An Investigation of in Vivo Wound Healing Activity of Biologically Synthesized Silver

Supplementary Information

An investigation of in vivo wound healing activity of biologically synthesized silver nanoparticles

Abhishek Kaler1, Amit Kumar Mittal1, Mahesh Katariya2#, Harshad Harde2#, Ashish Kumar Agrawal2#, Sanyog Jain2 and Uttam Chand Banerjee1*

1Department of Pharmaceutical Technology (Biotechnology)

National Institute of Pharmaceutical Education and Research

Sector-67, SAS Nagar-160062, Punjab, INDIA

2Centre for Pharmaceutical Nanotechnology

Department of Pharmaceutics

National Institute of Pharmaceutical Education and Research

Sector-67, SAS Nagar-160062, Punjab, INDIA

*Corresponding author. Tel:+91-172-2214682-687, Extn 2142; fax: +91-172-2214692

E-mail: [email protected]

# These authors contributed equally to the manuscript

1 List of Tables

Table S-1. Microorganisms screened for the synthesis of silver nanoparticles

Table S-2. Reaction of microbial cells with silver nitrate (1mM) in deionised water

Table S-3. Stability study of biologically synthesized nanosilver gel

Figure S-1. Characterization of synthesised silver nanoparticles

Figure S-2. Effect of gelation on the characterization of particle size by TEM

2 Table S-1

Sr. Microorganism Growth

No. medium 1. Lactobacillus fermentum MTCC 903 MRS medium 2. Lactobacillus plantarum MTCC 1746 NB medium 3. Lactobacillus rhamnosus MTCC 1408 MRS medium 4. Lactobacillus bulgaricus NCIM 2056 MRS medium 5. Lactobacillus bulgaricus NCIM 2057 MRS medium 6. Lactobacillus brevis NCIM 2356 MRS medium 7. Bacillus coagulans MTCC 492 NB medium 8. Burkholderia cenocepacia MTCC 5427 NB medium 9. Geotrichum candidum NCIM 980 YPD medium 10. Saccharomyces cerevisiae TYPE I (Dried powder purchased Lyophilized

from SIGMA) 11. Saccharomyces cerevisiae TYPE II (Dried powder purchased Lyophilized

from SIGMA) 12. Pseudomonas putida MTCC 102 NB medium 13. Phanerochaete.chrysosporium MTCC 787 NB medium 14. Escherichia coli MTCC 433 NB medium 15. Bacillus megaterium MTCC 428 NB medium 16. Phaffia rhodozyma NB medium 17. Rhodococcus erythropolis MTCC 1526 NB medium 18. Acinetobacter baumannii MTCC 1425 YPD medium 19 Saccharomyces boulardii (Probiotic) YPD medium 20 Pichia anomala MTCC 237 YPD medium 21 Rhodotorula bogariensis MTCC 2764 NA medium 22 Bacillus stereothermophillus NA medium

3 Table S-2

Microorganism Change in color of λmax (nm) of Absorbance

cell mass cell

supernatant Lactobacillus fermentum MTCC No change No peak Nil

903 Lactobacillus plantarum MTCC Dark brown No peak Nil

1746 Lactobacillus rhamnosus MTCC No change No peak Nil

1408 Lactobacillus bulgaricus NCIM No change No peak Nil

2056 Lactobacillus bulgaricus NCIM No change No peak Nil

2057 Lactobacillus brevis NCIM 2356 No change No peak Nil Bacillus coagulans MTCC 492 No change No peak Nil Burkholderia cenocepacia MTCC No change No peak Nil

5427 Geotrichum candidum NCIM 980 Light brown 420 1.233 Saccharomyces cerevisiae TYPE No change 400.6 0.945

I (Dried powder purchased from

SIGMA) Saccharomyces cerevisiae TYPE No change 426.1 1.026

II (Dried powder purchased from

SIGMA) Pseudomonas putida MTCC 102 No change No peak Nil Phanerochaete chrysosporium No change No peak Nil

MTCC 787 Escherichia coli (MTCC 433) No change No peak Nil Bacillus megaterium MTCC 428 No change No peak Nil Phaffia rhodozyme No change No peak Nil Rhodococcus erythropolis MTCC No change No peak Nil

1526 Acinetobacter baumannii MTCC No change No peak Nil

4 1425 Saccharomyces boulardii No change 417 0.715

(Probiotic) Pichhia anomala MTCC 237 No change No peak Nil Rhodotorula bogariensis MTCC No change No peak Nil

2764 Bacillus stereothermophillus No change No peak Nil

5 Table S-3 a) Characteristics of Ag-NPs gel

Storage 2-8°C 25°C/60% RH 40°C/75% RH conditions Particle size PDI Viscosity Spreadability Particle size PDI Viscosity Spreadability Particle size PDI Viscosity Spreadability (nm) (Pa.s) (cm²) (nm) ( Pa.s) (cm²) (nm) ( Pa.s) (cm²)

Initial 7±4 0.32±0.01 24.63±0.35 11.52±1.52 7±4 0.32±0.01 24.63±0.35 11.52±1.52 7±4 0.32±0.01 24.63±0.35 11.52±1.52

1st month 8±3 0.33±0.05 24.87±0.41 11.32±1.3 7±3 0.32±0.01 24.75±0.38 11.39±0.4 8±4 0.36±0.025 24.74±0.26 11.7±0.8

2nd month 7±3 0.31±0.03 25.12±0.66 11.29±1.4 7±3 0.33±0.01 24.93±0.43 11.18±0.8 11±3 0.37±0.034 24.89±0.56 11.9±0.6

3rd month 9±5 0.31±0.03 25.42±0.32 11.17±1.11 7±4 0.35±0.02 25.36±0.37 12.1±1.2 Aggregation 25.32±0.61 12.3±1.0

6 Figure S-1

TEM micrograph EDAX

UV-Vis spectrum

7 Figure S-2

TEM image of silver nanoparticles before TEM image of silver nanoparticles after gelation gelation