Wound Healing and Antimicrobial Property of Phytofabricated …
479
3 Results and Discussion
The average particle size calculated was found to be 3–10 nm using Dynamic Light
Scattering measurements (DLS). It was observed that 45 °C was the optimal temperature for the conversion of silver metal to silver nanoparticle and the SPR peaks
became sharper with an increase in optical density of the yellowish-brown solution
after 30 min of incubation depending on increasing temperature from 25 to 45 °C.
The SPR peak at 433 nm indicates the formation of silver nanoparticles. Further
increase in temperature (up to 65 °C) showed a clear shift of SPR peak from 433 to
455 nm and an indication for the increase in AgNPs size (Banerjee and Nath 2015).
Depending on the size of the nanoparticle, the sharpness in the absorbance peak can
be determined, as at 45 °C temperature, the particle size may be smaller that results
in sharpness of the plasmon resonance band of silver nanoparticles (Goia 2004).
Increasing temperature up to 65 °C induced the crystal growth formation around the
nucleus that results in decreasing the absorption as well as the intensity of synthesized
nanoparticle.
The presence of spherical shaped particles within the size range of <5 nm was
shown by AFM analysis. FTIR analysis indicated the involvement of carboxyl
(−C=O), hydroxyl (−OH), and amine (−NH) functional groups of the phytochemicals in capping and stabilizing silver nanoparticles. The rapid electrokinetic behavior
of the silver was evaluated using zeta potential (approximately −23.2 mV) confirmed
its stability (Banerjee and Nath 2015). According to the GC–MS analysis, different
phytochemicals or hydrocarbon groups are present on the surface of the synthesized
nanoparticle. The hydrocarbons present on the synthesized silver nanoparticle are
the capping agent of silver nanoparticles. These capping agents prevent the agglomeration of the particle and thus the synthesized nanoparticle was stabilized (Starlin
et al. 2012).
Biocompatibility study by DAPI staining revealed no change in nuclear
morphology and membrane integrity. The result of comet assay was also similar
in both treated and control groups of cells and it produced no evidence of release
of nuclear material and no disruption of the cellular membranes. Intracellular ROS
levels were not altered significantly when compared with that of the control. Results
of the tests conducted at the cellular and genetic levels indicated that the synthesized
silver nanoparticle is not potentially toxic to induce any significant or harmful change
(data not shown).
Without any treatment, the healing process continues but becomes accelerated by
silver nanoparticles in case of both diabetic and normal mice (Fig. 1). Histological
studies show that re-epithelialization almost complete within 15th day when it was
treated with AgNPs (Fig. 2). For wound healing process, fibroblast cells are important. Scanning electron microscopy images (Fig. 3) show that proper fibroblast cells
are formed by the treatment of AgNPs in relation to standard synthetic drug, SSD.
Results show that AgNPs are much more effective for healing of thermal wound.
The results revealed that the percent of wound contraction was 90% at 15th day
of treatment with the synthesized green silver nanoparticles of normal as well as
479
3 Results and Discussion
The average particle size calculated was found to be 3–10 nm using Dynamic Light
Scattering measurements (DLS). It was observed that 45 °C was the optimal temperature for the conversion of silver metal to silver nanoparticle and the SPR peaks
became sharper with an increase in optical density of the yellowish-brown solution
after 30 min of incubation depending on increasing temperature from 25 to 45 °C.
The SPR peak at 433 nm indicates the formation of silver nanoparticles. Further
increase in temperature (up to 65 °C) showed a clear shift of SPR peak from 433 to
455 nm and an indication for the increase in AgNPs size (Banerjee and Nath 2015).
Depending on the size of the nanoparticle, the sharpness in the absorbance peak can
be determined, as at 45 °C temperature, the particle size may be smaller that results
in sharpness of the plasmon resonance band of silver nanoparticles (Goia 2004).
Increasing temperature up to 65 °C induced the crystal growth formation around the
nucleus that results in decreasing the absorption as well as the intensity of synthesized
nanoparticle.
The presence of spherical shaped particles within the size range of <5 nm was
shown by AFM analysis. FTIR analysis indicated the involvement of carboxyl
(−C=O), hydroxyl (−OH), and amine (−NH) functional groups of the phytochemicals in capping and stabilizing silver nanoparticles. The rapid electrokinetic behavior
of the silver was evaluated using zeta potential (approximately −23.2 mV) confirmed
its stability (Banerjee and Nath 2015). According to the GC–MS analysis, different
phytochemicals or hydrocarbon groups are present on the surface of the synthesized
nanoparticle. The hydrocarbons present on the synthesized silver nanoparticle are
the capping agent of silver nanoparticles. These capping agents prevent the agglomeration of the particle and thus the synthesized nanoparticle was stabilized (Starlin
et al. 2012).
Biocompatibility study by DAPI staining revealed no change in nuclear
morphology and membrane integrity. The result of comet assay was also similar
in both treated and control groups of cells and it produced no evidence of release
of nuclear material and no disruption of the cellular membranes. Intracellular ROS
levels were not altered significantly when compared with that of the control. Results
of the tests conducted at the cellular and genetic levels indicated that the synthesized
silver nanoparticle is not potentially toxic to induce any significant or harmful change
(data not shown).
Without any treatment, the healing process continues but becomes accelerated by
silver nanoparticles in case of both diabetic and normal mice (Fig. 1). Histological
studies show that re-epithelialization almost complete within 15th day when it was
treated with AgNPs (Fig. 2). For wound healing process, fibroblast cells are important. Scanning electron microscopy images (Fig. 3) show that proper fibroblast cells
are formed by the treatment of AgNPs in relation to standard synthetic drug, SSD.
Results show that AgNPs are much more effective for healing of thermal wound.
The results revealed that the percent of wound contraction was 90% at 15th day
of treatment with the synthesized green silver nanoparticles of normal as well as