scholarly journals Effect of Antimicrobial Peptides HNP-1 and hBD-1 on Staphylococcus aureus Clinical Strains in vitro and in vivo

2019 ◽  
Author(s):  
Albert Bolatchiev ◽  
Vladimir Baturin ◽  
Igor Bazikov ◽  
Alexander Maltsev ◽  
Elena Kunitsina

ABSTRACTThe aims of this study were: (1) To investigate the activity of recombinant AMPs HNP-1 and hBD-1 in combination with cefotaxime against Staphylococcus aureus clinical strains (MSSA and MRSA) in vitro using checkerboard method; (2) To investigate the activity of HNP-1 and hBD-1 encapsulated in silicon nanoparticles (niosomes) in the treatment of MRSA-infected wound in rats. For this S. aureus clinical strains (MSSA and MRSA) were isolated from patients with diabetic foot infection. Cefotaxime, recombinant HNP-1 and hBD-1 (in all possible combinations with each other) were used for testing by the checkerboard method. Two niosomal topical gels with HNP-1/hBD-1 were prepared to treat MRSA-infected wounds in rats. Gels were administered once a day, the control group – without treatment. Wound healing rate was calculated on the 4th, 9th and 16th days of the experiment and compared using one-way ANOVA with Bonferroni correction. MIC of HNP-1 for MSSA and MRSA was the same – 1 mg/L. MIC of hBD-1 for MSSA and MRSA was also the same – 0.5 mg/L. Topical gels with niosomal HNP-1 (or hBD-1) showed a significantly faster wound healing in comparison with the control. The data obtained open up prospects for use of AMPs encapsulated in silica nanoparticles for the development of new antibiotics.

Author(s):  
Joon M. Jung ◽  
Hae K. Yoon ◽  
Chang J. Jung ◽  
Soo Y. Jo ◽  
Sang G. Hwang ◽  
...  

Cold plasma can be beneficial for promoting skin wound healing and has a high potential of being effectively used in treating various wounds. Our aim was to verify the effect of cold plasma in accelerating wound healing and investigate its underlying mechanism in vitro and in vivo. For the in vivo experiments, 2 full-thickness dermal wounds were created in each mouse (n = 30). While one wound was exposed to 2 daily plasma treatments for 3 min, the other wound served as a control. The wounds were evaluated by imaging and histological analyses at 4, 7, and 11 days post the wound infliction process. Immunohistochemical studies were also performed at the same time points. In vitro proliferation and scratch assay using HaCaT keratinocytes and fibroblasts were performed. The expression levels of wound healing–related genes were analyzed by real-time polymerase chain reaction and western blot analysis. On day 7, the wound healing rates were 53.94% and 63.58% for the control group and the plasma-treated group, respectively. On day 11, these rates were 76.05% and 93.44% for the control and plasma-treated groups, respectively, and the difference between them was significant ( P = .039). Histological analysis demonstrated that plasma treatment promotes the formation of epidermal keratin and granular layers. Immunohistochemical studies also revealed that collagen 1, collagen 3, and alpha-smooth muscle actin appeared more abundantly in the plasma-treated group than in the control group. In vitro, the proliferation of keratinocytes was promoted by plasma exposure. Scratch assay showed that fibroblast exposure to plasma increased their migration. The expression levels of collagen 1, collagen 3, and alpha-smooth muscle actin were elevated upon plasma treatment. In conclusion, cold plasma can accelerate skin wound healing and is well tolerated.


2013 ◽  
Vol 2 (1) ◽  
pp. 05 ◽  
Author(s):  
Abdul Razak ◽  
Aziz Djamal ◽  
Gusti Revilla

AbstrakJeruk Nipis (Citrus aurantifolia S.) merupakan salah satu tanaman obat keluarga yang banyak terdapat ditengah masyarkat dan banyak digunakan sebagai ramuan tradisional. Bagian yang sering digunakan adalah air perasannya, dengan salah satu manfaat dapat digunakan untuk menghilangkan jerawat serta penyembuhan luka agar tidak terjadi abses. Jerawat dan abses pada luka merupakan salah satu infeksi yang disebabkan oleh bakteri Staphylococcus aureus.Tujuan Penelitian ini adalah untuk mengetahui daya hambat air perasan buah jeruk nipis (Citrus aurantifolia S.) terhadap pertumbuhan bakteri Staphylococcus aureus secara invitro. Penelitian dilakukan dengan metoda eksperimental laboratorium dengan desain postest only control group design yang dilakukan di Laboratorium Mikrobiologi Fakultas Kedokteran Universitas Andalas.Hasil penelitian menunjukan bahwa air perasan buah jeruk nipis memiliki daya hambat terhadap pertumbuhan bakteri Staphylococcus aureus dengan berbagai konsentrasi yaitu 25%, 50%, 75%, dan 100% dan terdapat pengaruh lama kontak terhadap pertumbuhan bakteri dimana bakteri tidak tumbuh seteleh kontak 5 menit pertama dan diikuti menit-menit berikutnya dengan air perasan buah jeruk nipis konsentrasi 100%. Jadi, semakin tinggi konsentrasi air perasan buah jeruk nipis dan semakin lama kontak dengan bakteri Staphylococcus aureus maka daya hambatnya semakin baik.Kata kunci: Uji Daya Hambat, Air Perasan Buah Jeruk Nipis, Staphylococcus aureus.Abstract Lime (Citrus aurantifolia S.) is kind of family’s herbal medicine, most using in the community is widely used as a traditional herb. The most common used part is the lime fruit squeeze with one of the function is used for removing acne and wound healing to prevent the form of abscess. Pimples and abscesses of the wound is one of the infections caused by the bacterium Staphylococcus aureus.The purpose of this study was to determine the inhibition of lime fruit (Citrus aurantifolia S.) squeeze towards the growth of the bacteria Staphylococcus aureus in vitro condition. The study was conducted with laboratory experimental methods to the design of control group design postest only performed at the Laboratory of Microbiology Faculty of Medicine, University of Andalas.The results showed that the lime fruit (Citrus aurantifolia S.) squeeze has the ability to inhibite the bacterial growth of Staphylococcus aureus with various concentrations of 25%, 50%, 75%, and 100% and there is the effect of contact time on the growth of bacteria which the bacteria do not grow after contact the first 5 minutes and the next minute followed by lime fruit squeeze with 100% concentration lime fruit squeeze. Thus, the higher the concentration of lime fruit squeeze and the longer the contact with the bacteria Staphylococcus aureus is the better towards.Keywords:Inhibition test, The Lime Fruit Squeeze, Staphylococcus Aureus.


Author(s):  
Baowei Yang ◽  
Yicheng Mei ◽  
Qianhui Li ◽  
Mengyuan Zhang ◽  
Huiling Tang ◽  
...  

There is an urgent need for new antibiotics and alternative strategies to combat bacterial pathogens. Molecular docking, antibacterial evaluation in vitro and in vivo, cytotoxicity assessment and enzyme inhibition analyses were performed. Compound 12 exhibited antimicrobial activity against Staphylococcus aureus (MIC: 4 μg/ml), various clinically isolated strains of MRSA (MIC: 4–16 μg/ml) and Acinetobacter baumannii (MIC: 4 μg/ml) when combined with subinhibitory concentrations of colistin B. Compound 12 (20 mg/kg) yielded mild improvement in survival of methicillin-resistant Staphylococcus aureus (MRSA)-infected mice. Additionally, enzyme inhibition tests showed that compound 12 exhibited inhibitory effects against S. aureus dihydrofolate reductase (105.1 μg/ml) and DNA gyrase (122.8 μg/ml). Compound 12 is a promising antibacterial candidate for further development.


2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Angela Mutuku ◽  
Lizzy Mwamburi ◽  
Lucia Keter ◽  
Joyce Ondicho ◽  
Richard Korir ◽  
...  

Abstract Background Medicinal plants have been used in the treatment of various ailments in most developing countries. Oral infections are the most prevalent diseases in man. The Rhus family has been found to have antimicrobial, antimalarial, and anti-inflammatory properties. Few studies have been done on Rhus vulgaris Meikle. A study was conducted to determine the effect of Rhus vulgaris Meikle stem bark extracts against selected oral pathogenic microorganisms and the safety of the extracts in vitro and in vivo. Methods Methanol:dichloromethane (1:1), methanol and aqueous extracts were tested for bacteriostatic and bactericidal effects against Methicillin Resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus mutans and Candida albicans. Cytotoxicity of the active extracts was determined using Vero E6 cell lines while safety was evaluated in mice and rats. Phytochemical screening was performed on the methanol extracts. One-way ANOVA and Tukey’s multiple comparisons tests were performed using IBM SPSS statistics 20.0 for antimicrobial assay and acute toxicity testing. One-way ANOVA and Dunnett’s multiple comparison tests were conducted using GraphPad Prism 8.0 for cytotoxicity assay. Results Methanol extract of Rhus vulgaris showed significant antimicrobial activity against MRSA (12.00 ± 0.00 mm; p-value of < 0.005; Minimum Inhibitory Concentration of 0.391 mg/ml; Minimum Bactericidal Concentration of 1.563 mg/ml). The extract were not cytotoxic at 100 μg/ml which was the highest tested concentration. In acute dermal irritation testing, the methanol extract resulted in mild irritation with erythema and flaking that cleared within 8 days. There were no observable adverse effects from oral administration of the extracts (acute oral toxicity testing) at concentrations of 50 mg/kg, 300 mg/kg and 2000 mg/kg. Tannins, saponins, flavonoids, terpenoids, glycosides, alkaloids and phenols were detected in the methanol extract. Conclusions Antimicrobial activity of R. vulgaris extracts supports its traditional use as a toothbrush. Cytotoxicity demonstrated by the extracts as well as the mild skin irritation warrants further study before R. vulgaris can be recommended for the development of effective and safe mouthwashes.


Author(s):  
Xuan Zhao ◽  
Xin Zuo ◽  
Jing Zhong ◽  
Bowen Wang ◽  
Saiqun Li ◽  
...  

Ocular chemical burns are potentially blinding ocular injuries and require urgent management. Amniotic membrane (AM) transplantation is an effective surgical treatment, one of the reasons is because AM is a rich source of growth factors that can promote epithelialization and wound healing. However, growth factors will be gradually lost and insufficient after preparation process and long-time storage, leading to unsatisfactory therapeutic effects. Herein, we present a modified AM (AM-HEP) for the supplement and sustained release of growth factor by surface grafting heparin for treatment of ocular chemical burns. Heparin grafting rate and stability, microstructure, physical property, and sustained release of epithelial growth factor (EGF) of AM-HEP were characterized. Biocompatibility and ability to promote corneal epithelial cell growth and migration were evaluated and compared with a biological amnion, which is available on the market in vitro. The therapeutic effects of AM-HEP combined with EGF (AM-HEP@EGF) in vivo had been evaluated in a model of mouse corneal alkali burn. The results indicated that heparin was introduced into AM and maintain stability over 3 weeks at 37°C. The modification process of AM-HEP did not affect microstructure and physical property after comparing with non-modified AM. EGF could be combined quickly and effectively with AM-HEP; the sustained release could last for more than 14 days. AM-HEP@EGF could significantly promote corneal epithelial cell growth and migration, compared with non-modified AM and control group. Faster corneal epithelialization was observed with the transplantation of AM-HEP@EGF in vivo, compared with the untreated control group. The corneas in the AM-HEP@EGF group have less inflammation and were more transparent than those in the control group. The results from in vitro and in vivo experiments demonstrated that AM-HEP@EGF could significantly enhance the therapeutic effects. Taken together, AM-HEP@EGF is exhibited to be a potent clinical application in corneal alkali burns through accelerating corneal epithelial wound healing.


2019 ◽  
Vol 8 (9) ◽  
pp. 1486 ◽  
Author(s):  
Barbara De Angelis ◽  
Margarida Fernandes Lopes Morais D’Autilio ◽  
Fabrizio Orlandi ◽  
Giampiero Pepe ◽  
Simone Garcovich ◽  
...  

Chronic ulcers are characterized by loss of substance without a normal tendency towards spontaneous healing. The Wound Bed Preparation Guideline advises that after diagnosis, the expert should correct the biological state of the ulcer micro-environment based on TIME principles (Tissue, Infection, Moisture balance, Epidermal). There are many ways to treat such ulcers, for example through use of advanced dressings, negative pressure, surgical toilets, dermal substitutes, autologous skin grafting, and free or local flaps. In vitro and in vivo pre-clinical models hold widely acknowledged potential yet complex limitations. Tissue bioengineering could be an ideal approach to foster innovative strategies in wound healing. Our observational study reports on an in vitro and in vivo evaluation of a bio-functionalized scaffold composed of platelet-rich plasma (PRP) and hyaluronic acid (HA) used in 182 patients affected by chronic ulcers (diabetic and vascular), comparing the results with a control group of 182 patients treated with traditional dressings (HA alone). After 30 days the patients who had undergone the combined treatment (PRP + HA), showed 96.8% ± 1.5% re-epithelialization, as compared to 78.4% ± 4.4% in the control group (HA only). Within 80 days, they had 98.4% ± 1.3% re-epithelialization as compared to 87.8% ± 4.1% in the control group (HA only; p < 0.05). No local recurrence was observed during the follow-up period. PRP + HA treatment showed stronger regenerative potential in terms of epidermal proliferation and dermal renewal compared with HA alone.


2014 ◽  
Vol 34 (3) ◽  
pp. 260-265 ◽  
Author(s):  
F Yesildal ◽  
FN Aydin ◽  
S Deveci ◽  
S Tekin ◽  
I Aydin ◽  
...  

Angiogenesis is the process of generating new blood vessels from preexisting vessels and is considered essential in many pathological conditions. The purpose of the present study is to evaluate the effect of aspartame on angiogenesis in vivo chick chorioallantoic membrane (CAM) and wound-healing models as well as in vitro 2,3-bis-2 H-tetrazolium-5-carboxanilide (XTT) and tube formation assays. In CAM assay, aspartame increased angiogenesis in a concentration-dependent manner. Compared with the control group, aspartame has significantly increased vessel proliferation ( p < 0.001). In addition, in vivo rat model of skin wound-healing study showed that aspartame group had better healing than control group, and this was statistically significant at p < 0.05. There was a slight proliferative effect of aspartame on human umbilical vein endothelial cells on XTT assay in vitro, but it was not statistically significant; and there was no antiangiogenic effect of aspartame on tube formation assay in vitro. These results provide evidence that aspartame induces angiogenesis in vitro and in vivo; so regular use may have undesirable effect on susceptible cases.


2018 ◽  
Vol 4 (2) ◽  
Author(s):  
Andita Fitriani ◽  
Erni Setiyorini ◽  
Farach Khanifah

Pendahuluan : Bakteri Staphylococcus aureus merupakan bakteri yang paling sering menyebabkan infeksi. Pemberian antibiotik merupakan upaya pengendalian terhadap infeksi yang dapat menyebabkan resisten. Bakteri Staphylococcus aureus telah resisten terhadap antibiotik ampisilin, amoksisilin-asam klavulanat, amoksisilin, penisilin G, sulbenisilin, kloramfenikol dan siprofloksasin sehingga penanganan terhadap infeksi Staphylococcus aureus relatif sulit. Daun Srikaya diketahui mengandung senyawa flavonoid, saponin dan tannin yang memiliki efek antimikroba. Metode Penelitian : Dalam penelitian ini ditentukan Kadar Hambat Minimum (KHM) dengan menggunakan metode dilusi padat.Penelitian ini merupakan penelitian eksperimen analitik dengan post test only control group design. Sampel dalam penelitian ini adalah bakteri Staphylococcus aureus yang merupakan stok kultur milik Balai Besar Laboratorium Kesehatan (BBLK) Surabaya. Variabel independen dalam penelitian ini adalah ekstrak daun srikaya (Annona squamosa L.) dengan konsentrasi 3%, 6%, 12% dan 24%. Variabel dependen dalam penelitian ini adalah bakteri Staphylococcus aureus. Data dianalisis dengan uji one way ANOVA dilanjutkan uji Post Hoc LSD dengan nilai probabilitas (p)<0,05. Hasil Penelitian : Hasil penelitian menunjukkan bahwa terdapat penurunan jumlah koloni bakteri Staphylococcus aureus yang berbanding terbalik dengan peningkatan konsentrasi ekstrak daun srikaya mulai dari konsentrasi 3% hingga 24%.Kesimpulan : Kesimpulan pada penelitian ini yaitu ekstrak daun srikaya mempunyai efek antimikroba terhadap pertumbuhan Staphylococcus aureus dengan KHM terletak pada konsentrasi dua kali lipat dari konsentrasi 24%. Saran : Sebagai referensi bakteri yang dapat menyebabkan infeksi dan dapat menangsninys dengan antimikroba alami yang minimefek samping disbanding dengan BKOKata Kunci: Antimikroba, Ekstrak Daun Srikaya, , Kadar Hambat Minimum (KHM), Staphylococcus aureus


Biomedicines ◽  
2020 ◽  
Vol 8 (10) ◽  
pp. 386
Author(s):  
Anika Haq ◽  
Suneel Kumar ◽  
Yong Mao ◽  
Francois Berthiaume ◽  
Bozena Michniak-Kohn

The purpose of this study was to synthesize and characterize novel biocompatible topical polymeric film and hydrogel systems that have the potential to deliver the antibacterial agent thymoquinone (TQ) directly to the skin target site to manage the local wound infection and thereby wound healing. The polyvinyl pyrrolidone (PVP) matrix-type films containing TQ were prepared by the solvent casting method. In vitro skin permeation studies on human cadaver skin produced a mean flux of 2.3 µg TQ/cm2/h. Human keratinocyte monolayers subjected to a scratch wound (an in vitro wound healing assay) showed 85% wound closure at day 6 in the TQ group (100 ng/mL TQ) as compared to 50% in the vehicle control group (p = 0.0001). In a zone-of-inhibition (ZOI) assay, TQ-containing films and hydrogels completely wiped out Staphylococcus aureus in 10 cm diameter Tryptic Soy Agar plates while 500 µg/mL gentamicin containing filters gave 10 mm of ZOI. In an ex vivo model, TQ-containing films eradicated bacterial colonization on human cadaver skin. Furthermore, in a full-thickness wound infection model in mice, TQ-containing films showed significant activity in controlling Staphylococcus aureus infection, thereby disinfecting the skin wound. In summary, TQ-containing PVP films and hydrogels developed in this study have the potential to treat and manage wound infections.


2020 ◽  
Vol 13 (5) ◽  
pp. 865-871
Author(s):  
Yos Adi Prakoso ◽  
Chylen Setiyo Rini ◽  
Asih Rahayu ◽  
Miarsono Sigit ◽  
Dyah Widhowati

Background and Aim: Antimicrobial resistance is a global problem caused by extensive utilization of antibiotics that promote gene resistant among bacteria, including Staphylococcus aureus. This study aimed to analyze the potential effects of celery (Apium graveolens) extract as an antioxidant and antimicrobial agent against methicillin-resistant S. aureus (MRSA), in vitro and in vivo. Materials and Methods: Celery was extracted and tested against a MRSA isolate in vitro. The minimum inhibitory concentration (MIC) against MRSA for the celery extract (CE) was determined to be 0.1% and it was formulated into a cream. A total of 30 female Sprague Dawley rats were divided into five groups: Group 1, negative control; Group 2, positive control; Group 3, treated with 0.05% CE cream; Group 4, 0.1% CE cream; and Group 5, 0.2% CE cream. All animals in the groups were exposed to a full-thickness skin biopsy on the dorsal portion, and they were infected with 30 μL of 105 colony-forming units of the MRSA isolate. The treatment was administered twice a day for 7 days. The skin samples were collected on days 3 and 7 after the treatment. The skin tissue was examined histologically using hematoxylin and eosin, Gram staining, and immunohistochemistry against cytokeratin (CK)-17. Results: Results showed that 0.2% of CE cream was the best treatment for wounds infected with MRSA. CE (0.2%) cream increased skin reepithelialization, fibroblast proliferation, and CK-17 expression; it also decreased the percentage of wound area, inflammatory cell infiltration, and bacterial colonization in skin wound tissue compared to the other treatments (p≤0.05). Conclusion: This study demonstrated that celery could be utilized as an alternative herbal therapy against MRSA-associated skin infections.


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