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Research Article

Suspension of zinc oxide nanoparticles (ZnO-NP) as an intraoperative wound irrigation to prevent infection after fracture fixation

[version 1; peer review: 1 approved, 1 approved with reservations]
Krisna Yuarno Phatama
https://orcid.org/0000-0003-1050-7561
1,2, Respati S. Dradjat
https://orcid.org/0000-0001-8880-019X
2, Edi Mustamsir2, [...] Dwi Yuni Nurhidayati3, Dewi Santosaningsih3, Dwikora Novembri Utomo4, Mohamad Hidayat2
Krisna Yuarno Phatama
https://orcid.org/0000-0003-1050-7561
1,2, Respati S. Dradjat
https://orcid.org/0000-0001-8880-019X
2, [...] Edi Mustamsir2, Dwi Yuni Nurhidayati3, Dewi Santosaningsih3, Dwikora Novembri Utomo4, Mohamad Hidayat2
PUBLISHED 26 May 2023
Author details Author details
OPEN PEER REVIEW
REVIEWER STATUS

This article is included in the QUVAE Research and Publications gateway.

This article is included in the Nanoscience & Nanotechnology gateway.

Abstract

Background: Infection after fracture fixation (IAFF) associated with an implant in orthopaedic surgery is a feared complication, leading to non-union, loss of function, amputation, and even mortality and morbidity to the patient. A biofilm formation on the implant surface increases the difficulty of treatment. Therefore, surgical infection prevention with an effective antiseptic solution is required. This study aims to reveal more about the antibacterial effect of ZnO-NP suspension, particularly Staphylococcus aureus and its biofilm, in preventing surgical infection.
Method: An in vitro experimental study with a posttest-only control group design was used to see the antimicrobial activity and inhibitory effect of Staphylococcus aureus biofilm formation between a 20% zinc oxide nanoparticle (ZnO-NP) suspension (20 mg/mL) compared to 0.3% povidone-iodine solution. The statistical result was tested using an independent t-test antibacterial activity. Meanwhile biofilm identification was evaluated using Mann-Whitney & Kruskal Wallis test for each group.
Result: 20% ZnO-NP suspension has a minimum inhibitory concentration at 4 g/mL and a minimum bactericidal concentration at 16 g/mL, same as 0.3% povidone-iodine administration and its statistically significant (p-value 0.001). Minimum biofilm inhibitory concentration was seen at a concentration of 20% ZnO-NP suspension of 2 μg, while at higher doses, it showed lysis of bacterial cells.
Conclusion: 20% ZnO-NP suspension is a promising solution for preventing surgical infection due to its antibacterial and antibiofilm effects.

Keywords

ZnO-NP suspension, Wound Irrigation, Staphylococcus aureus, Biofilm, Infection After Fracture Fixation

Corresponding author: Krisna Yuarno Phatama
Competing interests: No competing interests were disclosed.
Grant information: The author(s) declared that no grants were involved in supporting this work.
Copyright:  © 2023 Phatama KY et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
How to cite: Phatama KY, Dradjat RS, Mustamsir E et al. Suspension of zinc oxide nanoparticles (ZnO-NP) as an intraoperative wound irrigation to prevent infection after fracture fixation [version 1; peer review: 1 approved, 1 approved with reservations]. F1000Research 2023, 12:554 (https://doi.org/10.12688/f1000research.132374.1) First published: 26 May 2023, 12:554 (https://doi.org/10.12688/f1000research.132374.1) Latest published: 28 Sep 2023, 12:554 (https://doi.org/10.12688/f1000research.132374.2)

Introduction

Infection after fracture fixation (IAFF) related to an implant in orthopaedic surgery is a commonly feared complication, resulting in non-union, poor functional outcome until loss of function, amputation, and even mortality and morbidity to the patient.1,2 This complication also carries a meaningful socioeconomic burden because treatment takes a long time and has high cost.3 Staphylococcus aureus is the most common cause of surgical site infection.4 Surgical site infection occurs because the microorganisms and dead tissue debris attached to the wound surface form biofilms. The formation of biofilm and the resistance of the microorganisms that form it to antimicrobials cause this process to continue into IAFF. Furthermore, the biofilm formation on the implant surface increases the difficulty of treatment.5 Therefore, the prevention of surgical infection becomes essential.

Intraoperative wound irrigation is one of the most important procedures for infection prevention. Normal saline is the most popular option because it is safe and acts as a diluent agent. However, normal saline does not have an antiseptic effect, so it cannot eradicate bacteria.6–8 Another option is povidone-iodine as an effective antiseptic. Unfortunately, an in vitro study showed that povidone-iodine could be toxic even at small concentrations and exposure duration of 1 minute.9 Because of the importance of intraoperative wound irrigation for infection prevention, a safe (non-toxic) and effective antiseptic solution is required.

Zinc oxide, especially in the form of a nanoparticle (ZnO-NP) suspension, appears to be very promising as a choice of intraoperative wound irrigation compared to non-antiseptic normal saline and toxic povidone-iodine.10 ZnO-NP is an effective antimicrobial material that damages the cell membrane of bacteria. Conversely, ZnO-NP is categorized as a safe substance to consume by the American Food and Drug Administration (FDA).11 This study aims to reveal more about the antibacterial effect of ZnO-NP suspension, particularly Staphylococcus aureus and its biofilm, in preventing surgical infection.

Methods

This research was conducted between June and December 2022 at the Laboratory of Clinical Microbiology, Faculty of Medicine, Universitas Brawijaya – Clinical Microbiology Installation of Dr. Saiful Anwar General Hospital.

Research design

An in vitro experimental study with a posttest-only control group design was used to see the antimicrobial activity and inhibitory effect of Staphylococcus aureus biofilm formation between a 20% ZnO-NP suspension (20 mg/mL) and compared to 0.3% povidone-iodine solution as the positive control.

Sample

The sample of this study is an isolate of Staphylococcus aureus bacteria obtained from the clinical isolate in the Clinical Microbiology Laboratory, Faculty of Medicine, Universitas Brawijaya. The isolate was stored in temperature -20°C in Tryptic Soy Broth media (TSB – Merck 105459).

Research procedure

Identification of Staphylococcus aureus bacteria

The procedure used for the identification of Staphylococcus aureus was Gram staining. In this study, the isolate used was Staphylococcus aureus. Furthermore, the identification and test of antimicrobial sensitivity with the Vitek® 2 system method were carried out. The procedure used for gram staining is listed below:

  • 1. Glass objects (GEA 7105) were cleaned with cotton swab (Onemed size S) and passed over fire to remove fat or other substances, then left to cool for a while.

  • 2. The bacterial preparation was made on a glass object with one inoculation, allowed to dry in air, then fixed on a Bunsen lamp.

  • 3. The preparation was poured with crystal violet solution (Sigma-aldrich V5265), after one minute, the preparation was rinsed with water.

  • 4. A Lugol solution (MERCK 1.09261.1000) was poured into the preparation, after one minute, the preparation was rinsed with water.

  • 5. A 90% alcohol was poured into the preparation and rinsed with water after 5-10 seconds.

  • 6. After that, a safranin (Sigma-aldrich 94635) was poured into the preparation, after 30 seconds, rinsed with water.

  • 7. The preparation was dried with blotting paper (Ahlstrom-Munksjö Grade 240), then viewed under a light microscope (Olympus CX-21, USA) with 1000× magnification. It shows round purple and clustered bacteria identified as gram-positive cocci.

After gram staining, the preparation was identified as gram-positive cocci, an antimicrobial sensitivity was performed using the Vitek® 2 system (Biomérieux, USA). It was a system to identify bacteria isolates using GP ID cards with faster and accurate bacterial identification.

Bacterial inoculation in nutrient agar plate (NAP)

Staphylococcus aureus bacterial suspension was inoculated and placed in an incubator (Digisystem DSI-500D) that has been set for 24 hours at 37°C on Mueller-Hinton agar media (Sigma-aldrich 70191) to obtain separate colonies with four suspensions. The targeted bacterial concentration was 5 × 105 colony forming unit per millilitre (CFU/mL).

Antimicrobial susceptibility test of ZnO-NP suspension and 0.3% povidone-iodine solution using broth dilution method

The broth dilution method was used to assess the antibacterial activity, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), of various antiseptic solutions. The principle of the broth dilution method is to expose the bacteria to be tested with a certain substance (in this study, an antiseptic solution). In general, the concentration of the solution used is a serial dilution multiple of two (2, 4, 6, 8, 10, 12, 14, 16 (μg/mL)). The first MIC value is the lowest ZnO-NP concentration, indicating a clear suspension. All 32 samples bacterial suspensions of various concentrations (2, 4, 6, 8, 10, 12, 14, 16 (μg/mL)) of ZnO-NP with four replication of each, were inoculated on Mueller Hinton agar and incubated at 37°C for 24 hours in an incubator that has been set (Digisystem DSI-500D) to determine the MBC value of ZnO-NP against Staphylococcus aureus. The MBC value is obtained when all bacteria are killed.

Biofilm identification

Biofilm formation was evaluated by inoculating Staphylococcus aureus bacteria cultured overnight plus tryptic soy broth (TSB – Merck 105459) into microwell in congo red agar plate. The congo red agar was made by infusing the congo red dye into mannitol salt agar (oxoid). After incubating for 24 hours at 37°C, the medium was rinsed twice using phosphate-buffered saline (PBS – Sigma-aldrich P4474) and then dried. The formation of biofilms is considered positive when a layer of violet crystalline colour appears on the walls and base of the tube, and the observation can be done by naked eye. The formation of rings at the level of the surface of a liquid is not considered meaningful in the formation of biofilms. MBIC from ZnO-NP suspension was assessed by evaluating biofilms formed on the surface of the microwell wall. Using the same method as the method for assessing the formation of biofilms, before incubation, a certain concentration of ZnO-NP suspension is added (1, 2, 3 μg/mL). Each concentration represents the intervention groups, rather than positive and negative control. A 1 μg/mL concentration of ZnO-NP suspension for group micro 1, 2 μg/mL concentration of ZnO-NP suspension for group micro 2 and 3 μg/mL concentration of ZnO-NP suspension for group micro 3. The ZnO-NP concentration is distinguished for the categorization of intervention groups. Then after the incubation is completed, staining is carried out, and the formed biofilm is evaluated using a scanning electron microscope (SEM - SU8600 Hitachi Europe) and based on optical density (OD) determined by an ELISA reader (Accuris Smartreader 96-T) with a wavelength of 630 nm (OD 630 nm). The lowest concentration that causes inhibition of biofilms on the surface of the microwell wall is considered to be MBIC. Meanwhile, concentrations without biofilm growth are considered minimum biofilm eradication concentration (MBEC) values.

Biofilm assay

Isolates from the microbiology lab, with a glucose TSB of 1% (Merck 105459). 200 μl or 0.2 ml of diluted suspension was added to 96 sterile microtiter wells. A total 20 wells were filled, 4 to each group negative control (A1-A4), positive control (A5-A8), micro 1 (A9-A12), micro 2 (B1-B4) and micro 3 (B5-8). After that, a suspension of ZnO-NP is added: 1 μg/mL to group micro 1, 2 μg/mL to group micro 2, and 3 μg/mL to group micro 3. Then incubated for 24 hours at a temperature of 37°C; after incubation, the suspension was removed, and the microtiter well was washed 3× with 0.2 ml sterile PBS pH 7.2 to remove planktonic cells (freeform). The biofilm formed at the wall or bottom of the well is stained with 1% violet crystals and left for 20 minutes. After that, 1 molar 200 μg hydrochloric acid (HCl) in concentrated isopropanol was added. The excess violet crystals are washed with water, the microtiter wells are then dried in an inverted position. Biofilm formation was assessed based on OD determined by an ELISA reader with a wavelength of 630 nm (OD 630 nm).

Data analysis

To evaluate the effect of various ZnO-NP suspension concentrations on the growth of colonies of Staphylococcus aureus bacteria, SPSS 25 (IBM Corp. 2017) (RRID:SCR_002865) for Windows statistical test program was used. Data distribution was tested for normality using the Shapiro-Wilk test, while homogeneity was tested using the Levene test. A comparison of the average number of colonies between 0.3% povidone-iodine and 20% ZnO-NP suspension has been done using an independent t-test.

The biofilm identification and assay result shows that data was not normally distributed and heterogenous; therefore, a Mann-Whitney test was used to determine the correlation between variables. The results of comparisons between groups were carried out with the Mann-Whitney test. It is statistically significance if the p-value < 0.05. All the statistical result from normality and homogeneity test to statistical data will be included in the underlying data.25

Ethical considerations

All protocols were approved by the Health Research Ethics Commission of General Hospital Dr. Saiful Anwar Malang with ethical approval number 400/104/K.3/302/2022.

Results

The experiment’s broth dilution method results showed that the MIC was at a concentration of 4 g/mL, where the black line began to appear and became clear in the test tube for ZnO-NP suspension. While the results of the colony forming unit (CFU) calculation and linear regression from serially increasing the dose of ZnO-NP at a concentration of 4 g/mL are shown in Table 1, and calculation results of methicillin-resistant Staphylococcus aureus (MRSA) colonies on Mueller–Hinton media with exposure to ZnO-NP suspension compared with 0.3% povidone-iodine are shown in Table 2.

Table 1. Results of CFU calculation and linear regression from serially increasing dose of ZnO-NP.

DosageReplication 1 (CFU/mL)Replication 2 (CFU/mL)Replication 3 (CFU/mL)Mean (CFU/mL)SD
02.3 × 1082.6 × 1082.9 × 1082.6 × 1082.8 × 107
21.6 × 1082.0 × 1081.4 × 1081.7 × 1083.0 × 107
42.6 × 1052.0 × 1051.8 × 1052.1 × 1054.0 × 104
61.1 × 1051.1 × 1059.7 × 1041.1 × 1059.2 × 103
86.6 × 1047.1 × 1047.1 × 1046.9 × 1043.1 × 103
103.5 × 1041.8E × 1046.9 × 1044.1 × 1042.6 × 104
122.8 × 1032.8 × 1032.8 × 1032.8 × 1030
147.1 × 1027.1 × 1022.8 × 1031.4 × 1031.2 × 103
1600000

Table 2. Calculation results of the number of MRSA colonies with exposure to ZnO-NP compared with 0.3% Povidone-Iodine (PI).

DosageCFU 1CFU 2CFU 3CFU 4MeanSD
02.2 × 1082.7 × 1082.7 × 1082.8 × 1082.6 × 1082.7 × 107
21.8 × 1081.9 × 1081.8 × 1082.0 × 1081.9 × 1089.4 × 106
42.4 × 1052.4 × 1052.8 × 1052.5 × 1052.5 × 1051.9 × 104
69.8 × 1049.3 × 1048.7 × 1049.2 × 1049.2 × 1044.6 × 103
85.7 × 1044.2 × 1044.4 × 1042.5 × 1044.2 × 1041.3 × 104
103.0 × 1041.3 × 1043.3 × 1043.0 × 1042.6 × 1049.2 × 103
123.5 × 1033.5 × 1032.1 × 1035.0 × 1033.5 × 1031.2 × 103
147.1 × 1027.1 × 1027.1 × 1027.1 × 1027.1 × 1020.0
160.00.00.00.00.00.0
PI0.00.00.00.00.00.0

The results of bacterial inoculation on Mueller–Hinton above show that the larger the dose, will decrease the number of bacteria (CFU). The dosage of MIC (4 g/mL) showed a decrease in CFU of as much as 99.3%, and a dose of 16 g/mL could eradicate all bacteria. In addition, the dose of 16 g/mL is the MBC. After all those procedures, the average number of colonies was compared. The independent t-test showed a significant difference between the number of colonies formed on the ZnO-NP 4 g/mL administration and 0.3% povidone-iodine (p-value = 0.001). Using 0.3% povidone-iodine causes bacteria not to grow at all. Similarly, administering ZnO-NP solution with a 16 g/mL concentration also caused no bacterial growth. Thus, the MBC concentration of ZnO-NP suspension had the same effect as 0.3% povidone-iodine administration. Biofilm identification was carried out in Congo Red agar to obtain a biofilm image with a picture of a black colony.

Analysis of the effect of ZnO-NP suspension in inhibiting the growth of biofilm-producing Staphylococcus aureus by SEM examination. SEM examination was carried out to qualitatively determine biofilms’ formation in Staphylococcus aureus bacteria. The formation of biofilms can be seen from the matrix extracellular of the biofilms formed and seen in SEM. SEM examinations were performed on negative, positive, and treatment groups (Figures 1 and 2).

0b6afd40-9779-4f80-8f32-473b266dd74b_figure1.gif

Figure 1. Scanning Electron Microscope (SEM) examination of the negative control group (left) and positive control group (right).

0b6afd40-9779-4f80-8f32-473b266dd74b_figure2.gif

Figure 2. Scanning Electron Microscope (SEM) examination of the treatment control group with 1 μgr ZnO-NP (A), 2 μgr ZnO-NP (B) and 3 μgr ZnO-NP (C).

This study found that the picture of biofilm formation in the treatment group was less compared to the control group (+). Coupled with the increased dose of ZnO-NP application to colonies of Staphylococcus aureus bacteria, there is less and less biofilm formation. MBIC was seen at a concentration of ZnO-NP suspension of 2 μg, while at higher doses, it showed lysis of bacterial cells.

Descriptive analysis is used to find out an overview of the purchasing variables. This descriptive analysis is expected to provide an overview of the state of patient data. To find out the description for each variable can be seen in Table 3.

Table 3. Descriptive statistics.

NMean ± SD
Negative control4‐0.547 ± 0.056
Positive control4‐0.301 ± 0.039
Group 1 μg4‐0.252 ± 0.011
Group 2 μg4‐0.247 ± 0.026
Group 3 μg4‐0.230 ± 0.009

The Mann-Whitney test was used to determine significant differences between the treatments with ordinal data scales or data that do not meet the assumptions of normality and homogeneity. Further analysis was carried out using Kruskal Wallis to test whether significant differences exist between one treatment group and another.

Based on the results of the Mann Whitney’s analysis in Table 4, it was found that the p-value for OD was 0.003, the result of the p-value < 0.05 for the OD parameter, which means that there is a significant difference between the treatments at an error rate of 5%. Testing to determine the differences between each group used the Kruskal-Wallis test. The results of the comparison between each group were carried out in Table 5, showing a significant difference in the average group if the p-value < 0.05.

Table 4. Mann Whitney test.

ParametersSig.pInformation
Log optical density17.9810.003Significant

Table 5. Kruskal Wallis log optical density test.

Group comparisonpInformation
Negative controlPositive control0.021Significant
Group 1 μg0.020Significant
Group 2 μg0.021Significant
Group 3 μg0.021Significant
Positive controlGroup 1 μg0.042Significant
Group 2 μg0.043Significant
Group 3 μg0.021Significant
Group 1 μgGroup 2 μg0.561Not significant
Group 3 μg0.020Significant
Group 2 μgGroup 3 μg0.248Not significant

Discussion

Staphylococcus aureus is a gram-positive bacteria that can cause various diseases. Infection by this pathogen is common both in the community and in hospitals. Staphylococcus aureus is also a bacterium that often causes infections in bones and joints.12 Zinc oxide nanoparticle (ZnO-NP) is a white to yellowish-white crystalline powder which are soluble in water. The most common crystal structures are wurtzite (hexagonal) and zinc alloys. The physico-chemical properties of ZnO-NP contribute to its antibacterial activity.13

This study proves that ZnO-NP suspension has antibacterial activity with MIC at 4 g/mL, effectively killing 99.3% of bacteria. In a maximal dose of 16 g/mL, ZnONP suspension killed 100% bacteria. Based on Table 2 above, in 2 g/mL, CFU count 1.9 × 108; in 4 g/mL, the CFU decreases to 2.5 × 105. For dose 16 g/mL, the CFU is 0, the same as 0.3% povidone-iodine. This study’s MBC had a killing power equivalent to that of diluted 0.3% povidone-iodine. A diluted povidone-iodine solution has been recommended for wound irrigation.14 This study shows that ZnO-NP is a potential antiseptic agent and can provide an effect equivalent to other recommended antiseptic solutions.15 On the other hand, there are significant differences in MBC values from those found in this study compared to previous studies. In this study, the MBC value obtained from the experiment was 16 g/mL, this was 7.81 g/mL in previous studies, reaching 50% of this study.16 In contrast, the studies above on poultry feedstuffs showed MBC as high as 100 g/mL. This difference could be caused by differences in the isolates used.

There are several antibacterial mechanism of ZnO-NP: (1) the release of ROS reactive oxygen species (ROS) generated on the surface of nanomaterials can attack the bacterial cell wall, and the antibacterial activity is further enhanced by increasing the surface area17; (2) another possible mechanism for the antibacterial activity of ZnO is the release of Zn2+ ions which can damage cell membranes and penetrate intracellular contents17; (3) Contact between bacterial cells and particles causes changes in the microenvironment within the contact area of organisms and particles. Brayner et al. also showed that the bacterial cell wall was damaged and disorganized after contact with ZnO-NP nanoparticles. The abrasive ZnO-NP causes an increase in membrane permeability leading to the subsequent cellular internalization of the nanoparticles18 and; (4) other mechanisms. It is also believed that the antimicrobial activity of ZnO may be related to its photocatalytic activity. By absorbing UV light which activates its interaction with bacteria, suspension of ZnO nanoparticles can produce ROS such as H2O2, which has a phototoxic effect on bacteria.19

ZnO-NP has been developed for research purposes and health-related applications. Not only does it have antibacterial activity, but ZnO-NP inhibits biofilm formation.19 Based on SEM and OD in this study, MBIC is at a dose of 2 μg/mL and is statistically significant. While bacteriolysis effectively started at a dose of 3 μg/mL. Research conducted by Abdelraheem also found the same thing. ZnO-NP was found to suppress the expression of the ica A, ica D, and fnb A genes, the main genes forming biofilms in Staphylococcus aureus.19 Based on the study’s results, it was used as a background to assess the ability of ZnO-NP to inhibit the formation of biofilms against Staphylococcus aureus bacteria. In research by Jasim and Abdelraheem, it was also explained that ZnO-NP is effective in inhibiting the formation of biofilms in Vancomycin and Linezolid-resistant Staphylococcus aureus so that it can also be used on multi-resistant Staphylococcus aureus against antibiotics.19,20 In another study by Abdulkareem, ZnO-NP was also used in dental implants and had benefits in antimicrobials and inhibition of biofilm formation.21

ZnO-NP has several mechanisms that inhibit biofilm formation: (1) inhibiting the adhesion and proliferation stages; (2) changing the formation of amyloid fibrils; and (3) inhibiting the Quorum sensing mechanism, which is not yet known in detail. The workings of ZnO-NP suspension are thought to have started by degrading biofilms with an effective dose of 2 μg/mL, and then after the biofilm is destroyed, the bacterial cells die with a higher dose. Without biofilms, bacteria become more susceptible to antibacterials agents.22–24

Conclusions

20% ZnO-NP suspension is a promising solution for preventing surgical infection due to its antibacterial and antibiofilm effects. It has a minimum inhibitory concentration (MIC) for Staphylococcus aureus at 4 g/mL and a minimum bactericidal concentration (MBC) at 16 g/mL. A higher suspension dose effectively eradicates Staphylococcus aureus bacteria and its biofilm, which has minimal biofilm inhibitory concentration (MBIC) at a dose of 2 μg/mL. This study has limitations because it does not evaluate the expression of genes that form Staphylococcus aureus biofilm formation; therefore, further research is needed.

Author contribution

Krisna Yuarno Phatama: Conceptualization, Writing – Original Draft Preparation, Validation, Project Administration, Methodology

Respati S. Dradjat: Conceptualization, Writing – Original Draft Preparation, Supervision

Edi Mustamsir: Conceptualization, Writing – Original Draft Preparation, Supervision

Dwi Yuni Nurhidayati: Supervision, Validation, Data Curation, Resources

Dewi Santosaningsih: Supervision, Validation, Data Curation, Resources

Dwikora Novembri Utomo: Writing – Review & Editing, Supervision, Validation

Mohamad Hidayat: Writing – Review & Editing, Supervision, Validation

Data availability

Underlying data

Some data underlying the results are available as part of the article, and the others are available on the list of additional resources below.

Zenodo: Suspension of Zinc Oxide Nanoparticles (ZnO-NP) as an Intraoperative Wound Irrigation to Prevent Infection After Fracture Fixation, https://doi.org/10.5281/zenodo.7765281. 25

This project contains the following underlying data:

  • - Raw Data Fixed.xlsx

  • - Descriptive ZnO-NP & PI CFU.xlsx (Descriptive comparative CFU between ZnO-NP and PI)

  • - Descriptive OD.xlsx (Descriptive Optical Density of Biofilm)

  • - Normality & Homogeneity Test.xlsx

  • - Statistical Test.xlsx (Mann-whitney & Kruskal-wallis test)

  • - Control Negative - UMM4247(x18k).jpg (SEM)

  • - Control Negative - UMM4247(x18k).txt (SEM)

  • - Control Negative - UMM4248(x18k).jpg (SEM)

  • - Control Negative - UMM4248(x18k).txt (SEM)

  • - Control Positive - UMM4249(x18k).jpg (SEM)

  • - Control Positive - UMM4249(x18k).txt (SEM)

  • - Control Positive - UMM4250(x18k).jpg (SEM)

  • - Control Positive - UMM4250(x18k).txt (SEM)

  • - Micro 1 - UMM4251(x18k).jpg (SEM)

  • - Micro 1 - UMM4251(x18k).txt (SEM)

  • - Micro 1 - UMM4252(x18k).jpg (SEM)

  • - Micro 1 - UMM4252(x18k).txt (SEM)

  • - Micro 1 - UMM4253(x18k).jpg (SEM)

  • - Micro 1 - UMM4253(x18k).txt (SEM)

  • - Micro 1 - UMM4254(x18k).jpg (SEM)

  • - Micro 1 - UMM4254(x18k).txt (SEM)

  • - Micro 2 - UMM4255(x18k).jpg (SEM)

  • - Micro 2 - UMM4255(x18k).txt (SEM)

  • - Micro 2 - UMM4256(x18k).jpg (SEM)

  • - Micro 2 - UMM4256(x18k).txt (SEM)

  • - Micro 2 - UMM4257(x18k).jpg (SEM)

  • - Micro 2 - UMM4257(x18k).txt (SEM)

  • - Micro 2 - UMM4258(x18k).jpg (SEM)

  • - Micro 2 - UMM4258(x18k).txt (SEM)

  • - Micro 3 - UMM4263(x18k).jpg (SEM)

  • - Micro 3 - UMM4263(x18k).txt (SEM)

  • - Micro 3 - UMM4265(x18k).jpg (SEM)

  • - Micro 3 - UMM4265(x18k).txt (SEM)

  • - Micro 3 - UMM4266(x18k).jpg (SEM)

  • - Micro 3 - UMM4266(x18k).txt (SEM)

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

Reporting guidelines

Zenodo: Suspension of Zinc Oxide Nanoparticles (ZnO-NP) as an Intraoperative Wound Irrigation to Prevent Infection After Fracture Fixation, https://doi.org/10.5281/zenodo.7765281. 25

This project contains the following reporting guidelines:

  • - Author Checklist.pdf (ARRIVE checklist)

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

References

  • 1.  Steinmetz S, Wernly D, Moerenhout K, et al.: Infection after fracture fixation. EFORT Open Rev. 2019; 4: 468–475. PubMed Abstract | Publisher Full Text | Free Full Text
  • 2.  Hak DJ, Fitzpatrick D, Bishop JA, et al.: Delayed union and nonunions: epidemiology, clinical issues, and financial aspects. Injury. 2014; 45: S3–S7. Publisher Full Text
  • 3.  Thakore R v, Greenberg SE, Shi H, et al.: Surgical site infection in orthopedic trauma: A case-control study evaluating risk factors and cost. J. Clin. Orthop. Trauma. 2015; 6: 220–226. PubMed Abstract | Publisher Full Text | Free Full Text
  • 4.  Tong SYC, Davis JS, Eichenberger E, et al.: Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin. Microbiol. Rev. 2015; 28: 603–661. PubMed Abstract | Publisher Full Text | Free Full Text
  • 5.  Almoudi MM, Hussein AS, Abu Hassan MI, et al.: A systematic review on antibacterial activity of zinc against Streptococcus mutans. Saudi Dent. J. 2018; 30: 283–291. Publisher Full Text
  • 6.  Tande AJ, Patel R: Prosthetic joint infection. Clin. Microbiol. Rev. 2014; 27: 302–345. Publisher Full Text
  • 7.  Suleiman LI, Mesko DR, Nam D: Intraoperative Considerations for Treatment/Prevention of Prosthetic Joint Infection. Curr. Rev. Musculoskelet. Med. 2018; 11: 401–408. PubMed Abstract | Publisher Full Text | Free Full Text
  • 8.  Bruins MJ, Juffer P, Wolfhagen MJHM, et al.: Salt Tolerance of Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus. J. Clin. Microbiol. 2007; 45: 682–683. Publisher Full Text
  • 9.  von Keudell A , Canseco JA, Gomoll AH: Deleterious Effects of Diluted Povidone–Iodine on Articular Cartilage. J. Arthroplast. 2013; 28: 918–921. Publisher Full Text
  • 10.  Xie Y, He Y, Irwin PL, et al.: Antibacterial activity and mechanism of action of zinc oxide nanoparticles against Campylobacter jejuni. Appl. Environ. Microbiol. 2011; 77: 2325–2331. PubMed Abstract | Publisher Full Text | Free Full Text
  • 11.  Aslam Z, Roome T, Razzak A, et al.: Investigation of wound healing potential of photo-active curcumin-ZnO-nanoconjugates in excisional wound model. Photodiagn. Photodyn. Ther. 2022; 39: 102956. PubMed Abstract | Publisher Full Text
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Phatama KY, Dradjat RS, Mustamsir E et al. Suspension of zinc oxide nanoparticles (ZnO-NP) as an intraoperative wound irrigation to prevent infection after fracture fixation [version 1; peer review: 1 approved, 1 approved with reservations]. F1000Research 2023, 12:554 (https://doi.org/10.12688/f1000research.132374.1)
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ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested
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Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions
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Reviewer Report 19 Apr 2024
Nicolaas C. Budhiparama, Arthroplasty & Sports Medicine, Medistra Hospital, Nicolaas Institute of Constructive Orthopaedics Research and Education Foundation, Jakarta, Indonesia 
Approved with Reservations
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https://doi.org/10.5256/f1000research.145288.r175829
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Well-written paper with good methods and discussion allowing future replication of the research. There are several recently published papers (systematic review and RCTs) on a similar topic (antibacterial use of zinc oxide); therefore, the author can provide ... Continue reading
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Budhiparama NC. Reviewer Report For: Suspension of zinc oxide nanoparticles (ZnO-NP) as an intraoperative wound irrigation to prevent infection after fracture fixation [version 1; peer review: 1 approved, 1 approved with reservations]. F1000Research 2023, 12:554 (https://doi.org/10.5256/f1000research.145288.r175829)
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Reviewer Report 20 Jul 2023
Ismail Hadisoebroto dilogo, Department of Orthopaedic and Traumatology, Dr. Cipto Mangunkusumo National Hospital, Jakarta, Indonesia 
Approved
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https://doi.org/10.5256/f1000research.145288.r175827
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  • Overall, this manuscript is already well-written and shows its purposes thoroughly.
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dilogo IH. Reviewer Report For: Suspension of zinc oxide nanoparticles (ZnO-NP) as an intraoperative wound irrigation to prevent infection after fracture fixation [version 1; peer review: 1 approved, 1 approved with reservations]. F1000Research 2023, 12:554 (https://doi.org/10.5256/f1000research.145288.r175827)
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  • Author Response 29 Nov 2023
    Krisna Yuarno Phatama, Doctoral Program in Medical Sciences, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia
    29 Nov 2023
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    Dear Reviewer,

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    Thank you for all the appreciation you have given to our manuscript. We would be glad if our research could be useful and can be followed ... Continue reading
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  • Author Response 29 Nov 2023
    Krisna Yuarno Phatama, Doctoral Program in Medical Sciences, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia
    29 Nov 2023
    Author Response
    Dear Reviewer,

    Greetings
    Thank you for all the appreciation you have given to our manuscript. We would be glad if our research could be useful and can be followed ... Continue reading
    Report a concern

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Alongside their report, reviewers assign a status to the article:

Approved
The paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations
A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approved
Fundamental flaws in the paper seriously undermine the findings and conclusions

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26 May 23 		read read
  1. Ismail Hadisoebroto dilogo, Dr. Cipto Mangunkusumo National Hospital, Jakarta, Indonesia
  2. Nicolaas C. Budhiparama, Nicolaas Institute of Constructive Orthopaedics Research and Education Foundation, Jakarta, Indonesia

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    Alongside their report, reviewers assign a status to the article:
    Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested
    Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
    Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions
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