Influence of Nanoparticles on Some Physiological and Therapeutic Characteristics of Phaseolus vulgaris L. on Induced Skin Injuries in Mice

          Phaseolus vulgaris seeds were surface sterilized using ethanol 90%, soaked in sterile distilled water for 60min, then soaked in copper and silver nanoparticles at different concentrations (25, 50 or 100mg/l) for 15 and 30min. The effect of nanoparticles on the plant physiological characteristics in vivo was studied. Callus was initiated from treated and non–treated seed embryos, then embryos were maintained and transferred to callus induction on Murashige and Skoog (MS) medium supplemented with combinations of plant growth regulators including Dichlorophynoxyacetic acid (2,4-D) at 2.5mg/l, Benzyl adenine (BA) at 0.5mg/l. The effect of nanoparticles on callus induction, callus fresh weight and callus dry weight were recorded. Aqueous and ethanol extracts of treated and non treated plant parts and tissues were prepared (seeds, seedlings and callus tissues), the presence and concentrations of some active constituents in the treated and non-treated plant tissue extracts were determined. The results concluded that alkaloids,flavonoids, glycosides, steroids, saponins, tannins and terpenes were present in the extracts. Results also indicated that the effective concentration of nanoparticles on studied physiological parameters of treated plant, callus induction, callus fresh and dry weight was 50mg/l silver nanoparticles for 30min. Determination of the carbohydrates, proline and total phenolics in ethanol and aqueous extracts of treated and non treated plant tissues were varied. Carbohydrates, proline and total phenolics increased significantly in ethanol extracts compared with aqueous extracts. Wounds in the skin of albino mice were induced, then different microorganisms were isolated from skin mice wounds, identified by using cultural media, microscopic, biochemical tests. Vitek 2 system was used for gram positive bacteria and API 20 kit for gram negative bacteria and yeast isolates for further identification. The isolated and characterized microorgansims were Staphylococcus aureus, Staphylococcus lentus, Klebsiella pneumoniae, Escherichia coli and yeast (Candida albicans). Antimicrobial activity of treated and non treated plant tissues extracted with ethanol (1mg/1ml) was observed in vitro using disc diffusion method, diameter of inhibition zones were recorded (mm) in comparison to the standard antibiotics (control) namely Amoxillin and Clavulanic acid, Ceftriaxone, Gentamicin (antibacterial) and Clotrimazole (antifungal). Treated and non-treated seeds of ethanol extracts exhibited a better antimicrobial activity in comparison with seedlings and callus tissues of ethanol extracts. Silver nanoparticles increased the efficacy against microorganisms resulting in the highest antibacterial activity followed by copper nanoparticles. On the other hand, the treated and non-treated tissues ethanol extracts also tested on albino mice skin wounds as an ointments for wounds healing and a significant increase occurred in the % of wound healing with a high potential for curing skin after 6-7 days of treatment in comparison with clotrimazole cream and piroxicam gel. Nanoparticles could be considered as a one of the promising compounds for improving therapeutic potential of medicinal plants because of their effects on plant tissues and have the ability to suppress of acute inflammation on induced skin injuries of albino mice.