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Poster marta michalska sionkowska
1. Surface properties of antibacterial materials based on collagen nad thymol.
Marta Michalska-Sionkowska1, Beata Kaczmarek2 Alina Sionkowska2, Maciej Walczak1
1 Nicolaus Copernicus University in Torun, Faculty Biology and Environment Protection, Department of Environmental Microbiology and Biotechnology,
Lwowska 1, Toruń, Poland
2Nicolaus Copernicus University, Faculty of Chemistry, Biomaterials and Cosmetics Department, Gagarina 7, 87-100 Toruń, Poland
INTRODUCTION
Collagen is a polymer that naturally occurs in the human body, and due to its characteristic properties such as biocompatibility, and non-
toxicity is commonly used in the biomedical and cosmetic field. Thymol is a monoterpene phenol which shows many biological activities, such
as antimicrobial, antioxidant, anti-inflammatory and local antiseptic [1]. Prepared collagen/thymol material showed antibacterial properties [2]. In
this study, surface properties of this material were analyzed.
MATERIALS AND METHODS
A solution of thymol (T) in ethanol was added to collagen with nonionic surfactant Polysorbate 80 (NS) to obtain better miscibility. Following
amount of thymol: 4; 1; 0.75; 0.5 and 0.25 mg per cm² of collagen film was added, not all concentrations are shown on the poster. The
concentrations were chosen based on our previous results. Materials were obtained by the solvent evaporation method. Thin film based on
collagen and nonionic surfactant was left as a control sample. Topographic images of materials were obtained using a multimode scanning probe
microscope with a Nanoscope IIIa controller (Digital Instruments, Santa Barbara, CA) operating in the tapping mode, in the air, at room
temperature. To determine the impact of thymol addition to collagen materials, pure collagen (Coll) film as well as collagen with a non-ionic
surfactant (NS) were taken as control samples. Tested films with 0.75 and 0.5 mg thymol were placed into the Staphylococcus aureus inoculated
flasks and left for 5 days in 21 °C. After the appropriate time, films were removed from flasks, washed in sterile PBS buffer and left to dry at
room temperature. Bacterial biofilm formation on the surface of all film samples was assessed using Scanning Electron Microscope.
RESULT AND DISCUSSION
CONCLUSION
The rough surface is proper for materials considering as wound dressing materials
because it allows cells to attach. Especially, this parameter is important for fibroblasts
and keratinocytes, involved in the wound healing process. Thymol addition to collagen
materials inhibits the biofilm formation by S. aureus. Collagen films modified by
thymol can be successfully used as antimicrobial materials for potential medical
application.
[1] K.R. Riella, R.R. Marinho, J.S. Santos, R.N. Pereira-Filho, J.C. Cardoso, R.L.C. Albuquerque- Junior, S.M. Thomazzi, Journal of Ethnopharmacology 143 (2012) 656-663
[2]M. Michalska-Sionkowska, M. Walczak, A.Sionkowska, Polymer Testing 63 (2017) 360-366
[3] S. Koosehgol, M. Ebrahimian-Hosseinabadi, M. Alizadeh, A. Zamanian, Materials Science and Engineering C 79 (2017) 66–75
[4] D. Altiok, E. Altiok, F. Tihminlioglu, Journal of Materials Science: Materials in Medicine 21 (2010) 2227–2236
CollColl/NSColl/NS/T0.25Coll/NS/T4
Examples of images in 2D and 3D of the topography of the material surface are
presented in Figure 2. Collagen film topography is more homogeneous compared to
other samples. Analyzing the structure of collagen after non-ionic surfactant addition,
microwaves on collagen surface are visible. Small addition of thymol results in a
slightly lower roughness (Rq= 49.5 nm; Ra= 40.5 nm) compared to control (Rq= 54.4
nm; Ra= 43.9 nm). Nevertheless, microwaves are transformed into larger waves.
On the surface of the material with the highest amount thymol, the bubble-like
structure is visible. It's happened because some amount of thymol evaporates during
materials preparation. Koosehgol et al. 2017 received similar results for
chitosan/polyethylene glycol fumarate/thymol hydrogels. They suggest that some part
of thymol evaporate, especially those that were faced to the air. The rest of the thymol
amount is captured between polymeric chains. Therefore, the present of thymol may
be confirmed by FTIR spectroscopy and bactericidal analysis [2]. Altiok et al. 2010
also notice, bubble-like structure and increase of Ra parameter on chitosan films with
thyme oil addition.
On the control surface S. aureus form biofilm, one can
see microcolonies distributed on the surface evenly (Fig.
1a). Microcolonies on the control surface (Coll/NS) were
smaller after 5 days but their numbers were higher
compared to the concentration of 0.5 mg of thymol
(Coll/NS/T0.5). For collagen films with 0.75 mg of thymol
a reduced biofilm formation was observed (Fig. 1c). On
the surface of the control film, cells into microcolonies
were not destroyed. The architecture of microcolonies on
Coll/NS/T0.5 was built by bacteria without any
morphological defect as well as by some other external
polymeric structures. Morphologically normal bacteria
cells were not observed on Coll/NS/T0.75. Polymeric
structures without cells were present on this surface only.
Fig.1 Biofilm formed by Staphylococcus aureus after 5 days on the surface on a-Coll/NS; b- Coll/NS/T0.5; c-
Coll/NS/T0.75 under magnification 2000 (1st line) and 20 000 times (2cd line).
Fig.2. 2D and 3D images of surface topography.
![Surface properties of antibacterial materials based on collagen nad thymol.
Marta Michalska-Sionkowska1, Beata Kaczmarek2 Alina Sionkowska2, Maciej Walczak1
1 Nicolaus Copernicus University in Torun, Faculty Biology and Environment Protection, Department of Environmental Microbiology and Biotechnology,
Lwowska 1, Toruń, Poland
2Nicolaus Copernicus University, Faculty of Chemistry, Biomaterials and Cosmetics Department, Gagarina 7, 87-100 Toruń, Poland
INTRODUCTION
Collagen is a polymer that naturally occurs in the human body, and due to its characteristic properties such as biocompatibility, and non-
toxicity is commonly used in the biomedical and cosmetic field. Thymol is a monoterpene phenol which shows many biological activities, such
as antimicrobial, antioxidant, anti-inflammatory and local antiseptic [1]. Prepared collagen/thymol material showed antibacterial properties [2]. In
this study, surface properties of this material were analyzed.
MATERIALS AND METHODS
A solution of thymol (T) in ethanol was added to collagen with nonionic surfactant Polysorbate 80 (NS) to obtain better miscibility. Following
amount of thymol: 4; 1; 0.75; 0.5 and 0.25 mg per cm² of collagen film was added, not all concentrations are shown on the poster. The
concentrations were chosen based on our previous results. Materials were obtained by the solvent evaporation method. Thin film based on
collagen and nonionic surfactant was left as a control sample. Topographic images of materials were obtained using a multimode scanning probe
microscope with a Nanoscope IIIa controller (Digital Instruments, Santa Barbara, CA) operating in the tapping mode, in the air, at room
temperature. To determine the impact of thymol addition to collagen materials, pure collagen (Coll) film as well as collagen with a non-ionic
surfactant (NS) were taken as control samples. Tested films with 0.75 and 0.5 mg thymol were placed into the Staphylococcus aureus inoculated
flasks and left for 5 days in 21 °C. After the appropriate time, films were removed from flasks, washed in sterile PBS buffer and left to dry at
room temperature. Bacterial biofilm formation on the surface of all film samples was assessed using Scanning Electron Microscope.
RESULT AND DISCUSSION
CONCLUSION
The rough surface is proper for materials considering as wound dressing materials
because it allows cells to attach. Especially, this parameter is important for fibroblasts
and keratinocytes, involved in the wound healing process. Thymol addition to collagen
materials inhibits the biofilm formation by S. aureus. Collagen films modified by
thymol can be successfully used as antimicrobial materials for potential medical
application.
[1] K.R. Riella, R.R. Marinho, J.S. Santos, R.N. Pereira-Filho, J.C. Cardoso, R.L.C. Albuquerque- Junior, S.M. Thomazzi, Journal of Ethnopharmacology 143 (2012) 656-663
[2]M. Michalska-Sionkowska, M. Walczak, A.Sionkowska, Polymer Testing 63 (2017) 360-366
[3] S. Koosehgol, M. Ebrahimian-Hosseinabadi, M. Alizadeh, A. Zamanian, Materials Science and Engineering C 79 (2017) 66–75
[4] D. Altiok, E. Altiok, F. Tihminlioglu, Journal of Materials Science: Materials in Medicine 21 (2010) 2227–2236
CollColl/NSColl/NS/T0.25Coll/NS/T4
Examples of images in 2D and 3D of the topography of the material surface are
presented in Figure 2. Collagen film topography is more homogeneous compared to
other samples. Analyzing the structure of collagen after non-ionic surfactant addition,
microwaves on collagen surface are visible. Small addition of thymol results in a
slightly lower roughness (Rq= 49.5 nm; Ra= 40.5 nm) compared to control (Rq= 54.4
nm; Ra= 43.9 nm). Nevertheless, microwaves are transformed into larger waves.
On the surface of the material with the highest amount thymol, the bubble-like
structure is visible. It's happened because some amount of thymol evaporates during
materials preparation. Koosehgol et al. 2017 received similar results for
chitosan/polyethylene glycol fumarate/thymol hydrogels. They suggest that some part
of thymol evaporate, especially those that were faced to the air. The rest of the thymol
amount is captured between polymeric chains. Therefore, the present of thymol may
be confirmed by FTIR spectroscopy and bactericidal analysis [2]. Altiok et al. 2010
also notice, bubble-like structure and increase of Ra parameter on chitosan films with
thyme oil addition.
On the control surface S. aureus form biofilm, one can
see microcolonies distributed on the surface evenly (Fig.
1a). Microcolonies on the control surface (Coll/NS) were
smaller after 5 days but their numbers were higher
compared to the concentration of 0.5 mg of thymol
(Coll/NS/T0.5). For collagen films with 0.75 mg of thymol
a reduced biofilm formation was observed (Fig. 1c). On
the surface of the control film, cells into microcolonies
were not destroyed. The architecture of microcolonies on
Coll/NS/T0.5 was built by bacteria without any
morphological defect as well as by some other external
polymeric structures. Morphologically normal bacteria
cells were not observed on Coll/NS/T0.75. Polymeric
structures without cells were present on this surface only.
Fig.1 Biofilm formed by Staphylococcus aureus after 5 days on the surface on a-Coll/NS; b- Coll/NS/T0.5; c-
Coll/NS/T0.75 under magnification 2000 (1st line) and 20 000 times (2cd line).
Fig.2. 2D and 3D images of surface topography.](https://image.slidesharecdn.com/xribqomvtlchtjl3lcbe-signature-1f13f8c56ce1ea493738e74b0969f1ff675723a88a6d8814ae1d6b10f073637a-poli-190427113557/85/Poster-marta-michalska-sionkowska-1-320.jpg)
