Satoshi Akaishi

Focal adhesion kinase links mechanical force to skin fibrosis via inflammatory signaling

Exuberant fibroproliferation is a common complication after injury for reasons that are not well understood. One key component of wound repair that is often overlooked is mechanical force, which regulates cell-matrix interactions through intracellular focal adhesion components, including focal adhesion kinase (FAK). Here we report that FAK is activated after cutaneous injury and that this process is potentiated by mechanical loading. Fibroblast-specific FAK knockout mice have substantially less inflammation and fibrosis than control mice in a model of hypertrophic scar formation. We show that FAK acts through extracellular-related kinase (ERK) to mechanically trigger the secretion of monocyte chemoattractant protein-1 (MCP-1, also known as CCL2), a potent chemokine that is linked to human fibrotic disorders. Similarly, MCP-1 knockout mice form minimal scars, indicating that inflammatory chemokine pathways are a major mechanism by which FAK mechanotransduction induces fibrosis. Small-molecule inhibition of FAK blocks these effects in human cells and reduces scar formation in vivo through attenuated MCP-1 signaling and inflammatory cell recruitment. These findings collectively indicate that physical force regulates fibrosis through inflammatory FAK–ERK–MCP-1 pathways and that molecular strategies targeting FAK can effectively uncouple mechanical force from pathologic scar formation.

Pushing Back: Wound Mechanotransduction in Repair and Regeneration

Human skin is a highly specialized mechanoresponsive interface separating our bodies from the external environment. It must constantly adapt to dynamic physical cues ranging from rapid expansion during embryonic and early postnatal development to ubiquitous external forces throughout life. Despite the suspected role of the physical environment in cutaneous processes, the fundamental molecular mechanisms responsible for how skin responds to force remain unclear. Intracellular pathways convert mechanical cues into biochemical responses (in a process known as mechanotransduction) via complex mechanoresponsive elements that often blur the distinction between physical and chemical signaling. For example, cellular focal adhesion components exhibit dual biochemical and scaffolding functions that are critically modulated by force. Moreover, the extracellular matrix itself is increasingly recognized to mechanically regulate the spatiotemporal distribution of soluble and matrix-bound ligands, underscoring the importance of bidirectional crosstalk between cells and their physical environment. It seems likely that a structural hierarchy exists to maintain both cells and matrix in mechanical homeostasis and that dysregulation of this architectural integrity may underlie or contribute to various skin disorders. An improved understanding of these interactions will facilitate the development of novel biophysical materials and mechanomodulatory approaches to augment wound repair and regeneration.

The relationship between skin stretching/contraction and pathologic scarring: The important role of mechanical forces in keloid generation

Keloids tend to occur on highly mobile sites with high tension. This study was designed to determine whether body surface areas exposed to large strain during normal activities correlate with areas that show high rates of keloid generation after wounding. Eight adult Japanese volunteers were enrolled to study the skin stretching/contraction rates of nine different body sites. Skin stretching/contraction was measured by marking eight points on each region and measuring the change in location of the marked points after typical movements. The distribution of 1,500 keloids on 483 Japanese patients was mapped. The parietal region and anterior lower leg were associated with the least stretching/contraction, while the suprapubic region had the highest stretching/contraction rate. With regard to keloid distribution, there were 733 on the anterior chest region (48.9%) and 403 on the scapular regions (26.9%). No keloids were reported on the scalp or anterior lower leg. Because these sites are rarely subjected to skin stretching/contraction, it appears that mechanical force is an important trigger that drives keloid generation even in patients who are genetically predisposed to keloids. Thus, mechanotransduction studies are useful for developing clinical approaches that reduce the skin tension around wounds or scars for the prevention and treatment of not only keloids but also hypertrophic scars.

The relationship between keloid growth pattern and stretching tension: visual analysis using the finite element method.

Background:Keloids grow and spread horizontally, like malignant tumors, for reasons that remain unknown. Yet, stretching tension is clearly associated with keloid generation, as keloids tend to occur on high tension sites such as the anterior chest and scapular region. Thus, we analyzed the relationship between keloid growth patterns and stretching tension using a visualized finite element study. Materials and Methods:Keloids, normal skin, and fat structures were reproduced using DISCUS software. The contours were transferred to ADINA analytical software to rebuild and mesh volumes. Results:(1) High tension was observed at the edges, and not in the entire region, of stretched keloids. (2) Keloid centers were regions of low tension, which helps to explain the healing that generally occurs in the central regions of keloids. (3) Expansion of a keloid occurred in the direction in which it was pulled. (4) The “crabs claw”-shaped invasion occurred in response to increased stretching tension. (5) Skin stiffness in the circumference of a keloid was associated with greatly increased tension. (6) Fat hardness and thickness did not influence the amount of tension. (7) Adhesion with subcutaneous hard tissue greatly increased the tension in the keloid. Conclusion:These stretching results have advanced understanding of keloid formation under various conditions. Our results suggest that stretching tension is an important condition associated with keloid growth.

Postoperative Radiation Protocol for Keloids and Hypertrophic Scars : Statistical Analysis of 370 Sites Followed for Over 18 Months

Background:Before 2002, keloids and intractable hypertrophic scars were treated at our facility with postoperative irradiation of 15 Gy (the traditional protocol). Analysis of the therapeutic outcomes of patients treated with this protocol showed that the recurrence rates of keloids and intractable hypertrophic scars in the anterior chest wall, as well as the scapular and suprapubic regions, were statistically higher than at other sites, while the recurrence rates in earlobes were lower. Thus, we customized doses for various sites. This report describes our trial of postoperative radiation therapy. Methods:Between January 2002 and September 2004, 109 patients with 121 keloid and intractable hypertrophic scar sites were treated with surgical excision following the new protocol: electron-beam irradiation at total doses of 10, 15, or 20 Gy, depending on the site. The recurrence rates and toxicities were historically followed in 218 patients with 249 keloid and intractable hypertrophic scar sites treated with the old protocol of surgical removal followed by irradiation at 15 Gy (without variation by site). The minimal follow-up time was 18 months. Statistical analysis was performed using Fisher exact probability test. Results:Total recurrence rates were 29.3% before 2002 and 14.0% after 2003. The recurrence rate in the anterior chest wall was statistically reduced. Outcomes of earlobe did not differ between irradiation with 15 Gy or 10 Gy. Conclusions:Keloids and intractable hypertrophic scars should be treated with dose protocols customized by site. Our results suggest that keloid and intractable hypertrophic scar sites with a high risk of recurrence should be treated with 20 Gy in 4 fractions over 4 days and that earlobe should be treated with 10 Gy in 2 fractions over 2 days.

Keloids and Hypertrophic Scars: Update and Future Directions

Summary: The development of cutaneous pathological scars, namely, hypertrophic scars (HSs) and keloids, involves complex pathways, and the exact mechanisms by which they are initiated, evolved, and regulated remain to be fully elucidated. The generally held concepts that keloids and HSs represent “aberrant wound healing” or that they are “characterized by hyalinized collagen bundles” have done little to promote their accurate clinicopathological classification or to stimulate research into the specific causes of these scars and effective preventative therapies. To overcome this barrier, we review here the most recent findings regarding the pathology and pathogenesis of keloids and HSs. The aberrations of HSs and keloids in terms of the inflammation, proliferation, and remodeling phases of the wound healing process are described. In particular, the significant roles that the extracellular matrix and the epidermal and dermal layers of skin play in scar pathogenesis are examined. Finally, the current hypotheses of pathological scar etiology that should be tested by basic and clinical investigators are detailed. Therapies that have been found to be effective are described, including several that evolved directly from the aforementioned etiology hypotheses. A better understanding of pathological scar etiology and manifestations will improve the clinical and histopathological classification and treatment of these important lesions.

Mechanosignaling pathways in cutaneous scarring

Mechanotransduction is the process by which physical forces are sensed and converted into biochemical signals that then result in cellular responses. The discovery and development of various molecular pathways involved in this process have revolutionized the fundamental and clinical understanding regarding the formation and progression of cutaneous scars. The aim of this review is to report the recent advances in scar mechanosignaling research. The mechanosignaling pathways that participate in the formation and growth of cutaneous scars can be divided into those whose role in mechanoresponsiveness has been proven (the TGF-β/Smad, integrin, and calcium ion pathways) and those who have a possible but as yet unproven role (such as MAPK and G protein, Wnt/β-catenin, TNF-α/NF-κB, and interleukins). During scar development, these cellular mechanosignaling pathways interact actively with the extracellular matrix. They also crosstalk extensively with the hypoxia, inflammation, and angiogenesis pathways. The elucidation of scar mechanosignaling pathways provides a new platform for understanding scar development. This better understanding will facilitate research into this promising field and may help to promote the development of pharmacological interventions that could ultimately prevent, reduce, or even reverse scar formation or progression.

In vivo Adipose Tissue Regeneration by Adipose-Derived Stromal Cells Isolated from GFP Transgenic Mice

We have previously demonstrated that pluripotent stem cells can be obtained from green fluorescence protein (GFP) transgenic mouse adipose tissue. In this study, we sought to determine whether adipose tissue regeneration can be induced in vivo using adipose-derived stromal cells (ASCs) from GFP mice. ASCs were isolated from inguinal fat pads of GFP mice, as described in our previous publication. After incubation in two passages in the control medium, the cells were incubated in the induction medium to induce adipogenesis. Induced ASCs were merged with fibrin glue, and the mixture was injected subcutaneously into the dorsum of athymic mice. Finally, specimens were harvested and analyzed morphologically and histologically. The regenerated tissue was macroscopically semitransparent and soft with slight angiogenesis. Fluorescence microscopy revealed that the specimens strongly emitted green fluorescence, suggesting that the transplanted ASCs had contributed to adipogenesis. Both hematoxylin and eosin and oil red O staining revealed that cells containing small lipid droplets had been regenerated histologically. These findings suggest that ASCs could contribute to adipose tissue regeneration in vivo. ASCs may be an ideal source for adipose tissue regeneration, which may in turn play an important role in augmentation surgery in surgically treated cancer or trauma patients.

Are keloid and hypertrophic scar different forms of the same disorder? A fibroproliferative skin disorder hypothesis based on keloid findings

Hypertrophic scars (HSs) and keloids are commonly seen as two different diseases by both clinicians and pathologists. However, as supported by histological evidence showing they share increased numbers of fibroblasts and accumulate collagen products, HS and keloid might be different forms of the same pathological entity, rather than separate conditions. To test this hypothesis, keloids from patients who underwent scar excisions (n = 20) in Nippon Medical School from 2005 to 2010 were examined histologically. The proportion and distribution of cellular and matrix collagen components were evaluated at the centre and periphery of each sample. In keloid samples, coexistence of hyalinised collagen, which is the most important pathognomonic characteristic of a keloid and dermal nodules that are considered to be characteristic of HS, was found. Moreover, hyalinised fibres appeared to initiate from the corner of the dermal nodules. Key features of inflammation such as microvessels, fibroblasts and inflammatory cells all decreased gradually from the periphery to the centre of keloids, indicative of reduced inflammation in the centre. Thus, we hypothesise that HS and keloid can be considered as successive stages of the same fibroproliferative skin disorder, with differing degrees of inflammation that might be affected by genetic predisposition.

Mechanical receptor-related mechanisms in scar management: a review and hypothesis.

Background: The physiopathogenesis of proliferative scarring in human skin is not well understood. Furthermore, knowledge of the precise mechanisms of action for physical treatment modalities is limited. Compression garments, occlusive/adhesive skin taping, and silicone gel sheets are applied to form an occlusion on the scar surface, reduce tension, and/or increase pressure on the scar itself. The mechanisms by which the external or superficial actions of these treatments cause remission of a protruding scar may be related to mechanoreceptor (nociceptor and cellular mechanoreceptor) responses. Methods: Basic research studies about mechanoreceptor-related (nociceptors and cellular mechanoreceptors, separately) events are reviewed and discussed based on proliferative scarring background. Scar management–related studies were corrected from the standpoint of mechanotransduction mechanisms. The methodologic quality of the clinical trials and basic studies was evaluated and reviewed. Results: It was suggested that many of the physical scar management methods, including compression therapy, silicone therapy, adhesive tape, and occlusive dressing therapy, are related to mechanotransduction mechanisms. Conclusions: A unifying perspective of basic research findings and clinical observations may be obtained by considering the mechanoreceptor-related events in scar management. Moreover, a precise understanding of the roles that cellular mechanoreceptors and mechanosensitive nociceptors play in proliferative scarring may lead to the development of innovative treatment strategies and new pharmacologic therapies targeting cellular mechanoreceptors and mechanosensitive nociceptors in fibroproliferative diseases.