Arch Dermatol Res
DOI 10.1007/s00403-015-1559-x
ORIGINAL PAPER
Biafine topical emulsion accelerates excisional and burn wound
healing in mice
Aimee E. Krausz1 • Brandon L. Adler1 • Angelo Landriscina1 • Jamie M. Rosen1
Tagai Musaev1 • Joshua D. Nosanchuk2 • Adam J. Friedman1,3
Received: 3 September 2014 / Revised: 28 February 2015 / Accepted: 10 March 2015
Ó Springer-Verlag Berlin Heidelberg 2015
Abstract Macrophages play a fundamental role in wound
healing; therefore, employing a strategy that enhances
macrophage recruitment would be ideal. It was previously
suggested that the mechanism by which BiafineÒ topical
emulsion improves wound healing is via enhanced mac-
rophage infiltration into the wound bed. The purpose of this
study was to confirm this observation through gross and
histologic assessments of wound healing using murine full-
thickness excisional and burn wound models, and compare
to common standards, Vaseline and silver sulfadiazine
(SSD). Full-thickness excisional and burn wounds were
created on two groups of 60 mice. In the excisional arm,
mice were divided into untreated control, Biafine, and
Vaseline groups. In the burn arm, mice were divided into
untreated control, Biafine, and SSD groups. Daily treat-
ments were administered and healing was measured over
time. Wound tissue was excised and stained to appropri-
ately visualize morphology, collagen, macrophages, and
neutrophils. Collagen deposition was measured and cell
counts were performed. Biafine enhanced wound healing in
murine full-thickness excisional and burn wounds com-
pared to control, and surpassed Vaseline and SSD in
A. E. Krausz and B. L. Adler contributed equally to this work.
& Adam J. Friedman
ajf0424@gmail.com
1
Division of Dermatology, Department of Medicine,
Montefiore Medical Center, Albert Einstein College of
Medicine, 111 E. 210th Street, Bronx, New York 10467,
USA
2
Department of Microbiology and Immunology, Albert
Einstein College of Medicine, Bronx, New York, USA
3
Department of Physiology and Biophysics, Albert Einstein
College of Medicine, Bronx, New York, USA
•
respective wound types. Biafine treatment accelerated
wound closure clinically, with greater epidermal/dermal
maturity, granulation tissue formation, and collagen quality
and arrangement compared to other groups histologically.
Biafine application was associated with greater macro-
phage and lower neutrophil infiltration at earlier stages of
healing when compared to other study groups. In conclu-
sion, Biafine can be considered an alternative topical
therapy for full-thickness excisional and burn wounds,
owing to its advantageous biologically based wound heal-
ing properties.
Keywords Biafine
Trolamine
Burn wound
Excisional
wound
Wound healing
Macrophage
Introduction
Wound healing is a complex process divided into three
discrete but overlapping phases: inflammation, prolif-
eration, and remodeling [18]. Numerous cell types, growth
factors, and cytokines must act in concert to enact orderly
wound repair. As the ‘‘orchestra leader’’ of wound healing,
the macrophage conducts and oversees essential events
during healing [13]. Macrophages arrive 48–96 h following
injury and serve as the primary cellular effectors of wound
healing as the proportion of neutrophils in the wound bed
wanes and the fibroblast population begins to rise. Mac-
rophages sterilize the wound through phagocytosis and
generation of reactive radicals, and secrete enzymes that
contribute to wound debridement [23]. Nitric oxide (NO)
generated by macrophages regulates collagen production,
cellular proliferation, and wound contraction [24], and kills
invading pathogens [23]. Cytokines and growth factors
secreted by macrophages recruit and activate other
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macrophages and fibroblasts for participation in healing,
and regulate such diverse processes as angiogenesis, col-
lagen synthesis and fibroplasia, and extracellular matrix
synthesis [23]. With this understanding of the fundamental
role macrophages play in wound healing, utilizing a ther-
apy that positively influences this ‘‘director of healing’’
would be ideal.
BiafineÒ (Valeant Pharmaceuticals North America LLC,
Bridgewater, NJ) is a trolamine-containing oil-in-water
topical emulsion approved by the FDA for use in a variety
of wound healing settings, from minor abrasions to full
thickness wounds, as well as first- and second-degree burns
and radiation dermatitis [12]. In a human suction blister
model, Biafine was previously shown to promote wound
healing by enhancing macrophage recruitment and in-
creasing the ratio of interleukin (IL)-1 to IL-6, as deter-
mined using wound exudates collected 24 h post-wounding
[7]. IL-1 stimulates fibroblast growth and influences ex-
tracellular matrix composition via collagen production and
collagenase activation, while IL-6 accelerates epidermal
growth and inhibits fibroblast proliferation. By inducing
IL-1 release and reducing IL-6 secretion, it was believed
that Biafine contributed positively to granulation tissue
formation by fibroblasts [7].
Biafine was previously evaluated in several clinical
scenarios, including enhancement of wound healing fol-
lowing shave biopsy [11] and prevention of radiation
dermatitis [22]. However, the emphasis in these studies
was on clinical endpoints as opposed to elucidating the
mechanism of healing. To investigate the impact of Bi-
afine in the wound environment, murine full-thickness
excisional and thermal burn wound models were utilized
to compare treatment with Biafine to commonly used
standards, Vaseline and silver sulfadiazine (SSD) respec-
tively, for each type of injury. To further elucidate the
mechanism of and role for this device in wound care,
immunohistochemistry was utilized to evaluate temporal
cell infiltration.
Materials and methods
Excisional wound model
All animal experimentation procedures were approved by
the Institutional Animal Care and Use Committee at Albert
Einstein College of Medicine (Bronx, New York). For the
excisional wound experiment, 30 female Balb/c mice
(6–8 weeks; National Cancer Institute, Frederick, MD)
were divided into three treatment groups (control, Vase-
line, and Biafine) comprising 10 animals each. On day 0,
the dorsal hair of each mouse was shaved and the skin was
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Arch Dermatol Res
disinfected using ethanol. Punch biopsies (4 mm Punch
Biopsy, Miltex, York, PA) were utilized to create 4 equally
sized 4-mm diameter full-thickness excisional wounds on
the back of each mouse, as previously described [5, 6].
One significant challenge of using mice to evaluate
wound repair is that in these animals wound contraction
originates outside of tissue, whereas in humans, re-ep-
ithelialization and granulation tissue formation occur
within the wound space. Parallel wounds exert traction on
each other, limiting this extracutaneous wound healing, and
allowing wound healing to occur through the processes of
granulation and re-epithelialization. In brief, the back skin
was lifted, compressed on a sterile piece of cardboard, and
a 4-mm sterile surgical-grade punch biopsy was used to
create full-thickness wounds extending through the pan-
niculus. Daily application of 10 ll of treatment with
Vaseline or Biafine began on the day of wounding. Control
mice were not treated. The experiment described was re-
peated to increase the power of the study (n = 20 animals/
80 wounds per group total).
Burn wound model
Thirty female Balb/c mice were divided into three treat-
ment groups (control, SSD, and Biafine) comprising 10
animals each. The burn model employed has been previ-
ously described [15, 17]. In short, on day 0, the dorsal hair
of each mouse was shaved and the skin was disinfected
using ethanol. A calibrated 160 °C heated bar was applied
to the back of each mouse for 10 s to create 2 full-thickness
5-mm diameter burns per mouse. Daily application of 10 ll
of treatment with SSD or Biafine began on the day of
wounding. Control mice were not treated. As mentioned
above, the experiment was repeated once to increase the
power of the study (n = 20 animals/40 wounds per group
total).
Gross assessment of wounds
Daily photographs were taken in order to visually track
wound healing progress. Using ImageJ software (Na-
tional Institutes of Health, Bethesda, MD), the daily
change in wound area relative to initial wound area was
calculated.
Histologic and immunohistochemical assessment
of wounds
On days 3 and 7 following excisional wounding and days
7 and 16 following burn wounding, wounds were excised.
Tissue was fixed in 10 % formalin for at least 24 h,
processed, and embedded in paraffin. Four-micron vertical
Arch Dermatol Res

Fig. 1 Biafine accelerates full-

thickness excisional wound
closure. a Representative
images of excisional wound
healing from days 0–10. Topical
administration of Biafine
decreased wound size and
qualitatively enhanced healing
compared to untreated control
and Vaseline groups. Scale bar
4 mm. b Wound size analysis
(relative area versus initial area)
revealed statistically significant
acceleration of excisional
wound healing in mice treated
with Biafine compared to
untreated control and Vaseline.
Time points are averages of 10
measurements. Statistical
analysis conducted using 2-way
ANOVA. Error bars denote
SEM

sections were fixed to glass slides and stained with he-
matoxylin and eosin (H&E), Masson’s trichrome, Iba-1,
and myeloperoxidase (MPO) to visualize morphology,
collagen deposition, macrophages, and neutrophils, re-
spectively. The slides were inspected using light mi-
croscopy, and images were captured digitally without
further processing. In order to blind the histologic inter-
pretation, slides were numbered without indication of
cohort. Collagen deposition was measured by intensity
using ImageJ and cell counts for Iba-1 and MPO were
performed. Ten high-power fields (HPFs; 409) were
evaluated per section.
Statistical analysis
All data were subjected to statistical analysis using
GraphPad Prism 6.4 (GraphPad Software, La Jolla, CA).
p values of \0.05 were considered significant.
Results
The effect of Biafine, Vaseline, and no treatment on exci-
sional wound healing in mice was evaluated. Topical ap-
plication of Biafine onto the wound bed significantly
decreased wound size compared to other groups (Fig. 1).
At day 2, control wounds and Biafine-treated wounds had
closed 1.99 and 2.5 %, respectively, compared to day 0,
while there was an 11.0 % increase in the size of Vaseline-
treated wounds. At day 6, treatment with Biafine resulted in
60.1 % closure relative to day 0, compared to 37.4 %
closure in Vaseline-treated wounds and 27.0 % closure in
control wounds. At day 10, there was near complete clo-
sure of Biafine-treated wounds (94.6 %), compared to
89.6 % closure in Vaseline-treated wounds and 85.3 %
closure in control wounds.
Histologic evaluation of excisional wound sections mir-
rored the clinical progression of the wounds in each treatment
group. Sections from day 3 of untreated controls demonstrated
a serous, neutrophil-rich hemorrhagic crust overlying imma-
ture inflammatory granulation tissue and poorly re-epithe-
lialized epidermis (Fig. 2a), which was found to be complete
though irregular by day 7 along with persistent inflammatory
infiltrate in the dermis (Fig. 2d). Similar findings were ap-
preciated in the Vaseline-treated wounds on both days 3 and 7
(Fig. 2b, e). Biafine-treated excisional wounds on day 3 were
histologically more advanced, demonstrating epidermal and
dermal maturation similar to that seen on day 7 in the other
cohorts (Fig. 2c). By day 7, dermal inflammation was mini-
mized, allowing for better visualization of freshly deposited

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Fig. 2 Biafine diminishes inflammation and enhances maturation in
excisional wounds. Histologic evaluation of untreated control exci-
sional wound sections from day 3 revealed serous, neutrophil-rich
hemorrhagic crust overlying immature inflammatory granulation
tissue and poorly re-epithelialized epidermis, found to be complete
though irregular by day 7 along with persistent dermal inflammatory
infiltrate. Vaseline-treated wounds displayed similar findings on both
collagen, and wound contraction was noted as compared to the
other groups (Fig. 2f).
Using Masson’s trichrome staining, control and Vase-
line-treated wounds displayed minimal formation of new
collagen on day 3 (Fig. 3). There was more collagen for-
mation in the Vaseline-treated group as compared to con-
trol (Fig. 3g), with necrosis (red) being the predominant
feature on examination in both groups (Fig. 3a, b). By day
7, pale blue staining was seen throughout the dermis in
both groups (Fig. 3d, e), indicating early deposition of new
collagen, once again more so in the Vaseline-treated group
based on staining intensity (Fig. 3g). In contrast, Biafine-
treated wounds evinced a similar pale blue staining pattern,
but on day 3 (Fig. 3c), with brighter blue, confluent bun-
dles in the dermis by day 7 (Fig. 3f). Masson’s trichrome
staining revealed increased collagen intensity (in arbitrary
units, A.U.) in Biafine wounds compared to other groups
on both day 3 (p B 0.01) and day 7 (p B 0.001), with a
larger margin of increase on day 7 (Fig. 3g).
Immunohistochemical staining on day 3 revealed fewer
neutrophils in Biafine-treated wound tissue compared to
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Arch Dermatol Res
days 3 and 7. On day 3, Biafine-treated excisional wounds were
histologically more advanced, with epidermal and dermal maturation
similar to that seen on day 7 in the other groups. By day 7, dermal
inflammation was minimal, permitting easier visualization of newly
deposited collagen, and wound contraction was noted in comparison
to the other cohorts. Scale bar 100 lm
control and Vaseline-treated tissue based on high-power
field cell counts (p B 0.0001; Fig. 4). Vaseline-treated
tissue samples contained fewer neutrophils than control
(p B 0.01), but more than Biafine (p B 0.0001, Fig. 4d).
Immunostaining for macrophages on day 3 revealed a
significant difference in dermal macrophage infiltration in
Biafine-treated tissue compared to control and Vaseline-
treated samples (p B 0.0001, Fig. 5). On day 7, the number
of macrophages decreased to below that of other groups
(p B 0.0001). Vaseline-treated tissue contained quantities
of macrophages intermediate between control and Biafine
groups on both days (p B 0.0001).
The effect of Biafine, SSD, and no treatment on burn
wound healing in mice was evaluated. Topical applica-
tion of Biafine onto the wound bed significantly de-
creased eschar size compared to other treatment groups
(Fig. 6). In the days immediately following burn injury,
eschar expansion occurred in control and SSD groups,
corresponding to progressive inflammation, edema, and
tissue necrosis. Notably, Biafine attenuated this phe-
nomenon of expansion, and at day 3 Biafine-treated
Arch Dermatol Res
Fig. 3 Biafine enhances collagen deposition in excisional wounds.
Visualized using Masson’s trichrome staining, control and Vaseline-
treated wounds demonstrated minimal new collagen deposition on
day 3. There was greater collagen formation in the Vaseline-treated
group as compared to control, and necrosis (red) was the predominant
feature on examination in both groups. By day 7, pale blue staining
was evident throughout the dermis in both groups, signaling early
deposition of new collagen, once again more so in the Vaseline-
treated group based on staining intensity (g right graph). In contrast,
wounds measured 94.5 % compared to day 0, in contrast
to enlarged burns measuring 123.9 % in control wounds
and 155.9 % in SSD wounds. At day 15, treatment with
Biafine resulted in 95.7 % closure relative to day 0,
compared to 67.4 % closure in SSD-treated wounds and
61.1 % closure in control wounds.
Morphologic inspection of burn wound sections from
day 7 revealed absence of epidermal re-epithelialization in
all cohorts, however the maturing dermis/granulation tissue
Biafine-treated wounds showed a similar pale blue staining pattern,
but on day 3, with brighter blue, confluent bundles in the dermis by
day 7. Masson’s trichrome staining revealed increased collagen
intensity (in arbitrary units, A.U.) in Biafine wounds compared to
other groups on both day 3 (g left graph) and day 7 (g right graph),
with a larger margin of increase on day 7. Statistical analysis
conducted using Student’s t test. Error bars denote SEM. Left graph
*p = 0.006, **p = 0.008, ***p = 0.014. Right graph *p B 0.001.
Scale bar 100 lm
in the Biafine-treated group was more organized and de-
veloped than untreated control and SSD (Fig. 7). Interest-
ingly, the SSD-treated burns revealed minimal dermal
reconstruction, but rather a pink, homogenized collection
of thermally damaged structures with less repair than the
untreated control group (Fig. 7b). By day 16, Biafine-
treated burns possessed a fully re-epithelialized epidermis
and minimally inflammatory dermis with evidence of new,
parallel-oriented collagen deposition (Fig. 7f), as compared
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Fig. 4 Biafine decreases neutrophil infiltration in the excisional
wound bed. Immunohistochemical staining on day 3 revealed fewer
neutrophils in Biafine-treated wound tissue compared to control and
Vaseline-treated tissue based on high-power field cell counts.
to SSD and untreated, both of which demonstrated a per-
sistent crust overlying an incomplete epidermis (Fig. 7d,
e).
Examination of collagen deposition showed that control
and SSD-treated wounds displayed minimal collagen
staining on day 7, though there was significantly more
within the SSD control wound bed as compared to un-
treated control (Fig. 8a, b). On day 16, while an increase in
collagen staining was noted, untreated control demon-
strated a greater intensity than SSD, though the stained
collagen in both was haphazard in it orientation (Fig. 8d,
e). On the other hand, Biafine-treated wounds displayed
increased intensity of collagen in the wound bed compared
to other groups on day 7 (p B 0.0001), likely representa-
tive of previous and newly deposited collagen (Fig. 8c). On
day 16, parallel-oriented brightly blue staining collagen
was appreciated (Fig. 8f), with statistically significant in-
creased intensity as compared to controls (p B 0.0001,
Fig. 8g).
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Arch Dermatol Res
Vaseline-treated tissue samples contained fewer neutrophils than
control, but more than Biafine. Statistical analysis conducted using
Student’s t test. Error bars denote SEM. *p \ 0.0001, **p = 0.008.
Scale bar 100 lm
Immunohistochemical staining on day 7 revealed fewer
neutrophils in Biafine-treated wound tissue compared to
control or SSD tissue (p B 0.0001; Fig. 9). SSD-treated
tissue contained the highest number of neutrophils, above
control levels (p B 0.0001). There were more macrophages
in Biafine-treated tissue compared to other groups
(p B 0.0001; Fig. 10), with no difference in number of
macrophages between SSD and control (p [ 0.05).
Discussion
In this study, we observed that Biafine-treated excisional
and burn wounds demonstrated enhanced healing clinically
and histologically. Biafine-treated wounds displayed ac-
celerated closure compared to control wounds as well as
commonly used standard therapies (Vaseline and SSD,
respectively). There was greater augmentation of healing
by Biafine in burn wounds versus excisional wounds, but in
Arch Dermatol Res
Fig. 5 Biafine enhances macrophage infiltration in the excisional
wound bed. Immunostaining for macrophages on day 3 revealed a
significant difference in dermal macrophage infiltration in Biafine-
treated tissue compared to control and Vaseline-treated groups. On
day 7, macrophages in Biafine tissue decreased to below other groups.
both experimental arms healing was statistically significant
compared to other groups. We discovered positive histo-
logic differences in the maturity of the epidermis and
dermis between Biafine and other groups. Biafine enhanced
the quality and arrangement of newly deposited collagen.
Consistent with previous findings, Biafine treatment was
associated with a decreased proportion of neutrophils and
increased number of macrophages as assessed by im-
munostaining [7].
As wound healing’s first responders, neutrophils arrive
at the wound site nearly immediately, their numbers
peaking at 24 h. Trafficking of neutrophils involves
Vaseline-treated tissue contained quantities of macrophages interme-
diate between control and Biafine cohorts on both days. Statistical
analysis conducted using Student’s t test. Error bars denote SEM.
*p \ 0.0001. Scale bar 100 lm
attachment to selectin receptors on endothelial surfaces,
followed by diapedesis (movement through the vessel wall)
and subsequent adherence to integrin extracellular matrix
receptors [3]. Neutrophils function mainly to phagocytose
invading pathogens and debride the wound bed during the
inflammatory phase of healing, and disappear following a
combination of cell signaling, apoptosis, and phagocytosis
by macrophages [3, 23]. Nevertheless, neutrophils are not
required for successful wound healing, and wounds devoid
of neutrophils do not display decreased rates of wound
debridement or extent of repair compared to normal
wounds [20].
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 Arch Dermatol Res

Fig. 6 Biafine accelerates full-

thickness thermal burn wound
closure. a Representative
images of burn wound healing
from days 0–14. Topical
administration of Biafine
decreased wound size and
qualitatively enhanced healing
compared to untreated control
and silver sulfadiazine groups.
Scale bar 4 mm. b Wound size
analysis (relative area versus
initial area) revealed
statistically significant
acceleration of burn wound
healing in mice treated with
Biafine compared to untreated
control and silver sulfadiazine.
Time points are averages of ten
measurements. Statistical
analysis conducted using 2-way
ANOVA. Error bars denote
SEM

We found that Biafine shifted wound healing toward a
more macrophage-dominant process earlier than what was
observed with the other experimental groups. We believe
this resulted from direct enhancement of macrophage re-
cruitment, as previously suggested [7]. A crucial role of
macrophages during the wound healing process is gen-
eration of nitric oxide. NO is a short-lived free radical
molecule that serves diverse physiologic functions, in-
cluding regulation of wound healing. In macrophages and
other cells, NO is produced from L-arginine when in-
ducible nitric oxide synthase is upregulated in the pres-
ence of cytokines and microbial products [3]. NO’s many
wound healing functions include antimicrobial activity,
vasodilation, neovascularization, collagen deposition, and
wound contraction [3, 24]. Given these findings, many
NO-releasing technologies have been piloted for wound
healing [4], with findings of improved healing accompa-
nied by increased fibroblast migration, collagen deposi-
tion, and angiogenesis [9]. On the cellular level, NO was
previously shown to inhibit neutrophil transmigration,
rolling, and adhesion via effects on cellular adhesion
molecules such as selectins [8–10, 14]. Therefore, in ad-
dition to direct effects on macrophage chemotaxis, Biafine
likely indirectly inhibited neutrophil accumulation
through the effects of heightened macrophage-generated
NO. The greater number of macrophages may also have
phagocytosed apoptotic and senescent neutrophils more
rapidly than expected.

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Arch Dermatol Res
Fig. 7 Biafine diminishes inflammation and enhances maturation in
burn wounds. Morphologic examination of burn wound sections from
day 7 revealed absence of epidermal re-epithelialization in all
experimental groups. Nonetheless, the maturing dermis/granulation
tissue in Biafine-treated wounds was more organized and developed
than untreated control and silver sulfadiazine (SSD). SSD-treated
burns revealed minimal dermal reconstruction, with a pink,
There are numerous benefits to decreasing the neutrophil
population and converting wound healing to a more mac-
rophage-centric process earlier in the inflammatory phase
of wound healing. In chronic wounds, ongoing inflamma-
tion leads to persistence of neutrophils that secrete colla-
gen-degrading matrix metalloproteinases in excess of the
collagen-producing capability of fibroblasts, thereby de-
laying wound healing [16]. The neutrophilic dermatoses
represent an extreme example of this concept, embracing
conditions such as pyoderma gangrenosum, which features
a diffuse dermal neutrophilic infiltrate and clinically
manifests as a progressive, non-healing suppurative ulcer
[21]. Chronic wounds represent a tremendous economic
and management burden for the US healthcare system [19].
Reduction in neutrophils attenuates the early inflammatory
phase of wound healing, shifting instead toward prolif-
eration and remodeling accomplished by macrophages and
fibroblasts. When induced to enter the wound site earlier in
homogenized collection of thermally damaged structures with less
repair than untreated control wounds. By day 16, Biafine-treated
burns demonstrated a fully re-epithelialized epidermis and minimally
inflammatory dermis with evidence of new, parallel-oriented collagen
deposition, as compared to SSD and control, both of which showed a
persistent crust overlying an incomplete epidermis. Scale bar 100 lm
healing, macrophages can initiate prompter angiogenesis,
fibroblast migration, and collagen synthesis, leading to
earlier re-epithelialization and more rapid wound resolu-
tion [23]. This modulation of the wound healing milieu
could benefit not only patients, but providers, the health-
care system, and the economy overall.
Interestingly, we found that use of SSD enhanced
inflammatory wound expansion soon after burn wound-
ing, and did not significantly accelerate wound closure
compared to control wounds. Although SSD is the gold
standard for topical burn wound therapy and antimicro-
bial prophylaxis, systematic reviews have discovered
associations with delayed wound healing as well as in-
creased incidence of infection [1, 2]. In the excisional
wound model employed in this study, Vaseline treatment
did not enhance wound closure time compared to con-
trol, but did significantly increase collagen deposition
and macrophage count, and decrease the number of
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Fig. 8 Biafine enhances collagen deposition in burn wounds.
Inspection of collagen deposition revealed that control and silver
sulfadiazine (SSD)-treated wounds displayed minimal collagen
staining on day 7 (g left graph). On day 16, untreated control
collagen had a greater intensity than SSD (g right graph), though the
stained collagen in both was haphazard in orientation. In contrast,
neutrophils, compared to control. However, the magni-
tude of these changes was significantly less than that
observed with Biafine treatment.
In conclusion, Biafine enhanced wound healing in
murine full-thickness excisional and burn wounds com-
pared to control wounds, and outperformed Vaseline and
silver sulfadiazine in the respective wound types.
Clinically, Biafine-treated wounds displayed enhanced
reepithelialization and accelerated closure. Histologically,
Biafine-treated wounds demonstrated greater epider-
mal/dermal maturity, granulation tissue formation, and
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Arch Dermatol Res
Biafine-treated wounds demonstrated increased intensity of collagen
in the wound bed compared to other groups on day 7. On day 16,
parallel-oriented brightly blue staining collagen was appreciated, with
statistically significant increased intensity as compared to controls.
Statistical analysis conducted using Student’s t test. Error bars denote
SEM. *p B 0.001, **p = 0.072. Scale bar 100 lm
collagen quantity and quality. Biafine treatment was asso-
ciated with earlier and greater macrophage infiltration and
lower neutrophil presence at earlier stages of healing, with
major implications for both acute and chronic wound
healing. Although larger clinical trials would help fully
characterize the healing efficacy of this trolamine-con-
taining oil-in-water topical emulsion, the present study
gives a biological basis for the use of Biafine as an alter-
native topical therapy for full-thickness excisional and burn
wounds, owing to its advantageous wound healing
properties.
Arch Dermatol Res

Fig. 9 Biafine decreases neutrophil infiltration in the burn wound bed. (SSD) tissue. SSD-treated tissue contained the highest number of
Immunohistochemical staining on day 7 revealed fewer neutrophils in neutrophils, above control levels. Statistical analysis conducted using
Biafine-treated wound tissue compared to control or silver sulfadiazine Student’s t test. Error bars denote SEM. *p \ 0.0001. Scale bar 100 lm

Fig. 10 Biafine enhances macrophage infiltration in the burn wound no difference in number of macrophages between silver sulfadiazine
bed. Immunohistochemical staining on day 7 revealed a greater number and control. Statistical analysis conducted using Student’s t test. Error
of macrophages in Biafine-treated tissue compared to other groups, with bars denote SEM. *p \ 0.0001, **p = 0.598. Scale bar 100 lm

123

Acknowledgments This study was funded by Valeant Pharmaceu-
ticals North America LLC.
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