545 Modern Skincare
years, phages have been investigated as an antibiotic alternative for a variety of different
applications across the globe, prompting careful consideration of how to expand access
to their use. This expansion has continued into the skin care industry, where there is a
desire to leverage a new appreciation of the skin microbiome into thoughtful microbiota
modulatory active ingredients.
The studies described herein demonstrate the efficacy of this approach in mitigating acne
vulgaris via topical application of a cocktail of phage targeting C. acnes. The phage cocktail
performed exceptionally at the in vitro, ex vivo, and in vivo level with an excellent safety
profile. Examinations in the laboratory saw rapid clearance of bacteria in both planktonic
and biofilm conditions. Furthermore, the diminishment of the bacterial levels was rapid,
occurring within a 24 hour timeframe. In three-dimensional tissue modeling blemished
skin, the phage succeeded in reducing bacterial levels, a key inflammatory molecule, and once
again showed no associated tissue toxicity. Finally, a limited pilot clinical study produced
evidence of improved skin conditions within 7 days, where sebum was reduced, the cognate
bacteria were diminished, and no significant fluctuation in neighboring bacterial groups
was observed. Like all the prior experiments, there was no reported adverse reactions. This
latter point has been emphasized as topically deploying bacteriophages (literally viruses that
consume bacteria) as a skincare microbiome modulatory ingredient is likely to be heavily
scrutinized for its safety profile. Their relative safety is unsurprising to microbiologists as
phages are the largest constituent of the planetary biosphere. Indeed, at any given time, there
are approximately 1031 phage particles on the planet, and as such, phages are quite literally
part of every environmental niche.64 And with the notion that phages are everywhere, they
are also naturally found on the skin. So, when phages are introduced to the skin as a bio-
active topical ingredient, it is not a foreign material to the tissue.
One particular aspect of the findings from these studies that deserves to be highlighted is the
impact that the phage cocktail had against C. acnes biofilms (Figure 4). Biofilms represent
the closest that prokaryotic microorganisms come to forming complex multicellular tissues
observed in higher eukaryotic organisms. Existing in a biofilm relative to a unicellular
planktonic state offers a variety advantages to such microbes, particularly a demonstrable
resistance to various antimicrobial measures.47 C. acnes has been established as a biofilm-
forming bacterial species, and its capacity to form these biological structures has been
suggested as a mechanism to circumvent topical antibiotics and as means to insulate the
anaerobic microbe from the oxygen rich environment outside the pilosebaceous unit where
it takes up residence in the skin.7,47–50 As such, it is highly fortunate that in the context of
multiple biofilm-forming pathogenic bacteria, phages (particularly a cocktail of multiple
distinct phages) have reproducibly demonstrated biofilm inhibition and disruption
efficacy.51–53 Consistent with those trends, the C. acnes phage cocktail utilized here showed
a dose-dependent reduction in C. acnes biofilms (Figure 4).
As the skin care industry considers utilizing this technological platform based on the
naturally occurring predators of bacteria, there are two different approaches that they can
embrace. The first involves the creation of a standardized cocktail of phages that has as broad
coverage as possible of the different strains of its cognate bacterial species—a “one-size fits
all” model. The alternative requires considerably greater investment in infrastructure and
time. This involves the development and delivery of “personalized” phage cocktails. For the
latter approach, an individual would submit a swab sample of the region of their skin for
which they would like to receive a phage-infused formulation. The sample would have the
strain diversity of the intended target bacteria in that skin region determined by genomic
546 JOURNAL OF COSMETIC SCIENCE
sequencing. The phage provider would subsequently compile a cocktail of phage that offers
the best coverage against the bacterial strains identified at that skin location the individual
wants to target for bacterial reduction through phage lytic action. The former option is
simpler and requires less of an orchestrated framework but runs the risk that the phages
in the cocktail used for the topical formulation do not provide the best coverage for the
strains of the target bacteria that are present on every potential user of the material. The
latter option provides greater specificity for each end user, but certainly necessitates costlier
initial and ongoing investment.
CONCLUSIONS
In conclusion, these findings have demonstrated the successful adaptation of a century-
old technology to modern skin microbiome modulatory efforts. In contrast to current
microbiome interventions and topical antibiotic use, phage provide a measure of precision
with their species-specificity not afforded by alternative approaches. This allows for finished
formulations infused with phage cocktails to assert “microbiome selectivity.”
The evidence presented here reinforces the above statements with a phage-based blemish
solution investigated from the laboratory setting through to pilot clinical study. The C.
acnes phage cocktail exhibited lytic activity against its cognate bacteria in both planktonic
and biofilm settings. This was further extended to a three-dimensional skin model of
blemish-prone skin, where the formulas containing the phage cocktail countered C. acnes
growth and mitigated the levels of a pro-inflammatory factor. In a pilot clinical study,
efficacy was observed for diminishing sebum and C. acnes levels on the skin, which did not
appear to affect any neighboring bacterial groups.
This adaptation of a more than century old technology can be utilized to selectively
diminish other bacterial species on the skin that have roles in various skin conditions.
Given the desire to maintain a delicate balance in the skin microbiome, an approach that
allows for targeted precision modulation is highly advantageous.
MATERIALS AND METHODS
BACTERIA AND BACTERIOPHAGES
Cutibacterium acnes (29399) purchased from American Type Culture Collection (ATCC®,
Manassas, VA, USA) was cultured for 48 hours in Luria broth (LB, Thermo Fisher Scientific,
Waltham, MA, USA) under anaerobic conditions at 37°C with slight agitation. Three
distinct phages targeted to C. acnes were isolated from natural sources via screening on
lawns of the target bacterial species looking for plaque forming units (PFUs) via standard
double-layer agar assay.
MAMMALIAN CELL CULTURE
Normal human keratinocytes (nHEK) were obtained from Thermo Fisher Scientific, normal
human dermal fibroblasts (NHDF) were obtained from ATCC®, and immortalized human
keratinocyte cell line (HaCaT) were obtained from AddexBio (San Diego, CA, USA).
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