peptide if compared to 177Lu-PSMA-617.
 Conclusion. Modified peptides with a PSMA-tropic peptide inserted into the structure of U5-Sth1a toxin demonstrate high stability both in saline and in blood plasma, as well as good binding to cell cultures and tumors.

The potential of carrot extract as a sunscreen to prevent apoptosis in white mice (Mus musculus) fibroblast cell cultures exposed to UVB light

Introduction: UVB rays are one of the main external factors that play a role in the aging process. Apoptosis of fibroblasts affects skin aging, and the decline in fibroblasts is also caused by the reduced number and ability of growth factors and other hormones. Sunscreen has been used for a long time to protect the skin from the harmful effects of solar radiation, especially UVB rays. Antioxidant activity such as β-carotene and vitamin E from carrots can counteract free radicals and oxidative stress caused by UV radiation. This study aims to test whether in vitro administration of carrot extract can be used as a sunscreen to prevent apoptosis in mouse fibroblast cell cultures exposed to UVB light.
 Method: This research is a laboratory experimental study, using a Randomized Post-test Only Control Group Design. The research sample was fibroblast cell culture derived from the back skin of white mice (Mus Musculus) isolated with Collagenase 1 (primary culture). Observation of fibroblast apoptosis using propidium iodide staining. Data were analyzed using Shapiro-Wilk test, Levene's test, One Way Anova and continued with LSD test. The confidence level in this study is 95%.
 Result: There was a significant difference between the control group and the carrot extract treatment group, but no significant difference was found between the SPF15 group and the carrot group (p>0.01).
 Conclusion: Carrot extract can provide a protective effect on the skin (fibroblasts) and prevent apoptosis.

Rapid synthesis of biocompatible bilayer hydrogels via frontal polymerization

AbstractHydrogels have been attracting much attention as smart soft materials. The exploration of methods allowing hydrogels to have anisotropic composition, structure, and properties would benefit the fabrication of smart materials with versatile multi‐functions. Here we report the rapid frontal polymerization (FP) of bilayer hydrogels possessing good water swelling, mechanical strength, self‐healing, biocompatibility, and antibacterial capabilities. The bilayer hydrogels, consisting of poly(AM‐co‐AMPS) (G‐1) and poly(AM‐co‐HPA) (G‐2) (AM, acrylamide; AMPS, 2‐acrylamide‐2‐methylpropanesulfonic acid; HPA, hydroxypropyl acrylate), are synthesized in one step within 6 minutes (min) by horizontal FP. These polymer hydrogels have hierarchical porous morphology with pore sizes of several micrometers and several hundreds of micrometers respectively, equilibrium swelling ratio up to 3371% in water, tensile strength up to 0.204 MPa, and self‐healing ability. The good cytocompatibility and low toxicity of the hydrogels are demonstrated through cell culture of L929 fibroblasts in vitro experiments. The tests of hydrogels with Escherichia coli and Staphylococcus aureus reveal their antibacterial property. The results demonstrate that biocompatible anisotropic hydrogels with versatile functions could be fabricated via the rapid and energy‐saving approach of FP.

Evaluation of NACA and diNACA in human cystinosis fibroblast cell cultures as potential treatments for cystinosis

BackgroundCystinosis is a rare autosomal recessive lysosomal storage disease, associated with high morbidity and mortality. Mutations in the CTNS gene disable a membrane protein responsible for the transport of cystine out of the lysosome. Loss of transporter function leads to intralysosomal cystine accumulation and long-term damage to various tissues and organs, including the kidneys, eyes, liver, muscles, pancreas, and brain. The only cystine-depletion therapy for treatment of cystinosis is cysteamine which requires frequent administration of high doses and often causes gastrointestinal pain as well as pungent sulfurous odor in patients. The current in vitro study evaluated antioxidants, N-acetylcysteine amide (NACA; NPI-001) and (2R,2R′)-3,3′-disulfanediyl bis(2-acetamidopropanamide) (diNACA; NPI-002), as potential treatments for cystinosis.MethodsCytotoxicity of cysteamine, NACA and diNACA was evaluated in cultured human cystinotic fibroblasts (HCFs). HCFs were cultured in 96 well plates incubated for 0–72 h in the presence of 25, 50 or 75 μM each of either cysteamine, NACA or diNACA along with an untreated control. Media was removed and cell viability assessed. Next, cystine-depleting activities of cysteamine, NACA and diNACA were screened in HCFs cell culture utilizing an inexpensive, proven colorimetric assay. HCFs were seeded and allowed to reach approximately 80% confluence before the addition of the test articles: 50 μM of either cysteamine, NACA or diNACA in media along with an untreated control. HCFs were incubated, harvested, and cystine was reduced to cysteine, the concentration of which was then determined per quantity of protein compared to a cysteine standard. Statistically significant cystine depletion was determined by paired t-test versus untreated control (p < 0.05).ResultsNeither cysteamine, NACA nor diNACA at 25, 50 or 75 μM caused cytotoxicity in HCFs. Treatment with all tested concentrations (25, 50 or 75 µM) of either NACA or diNACA at 48 or 72 h resulted in statistically significant increases in cell viability, relative to untreated control, whereas the higher concentrations (50 or 75 µM) of cysteamine achieved statistical significance at both timepoints but not the lowest concentration (25 µM). All test articles depleted cystine from HCFs compared to control. NACA depletion of cystine was statistically superior to cysteamine at 6, 24 and 48 h and numerically greater at 72 h. DiNACA depletion of cystine was statistically superior to cysteamine at 6 and 48 h, slightly numerically greater at 24 h and slightly less at 72 h.ConclusionsNACA and diNACA were non cytotoxic to HCFs and significantly increased cell viability. Cystine reduction was determined as percent of control after incubation with 50 µM of NACA, diNACA or cysteamine in HCFs cell culture for 6, 24, 48 and 72 h. Of the three test articles, NACA exhibited most rapid and greatest potency in cystine reduction. Rank order potency for cystine reduction over time was observed, NACA > diNACA ≥ cysteamine. Therefore, further study of NACA and diNACA as potential treatments for cystinosis is warranted.

Solution blow spinning of highly deacetylated chitosan nanofiber scaffolds for dermal wound healing.

Biocompatible fibrous scaffolds based on highly deacetylated chitosan were fabricated using high-throughput solution blow spinning. Scanning electron microscopy analysis revealed that the chitosan nanofiber scaffolds had ultrafine and continuous fibers (300-1200nm) with highly interconnected porous structures (30-75% porosity), mimicking some aspects of the native extracellular matrix in skin tissue. Post-treatment of as-spun nanofibers with aqueous potassium carbonate solution resulted in a fibrous scaffold with a high chitosan content that retained its fibrous structural integrity for cell culture. Analysis of the mechanical properties of the chitosan nanofiber scaffolds in both dry and wet conditions showed that their strength and durability were sufficient for wound dressing applications. Significantly, the wet scaffold underwent remarkable elastic deformation during stretch such that the elongation at break dramatically increased to up to 44% of its original length, showing wavy fiber morphology near the break site. The culture of normal human dermal fibroblast cells onto scaffolds for 1-14days demonstrated that the scaffolds were highly compatible and a suitable platform for cell adhesion, viability, and proliferation. Secretion profiles of wound healing-related proteins to the cell culture medium demonstrated that chitosan fibers were a promising scaffold for wound healing applications. Overall, the dense fibrous network with high porosity of the chitosan nanofiber scaffold and their mechanical properties indicate that they could be used to design and fabricate new materials that mimic the epidermis layer of natural skin.

Detection and Isolation of Camelpox Virus in Wasit Province, Iraq.

Camels are susceptible to a variety of infectious diseases, such as trypanosomiasis, anthrax, hemorrhagic septicemia, brucellosis, mange, and pox, which can also affect other farm animal species. Camelpox is one of the most infectious skin diseases, which is caused by the Camelpox virus (CPV), a member of the Orthopoxvirus genus in the Poxviridae family. This study mainly aimed to detect and isolate CPV affecting camels in Wasit province, Iraq. Initially, the study focused principally on clinical manifestations of the disease in affected animals. Afterward, 110 skin samples were collected from infected animals under strict aseptic conditions. They were to be subjected to inoculation into the local embryonated chicken eggs to isolate the virus from the chicken embryo fibroblast cell culture. Finally, the isolates were confirmed using the molecular polymerase chain reaction (PCR) assay. Camelpox virus isolates appeared as several necrotic pock lesions on the Chorioallantoic Membrane (CAM) with cell clustering and sloughing or detachment from the monolayer. Molecular testing was conducted using the PCR assay by targeting the ATIP gene at 881bp. The findings demonstrated that all the investigated isolates were poxvirus positive. In addition, it was found that Camelpox disease is significantly endemic in Wasit province, Iraq. The visualization of the characteristic features of the virus also revealed it can easily adapt to replication in the CAM and cell culture.

Green synthesis of bioinspired chitosan-ZnO-based polysaccharide gums hydrogels with propolis extract as novel functional natural biomaterials

A facile, green synthesis methodology to obtain zinc oxide nanoparticles using three polysaccharide gums (Acacia gum, Guar gum and Xanthan gum) of biological origin was developed. Subsequently, biosynthesized zinc oxide nanoparticles were incorporated into a sustainable chitosan hydrogel matrix functionalized with propolis extract. This study has revealed that the selected polysaccharides as chelates represents a suitable approach to synthesize ZnO nanoparticles of particular interest with controlled morphology. The formation of ZnO nanoparticles using polysaccharide gums was confirmed by FTIR, XRD, UV–Vis spectroscopy, thermal analysis, SEM, Raman and photoluminescence spectroscopies. The rheological behaviour of obtained hydrogels was evaluated. The AFM studies demonstrate that all synthesized chitosan incorporated ZnO composites hydrogels functionalized with propolis extract exhibit corrugated topographies. The present study highlights the possible incorporation of various guest molecules into hydrogel matrix due to its tuneable morphologies. The obtained hydrogel composites were cytocompatible in L929 fibroblast cell culture, in a range of concentrations between 50 and 1000 μg/mL, as assessed by MTT, LDH and Live/Dead double staining assays. By enhancing the biological properties, these novel green hydrogels show attractive superior performance in a wide concentration range to develop future in vivo suitable natural platforms as effective delivery systems of pharmacologic agents for biomedical applications.

Poly(2-hydroxyethyl methacrylate-co-quaternary ammonium salt chitosan) hydrogel: A potential contact lens material with tear protein deposition resistance and antimicrobial activity.

Tear protein deposition resistance and antimicrobial property are two challenges of conventional poly(2-hydroxyethyl methacrylate) (pHEMA) contact lenses. In this work, we developed a poly(2-hydroxyethyl methacrylate-co-quaternary ammonium salt chitosan) hydrogel, named as p(HEMA-co-mHACC) hydrogel, using acryloyl HACC (mHACC) as a macromolecular crosslinker. With increasing the acryloyl substitution degree (14-29%) or mHACC content (2-11%), the hydrogel showed an enhanced tensile strength (432-986kPa) and Young's modulus (360-1158kPa), a decreased elongation at break (242-84%), and an increased visible light transmittance (0-95%). At an optimal acryloyl substitution degree of 26%, with the increase of mHACC content from 2% to 11%, p(HEMA-co-mHACC) hydrogel presented a decreased water contact angle from 84.6 to 55.3 degree, an increased equilibrium water content from 38% to 45%, and an enhanced oxygen permeability from 8.5 to 13.5 barrer. Due to the enhancement in surface hydrophilicity and electropositivity, p(HEMA-co-mHACC) hydrogel remarkably reduced the deposition of lysozyme, but little affected the adsorption of BSA, depending on the hydrophilic/hydrophobic and electrostatic interactions. The antimicrobial test against Staphylococcus aureus and Escherichia coli showed that p(HEMA-co-mHACC) hydrogel presented an 8-32 times higher germicidal ability than pHEMA hydrogel, indicative of a better antimicrobial activity. The in vitro cell culture of mouse NIH3T3 fibroblasts and immortalized human keratinocytes showed that p(HEMA-co-mHACC) hydrogel was non-toxic. Thus, p(HEMA-co-mHACC) hydrogel with tear protein deposition resistance and antimicrobial activity is a potential candidate for contact lenses.

Electrochemical and In Vitro Biological Evaluation of Bio-Active Coatings Deposited by Magnetron Sputtering onto Biocompatible Mg-0.8Ca Alloy.

The use of resorbable magnesium alloys in the design of implants represents a new direction in the healthcare domain. Two main research avenues are currently explored for developing or improving metallic biomaterials: (i) increase of their corrosion resistance by designed compositional and structural modifications, and (ii) functionalization of their surfaces by coating with ceramic or polymeric layers. The main objective of this work was to comparatively assess bio-functional coatings (i.e., highly-crystallized hydroxyapatite and silica-rich glass) deposited by radio-frequency magnetron sputtering (RF-MS) on a biodegradable Mg-0.8Ca alloy (0.8 wt.% of Ca). After probing their morphology (by scanning electron microscopy) and structure (by Fourier transform infrared spectroscopy and grazing incidence X-ray diffraction), the corrosion resistance of the RF-MS coated Mg-0.8Ca substrates was electrochemically tested (in synthetic biological media with different degrees of biomimicry), and their cytocompatibility was assessed in osteoblast and fibroblast cell cultures. By collective assessment, the most promising performances, in terms of mass loss (~7% after 12 days), hydrogen release rate (~6 mL/cm2 after 12 days), electrochemical corrosion parameters and cytocompatibility, were obtained for the crystalline HA coating.

Liposomal Formulations to Improve Antioxidant Power of Myrtle Berry Extract for Potential Skin Application

Many substances in plant extracts are known for their biological activities. These substances act in different ways, exerting overall protective effects against many diseases, especially skin disorders. However, plant extracts’ health benefits are often limited by low bioavailability. To overcome these limitations, drug delivery systems can be employed. In this study, we evaluated the antioxidant power of an ethanolic extract from Myrtus communis L. (myrtle) berries through colorimetric tests (DPPH and FRAP). The antioxidant activity was also verified by using fibroblast cell culture through cellular Reactive Oxygen Species (ROS) levels measurements. Moreover, the myrtle extract was formulated in phospholipid vesicles to improve its bioavailability and applicability. Myrtle liposomes were characterized by size, surface charge, storage stability, and entrapment efficiency; visualized by using cryo-TEM images; and assayed for cytocompatibility and anti-ROS activity. Our results suggest that myrtle liposomes were cytocompatible and improved the extract’s antioxidant power in fibroblasts, suggesting a potential skin application for these formulations and confirming that nanotechnologies could be a valid tool to enhance plant extracts’ potentialities.

Comparative study on the expression profile of Aquaporin5 (AQP5) gene in skinfibroblast cells of native goats enabling their water utilization efficiency

Lack of availability of good quality and quantity of water affects the physiology of goats. Goats may adapt to thrive well in water deficit areas by making alterations in the expressions of aquaporin (AQP) genes for their efficient water use. The present study was carried out in skin fibroblast cells of Barbari and Sirohi goats. Under in vitro conditions, during different temperatures and osmotic treatments, variations in relative expression of AQP3 genes were observed. At higher temperatures the relative expression of AQP3 genes in Sirohi breed was found to be significantly higher (p&lt;0.01) than Barbari breed. However, during hyperosmotic concentrations at normal temperatures the relative expression of AQP3 mRNA was significantly ((p&lt;0.05) increased in both the breeds. During a high temperature combination with hyperosmotic concentration in cell culture of skin fibroblast cell the relative expression of AQP3 mRNA was increased in Sirohi breed (p&lt;0.01) whereas it decreased in Barbari goat’s breed. The variations in AQP3 gene expression during water stress condition and during hyperosmotic concentrations of skin fibroblasts cell culture suggests the positive involvement of AQP3 gene in maintaining the water balance in the body.