REVIEW
Connective Tissue and Fibroblast
Senescence in Skin Aging
Meinhard Wlaschek1, Pallab Maity1, Evgenia Makrantonaki1,2 and Karin Scharffetter-Kochanek1
There is increasing evidence that skin aging is signif-
icantly enforced by the accumulation of senescent
dermal fibroblasts. Various stressors damaging mac-
romolecules inside and outside fibroblasts are
responsible. In addition, NK cells fail to adequately
remove senescent (SEN) fibroblasts from tissues. SEN
fibroblasts by the release of the proinflammatory,
tissue degrading senescent-associated secretory
phenotype factors and vesicles with distinct cargo
impact on their endogenous niche and spread
senescence and skin aging. In this review, we will
further discuss less noticed facets, including the
plasticity of distinct dermal fibroblast phenotypes, the
underestimated impact of the extracellular matrix it-
self, and the depletion of fibroblast subsets on skin
homeostasis and aging.
Journal of Investigative Dermatology (2021) 141, 985e992; doi:10.1016/
j.jid.2020.11.010
Introduction
Dermal connective tissue connects all histogenetically
distinct cells and tissues of the skin to a functional organ.
Fibroblasts constitute the principal component of the con-
nective tissue. As such, fibroblasts occur—apart from the
skin—in virtually every tissue and organ of the body. The
capacity of fibroblasts to synthesize and organize the extra-
cellular matrix (ECM) and communicate with the adjacent
stem cell (SC) niche and quiescent, mitotic, and postmitotic
cells and tissues of distinct origin makes them, so far, an
underestimated central component in skin homeostasis and
aging. In this review, we will summarize the advanced un-
derstanding of the role of cellular senescence for stromal
aging in general as well as for skin aging in particular.
Wherever possible, we will differentiate between data that
are relevant for aging in general and stromal aging and—most
importantly—the ones that have truly been shown to play a
role in murine and human skin aging. This review
1
Department of Dermatology and Allergic Diseases, Ulm University, Ulm,
Germany; and 2Dermatology Center Wildeshausen, Wildeshausen,
Germany
Correspondence: Karin Scharffetter-Kochanek, Department of Dermatology
and Allergic Diseases, Ulm University, Albert-Einstein-Allee 23, 89081 Ulm,
Germany. E-mail: karin.scharffetter-kochanek@uniklinik-ulm.de
Abbreviations: AP-1, activator protein-1; CDK, cell cycle kinase; DDR, DNA
damage response; ECM, extracellular matrix; MMP, matrix metalloproteinase;
MSC, mesenchymal stem cell; scRNA-seq, single cell RNA sequencing; SASP,
senescence-associated secretory phenotype; SEN, senescent
Received 27 August 2020; revised 28 October 2020; accepted 11 November
2020; corrected proof published online 7 February 2021
ª 2021 The Authors. Published by Elsevier, Inc. on behalf of the Society for Investigative Dermatology. This is an open access
article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
furthermore intends to provide insight into state-of-the-art
technology, which has undoubtedly advanced our under-
standing of fibroblast senescence driving skin and other organ
aging with the unique opportunity to develop and refine
modern preventive and therapeutic strategies.
Senescence of fibroblasts in skin aging
The role of senescence in skin aging. An emerging
hypothesis of skin aging postulates that mainly fibroblast
senescence drives skin decline and skin aging due to irre-
versible proliferation arrest and enhanced release of a
senescence-associated secretory phenotype (SASP). SASP,
through chemokines and proinflammatory factors, induces
chronic inflammation, reduces proliferation by impaired
release of essential GFs, and enhances the degradation of the
ECM (connective tissue) by enhanced activation of proteo-
lytic enzymes, including matrix-degrading metal-
loproteinases. Cellular senescence transiently occurs in
several physiological conditions during embryogenesis
(Muñoz-Espı́n and Serrano, 2014) and wound healing
(Demaria et al., 2014) shaping and regenerating organs. By
contrast, fibroblast senescence is mainly a persisting perma-
nent state and, in consequence, drives skin aging, aging in
general and aging-related diseases (Campisi et al., 2019,
2005; Demaria et al., 2015; Toutfaire et al., 2017; Treiber
et al., 2011; Velarde and Demaria, 2016; Wang and
Dreesen, 2018). Given that the fibroblasts confined to the
connective tissue stabilize every organ parenchyma, cellular
senescence of fibroblasts may play a more general role in
organ aging, in organismal aging, and likely in many aging-
related diseases (Figure 1). In fact, senescent (SEN) fibro-
blasts are increased in human skin, in the dermis (Dimri
et al., 1995; Ressler et al., 2006), and in many other organs
(Krishnamurthy et al., 2004; Tuttle et al., 2020). The most
important evidence for the role of SEN fibroblasts in enforc-
ing skin aging is summarized in Table 1. Permanent senes-
cence can be induced in quiescent (nonreplicating)
fibroblasts in the skin by cell-autonomous and exogenous
stressors leading to the shortening of the chromosomal
termini, termed telomeres (Bodnar et al., 1998; Harley et al.,
1990; Yu et al., 1990), mitochondrial dysfunction (Berneburg
et al., 1997; Birch et al., 2018; Birch-Machin et al., 1998;
Passos et al., 2007), DNA damage, and subsequent activation
of DNA damage response (DDR) signaling pathways leading
to permanent or transient cell cycle arrest (d’Adda di
Fagagna, 2008; Hoeijmakers, 2009) and metabolic reprog-
ramming (da Silva and Schumacher, 2019; Figure 2). In this
regard, also the accumulation of oxidized proteins due to the
loss of the proteasomal capacity to remove oxidized and
misfolded protein activates DDR pathways and thus leads to
the induction of cellular senescence (Chondrogianni and
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Fibroblast Senescence and Skin Aging
Figure 1. SEN fibroblasts drive tissue,
organ, and organismal aging. SEN
fibroblasts imprint their environment
with a proinflammatory SASP, soluble
factors released upon the installment
of the senescence program, thereby
affecting distinct tissue in the skin and
various organs, here apart from the
skin, bone, and muscle among others.
This drives organ aging, aging-related
diseases, and likely organismal aging.
This hypothesis needs to be further
proven because it bears many
promising therapeutic and preventive
opportunities. This review gathers
further evidence. SASP, senescence-
associated secretory phenotype; SEN,
senescent.
Gonos, 2010) and to the release of soluble senescence-
spreading factors (Catalgol and Grune, 2009; Eckhart et al.,
2019). Apart from many other stressors (summarized in
Campisi [2005]; Gorgoulis et al. [2019]), mitochondrial
dysfunction in fibroblasts with increasing concentrations of
ROS (Gu et al., 2020; Rinnerthaler et al., 2015; Treiber et al.,
2011) either leads to DNA damage of fibroblasts in the skin
and other organs (for review see Demaria et al. [2015];
Treiber et al. [2011]) or enhances activator protein-1 (AP-1)‒
and NF-kB‒dependent signaling pathways among others
related to inflammation (Meyer et al., 2017). In fact, mito-
chondrial dysfunction is a major driver in both intrinsic and
extrinsic skin aging and possibly constitutes the common link
between both of them (Krutmann and Schroeder, 2009).
To further explore the link between ROS and fibroblast
senescence, we earlier generated a mouse model with a
fibroblast specific deficiency for SOD2. Hence, mitochon-
drial superoxide anions cannot be detoxified and, in conse-
quence, accumulate in mitochondria of fibroblasts. These
fibroblast-specific, SOD2-deficient mice develop a severely
accelerated skin aging phenotype with a marked reduction in
collagen and dermal thickness, lack of resilience, enhanced
DNA damage, and the induction and accumulation of SEN
fibroblasts in the skin, bone, and muscle leading to skin
atrophy, osteoporosis, and sarcopenia (Treiber et al., 2011).
These data show that mitochondrial dysfunction and
enhanced superoxide anion concentrations lead to fibroblast
senescence accelerating aging phenotype in connective tis-
sue‒rich organs and reduce the life span of fibroblast-specific
SOD2-deficient mice. Of note, the transcription factor Nrf2,
which is a key regulator of the antioxidant system if overex-
pressed in murine skin fibroblasts, induces skin aging
(Hiebert et al., 2018). Thus, an imbalance of ROS detoxifi-
cation enforces tissue decline and aging. The activation of the
specific signaling pathways promoting the overexpression of
cell cycle kinase (CDK) inhibitor proteins such as p16, p21,
986 Journal of Investigative Dermatology (2021), Volume 141
p15, or p27, which induce and maintain the state of senes-
cence, clearly depends on the specific triggers of senescence,
including ROS imbalance (for review, see Lozano-Torres
et al. [2019]). In case of their inhibition, CDKs cannot
phosphorylate Rb, which physiologically sequesters the
transcription factor E2F. E2F is a crucial protein enforcing the
expression of genes that are mandatory for cell cycle pro-
gression. Its inhibition leads to cellular senescence.
SASP and senescence spreading in skin aging. Clearly by
their nonproliferating state with increased proteolytic activity
and suppressed function to deposit components of the ECM,
long-term SEN fibroblasts enforce tissue decline and skin
aging. In addition, SEN fibroblasts have the unique property
not to die. They hardly can undergo apoptosis, cannot be
sufficiently removed by the adaptive immune system, and, in
consequence, hang around in the connective tissue for a long
time with enhanced release of a variety of proinflammatory,
matrix-degrading chemokines, cytokines, and proteases and
a concomitantly suppressed release of essential GFs, collec-
tively referred to as SASP (Ghosh and Capell, 2016). The
SASP released from SEN fibroblasts is a typical DDR and can
occur either transiently as in wound healing and develop-
ment or persistently as in aging (Campisi and d’Adda di
Fagagna, 2007; Rodier et al., 2009). The SASP differs
depending on the studied cell type, tissue, overall context,
and likely the senescence-inducing agent, although these
differences have not been systematically studied (Demaria
et al., 2014; Ghosh and Capell, 2016; Meyer et al., 2017;
Muñoz-Espı́n et al., 2013; Rodier et al., 2009). Recently,
Basisty et al. (2020) reported a SASP Atlas. This represents a
comprehensive proteomic database of soluble and exosomal
SASP factors, which are enforced by a variety of senescence
inducers in skin fibroblasts. Each profile consists of hundreds
of largely distinct proteins but also includes a subset of pro-
teins elevated in all SASPs (Basisty et al., 2020). SASP factors
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Fibroblast Senescence and Skin Aging

Table 1. Evidence for the Role of Fibroblast Senescence in Skin Aging

Experimental Evidence for a Role of Fibroblast Senescence in Skin Aging Reference

p16INK4a-positive SEN cells increase in the dermis of human skin and the number correlates with wrinkle formation and Ressler et al. (2006);
typical morphologic features of aging of the elastic fibers in human skin. Waaijer et al. (2016)
scRNA-seq of young versus old human skin reveals that aged fibroblasts show a functional decline, loss of priming into Salzer et al. (2018);
distinct subtypes, and a propensity for adipogenic traits with disturbed dermal‒epidermal interactions. Solé-Boldo et al. (2020)
In situ aged human dermal fibroblasts in human skin reveal in part a proteome profile identical to senescence-associated Waldera-Lupa et al. (2014);
changes. Waldera Lupa et al. (2015)
A proteome atlas depicts that in vitro‒induced senescence of human fibroblasts results in the secretion of factors enforcing Basisty et al. (2020);
skin aging, inflammaging, as well as matrix degradation. Coppé et al. (2010)
Fibroblast-specific genetic ablation of SOD2 enhanced the number of SEN (p16INK4a-positive) fibroblasts in the skin and Treiber et al. (2011)
other organs in a murine model, and this correlates with severe skin aging, osteoporosis, and sarcopenia.
Transplantation of human SEN fibroblasts into the skin of young NSG mice resulted in the spreading of senescence with the da Silva et al. (2019)
expression of senescence markers in the dermis 3 weeks after transplantation.
Organotypic human skin culture models constructed with human SEN fibroblasts show hallmarks of skin aging with disrupted Lämmermann et al., (2018);
epidermal differentiation and function. This can partly be reversed by plant extracts. Weinmüllner et al. (2020)
In a perforin-deficient mouse with reduced removal capacity of SEN cells by NK cells, SEN cells accumulate in various organs Ovadya et al. (2018)
and, importantly, in the dermis of the skin leading to structural changes in skin aging.
Topical treatment of human skin with rapamycin in vivo resulted in the reduction of p16INK4a-positive cells in combination Chung et al. (2019)
with a decrease in fine wrinkles, reduced sagging, and an increase in dermal volume.
Abbreviations: scRNA-seq, single-cell RNA sequencing; SEN, senescent.

released from SEN fibroblasts are responsible not only for
skin aging and aging of other organs but also for a variety of
age-related diseases (summarized in Childs et al. [2015]); for
Figure 2. Cell-autonomous and external cues determine fibroblast
senescence and skin aging. Senescence-promoting and -suppressing
mechanisms depend on the quality control and its disturbances of cell‒cell
and cell‒matrix interactions, cellular location, and the release of a specific
SASP from fibroblasts. Malfunction drives and spreads fibroblasts senescence,
skin aging, likely the aging of other organs, aging-related diseases, and
organismal aging. There is an intimate relationship between autonomous
(mitochondrial dysfunction, enhanced concentrations of ROS, DNA damage,
telomere attrition, epigenetic marks, loss of proteostasis, others) and external
(cell nonautonomous) cues. For further details, see the main manuscript.
SASP, senescence-associated secretory phenotype; SEN, senescent.
the progression of malignant tumors, including squamous
cell carcinomas (reviewed in Coppé et al. [2010]; Farsam
et al. [2016]; Liakou et al. [2016]; Mavrogonatou et al.
[2019]); and for the nonhealing state of wounds (Sindrilaru
et al., 2011). The SASP derived from human fibroblasts
consists, among many other factors, of increased concentra-
tions of IL-1, IL-6, IL-8, IL-18, matrix metalloproteinases
(MMPs), and a variety of chemokines (Coppé et al., 2010). In
addition, lipids (Ni et al., 2016) and micro RNA (Terlecki-
Zaniewicz et al., 2018) packaged in extracellular vesicles
direct and modulate basic biological functions such as
migration, proliferation, and senescence (da Silva et al.,
2019; Gonzalez-Meljem et al., 2018; Herranz and Gil,
2018; Malaquin et al., 2016; Weilner et al., 2013) of neigh-
boring cells (Faget et al., 2019; Lee and Schmitt, 2019), im-
mune cells (Prata et al., 2018), and other skin cells.
Interestingly, in vivo reprogramming and the transplantation
of mesenchymal stem cells (MSCs) suppress SASP factors
(Taguchi and Yamada, 2017; Vander Beken et al., 2019).
Interrelated paracrine loops of some of these factors such as
IL-1 and IL-6 have earlier been described to induce c-Jun
N-terminal kinase and AP-1 and, in consequence, matrix-
degrading metalloproteases as a response of human fibro-
blasts to UV irradiation and in part also for intrinsic aging
in vitro and in vivo in healthy humans (Fisher et al., 2009;
Schneider et al., 2017; Urbanski et al., 1990; Wlaschek et al.,
1994) and in intrinsic aging. IL-6, in fact, increases in elderly
adults and contributes to wrinkle formation in human skin
(Kim et al., 2012) and other age-related diseases (Beyer et al.,
2012; Cohen et al., 1997; Jenny et al., 2002) and increased
mortality (Lee et al., 2012). Senescence spreading from SEN
human fibroblasts to adjacent non-SEN human fibroblasts
in vitro and SEN hepatocytes to non-SEN hepatocytes in the
liver in vivo has earlier been shown (Acosta et al., 2013;
Nelson et al., 2018, 2012). It is of relevance to interrupt the
spreading of senescence and the SASP. In an independent
recent systems biology approach employing a Boolean
network technology, we were able to mechanistically show

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Fibroblast Senescence and Skin Aging
that DNA damage‒mediated SASP triggering of IL-6 and IL-8
is mainly relayed through NF-kB in human dermal fibro-
blasts. This computational study also identified and experi-
mentally verified NF-kB essential modulator as a promising
inhibitor among other inhibitors (Meyer et al., 2017).
SEN fibroblasts with an aberrant SASP disrupt dermal‒
epidermal interactions. Recently, Liu et al., (2019) made
the seminal observation that weakening of collagen type
XVII, which serves to anchor murine epidermal progenitor
cells in the hemidesmosome and, in consequence, within the
basement membrane of the skin, results in being replaced by
another progenitor cell that produces intact collagen XVII
(Liu et al., 2019). Enhanced disintegrin and metalloproteinase
and MMPs released by SEN fibroblasts in the papillary dermis
or shedding of collagen XVII induced by the SASP factor IL-
1b released by SEN fibroblasts (Franzke et al., 2002) may
contribute to the reduction of collagen XVII, with possibly
constant replacement of epidermal progenitor cells, and this
most likely will contribute to epidermal SC exhaustion. In
addition, IGF-1 mainly released by dermal fibroblasts is
essential for MSC niches and the balanced regulation of
epidermal cell proliferation and differentiation. Keratinocytes
themselves cannot synthesize IGF-1 but do express the IGF-1
receptor. IGF-1 belongs to the SASP factors, and the IGF-1
released from SEN fibroblasts is significantly reduced. This
is due to enhanced superoxide anion production in the
mitochondria of human and murine dermal fibroblasts
in vitro and in vivo (Singh et al., 2015). In fact, earlier, we
could show that superoxide anions at increased concentra-
tions in murine and human skin fibroblasts suppress IGF-1
signaling at the level of decreased IGF-1 receptor activation
(Singh et al., 2015). This deprived IGF-1 release, and the
inhibition of relaying the IGF-1 signal suppresses collagen
synthesis in the dermis and led to epidermal atrophy with
enhanced accumulation of the DNA damage‒induced
phosphorylated histone protein, gH2AX, and p16INK4A-posi-
tive epidermal cells. The essential role of IGF-1 in human
skin, muscle, and bone was earlier reported (Anisimov and
Bartke, 2013; Gallagher and LeRoith, 2011). Reduced IGF-
1 expression in the dermis of elderly humans and reduced
IGF-1 serum concentrations directly correlate with tissue
decline in human skin aging and osteoporosis (Hassan et al.,
2015; Lamberts et al., 1997). Of note, the age of human in-
dividuals from whom dermal human fibroblasts have been
isolated impacts on keratinocyte differentiation and wound
healing (Hausmann et al., 2019). These data clearly show
that SEN and aged fibroblasts play a major role in dermal‒
epidermal interactions and overall skin tissue decline.
The development of senolytic agents. Employing a genetic
approach that selectively induced apoptosis of p16INK4A-
positive SEN cells in murine models of aging-related diseases
led to impressive improvement (Baker et al., 2011), in
particular of osteoarthritis, atherosclerosis, cataract, sarco-
penia, and dementia (summarized in Childs et al. [2017]).
Currently, it is unknown whether the genetic depletion of
SEN cells from the dermis would delay aging. The depletion
of p16INK4A SEN cells significantly improves organ function
and thus extends both health span and overall life span
988 Journal of Investigative Dermatology (2021), Volume 141
(Baker et al., 2016; Soto-Gamez and Demaria, 2017). The
genetic ablation of SEN cells was an encouraging proof for
the causal role of SEN cells in aging-associated disorders.
The next step was to screen for small pharmacologically
active molecules. Employing bioinformatics and small
interfering RNA technology, anti-apoptotic pathways active
in SEN cells were uncovered as senolytic targets, including
the anti-apoptotic proteins BCL-2, BCL-W, and BCL-XL; the
transcription factors p53 and p21; HIF1a; phosphatidyli-
nositol-4,5-bisphosphate 3-kinase and protein kinase B; the
serine protein inhibitors (serpins); and the heat shock pro-
tein 90, among others (Fuhrmann-Stroissnigg et al., 2017;
Zhu et al., 2015). With this advanced understanding, the
rational design of drugs became possible. In this context,
navitoclax (ABT-263) is a specific inhibitor of BCL-2, BCL-
XL, and BCL-W, enforcing the killing of SEN human WI-38
fibroblasts, human IMR-90 lung fibroblasts, and many other
cell types. Interestingly, this senolytic action is independent
of the senescence-inducing trigger. Navitoclax is endowed
with the capacity to rejuvenate SEN hematopoietic stem
cells and SEN muscle stem cells (Chang et al., 2016; Kim
et al., 2017). Importantly, the BCL-2 inhibitor ABT-737
reduced the population of SEN stem cells in the epidermis
in a mouse model of inducible senescence in the basal layer
of the epidermis of the skin (Yosef et al., 2016). Currently,
the combination of two senolytic agents (such as the
tyrosinase inhibitor dasatinib and quercetin, a singlet
oxygen-quenching agent) has entered clinical trials. One
study is successfully completed now for primary pulmonary
fibrosis, a severe lung disease with a proven accumulation
of SEN cells and life-threatening fibrosis destroying the lung
structure (Justice et al., 2019). The other disease treated with
a senolytic approach in a clinical trial is human diabetic
kidney disease (Hickson et al., 2019). Of note, the com-
bined therapy with quercetin and dasatinib resulted in the
suppression of the low-grade inflammation with the reduc-
tion of CD68þ macrophages in the subcutaneous fat tissue
beneath the dermis in human individuals (Ellison-Hughes,
2020). Of note, a plant extract of goldenrod displayed a
moderate senolytic activity and a substantial suppression of the
SASP with some rejuvenating effects on human skin equivalents
(Lämmermann et al., 2018). Collectively, senolytics hold sub-
stantial promise for the successful treatment and delay of a
variety of aging-related diseases that are due to SASP-induced
low-grade inflammation.
Fibroblast heterogeneity and its potential role in aging
By contrast to the well-established lineage hierarchy in the
hematopoietic system (Zhang et al., 2018), the existence of a
hierarchy of fibroblast lineage is largely unresolved. Single-
cell RNA sequencing (scRNA-seq) analysis recently uncov-
ered an enormous variety of fibroblast subsets, each with a
typical and reproducible transcriptional fingerprint (Driskell
and Watt, 2015; Jahoda and Gilmore, 2016; Korosec et al.,
2019; LeBleu and Neilson, 2020; Lynch and Watt, 2018;
Philippeos et al., 2018; Sacco et al., 2019; Sahai et al., 2020;
Salzer et al., 2018). Accordingly, a lineage hierarchy of these
newly described fibroblast subsets is slowly emerging, and it
is possible—although currently not understood—that distinct
fibroblast subsets are more prone to senescence than others.

Using transplantation assays and lineage tracing, Driskell and
Watt (2015, 2013) showed that murine skin fibroblasts arise
from two distinct embryonic lineages. Similarly, Jiang et al.
(2018) characterized embryonic fibroblast lineages on the
basis of their history of expressing the EN-1 gene. EN-1 is a
transcription factor expressed in a small subset of early em-
bryonic cells and switched off permanently later in embryo-
genesis (Rinkevich et al., 2015). Single-cell fate mapping, life
three-dimensional confocal imaging showed that wound
healing and regeneration are driven by those fibroblast de-
scendants originating from cells that during embryogenesis
did not express the EN-1 gene (engrailed history-naive cells).
By contrast, EN-1 history‒positive fibroblasts deriving from
embryonic cells, which transiently express EN-1 during
embryogenesis, are responsible for scarring. Transplantation
of EN-1‒naive cells prevents scarring. Interestingly, using
scRNA-seq of normal human skin, Vorstandlechner et al.
(2020) made the seminal observation that the tran-
scriptomic heterogeneity of fibroblast subsets in the adult
human skin is more extended than previously anticipated in
the earlier described two lineage models during embryo-
genesis. Studies based on scRNA-seq of skin from photo-
protected young and old human individuals (Dubois et al.,
2020; Solé-Boldo et al., 2020) and from mice (Salzer et al.,
2018) highlight the age-related loss of fibroblast cellular
identity with alteration in functional and spatial gene
expression signatures. Using population and single-cell
transcriptomics as well as long-term lineage tracing, Beni-
tah’s group (Salzer et al., 2018) convincingly demonstrates
that the identity of old fibroblasts becomes undefined in a
murine model where fibroblast signatures present in young
skin are no longer clearly demarcated in old skin. In addition,
old fibroblasts not only reduce the expression of genes
involved in the formation of the ECM but also gain adipo-
genic traits. These potential tissue devastating changes can,
however, in part be restored by long-term caloric restriction.
By contrast, a high-fat diet potentiates them. Apparently,
tissue decline and aging can also occur owing to the loss of
fibroblast identity and the adoption of adipogenic traits, and
this can be reversed by metabolic changes with caloric re-
striction. This exciting observation supports the beneficial
effects of caloric restriction, which in part has already been
translated for clinical application (Longo et al., 2015).
Fibroblast plasticity and cues from the ECM
There is growing evidence that fibroblasts are not only
regulated by their developmental origin and position with
retained memory and transcriptomic signature indicating
where they come from (Sacco et al., 2019). In addition, their
morphologic and functional plasticity is further amplified by
epigenetic marks, by transitions from distinct cells to a
fibroblast phenotype, by cell‒cell interactions, and impor-
tantly, by the ECM itself (for review, see Fedintsev and
Moskalev [2020]; LeBleu and Neilson [2020]). In fact, there
is emerging evidence that stochastic nonenzymatic modifi-
cation of the long-lived ECM molecules and other mac-
romolecules have a previously underestimated impact on
skin, tissue, and organ aging (Fedintsev and Moskalev, 2020).
Postulated first by Björksten in 1942, this hypothesis predicts
that crosslinking of macromolecules such as collagen and
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Fibroblast Senescence and Skin Aging
elastin among many others (Bjorksten and Tenhu, 1990) af-
fects the physicomechanical properties (stiffness) of ECM
proteins, and this likely is responsible for disrupted mecha-
nosensing causing cell and tissue dysfunction and aging
(Segel et al., 2019; Wang et al., 2019). Earlier, Sun et al.
(2011) placed MSCs from old mice on the ECM from young
mice and observed an impressive 16-fold increase in their
proliferation. Although it is unclear whether enhanced stiff-
ness of the ECM from old mice is responsible for this, reju-
venated MSCs showed a decrease in intracellular ROS
concentrations and increased adenosine triphosphate levels.
Interestingly, aging-dependent collagen fragmentation by
enhanced activity of distinct MMPs impaired fibroblast‒ECM
interactions with reduced mechanical forces, resulting in
fibroblast reprogramming with dampened collagen synthesis
and enhanced proteolysis amplifying a vicious cycle of tissue
decline (Fisher et al., 2009). Injection of hyaluronic acid
fillers into the human skin of elderly adults helped to restore
the functional capacity of aged human fibroblasts in the skin
with increased collagen synthesis (Quan et al., 2013). These
data may imply that the quality of the ECM impacts fibroblast
function and senescence.
Position stability and depletion of fibroblast may drive skin
aging
Tracking the same skin fibroblasts in live mice, Marsh et al.
(2018) show that fibroblast position is stable over time and
that this stability is maintained also in the case that neigh-
boring fibroblasts are experimentally depleted by laser abla-
tion. Interestingly, fibroblast membranes are apparently
highly dynamic in skin maintenance and homeostasis. This
also holds true during aging, with clustered fibroblast
depletion in aging skin (Marsh et al., 2018). Fibroblast
membranes surprisingly spread and extend to fill the space of
lost adjacent fibroblasts. This extension of cell membranes
occurs in a Rac1-dependent manner. These data imply that
positional stability and cell occupancy prime events that are
key for the homeostasis of skin fibroblasts in vivo throughout
the lifespan of mice. This network of fibroblasts may also
provide interaction substrates for recruited immune cells and,
in addition, may facilitate paracrine communication between
both immediate and more distant neighbors (Adler et al.,
2018; Inaba et al., 2015; Sanders et al., 2013; Zhou et al.,
2018) and if disrupted—possibly by the release of a persis-
tent and proinflammatory SASP—may even drive skin aging.
The capacity of fibroblasts to spread their membranes may
imply that changing the cell size requires less energy
compared with the energy demand for proliferation and
migration but is sufficient to survey putative ECM zones
(Abdel-Haleem et al., 2017; Zanotelli et al., 2018). Interest-
ingly, although the number of SEN cells accumulates in hu-
man skin during aging (Ressler et al., 2006), there is evidence
that older human individuals when compared with young
individuals had fewer fibroblasts and with a greater number
of longer membrane projections (Andrew et al., 1964e1965;
Novotny and Gnoth, 1991; Salzer et al., 2018). The cellular
depletion does not occur uniformly, but there are clustered
cell losses. Collectively, depletion of fibroblasts during aging
with the concomitant accumulation of SEN fibroblasts con-
tributes to skin aging.
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Fibroblast Senescence and Skin Aging
Perspectives
The unique opportunity to unveil the role of fibroblast subsets
and modifications of various ECM proteins during skin aging
will pave the way for future research directions and the
development of novel preventive and therapeutic strategies.
These are currently very dynamic research foci of aging in
general, and preventive and therapeutic strategies are
recently summarized in several reviews (Childs et al., 2017;
Choi et al., 2011; Fedintsev and Moskalev, 2020; Prata et al.,
2018; Toutfaire et al., 2017; Velarde and Demaria, 2016; Zhu
et al., 2015).
ORCIDs
Meinhard Wlaschek: http://orcid.org/0000-0001-5821-1140
Pallab Maity: http://orcid.org/0000-0001-9769-3533
Evgenia Makrantonaki: http://orcid.org/0000-0001-8522-2128
Karin Scharffetter-Kochanek: http://orcid.org/0000-0002-9655-685X
CONFLICT OF INTEREST
The authors state no conflict of interest.
AUTHOR CONTRIBUTIONS
Conceptualization: MW, PM, KSK; Writing - Original Draft Preparation: MW,
KSK; Writing - Review and Editing: MW, KSK, PM, EM
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