Simultaneous Application of High-Intensity Focused Electromagnetic and Synchronized Radiofrequency for Fat Disruption: Histological and Electron Microscopy Porcine Model Study.

BACKGROUND: Radiofrequency (RF) and high-intensity focused electromagnetic (HIFEM) technologies are used for noninvasive body shaping as standalone modalities. OBJECTIVE: To examine the effects of novel synchronized RF and HIFEM on subcutaneous adipose tissue in a porcine animal model. MATERIALS AND METHODS: Seven large white pigs aged 6 months received 3 abdominal treatments of simultaneous application of synchronized RF and HIFEM (30 minutes, once per week). Punch biopsies of treated and control subcutaneous tissue were collected at the baseline, 4 days, 2 weeks, 1 month, and 2 months. Specimens were examined by light and scanning electron microscopy. Adipocyte volume was analyzed. Fat tissue temperature was measured in situ (fiber optic probes) and superficially (thermal imager). RESULTS: Fat layer was heated to temperatures of 42 to 45°C. Signs of fat apoptosis (shape alternations and pyknotic nuclei) appeared at day 4 and peaked between 2 weeks and 1 month. Adipocyte volume decreased significantly (p < .001) by 31.1% at 2 weeks, 1 month (-23.6%), and 2 months (-22.0%). Control samples showed healthy adipocytes. Scanning electron microscopy micrographs corroborated histology findings, showing flattened, volume-depleted and disrupted adipocytes. CONCLUSION: Synchronized RF with HIFEM procedure resulted in a significant and sustained fat reduction with no adverse events.

Mechanism of nonthermal induction of apoptosis by high-intensity focused electromagnetic procedure: Biochemical investigation in a porcine model.

BACKGROUND: Multiple studies have reported adipose tissue reduction after the application of the High-Intensity Focused Electromagnetic (HIFEM) field technology, yet cellular level evidence of the mechanisms has remained scarce. OBJECTIVES: This study aims to verify or refute previous single-study histological evidence and further investigates the proposed mechanism of apoptotic induction. METHODS: The thigh of two Large White pigs was treated with HIFEM for 30 minutes. Fat punch biopsies were collected from the application area before, immediately after, and 8 hours post-treatment. Control samples were taken from the abdomen immediately after and 8 hours post-treatment. Samples were analyzed for pro-apoptotic DNA markers (BAX, BCL-2, TXNIP, MMP9, TNF-α), the levels of free fatty acids (FFA), and the pH levels of the adipose tissue. RESULTS: The levels of FFA in the treated adipose tissue increased on average by 127.1% immediately post-treatment and by 134.1% 8 hours post-treatment, indicating a rapid breakdown of lipids. The average recorded adipose pH changed from 7.30 ± 0.12 at baseline to 6.60 ± 0.07 immediately post-treatment (P = .001) and to 7.19 ± 0.12 8 hours post-treatment. The levels of BAX, TXNIP, MMP9, and TNF-α increased post-treatment while BCL-2 decreased. Control samples showed constant levels of pH and pro-apoptotic markers. The FFAs in the control samples were increased by 41.6%-51.4%. CONCLUSION: The changes in the levels of the pro-apoptotic markers conformed to the previously reported elevated fat apoptosis post-HIFEM treatments. These effects were accompanied by an increase in FFA levels, and by reduced pH levels, due to the increased acidity in the adipose tissue. Further research is required to explore the potential of nonthermal induction of apoptosis.

Induction of fat apoptosis by a non-thermal device: Mechanism of action of non-invasive high-intensity electromagnetic technology in a porcine model.

OBJECTIVES: While controlled thermal changes in subcutaneous tissue have been used to trigger apoptosis of fat cells and have been proven clinically efficacious, another mechanism of electromagnetic stress suggests that fat apoptosis could be achieved by a non-thermal manner as well. This animal model study investigates the use of a non-invasive high-intensity magnetic field device to induce apoptosis in fat cells. METHODS: Yorkshire pigs (N = 2) received one treatment (30 minutes) in the abdominal area using a High-Intensity Focused Electromagnetic (HIFEM) device. Punch biopsy samples of fat tissue and blood samples were collected at the baseline, 1 and 8 hours after the treatment. Biopsy samples were sectioned and evaluated for the levels of an apoptotic index (AI) by the TUNEL method. Statistical significance was examined using the rANOVA and Tukey's test (α 5%). Biopsy samples were also assessed for molecular biomarkers. Blood samples were evaluated to determine changes related to fat and muscle metabolism. Free fatty acids (FFA), triacylglycerol (TG), glycerol and glucose (Glu) were used as the main biomarkers of fat metabolism. Creatinine, creatinine kinase (CK), lactate dehydrogenase (LDH) and interleukin 6 (IL6) served as the main biomarkers to evaluate muscle metabolism. RESULTS: In treated pigs, a statistically significant increase in the apoptotic index (AI) (P = 1.17E-4) was observed. A significant difference was found between AI at baseline (AI = 18.75%) and 8-hours post-treatment (AI = 35.95%). Serum levels of fat and muscle metabolism indicated trends (FFA -0.32 mmol · l-1 , -28.1%; TG -0.24 mmol · l-1 , -51.8%; Glycerol -5.68 mg · l-1 , -54.8%; CK +67.58 µkat · l-1 , +227.8%; LDH +4.9 µkat · l-1 ,+35.4%) suggesting that both adipose and muscle tissue were affected by HIFEM treatment. No adverse events were noted to skin and surrounding tissue. CONCLUSIONS: Application of a high-intensity electromagnetic field in a porcine model results in adipocyte apoptosis. The analysis of serum levels suggests that HIFEM treatment influences fat and muscle metabolism. Lasers Surg. Med. 51:47-53, 2019. © 2018 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Efficacy of monopolar radiofrequency on skin collagen remodeling: a veterinary study.

The aesthetic market offers various radiofrequency treatments for the reduction of wrinkles and rhytids. Even though this not an uncommon aesthetic therapy, there is considerable lack of clinical evidence on the various energy delivery systems available (unipolar, bipolar, tripolar, multipolar, etc.). The purpose of this study was to demonstrate the efficacy of a monopolar radiofrequency device (Exilis Elite, BTL Industries Inc., Boston, MA, USA) on the skin collagen in an animal model. The study treatment was done on the abdominal area of the potbellied Vietnamese mini pigs in the Veterinary Research Institute facility. All pigs were treated once per week for 4 weeks. The treatment area was sized 20 × 10 cm. The surface temperature was kept in the therapeutic interval from 39 °C to 43 °C and the therapy lasted for 10 minutes after reaching the therapeutic temperature. Biopsy samples were taken before the therapy and at the 3-month follow-up. The histology samples were stained and magnified (×400) before computer processing. The collagen volume was calculated using the stereological analysis and the data were statistically processed (using the nonparametric two-sample t-test). The collagen content tissue increased from average of 9.0% before the therapy up to 25.9% after the 3-month follow-up period. The statistical comparison of 54 samples taken before and after the treatment acknowledged the significant difference (p = 0.018). The stereological analysis proved large-scale improvement of collagen in the treated area. We have observed that the monopolar radiofrequency therapy significantly increases collagen remodeling.

Operator independent focused high frequency ISM band for fat reduction: porcine model.

BACKGROUND: Selective fat reduction has been clearly shown for various methods and energy modalities including cryolipolysis and high intensity focused thermal ultrasound. Mathematical modeling of focused high frequency of the EM spectrum has indicated that selective heating of fat is possible using wavelengths not previous explored. The purpose of this study was to demonstrate in the porcine model that selective heating of fat is possible with a non-contact, operator independent device. METHODS: High frequencies of the Industrial, Scientific and Medical (ISM) RF band were utilized to reduce abdominal fat in a porcine model. Practical application of mathematical modeling allowed an auto-feedback loop to be developed to allow operator independent adjustment of energy to maintain subcutaneous fat at 45-46°C while overlying skin remained at 40-41°C. RESULTS: Treatments of three Vietnamese pigs were performed under anesthesia in a certified veterinary facility. Gross and microscopic histologic results demonstrated a marked reduction in adipocytes of the treated area after 4 treatments of a total of 30 minutes each, with incremental fat diminution after each treatment. A final 70% reduction of the abdominal fat layer was seen in the treated areas. Duplex ultrasound revealed a reduction of fat layer from 7.6 to 2.9 mm. Histologic evaluation revealed that epidermis, dermis, and adnexal structures such as hair follicles were unaffected by the treatment, while adipocytes were significantly affected. CONCLUSION: A new model of fat reduction using high frequency RF has been successfully achieved in a porcine model. This has very positive implications in the development of an operator independent, contact free device for reduction of fat in clinical practice.