European Union’s Horizon 2020 research and innovation program under grant agreement No. 952033

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European Union’s Horizon 2020 research and innovation program under grant agreement No. 952033

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Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application

Vukašinović-Sekulić, Maja; Đošić, Marija; Janković, Ana; Stevanović, Milena; Grujić, Svetlana; Matić-Bujagić, Ivana; Kojić, Vesna; Mišković-Stanković, Vesna

(Beograd : Savez hemijskih inženjera Srbije, 2024)

TY  - CONF
AU  - Vukašinović-Sekulić, Maja
AU  - Đošić, Marija
AU  - Janković, Ana
AU  - Stevanović, Milena
AU  - Grujić, Svetlana
AU  - Matić-Bujagić, Ivana
AU  - Kojić, Vesna
AU  - Mišković-Stanković, Vesna
PY  - 2024
UR  - https://ritnms.itnms.ac.rs/handle/123456789/1244
AB  - INTRODUCTION: Following orthopedic surgery, bacterial infection may lead to significant complications related to
inflammation in the peri-implant region, including the potential for implant loosening. To address this concern, the
surface of metallic implants was modified through the application of bioactive and antibacterial coatings, aiming to
mitigate these issues.
EXPERIMENTAL: HAP/PVA/CS/Gent coatings was single-step electrophoretically deposited from four-component
aqueous suspension containing 1 wt.% hydroxyapatite powder (HAP, particles < 200 nm particle size, Sigma-Aldrich),
0.1 wt.% poly(vinyl alcohol) (PVA, medium molecular weight 89 to 98 kDa, 99 % hydrolysed, Sigma-Aldrich), 0.05 wt%
chitosan powder (CS, medium molecular weight, 190 to 310kDa with 75 to 85 % deacetylation degree, Sigma-Aldrich), and
aqueous gentamicin sulfate solution (Gent, concentration 50 mg/ml, Sigma-Aldrich) on titanium plates (Sigma-Aldrich).
Antibacterial activity of HAP/PVA/CS/Gent coating was evaluated against Staphylococcus aureus TL (culture collection-
FTM, University of Belgrade, Serbia) and Escherichia coli ATCC 25922, while kinetics of antibacterial activity was monitored
according to our previously published data [1]. In vitro cytotoxicity assay was performed towards two fibroblast cell lines
of different origin-mouse origin cell line (L929 (ATCC CRL-6364)) and human lung origin cell line (MRC-5 (ATCC CCL-171)) [2].
To evaluate the statistical significance of the biological assay results (antibacterial activity, cytotoxicity and ALP activity),
one-way analysis of variance (ANOVA) followed by a multiple comparisons posthoc test was used.
RESULTS AND DISCUSSION: HAP/PVA/CS/Gent coating exhibited strong antibacterial effect against both S. aureus
and E. coli, especially pronounced against S. aureus, causing bactericidal effect. Cytotoxic effect of HAP/PVA/CS/Gent
coating was not pronounced in investigated MRC-5 and L929 cell lines. MRC-5 fibroblast cells in contact with
HAP/PVA/CS/Gent doubled alkaline phosphatase levels compared to their contact with the control samples
(HAP/PVA/CS), indicating good osteogenic properties.
CONCLUSIONS: Electrophoretically deposited HAP/PVA/CS/Gent bioceramic coatings on titanium, demonstrated
significant potential as implants in orthopedic practice, functioning as drug carriers. Not only do they possess
antibacterial properties, but they also exhibit no adverse effects on living tissue.
PB  - Beograd : Savez hemijskih inženjera Srbije
C3  - ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices
T1  - Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application
EP  - 59
IS  - 1s
SP  - 59
VL  - 78
ER  - 
@conference{
author = "Vukašinović-Sekulić, Maja and Đošić, Marija and Janković, Ana and Stevanović, Milena and Grujić, Svetlana and Matić-Bujagić, Ivana and Kojić, Vesna and Mišković-Stanković, Vesna",
year = "2024",
abstract = "INTRODUCTION: Following orthopedic surgery, bacterial infection may lead to significant complications related to
inflammation in the peri-implant region, including the potential for implant loosening. To address this concern, the
surface of metallic implants was modified through the application of bioactive and antibacterial coatings, aiming to
mitigate these issues.
EXPERIMENTAL: HAP/PVA/CS/Gent coatings was single-step electrophoretically deposited from four-component
aqueous suspension containing 1 wt.% hydroxyapatite powder (HAP, particles < 200 nm particle size, Sigma-Aldrich),
0.1 wt.% poly(vinyl alcohol) (PVA, medium molecular weight 89 to 98 kDa, 99 % hydrolysed, Sigma-Aldrich), 0.05 wt%
chitosan powder (CS, medium molecular weight, 190 to 310kDa with 75 to 85 % deacetylation degree, Sigma-Aldrich), and
aqueous gentamicin sulfate solution (Gent, concentration 50 mg/ml, Sigma-Aldrich) on titanium plates (Sigma-Aldrich).
Antibacterial activity of HAP/PVA/CS/Gent coating was evaluated against Staphylococcus aureus TL (culture collection-
FTM, University of Belgrade, Serbia) and Escherichia coli ATCC 25922, while kinetics of antibacterial activity was monitored
according to our previously published data [1]. In vitro cytotoxicity assay was performed towards two fibroblast cell lines
of different origin-mouse origin cell line (L929 (ATCC CRL-6364)) and human lung origin cell line (MRC-5 (ATCC CCL-171)) [2].
To evaluate the statistical significance of the biological assay results (antibacterial activity, cytotoxicity and ALP activity),
one-way analysis of variance (ANOVA) followed by a multiple comparisons posthoc test was used.
RESULTS AND DISCUSSION: HAP/PVA/CS/Gent coating exhibited strong antibacterial effect against both S. aureus
and E. coli, especially pronounced against S. aureus, causing bactericidal effect. Cytotoxic effect of HAP/PVA/CS/Gent
coating was not pronounced in investigated MRC-5 and L929 cell lines. MRC-5 fibroblast cells in contact with
HAP/PVA/CS/Gent doubled alkaline phosphatase levels compared to their contact with the control samples
(HAP/PVA/CS), indicating good osteogenic properties.
CONCLUSIONS: Electrophoretically deposited HAP/PVA/CS/Gent bioceramic coatings on titanium, demonstrated
significant potential as implants in orthopedic practice, functioning as drug carriers. Not only do they possess
antibacterial properties, but they also exhibit no adverse effects on living tissue.",
publisher = "Beograd : Savez hemijskih inženjera Srbije",
journal = "ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices",
title = "Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application",
pages = "59-59",
number = "1s",
volume = "78"
}
Vukašinović-Sekulić, M., Đošić, M., Janković, A., Stevanović, M., Grujić, S., Matić-Bujagić, I., Kojić, V.,& Mišković-Stanković, V.. (2024). Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application. in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices
Beograd : Savez hemijskih inženjera Srbije., 78(1s), 59-59.
Vukašinović-Sekulić M, Đošić M, Janković A, Stevanović M, Grujić S, Matić-Bujagić I, Kojić V, Mišković-Stanković V. Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application. in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices. 2024;78(1s):59-59..
Vukašinović-Sekulić, Maja, Đošić, Marija, Janković, Ana, Stevanović, Milena, Grujić, Svetlana, Matić-Bujagić, Ivana, Kojić, Vesna, Mišković-Stanković, Vesna, "Bioactivity of gentamicin-loaded hydroxyapatite/poly(vinyl alcohol)/chitosan composite coatings aimed for orthopedic application" in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices, 78, no. 1s (2024):59-59.

Electrochemically synthesized biomaterials

Janković, Ana; Đošić, Marija; Stevanović, Milena; Vukašinović-Sekulić, Vesna; Kojić, Vesna; Mišković-Stanković, Vesna

(Beograd : Savez hemijskih inženjera Srbije, 2024)

TY  - CONF
AU  - Janković, Ana
AU  - Đošić, Marija
AU  - Stevanović, Milena
AU  - Vukašinović-Sekulić, Vesna
AU  - Kojić, Vesna
AU  - Mišković-Stanković, Vesna
PY  - 2024
UR  - https://ritnms.itnms.ac.rs/handle/123456789/1242
AB  - INTRODUCTION: Electrochemical methods were employed for synthesizing composites intended for applications in
medicine as antibacterial coatings on titanium bone implants or as highly efficient antibacterial hydrogels for
accelerated wound healing.
EXPERIMENTAL: Hydroxyapatite-based coatings were single-step electrophoretically deposited from fourcomponent
aqueous suspension containing 1 wt% hydroxyapatite powder (HAP, particles < 200 nm particle size, Sigma-
Aldrich), 0.1 wt% poly(vinyl alcohol) (PVA, medium molecular weight, Sigma-Aldrich), 0.05 wt% chitosan powder (CS,
medium molecular weight, Sigma-Aldrich), and aqueous gentamicin sulfate solution (Gent, concentration 50 mg/ml,
Sigma-Aldrich) on titanium plates (Sigma-Aldrich). Antibacterial activity of HAP/PVA/CS/Gent coating was evaluated
against Staphylococcus aureus TL (culture collection-FTM, University of Belgrade, Serbia) and Escherichia coli ATCC
25922. For poly(vinyl alcohol)/chitosan (PVA/CHI) hydrogel synthesis poly(vinyl alcohol) powder (PVA, Sigma Aldrich,
USA), chitosan powder (CHI, Sigma Aldrich, USA), graphene (Graphene Supermarket, USA), glacial acetic acid (Beta
Hem), silver nitrate (MP Hemija), potassium nitrate (Centrohem, Serbia) were used. For antibacterial properties
evaluation, monobasic and dibasic (Sigma Aldrich, USA) potassium phosphates were used. Cell culture suspensions for
cytotoxicity tests were prepared using MTT tetrazolium salt, EDTA, fetal calf serum and antibiotic-antimycotic solution
(Sigma Aldrich, USA).
RESULTS AND DISCUSSION: Surface modification of titanium with innovative bioactive coatings enhances
biomineralization and reduces post-operation pain and infection risk. Using powerful single-step electrophoretic
deposition (EPD) at a constant voltage we have produced various hydroxyapatite-based composite bioceramic coatings
in combination with polymers (lignin, chitosan), graphene, and antibacterial agents (silver, gentamicin).
Electrochemical methods enable in situ synthesis of AgNPs inside polymer matrices, with the main advantage being
the complete absence of any chemical reducing agents that are often toxic and difficult to remove from the material.
The electrochemical reduction of Ag+ ions is achieved only using electrical current or pure hydrogen gas that is generated
at the cathode in aqueous electrolytes, which allows for obtaining completely green and non-toxic product.
Modifications in the synthesis process also enable the control of AgNPs properties, such as size and concentration.
CONCLUSIONS: Composite antibacterial coatings were obtained on titanium plates using EPD. Strong antibacterial
activity against E. coli and S. aureus was found. Biocompatibility was confirmed using in vitro MTT testing since a noncytotoxic
effect was shown towards healthy human peripheral blood mononuclear cells PBMC, fibroblast cell lines MRC-5
and L929, suggesting high potential for bone tissue engineering and medical applications. Hydrogels composed of physically
cross-linked poly(vinyl alcohol)/chitosan with embedded AgNPs were successfully developed for applications as antibacterial
wound dressings due to their highly favorable properties of silver release, antibacterial activity, and non-toxicity.
PB  - Beograd : Savez hemijskih inženjera Srbije
C3  - ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices
T1  - Electrochemically synthesized biomaterials
EP  - 57
IS  - 1s
SP  - 57
VL  - 78
ER  - 
@conference{
author = "Janković, Ana and Đošić, Marija and Stevanović, Milena and Vukašinović-Sekulić, Vesna and Kojić, Vesna and Mišković-Stanković, Vesna",
year = "2024",
abstract = "INTRODUCTION: Electrochemical methods were employed for synthesizing composites intended for applications in
medicine as antibacterial coatings on titanium bone implants or as highly efficient antibacterial hydrogels for
accelerated wound healing.
EXPERIMENTAL: Hydroxyapatite-based coatings were single-step electrophoretically deposited from fourcomponent
aqueous suspension containing 1 wt% hydroxyapatite powder (HAP, particles < 200 nm particle size, Sigma-
Aldrich), 0.1 wt% poly(vinyl alcohol) (PVA, medium molecular weight, Sigma-Aldrich), 0.05 wt% chitosan powder (CS,
medium molecular weight, Sigma-Aldrich), and aqueous gentamicin sulfate solution (Gent, concentration 50 mg/ml,
Sigma-Aldrich) on titanium plates (Sigma-Aldrich). Antibacterial activity of HAP/PVA/CS/Gent coating was evaluated
against Staphylococcus aureus TL (culture collection-FTM, University of Belgrade, Serbia) and Escherichia coli ATCC
25922. For poly(vinyl alcohol)/chitosan (PVA/CHI) hydrogel synthesis poly(vinyl alcohol) powder (PVA, Sigma Aldrich,
USA), chitosan powder (CHI, Sigma Aldrich, USA), graphene (Graphene Supermarket, USA), glacial acetic acid (Beta
Hem), silver nitrate (MP Hemija), potassium nitrate (Centrohem, Serbia) were used. For antibacterial properties
evaluation, monobasic and dibasic (Sigma Aldrich, USA) potassium phosphates were used. Cell culture suspensions for
cytotoxicity tests were prepared using MTT tetrazolium salt, EDTA, fetal calf serum and antibiotic-antimycotic solution
(Sigma Aldrich, USA).
RESULTS AND DISCUSSION: Surface modification of titanium with innovative bioactive coatings enhances
biomineralization and reduces post-operation pain and infection risk. Using powerful single-step electrophoretic
deposition (EPD) at a constant voltage we have produced various hydroxyapatite-based composite bioceramic coatings
in combination with polymers (lignin, chitosan), graphene, and antibacterial agents (silver, gentamicin).
Electrochemical methods enable in situ synthesis of AgNPs inside polymer matrices, with the main advantage being
the complete absence of any chemical reducing agents that are often toxic and difficult to remove from the material.
The electrochemical reduction of Ag+ ions is achieved only using electrical current or pure hydrogen gas that is generated
at the cathode in aqueous electrolytes, which allows for obtaining completely green and non-toxic product.
Modifications in the synthesis process also enable the control of AgNPs properties, such as size and concentration.
CONCLUSIONS: Composite antibacterial coatings were obtained on titanium plates using EPD. Strong antibacterial
activity against E. coli and S. aureus was found. Biocompatibility was confirmed using in vitro MTT testing since a noncytotoxic
effect was shown towards healthy human peripheral blood mononuclear cells PBMC, fibroblast cell lines MRC-5
and L929, suggesting high potential for bone tissue engineering and medical applications. Hydrogels composed of physically
cross-linked poly(vinyl alcohol)/chitosan with embedded AgNPs were successfully developed for applications as antibacterial
wound dressings due to their highly favorable properties of silver release, antibacterial activity, and non-toxicity.",
publisher = "Beograd : Savez hemijskih inženjera Srbije",
journal = "ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices",
title = "Electrochemically synthesized biomaterials",
pages = "57-57",
number = "1s",
volume = "78"
}
Janković, A., Đošić, M., Stevanović, M., Vukašinović-Sekulić, V., Kojić, V.,& Mišković-Stanković, V.. (2024). Electrochemically synthesized biomaterials. in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices
Beograd : Savez hemijskih inženjera Srbije., 78(1s), 57-57.
Janković A, Đošić M, Stevanović M, Vukašinović-Sekulić V, Kojić V, Mišković-Stanković V. Electrochemically synthesized biomaterials. in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices. 2024;78(1s):57-57..
Janković, Ana, Đošić, Marija, Stevanović, Milena, Vukašinović-Sekulić, Vesna, Kojić, Vesna, Mišković-Stanković, Vesna, "Electrochemically synthesized biomaterials" in ExcellMater Conference 2024 - Innovative Biomaterials for Novel Medical Devices, 78, no. 1s (2024):57-57.

Elektroforetski taložene kompozitne prevlake sa gentamicinom

Đošić, Marija; Janković, Ana; Stevanović, Milena; Vukašinović-Sekulić, Maja; Mišković-Stanković, Vesna

(Belgrade : Serbian Chemical Society, 2023)

TY  - CONF
AU  - Đošić, Marija
AU  - Janković, Ana
AU  - Stevanović, Milena
AU  - Vukašinović-Sekulić, Maja
AU  - Mišković-Stanković, Vesna
PY  - 2023
UR  - https://ritnms.itnms.ac.rs/handle/123456789/749
AB  - Uvođenje antibiotika u polimer/mineralnu kompozitnu prevlaku poboljšava antibakterijska svojstva prevlaka. Kompozitna prevlaka (hidroksiapatit - HAP, poli(vinilalkohol), hitozan, gentamicin) dobijena je elektroforetskim taloženjem iz četvorokomponentne vodene suspenzije. Primenom termogravimetrijske i rendgenske difrakcione analize pokazano je prisustvo karbonatno-supstituisanog HAP. Antibakterijska aktivnost kompozitne prevlake sa gentamicinom potvrđena je protiv soja S. aureus i E. coli, čineći ovu prevlaku pogodnom za potencijalnu primenu u biomedicini.
AB  - Antibiotic introduction in polymer/mineral coating can improve the antibacterial properties of coatings. Composite coating (hydroxyapatite, poly(vinyl alcohol), chitosan, gentamicin) was fabricated from a four-component aqueous suspension using electrophoretic deposition. Structural and morphological characterization of the coating was investigated by X-ray diffraction and thermogravimetric analysis proving carbonate-substituted HAP. The presence of gentamicin contributed to the antibacterial activity of composite coating against S. aureus and E. coli strains, representing a suitable material for possible application in biomedicine.
PB  - Belgrade : Serbian Chemical Society
C3  - 59. Savetovanje Srpskog hemijskog društva
T1  - Elektroforetski taložene kompozitne prevlake sa gentamicinom
T1  - Electrophoretically deposited gentamicin-loaded composite coating
EP  - 58
SP  - 58
ER  - 
@conference{
author = "Đošić, Marija and Janković, Ana and Stevanović, Milena and Vukašinović-Sekulić, Maja and Mišković-Stanković, Vesna",
year = "2023",
abstract = "Uvođenje antibiotika u polimer/mineralnu kompozitnu prevlaku poboljšava antibakterijska svojstva prevlaka. Kompozitna prevlaka (hidroksiapatit - HAP, poli(vinilalkohol), hitozan, gentamicin) dobijena je elektroforetskim taloženjem iz četvorokomponentne vodene suspenzije. Primenom termogravimetrijske i rendgenske difrakcione analize pokazano je prisustvo karbonatno-supstituisanog HAP. Antibakterijska aktivnost kompozitne prevlake sa gentamicinom potvrđena je protiv soja S. aureus i E. coli, čineći ovu prevlaku pogodnom za potencijalnu primenu u biomedicini., Antibiotic introduction in polymer/mineral coating can improve the antibacterial properties of coatings. Composite coating (hydroxyapatite, poly(vinyl alcohol), chitosan, gentamicin) was fabricated from a four-component aqueous suspension using electrophoretic deposition. Structural and morphological characterization of the coating was investigated by X-ray diffraction and thermogravimetric analysis proving carbonate-substituted HAP. The presence of gentamicin contributed to the antibacterial activity of composite coating against S. aureus and E. coli strains, representing a suitable material for possible application in biomedicine.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "59. Savetovanje Srpskog hemijskog društva",
title = "Elektroforetski taložene kompozitne prevlake sa gentamicinom, Electrophoretically deposited gentamicin-loaded composite coating",
pages = "58-58"
}
Đošić, M., Janković, A., Stevanović, M., Vukašinović-Sekulić, M.,& Mišković-Stanković, V.. (2023). Elektroforetski taložene kompozitne prevlake sa gentamicinom. in 59. Savetovanje Srpskog hemijskog društva
Belgrade : Serbian Chemical Society., 58-58.
Đošić M, Janković A, Stevanović M, Vukašinović-Sekulić M, Mišković-Stanković V. Elektroforetski taložene kompozitne prevlake sa gentamicinom. in 59. Savetovanje Srpskog hemijskog društva. 2023;:58-58..
Đošić, Marija, Janković, Ana, Stevanović, Milena, Vukašinović-Sekulić, Maja, Mišković-Stanković, Vesna, "Elektroforetski taložene kompozitne prevlake sa gentamicinom" in 59. Savetovanje Srpskog hemijskog društva (2023):58-58.