Characterization and Evaluation of the Antioxidant, Antidiabetic, Anti-Inflammatory, and Cytotoxic Activities of Silver Nanoparticles Synthesized Using Brachychiton populneus Leaf Extract

Round 1

Reviewer 1 Report

In this article, Naveed et. al. describes the synthesis of silver nanoparticles and characterization of it using UV-spectrophotometers, FTIR, SEM, and X-ray.  The author also presented anti-inflammatory and cytotoxicity studies in this manuscript. This is an interesting study, and the authors systematically presented the study in this manuscript and the control experiment, and the results were well corroborated with the data, so, in my opinion, this study will be very interesting for the Processes the readers.

Author Response

Dear Ms. Eileen Zhang

Section Managing Editor

Processes,

Dear Sir,

Please find the attached response to reviewer comments on our research article manuscript entitled “Characterization and evaluation of antioxidant, antidiabetic, anti-inflammatory, and cytotoxicity of silver nanoparticles synthesized using Brachychiton populneus leaves extract.

with manuscript id” “processes-1832040.

 

Reviewer 1

In this article, Naveed et. al. describes the synthesis of silver nanoparticles and characterization of it using UV-spectrophotometers, FTIR, SEM, and X-ray.  The author also presented anti-inflammatory and cytotoxicity studies in this manuscript. This is an interesting study, and the authors systematically presented the study in this manuscript and the control experiment, and the results were well corroborated with the data, so, in my opinion, this study will be very interesting for the Processes the readers.

AR: Thank you very much for your valuable comments. It means a lot to us.

 

 

 

 

Regards

 

Dr. Tariq Aziz (PhD, Postdoc)

Assistant Professor

Pak-Austria Fachhochschule Institute of Applied Sciences and Technology

 

Author Response File: Author Response.docx

Reviewer 2 Report

In this paper, the authors assessed physically and chemically properties of silver nanoparticles (AgNPs) synthesized using Brachychiton populneus leaf extract (BPL-AgNPs), and characterized biological activities of BP-AgNPs. Many studies on characteristics of AgNPs synthesized using biomaterials have been published. However, the AgNPs synthesized using Brachychiton populneus leaf extract seem to be novel, and I believe that the paper is interesting for the readers of the journal, Processes.

 

Comment #1: The abbreviation, AgNPs is not defined in the abstract section. Further, the abbreviation, AgNPs used in this study means both general AgNPs and AgNPs synthesized using Brachychiton populneus leaves extract. These AgNPs should be clearly distinguished in the manuscript. To use another abbreviation defined for AgNPs synthesized using Brachychiton populneus leaves extract may help the readers to understand.

 

Comment #2: Advantages BP-AgNPs in comparison with AgNPs synthesized using other biomaterials are not shown in this manuscript. The differences and characteristics of BP-AgNPs compared with other AgNPs synthesized using biomaterials should be discussed in the Discussion section.

 

Comment #3: Inhibitory activity of α-amylase is not able to demonstrate that the agent presents anti-diabetic activity. In addition, inhibitory activity of heat-denaturation of albumin is not able to demonstrate that the agent exhibit anti-inflammatory activity. Inhibitory activities of α-amylase and heat-induced protein denaturation are only show the possibilities that the agent presents anti-diabetic and anti-inflammatory activities respectively. Certainly, several papers describe Inhibitory activities of α-amylase and heat-induced protein denaturation as anti-diabetic and anti-inflammatory activities. However, these expression ways are not fair, and may cause misleading. The inhibitory activities of BP-AgNPs against starch degradation by α-amylase and heat-denaturation of albumin should be described separately the potential of BP-AgNPs for the anti-diabetic and the anti-inflammatory agent, and these potential should be discussed with proper references. For instance, the subtitle, "Anti-diabetic activity of silver nanoparticles (Alpha-amylase assay)" should be modified as "Inhibition of enzymatic activity of α-amylase by BP-AgNPs" or something.

 

Comment #4: The figure legend of Figure 5 was described "Figure 5: SEM micrograph of silver nanoparticles at (A). 2000x, (B). 500x, (C). 3000x and (D). 1000x magnification." However, each panel seems to indicate 3000x (A), 2000x (B), 1000x (C) and 500x (D) magnification.  Further, drawing quality of the figure does not seem to be high. The figure should be modified, and the sample is shown in attached the file.

 

Comment #5: Figure 7-9, the error bars were not able to find in the figures.

 

Comment #6: A unit, "μg" was indicated as "ug" in this manuscript. This should be corrected.

 

Comment #6: It is better to provide a table which summarizes the IC50 (50% inhibition concentration) values described in the results section. This may help the readers to understand the results.

Comments for author File: Comments.pdf

Author Response

Dear Ms. Eileen Zhang

Section Managing Editor

Processes,

Dear Sir,

Please find the attached response to reviewer comments on our research article manuscript entitled “Characterization and evaluation of antioxidant, antidiabetic, anti-inflammatory, and cytotoxicity of silver nanoparticles synthesized using Brachychiton populneus leaves extract.

with manuscript id” “processes-1832040.

 

Reviewer 2

In this paper, the authors assessed physically and chemically properties of silver nanoparticles (AgNPs) synthesized using Brachychiton populneus leaf extract (BPL-AgNPs), and characterized biological activities of BP-AgNPs. Many studies on characteristics of AgNPs synthesized using biomaterials have been published. However, the AgNPs synthesized using Brachychiton populneus leaf extract seem to be novel, and I believe that the paper is interesting for the readers of the journal, Processes.

 AR: Thank you very much for your comments. It means a lot to us.

Comment #1: The abbreviation, AgNPs is not defined in the abstract section. Further, the abbreviation, AgNPs used in this study means both general AgNPs and AgNPs synthesized using Brachychiton populneus leaves extract. These AgNPs should be clearly distinguished in the manuscript. To use another abbreviation defined for AgNPs synthesized using Brachychiton populneus leaves extract may help the readers to understand.

AR: Thank you very much for your comments. Here we mean to say the synthesized silver nanoparticles (AgNPs) using Brachychiton populneus leaves extract. It has been revised and modified accordingly. It has been highlighted in red color in the revised manuscript. Please see revised manuscript.

Comment #2: Advantages BP-AgNPs in comparison with AgNPs synthesized using other biomaterials are not shown in this manuscript. The differences and characteristics of BP-AgNPs compared with other AgNPs synthesized using biomaterials should be discussed in the Discussion section.

AR: Thank you very much for your comment. It has been revised and modified accordingly. It has been highlighted in red color in the revised manuscript. Please see revised manuscript.

 

 Comment #3: Inhibitory activity of α-amylase is not able to demonstrate that the agent presents anti-diabetic activity. In addition, inhibitory activity of heat-denaturation of albumin is not able to demonstrate that the agent exhibit anti-inflammatory activity. Inhibitory activities of α-amylase and heat-induced protein denaturation are only show the possibilities that the agent presents anti-diabetic and anti-inflammatory activities respectively. Certainly, several papers describe Inhibitory activities of α-amylase and heat-induced protein denaturation as anti-diabetic and anti-inflammatory activities. However, these expression ways are not fair, and may cause misleading. The inhibitory activities of BP-AgNPs against starch degradation by α-amylase and heat-denaturation of albumin should be described separately the potential of BP-AgNPs for the anti-diabetic and the anti-inflammatory agent, and these potential should be discussed with proper references. For instance, the subtitle, "Anti-diabetic activity of silver nanoparticles (Alpha-amylase assay)" should be modified as "Inhibition of enzymatic activity of α-amylase by BP-AgNPs" or something.

AR: Thank you for your comment. We do understand your point of view. Considering the above mentioned comment we have  modified and replaced Anti-diabetic activity of silver nanoparticles (Alpha-amylase assay) with Inhibition of enzymatic activity of  α-amylase by BP-AgNPs. Likewise Anti-inflammatory activity (egg albumin denaturation assay) is also modified and replaced with Inhibition of protein denaturation by BP-AgNPs. Please see revised manuscript.

 Comment #4: The figure legend of Figure 5 was described "Figure 5: SEM micrograph of silver nanoparticles at (A). 2000x, (B). 500x, (C). 3000x and (D). 1000x magnification." However, each panel seems to indicate 3000x (A), 2000x (B), 1000x (C) and 500x (D) magnification.  Further, drawing quality of the figure does not seem to be high. The figure should be modified, and the sample is shown in attached the file.

AR: Thank you very much for your comment. The figure legend has been modified in the revised manuscript and highlighted in red color. Furthermore the quality of SEM micrographs are also enhanced. Please see revised manuscript.

 Comment #5: Figure 7-9, the error bars were not able to find in the figures.

AR: Thank you very much for your comment. The Figures have been modified accordingly and highlighted in red color. Please see revised manuscript.

 Comment #6: A unit, "μg" was indicated as "ug" in this manuscript. This should be corrected.

AR: Thank you very much for your comment. It has been modified accordingly and highlighted in red color. Please see revised manuscript.

 Comment #6: It is better to provide a table which summarizes the IC50 (50% inhibition concentration) values described in the results section. This may help the readers to understand the results.

AR: Thank you very much for your comment. Table 6 has been added in the revised manuscript and highlighted in red color. Please see revised manuscript.

 

 

 

 

Regards

 

Dr. Tariq Aziz (PhD, Postdoc)

Assistant Professor

Pak-Austria Fachhochschule Institute of Applied Sciences and Technology

 

Author Response File: Author Response.docx

Reviewer 3 Report

The proposal of the article is attractive, however, there are some points to improve. In the case of the methodology, it would be better to know how much of the (dry) extract was used with the nanoparticle solution, so that it is possible to know how much each nanoparticle captures. This data would help to better discuss the results obtained.

In the results, I suggest incorporating the standard deviation or error into your calculations and graphs, so that it is possible to appreciate whether or not there really is a significant difference between the controls and their nanoparticles. This would also help to better discuss the results obtained.

Finally, the discussion is quite brief, they have enough results that they can take advantage of to carry out an adequate discussion that gives more support to what they have obtained. As I already mentioned, statistically compare their results, present more works with which they can make efficient comparisons between them and yours. Not only similar results, but better or lesser ones and analyze the reason for it.

Author Response

Dear Ms. Eileen Zhang

Section Managing Editor

Processes,

Dear Sir,

Please find the attached response to reviewer comments on our research article manuscript entitled “Characterization and evaluation of antioxidant, antidiabetic, anti-inflammatory, and cytotoxicity of silver nanoparticles synthesized using Brachychiton populneus leaves extract.

with manuscript id” “processes-1832040.

Reviewer 3

The proposal of the article is attractive, however, there are some points to improve. In the case of the methodology, it would be better to know how much of the (dry) extract was used with the nanoparticle solution, so that it is possible to know how much each nanoparticle captures. This data would help to better discuss the results obtained.

AR: Thank you very much for your comment. The 600ml of extract was prepared from 5g of dried leaves powder. Then 180ml of extract and 20ml of silver nitrate solution mixed together to form nanoparticle solution. After incubation and centrifugation pellet was dried and 20mg of pure silver nanoparticles were obtained.

In the results, I suggest incorporating the standard deviation or error into your calculations and graphs, so that it is possible to appreciate whether or not there really is a significant difference between the controls and their nanoparticles. This would also help to better discuss the results obtained.

AR: Thank you very much for your comment. It has been added to the revised manuscript accordingly and highlighted in red color. Please see revised manuscript.

Finally, the discussion is quite brief, they have enough results that they can take advantage of to carry out an adequate discussion that gives more support to what they have obtained. As I already mentioned, statistically compare their results, present more works with which they can make efficient comparisons between them and yours. Not only similar results, but better or lesser ones and analyze the reason for it.

AR: Thank you very much for your comment. It has been added to the revised manuscript accordingly and highlighted in red color. Please see revised manuscript.

Regards

 

Dr. Tariq Aziz (PhD, Postdoc)

Assistant Professor

Pak-Austria Fachhochschule Institute of Applied Sciences and Technology

Author Response File: Author Response.docx

Reviewer 4 Report

I think overall this is a great paper and is well written. I do have a few comments for the author to consider if improvements are considered. please review the attached pdf

Comments for author File: Comments.pdf

Author Response

Dear Ms. Eileen Zhang

Section Managing Editor

Processes,

Dear Sir,

Please find the attached response to reviewer comments on our research article manuscript entitled “Characterization and evaluation of antioxidant, antidiabetic, anti-inflammatory, and cytotoxicity of silver nanoparticles synthesized using Brachychiton populneus leaves extract.

with manuscript id” “processes-1832040.

 

                                 Reviewer 4

Review comments on Brachychiton populneus leave extract Sliver nanoparticles

Muhammad and team are able to synthesize silver nanoparticles around 15nm using Brachychiton populneus Leaves extract. They then proceeded to characterize the nanoparticles and did some biological studies which are interesting.

I think this is a fine paper and can be accepted. Using the leave extract did interestingly create some sliver nanoparticles that potentially have applications if used as coating. I do have several questions to the author and for the editor to consider, which potentially can make the paper even stronger.

1. What does color change means? I think the color change could potentially means the size difference of the silver nanoparticles, I think it might be meaningful to explain during the synthesis since the color is a form of judgement for the completion of the synthesis.

 

AR:  Thank you very much for your comment. During the synthesis process, the change in color indicates alterations in surface plasmon resonance of silver nitrate solution by the reduction process and led to nanoparticle synthesis. But later, if the nanoparticle solution does not stabilize, particles aggregate into larger sizes, which changes the color. Silver ions reduction and change in color are shown in figure 1 of the manuscript.

2. Surface coating using gel electrophoresis or hplc? I have read the synthesis procedure, it appears that the size of the particles are large enough to be centrifuged down and washed with water. I think it will be a lot convincing to use a HPLC or Gel electrophoresis to justify how pure the particles are and the IR signals are not from the free leaf extracts

 AR:  Thank you very much for your comment. There is no surface coating of silver nanoparticles carried out because coating alters          characteristics of particles. In present work Nanoparticles separated by centrifugation and washed with water three times to remove impurities from the surface and then pure particles are characterized. HPLC or gel electrophoresis are also effective to know the purity of particles but In general, centrifugation is considered a low-cost, straightforward, and appropriate method to isolate and purify AgNPs, especially to remove the residues from newly prepared suspensions (Kahnouji et al., 2019).

3. I also have a question on how are the leaf extracts attached or integrate to the silver nanoparticles, since it is explained that there are components of the leaf extract on the particles.

AR:  Thank you very much for your comment. When plant extract was mixed with silver nitrate solution, the plant metabolites or components comes in contact with silver ions and act as reducing agent to form nanoparticles. Therefore, they have these metabolites on their surface.

4. I am not sure how is the 15nm size being measured, since from figure 5 the scale bar is not clear and it seems like the particles are clumps and not homogeneously distributed.

AR:  Thank you very much for your comment. To estimate the particle sizes Image J software was used (Thiruvengadam and Bansod, 2020). The data obtained were used to draw the frequency distribution histogram using origin graphing and analysis software. Then data put on excel file to calculate average size 15nm of nanoparticles. The particles were uniformly distributed as SEM analysis revealed.

 

Regards

 

Dr. Tariq Aziz (PhD, Postdoc)

Assistant Professor

Pak-Austria Fachhochschule Institute of Applied Sciences and Technology