Workout Textile Spray Technology

In vivo: A group of 60 female volunteers (35-50 years old) with sedentary lifestyles were included and split into three different groups. The subjects in the first group performed a standardized exercise program under the supervision of a personal trainer in a gym, twice a week, and applied placebo for 56 days. The second group applied marine ingredient daily to the body (arms, abdomen and thighs) with no physical activity. The third group applied the marine ingredient daily and additionally, complied with the training program. The reducing efficacy of marine ingredient was assessed by the comparison of centimetric measurements of abdomen, thighs and arms contour before and after 28 days and 56 days of treatment.

• After 56 days, arm contour reduced up to -1.3 cm without exercise, and -2.4 cm with exercise
• After 56 days, abdomen contour decreased up to -2.8 cm without exercise, and -3.1 cm with exercise
• After 56 days, thigh girth reduced up to -2.1 cm without exercise, and -2.9 cm with exercise

STUDY 1: INDUCTION OF ADIPONECTIN RELEASE BY ADIPOCYTES

The efficacy of marine ingredient to enhance the secretion of adiponectin by adipocytes was assessed. Primary human subcutaneous preadipocytes were induced to mature towards adipocytes by incubation in differentiation medium for 8 days.

During adipogenesis, cells were left untreaded (control) or treated with marine ingredient concentrate.
Afterwards, culture supernatants were harvested and the levels of adiponectin secreted by the adipocytes were quantified by and Enzyme-Linked Immonosorbent Assay (ELISA).

The ELISA results were normalized with respect to the final amount of cells in each condition, which was assessed by Crystal Violet staining of the cells.

Results: The marine ingredient significantly increased the levels of adiponectin produced by human subcutaneous adipocytes by 68.3% with respect to the control

STUDY 2: ENHANCEMENT OF MITOCHONDRIAL ACTIVITY IN MUSCLE FIBERS

This study was carried out to evaluate to what extent the increased levels of adiponectin induced by marine ingredient in adipocytes translated into an enhancement of mitochondrial metabolic activity in skeletal muscle fibers. The activity of the pace-maker enzyme of the Krebs cycle citrate synthase was used as a quantitative marker of mitochondrial activity.

Differentiating human skeletal muscle cells were incubated for 48 hours either with the supernatants from non-treated adipocytes (control) or with the supernatants from the adipocytes that had been treated with the active ingredient and thus contained higher levels of adiponectin.

After the treatment, human skeletal muscle cells were lysed and citrate synthase activity was detected by a colorimentric assay. In the assay, one of the intermediates formed by the citrate synthase, reacts with a substance to generate a colored product, which is measured spectrophotometrically. The rate of the formation of the colored product is proportional to the activity of citrate synthase.

Results: The treatment with adipocytes supernatants containing high levels of adiponectin increased citrate synthase activity and therefore mitochondrial function by 47.9% in muscle cells.

STUDY 3: ATP PRODUCTION IN MUSCLE CELLS

To further verify the enhancement of mitochondrial metabolism of muscle cells by adiponectin released from adipocytes, the production of ATP was quantified.

Human skeletal muscle cells in the last days of the differentiation process were incubated with supernatants from adipocytes for 48 hours. The supernatants derived from adipocytes that had been treated or left untreated (control) for a period of 8 days.

After the incubation with adipocyte supernatants, skeletal muscle cells were lysed and the amount of ATP was quantified using a fluorescence assay. The results were normalized by the total protein concentration is each condition, which was assessed by the colorimetric bicinchoninic acid (BCA) assay.

Results: The adipocyte supernatant, rich in adiponectin due to treatment with the marine ingredient, significantly increased ATP production by skeletal muscle cells by 136%.

STUDY 4: SLOW MYOSIN LEVELS IN SKELETAL MUSCLE

The distinct types of muscle fibers contain different isoforms of myosin protein. Slow myosin heavy chain (MYH7) is expressed by slow (type1) muscle fibers.

The efficacy of adipocytes culture supernatants containing a high level of adiponectin after treatment to induce increased expression of type1 fibers myosin was assessed.

Human skeletal muscle cells in the last days of the differentiation process were incubated for 48 hours with supernatants derived from non-treated (control) adipocytes or from adipocytes that had been incubated with active ingredient.

Afterwards, immunofluorescence staining of the muscle cells was performed to stain slow myosin heavy chain protein with a fluorescent dye. Cells nuclei were marked with Diamidino-2- phenylindole (DAPI).

Images were captured using fluorescence microscopy and the intensity of fluorescence was quantified and normalized by the number of cells on each image.

Results: Increased levels of type I muscle myosin were induced by adipocytes supernatants rich in adiponectin after the treatment with marine ingredient.