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1.
Bratisl Lek Listy ; 119(6): 391-396, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29947241

RESUMEN

OBJECTIVE: Platelet-derived growth factor (PDGF-BB) is an important factor in the regeneration and wound healing. One of the problems is that there is not enough efficiency in the transmission or delivery of such factors in the wound site. In this study, alginate sulfate hydrogel was synthesized with recombinant PDGF-BB growth factor for achieving a quick method in wound healing. METHODS: In this study, Alginate sulfate hydrogel was made by photo-crosslinking method and methacrylate. Its toxicity was evaluated by MTT assay. Transforming growth factor was studied in releasing from synthesized alginate sulfate hydrogels and also, lack of toxicity was confirmed, and hydrogel was made with a recombinant human growth factor. Wounds were created with a diameter of 10 mm on the back of rats in order to check the wound healing. RESULTS: This study showed that alginate sulfate hydrogels-PDGF-BB were faster in wound healing than non-sulfate alginate hydrogels-PDGF-BB. Therefore, the controlled delivery of growth factor system can be a powerful idea for therapeutic applications for wound healing. CONCLUSION: Alginate sulfate hydrogel with recombinant growth factor 4 µg/cm2 (PDGF-BB) was very suitable for wound healing (Tab. 1, Fig. 3, Ref. 34).


Asunto(s)
Alginatos/farmacología , Inductores de la Angiogénesis/farmacología , Hidrogeles/farmacología , Proteínas Proto-Oncogénicas c-sis/farmacología , Proteínas Recombinantes/farmacología , Sulfatos/farmacología , Cicatrización de Heridas/efectos de los fármacos , Animales , Becaplermina , Ratas , Regeneración/efectos de los fármacos
2.
Cell Mol Biol (Noisy-le-grand) ; 62(12): 31-36, 2016 Oct 31.
Artículo en Inglés | MEDLINE | ID: mdl-27894397

RESUMEN

Incheh Broun hypersaline wetland is located near the border of Turkmenistan in thenorth of Iran. This wetland is notable because of salinity (280g/l) and alteration in pH range (2.8 to 6.8). Eastern part of wetland is affected by wastewater of iodine extraction factory.  Samples were taken from soil, water and salt. Totally, 400 bacterial strains were isolated of which 194 strains were Gram-positive bacilli, 184 strains were Gram-negative rod and 22 strains were Gram-positive cocci. According to phylogenetic analysis of 16S rRNA, selected strains were placed in three taxonomic phyla including Firmicutes, Actinobacteria and Gammaproteobacteria. Optimum growth was evaluated for salt and 22 strains were found to be moderate halophile and 33 strains were halotolerant. Production of lipase, amylase, gelatinase and protease was examined. Gram-positive bacilli were the main producers of hydrolytic enzymes. Gelatinase and protease were the most frequent enzymes. Gram-positive cocci were the main producers of lipase but they didn't produce amylase.


Asunto(s)
Bacterias/genética , Microbiología del Suelo , Bacterias/clasificación , Bacterias/aislamiento & purificación , ADN Bacteriano/aislamiento & purificación , ADN Bacteriano/metabolismo , Bacterias Gramnegativas/clasificación , Bacterias Gramnegativas/genética , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/genética , Bacterias Grampositivas/aislamiento & purificación , Concentración de Iones de Hidrógeno , Irán , Filogenia , ARN Ribosómico 16S/química , ARN Ribosómico 16S/genética , ARN Ribosómico 16S/metabolismo , Salinidad , Análisis de Secuencia de ADN , Eliminación de Residuos Líquidos , Humedales
3.
Cell Mol Biol (Noisy-le-grand) ; 62(8): 45-51, 2016 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-27545214

RESUMEN

The PDGF-BB plays a key role in several pathogenesis diseases and it is believed to be an important mediator for wound healing. The recombinant human PDGF-BB is safe and effective to stimulate the healing of chronic, full thickness and lower extremity diabetic neurotrophic ulcers. In the present study, we attempted to produce a PDGF-BB growth factor and also, evaluate its functionality in cell proliferation in yeast host Pichia pink. Pichia pink yeast was used as a host for evaluation of the rhPDGF-BB expression. The coding sequence of PDGF-BB protein was synthesized after optimization and packed into the pGEM. Recombinant proteins were produced and purified. The construct of pPinkα-HC-pdgf was confirmed by sequence, the PDGF-BB protein was expressed and purified with using a nickel affinity chromatography column and then characterized by SDS-PAGE electrophoresis. The biological activity of PDGF-BB was estimated with using human fibroblast cell line. The measurement of protein concentration was determined by Bradford and human PDGF-BB ELISA kit. Purified rhPDGF-BB showed similar biological activity (as the standard PDGF-BB) and suggested that the recombinant protein has a successful protein expression (as well as considerable biological activity in P. pink host). The exact amount of recombinant PDGF-BB concentrations were measured by specific ELISA test which it was about 30 µg/ml. Our study suggested that efficiency of biological activity of PDGF-BB protein may be related to its conformational similarity with standard type and also, it practically may be important in wound healing and tissue regeneration.


Asunto(s)
Pichia/metabolismo , Proteínas Proto-Oncogénicas c-sis/genética , Proteínas Recombinantes/genética , Células 3T3 , Animales , Becaplermina , Western Blotting , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Humanos , Ratones , Plásmidos/metabolismo , Reacción en Cadena de la Polimerasa , Proteínas Proto-Oncogénicas c-sis/aislamiento & purificación , Proteínas Proto-Oncogénicas c-sis/metabolismo , Proteínas Recombinantes/aislamiento & purificación , Estándares de Referencia , Reproducibilidad de los Resultados , Transformación Genética
4.
Probiotics Antimicrob Proteins ; 4(2): 133-9, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-26781855

RESUMEN

In recent years, the widespread use of antibiotics has caused many bacterial pathogens resistance to conventional antibiotics. Therefore, generation of new antibiotics to control and reduce the effects of these pathogens is urgently needed. Antimicrobial peptides and proteins are important members of the host defense system in eukaryotes. These peptides are potent, broad-spectrum antibiotics that demonstrate potential as novel and alternative therapeutic agents for the treatment of drug-resistant infections. Accordingly, we evaluated two hybrid peptides CM11 (WKLFKKILKVL-NH2) and CM15 (KWKLFKKIGAVLKVL-NH2) on five important pathogenic bacteria. These peptides are short cecropin-melittin hybrid peptides obtained through a sequence combination approach, which are highly effective to inhibit the growth of important pathogenic bacteria. The activity of these two cationic peptides (CM11 and CM15) in different concentrations (2-64 mg/L) was investigated against standard and clinical isolates of important hospital infection bacteria by measuring MIC, MBC, and bactericidal assay. These peptides demonstrated the same ranges of inhibitory values: The organisms in early 24 h were more susceptible to polycationic peptides (MIC: 8 mg/L and MBC 32 mg/L), but after 48 h the MIC and MBC remained constant for the CM11 peptide. Bactericidal assay showed that all bacteria strains did not have any growth in agar plates after 40 min. The result showed that these two peptides are more effective than other peptides.

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