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Current Organic Chemistry, 2013, 17, 125-131
125
Department of Science and Engineering of Oxidic Materials and Nanomaterials, Faculty of Applied Chemistry and Materials
Science, University Politehnica of Bucharest, Polizu Street no 1-7, 011061, Bucharest, Romania
INCDTP Leather & Footwear Res Inst, Collagen Dept, Bucharest 031215, Romania
Stefan Nicolau Institute of Virology, 285 Mihai Bravu Avenue, 030304, Bucharest, Romania
Department of Microbiology, Faculty of Biology, Universtity of Bucharest, Aleea Portocalelor no. 1-3, 060101, Bucharest, Romania
Abstract: The aims of this research were to obtain improved wound dressings based on collagen (COLL), polysaccharides (dextran=
DEX, diethylaminoethyl-cellulose= DEAEC), silica network and usnic acid, as quorum sensing inhibitor. FT-IR, SEM, interaction with
eukaryotic cells and a novel protocol to evaluate the antimicrobial activity of the new wound dressing, firstly reported in literature were
used for the characterization of fabricated wound dressings. The obtained wound dressings are not cytotoxic, do not influences the mesenchymal stem and exhibit good anti-biofilm properties. Taken together, these results are suggesting that the new systems can be safely
used for local applications on the lesional tissues.
1385-2728/13 $58.00+.00
rounded by a glycocalyx composed of a combination of an extracellular matrix that is produced by the microorganisms and the host
surrounding tissues [14]. The glycocalix contributes to the enhanced resistance of microorganisms to the host response as well as
to various antibiotic treatments [15, 16].
Polysaccharide-based hydrogels are non-cytotoxic and biodegradable [17]. From a structural point of view, polysaccharides
have reactive functional groups that can be modi ed to form hydrogels with speci c characteristics of interest [18]. Dextran is a hydrophilic natural polysaccharide and has attracted much attention
for use in controlled drug-delivery system because of its excellent
hydrophilic nature and biocompatibility [19, 20]. Cellulose, is a
highly interesting material due to its renewability, low price, high
availability, good mechanical properties and has safe characters
such as no taste and odorless, biodegradability, insolubility in water
and most organic solvents [21-23]. Cellulose and its derivatives are
regarded as one of the most popular polymeric materials to prepare
nanoparticles for drug delivery systems [24, 25].
Silica has increasingly attracted interest due to their unique
properties and potential applications in biotechnology and materials
science [26]. Due to its excellent biocompatibility, silica is also an
ideal candidate for biomedical applications such is the targeted drug
release [27, 28]. The porosity of silica, which efficiently encapsulates drugs at high concentrations assures the afore mentioned properties [29]. A surface enriched in silica in the presence of surface
Si-OH groups provides intrinsic hydrophilicity, thus allowing surface attachment of specific biomolecules and increasing target
specificity [30-32].
Usnic acid, a yellowgreen cortical pigment, is a derivative of
dibenzofuran produced by several lichen species, as a product of
fungal secondary metabolism [33]. The antibacterial activity of
usnic acid was recognized early against a number of planktonic
Gram-positive bacteria, to which ndings of anticancer, antiviral,
2013 Bentham Science Publishers
Grumezescu et al.
Materials
All chemicals used for the preparation of the compounds were
of reagent grade quality and were purchased from Sigma- Aldrich.
Collagen (300.000 Da; COLL) gel was obtained in the Leather and
Footwear Research Institute- Collagen Department starting from
calf hides by chemical and enzymatic extraction. The collagen gel
concentration was 2.54 % and pH=7 [36].
Fabrication of Wound Dressing Based Collagen, Polysaccharides and Silica
Wound dressing based collagen, polysaccharides and silica was
prepared as follow: 50 mL of polymeric suspension (1,27% dextran
(DEX); 1,27 % diethylaminoethylcellulose (DEAEC)) is added
onto the collagen gel (50 mL 2,54 %) and let to interact for 30 minutes. Polysaccharides (DEX and DEAEC) and collagen were mixed
in 0.5 M acetic acid by stirring and homogenizing several times.
Silica network was obtained from Na2SiO3 solution (50 mL; 1.27
%) dropped into COLL-DEX/DEAEC solutions until the gel pH=7.
COLL/DEX/SiO2 and COLL/DEAEC/SiO2 were divided in two
halfs, one being cross-linked (CL) with 0,5 % (w/v) glutaraldehyde
solution [37] and the other one being not cross-linked (NCL). Obtained CL and NCL gels were casted into glass Petri dishes (12.5
cm in diameter; 20 mL) to be lyophilized.
Characterization of Wound Dressing Based Collagen, Polysaccharides and Silica
FT-IR. A Nicolet 6700 FT-IR spectrometer (Thermo Nicolet,
Madison, WI) connected to software of the OMNIC operating system (Version 7.0 Thermo Nicolet) was used to obtain FT-IR spectra
of hybrid materials. The samples were placed in contact with attenuated total reflectance (ATR) on a multibounce plate of ZnSe
crystal at controlled ambient temperature (25oC). FT-IR spectra
were collected in the frequency range of 4,000650 cm-1 by coadding 32 scans and at a resolution of 4 cm-1 with strong apodization. All spectra were ratioed against a background of an air spectrum.
SEM. SEM analysis was performed on a HITACHI S2600N
electron microscope, at 15 and 25 keV, in primary electrons fascicle, on samples covered with a thin silver layer.
Isolation, Culture and Characterization of Human Bone Marrow Mesenchymal Stem Cells (MSCs)
Mesenchymal stem cells were isolated using Sirbu-Boeti
method [38] slightly modified. Briefly, MCSs were obtained by
centrifugation of bone marrow aspirate in Biocoll (Biochrom, density 1.077 g/mL). The cells from inner (containing mononuclear
127
Fig. (2). SEM micrographs of COLL/DEX/SiO 2 (CL): a,b; and (NCL): c,d;
inside the composite, and they could help to form a high-watercontent wound dressing. On the other hand, the COLL/DEX/SiO 2
(CL) showed morphology of continuous polymer matrix compared
to COLL/DEX/SiO2 (NCL) composite which displayed an
unconsolidated and fragile pattern. The reason might be that collagen fibers could enhance the formability of the composite. Some
lamentous fibers even could be seen in the SEM micrographs.
There were large numbers of SiO2 microcrystals on the wound
dressing surface.
Grumezescu et al.
Fig. (3). SEM micrographs of COLL/DEAC/SiO 2 (CL): a,b; and (NCL): c,d.
(a)
(c)
(b)
(d)
Fig. (4). Biocompatibility evaluation of COLL/DEAEC/SiO 2 (a,b) and COLL/DEX/SiO 2 (c,d) with mesenchymal stem cell (100X: a,c; 200X: b,d)
129
Fig. (5). The aspect of bacterial growth inhibition zones after the removal of the obtained specimens (T0): (a) Coll/DEX/SiO2 /UA (CL); (b)
Coll/DEX/SiO 2/UA (NCL); (c) UA; (d) Coll/DEAEC/SiO2 /UA (NCL); (e) Coll/DEAEC/SiO2 /UA (NCL).
Fig. (6). The aspect of bacterial growth inhibition zones after the removal of
the obtained specimens (T1): (a) Coll/DEX/SiO2/UA (CL); (b)
Coll/DEX/SiO 2/UA (NCL); (c) UA; (d) Coll/DEAEC/SiO 2/UA (NCL); (e)
Coll/DEAEC/SiO2 /UA (NCL);
Grumezescu et al.
Fig. (7). The aspect of bacterial growth inhibition zones after the removal of the obtained specimens (T2): (a) Coll/DEX/SiO2 /UA (CL); (b)
Coll/DEX/SiO 2/UA (NCL); (c) UA; (d) Coll/DEAEC/SiO2 /UA (NCL); (e) Coll/DEAEC/SiO2 /UA (NCL).
CONCLUSION
Preparation and characterization of COLL/DEX/SiO2/UA and
COLL/DEAEC/SiO2/UA wound dressing including the morphology
and their in vitro biological efficacy are reported. The FT-IR, SEM,
interaction with mesenchymal stem cell and a novel protocol to
evaluate wound dressing, firstly reported in literature were used for
the characterization of fabricated wound dressings. The results
demonstrated that the newly obtained materials are exhibiting structural and functional properties (bacterial killing, biofilm inhibition
and disruption, lack of cytotoxicity) that recommend them for further and safe applications in the biomedical field, as efficient
wound dressing systems.
CONFLICT OF INTEREST
The author(s) confirm that this article content has no conflicts
of interest.
ACKNOWLEDGEMENT
The results presented in this work were supported by the Human Resources 135/2010 grant (Contract no. 76/29.07.2010).
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