Expression of HSP70 in Human Skin After Cryolipolysis Treatment.

BACKGROUND: Cryolipolysis nonsurgically targets and reduces subcutaneous fat through controlled cooling of skin and underlying fatty tissue. Although skin changes after cryolipolysis treatment have been observed clinically, the mechanisms by which these occur are not well understood. OBJECTIVES: The aim of this study was to investigate the expression of heat shock protein 70 (HSP70) in the epidermal and dermal layers of human skin following cryolipolysis treatment. METHODS: Subjects (N = 11; average age, 41.8 years; average BMI, 29.59 kg/m2) were recruited to receive cryolipolysis treatment with a vacuum cooling cup applicator (-11°C/35 minutes) prior to abdominoplasty surgery. Treated and untreated abdominal tissue samples were harvested immediately after surgery (average follow-up, 15 days; range, 3 days to 5 weeks). Immunohistochemistry for HSP70 was performed on all samples. Slides were digitized and quantified in epidermal and dermal layers. RESULTS: There was higher epidermal and dermal HSP70 expression in cryolipolysis-treated pre-abdominoplasty samples vs untreated samples. There was a 1.32-fold increase of HSP70 expression in the epidermis (P < .05) and a 1.92-fold increase in the dermis (P < .04) compared with untreated samples. CONCLUSIONS: We found significant induction of HSP70 after cryolipolysis treatment in epidermal and dermal layers. HSP70 has potential therapeutic benefits and is recognized to have a role in skin protection and adaption after thermal stress. Although cryolipolysis is popular for subcutaneous fat reduction, cryolipolytic HSP induction in the skin may prove valuable for additional therapeutic applications, including skin wound healing, remodeling, rejuvenation, and photoprotection.

Molecular and Histological Evidence Detailing Clinically Observed Skin Improvement Following Cryolipolysis.

BACKGROUND: In addition to body contouring, there is anecdotal and clinical evidence of reduced laxity caused by skin tightening after cryolipolysis. However, it has not been established how cryolipolysis triggers dermal changes. OBJECTIVES: The aim of this study was to investigate the fundamental mechanisms behind clinically observed dermal changes by molecular and immunohistochemistry (IHC) analytical methods. METHODS: This feasibility study involved 7 subjects who received cryolipolysis treatment. Tissue samples were harvested from 3 days to 5 weeks after treatment. RNA-sequencing examined differential gene expression of major collagens. RNA in situ hybridization (RNA-ISH) investigated the distribution of 1 of the gene markers for collagen type I (COL1A1). IHC for procollagen type I, heat shock protein 47 (HSP47), transforming growth factor ß (TGF-ß), and tropoelastin was performed and quantified. RESULTS: Gene expression analysis highlighted a gradual upregulation of collagen mRNA genes. RNA-ISH confirmed upregulation of COL1A1 mRNA and showed a homogeneous distribution through the dermis. IHC showed increases in protein expression. Quantification revealed a 3.62-fold increase of procollagen type I (P < 0.0071), a 2.91-fold increase of TGF-ß (P < 0.041), a 1.54-fold increase of HSP47 (P < 0.007), and a 1.57-fold increase of tropoelastin (P < 0.39) compared with untreated areas. CONCLUSIONS: This study revealed significant induction of molecular and protein markers of type I collagen, which supports neocollagenesis and may play an essential role in clinically relevant skin improvement. A dermal remodeling process driven by increased TGF-ß and higher expression of HSP47 was observed. Overall, these data provide the first evidence of dermal remodeling and clarify the mechanism by which cryolipolysis may induce skin improvement.

Clinical Study Demonstrates that Electromagnetic Muscle Stimulation Does Not Cause Injury to Fat Cells.

BACKGROUND AND OBJECTIVES: A previous pre-clinical study on electromagnetic muscle stimulation (EMMS) suggested that fat cell apoptosis occurs following treatment in a porcine model. While EMMS can induce changes in muscle, the effect on fat tissue is not established. This clinical study sought to assess adipose tissue response to EMMS in comparison to cryolipolysis treatment. STUDY DESIGN/MATERIALS AND METHODS: Study subjects were recruited prior to abdominoplasty to receive body contouring treatments and subsequently to obtain tissue for histological analysis. Non-invasive abdominal treatments were delivered using a commercially available (n = 6) or prototype (n = 3) EMMS system or a cryolipolysis system (n = 2). Subjects received a single EMMS treatment (100% intensity for 30 minutes) or a single cryolipolysis treatment (-11°C for 35 minutes) to the abdomen. Superficial and deep (i.e., adjacent to muscle layer) subcutaneous adipose tissue was harvested at set timepoints post-treatment. The presence or absence of an inflammatory response was evaluated using standard hematoxylin and eosin (H&E) staining. As adipocytes that are destined to become apoptotic cannot be distinguished by traditional H&E staining during the early phases of injury, irreversible fat cell injury was assessed using perilipin immunofluorescence. RESULTS: Following H&E histological analysis at 3, 10, 11, and 17 days post-treatment, no EMMS-treated samples showed an inflammatory response in either the superficial or deep subcutaneous adipose tissue. For the cryolipolysis-treated adipose tissue, however, the H&E staining revealed a marked inflammatory response with an influx of neutrophils, lymphocytes, and macrophages at timepoints consistent with previous histological studies. Further, loss of perilipin staining provided clear visual evidence of irreversible fat cell injury in the cryolipolysis-treated adipose tissue. In contrast, the electromagnetic muscle stimulation-treated samples showed persistence of perilipin staining of adipose tissue indicating that all fat cells were viable. CONCLUSION: This study failed to demonstrate either fat cell injury or inflammatory response following EMMS treatment. While electromagnetic muscle stimulation may non-invasively induce muscle changes, this clinical study found no evidence of an impact injurious or otherwise on subcutaneous fat. © 2020 The Authors. Lasers in Surgery and Medicine published by Wiley Periodicals LLC.

Feasibility Study of Electromagnetic Muscle Stimulation and Cryolipolysis for Abdominal Contouring.

BACKGROUND: In addition to reducing subcutaneous fat for body contouring, some patients are interested in toning the underlying muscle layer. OBJECTIVE: This feasibility study evaluated the safety and efficacy of electromagnetic muscle stimulation (EMMS) alone, cryolipolysis alone, and cryolipolysis with EMMS for noninvasive contouring of abdomen. METHODS: Abdomens of 50 subjects were treated in a study with 3 cohorts: EMMS alone, Cryolipolysis alone, and Cryolipolysis + EMMS in combination. Electromagnetic muscle stimulation treatments were delivered in 4 sessions over 2 weeks. Cryolipolysis treatments were delivered in one session. Combination treatments consisted of one cryolipolysis and 4 EMMS visits. Efficacy was assessed by independent physician Global Aesthetic Improvement Scale (GAIS), circumferential measurement, Subject GAIS (SGAIS), and Body Satisfaction Questionnaire (BSQ). RESULTS: Safety was demonstrated for all study cohorts with no device- or procedure-related adverse events. Independent photo review showed greatest mean GAIS score for the Cryolipolysis + EMMS cohort followed by Cryolipolysis only, then EMMS only cohort. BSQ showed greatest average score increase for Cryolipolysis + EMMS cohort followed by Cryolipolysis only cohort, then EMMS only cohort. Mean circumferential reduction measurements were greatest for Cryolipolysis + EMMS cohort followed by Cryolipolysis only, and then EMMS only cohort. The mean SGAIS improvement score was equal for the Cryolipolysis only and Cryolipolysis + EMMS cohorts, followed by the EMMS only cohort. CONCLUSION: A multimodal approach using cryolipolysis and EMMS was safe and demonstrated enhanced body contouring efficacy for this feasibility study.