c collagen plus collagenase inhibitor (Ci-I) and collagenases (c-C) were added to the mixture. The mixture was incubated at 37°C for 30 min. After washing with PBS, the medium was centrifuged at 13,000 × g for 10 min at 4° C. Protein concentration was determined by HPLC using a Bio-Rad Mass Spectrometer (Bio-Raman, Hercules, CA). The protein concentration of the collagen-containing medium (10 μg/ml) was measured by using the BioRad Protein Assay Kit (Thermo Scientific, Waltham, MA).
The collagen was extracted from the skin using an EtOH solution (1:1,200) in a 1:2 ratio of EtOAc to EtH 2 O. A 1% (v/v) solution of collagen (0.1% Tween 20) (Sigma-Aldrich, St. Louis, MO) containing 0.5% SDS was added. This solution was then incubation at room temperature for 1 h at RT.
, and the protein content of skin was calculated by the following formula:
. In addition, a collagen inhibitor was applied to a small amount of tissue (5 μg) to prevent the formation of a gel. To prevent formation, collagen inhibitors were applied in the same manner as described above. For the determination of protein concentrations, protein was separated by centrifuge at 15,500 ×g for 5 min and then the supernatant was collected. Total protein (T) concentration (μg/mL) of each sample was analyzed by a HPTLC system (Roche Diagnostics, Inc., Rockford, IL). For each protein, T was quantified by measuring the absorbance at 490 nm.
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The benefits of collagen are so great that it’s been shown to help reduce the risk of heart disease, stroke, and cancer. It’s also been found to reduce wrinkles, improve skin elasticity, reduce skin dryness, help prevent wrinkles and wrinkles-related skin cancer, as well as reduce inflammation and improve the appearance of wrinkles. The benefits are even more impressive when you consider that collagen is also a powerful antioxidant. In fact, collagen has been linked to a number of health benefits, including:
,
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) and the protein content of the diet was determined by the method of Krieger et al. (19). The protein concentration of each diet group was calculated by using the formula: (protein + carbohydrate + fat) × (carbohydrate + protein) = protein.
The study was approved by a local ethics committee and all participants gave written informed consent. The study protocol was written by Dr. K.A. and approved in accordance with the Declaration of Helsinki. All participants provided written consent to participate in the study. Written informed consents were obtained from all subjects. Informed consent was obtained for all patients who were not taking any medications. Patients were excluded if they had a history of cardiovascular disease, diabetes mellitus, or any other medical condition that could affect the ability to perform the dietary intervention. A total of 12 patients were enrolled in this study, and they were randomly assigned to the intervention group and to a control group. Each patient was followed for a total duration of 6 months. After the 6-month follow-up period, the patients in both groups were followed up for another 6–8 months to determine if the changes in body composition and body weight were maintained. At the end of this period of follow up, all the subjects were asked to complete a questionnaire to assess their body fat percentage and their changes over time. Body fat was measured by dual-energy X-ray absorptiometry (DXA) using a standard DXA system (DxA Systems, Inc., San Diego, CA).
, where the mean body mass index (BMI) was 25.0 kg/m2 and was divided into the following categories: <25.00 kg; 25–29.99 kg, 30–34.9 kg and ≥35.5 kg. For the purpose of analysis, we used the BMI as the reference category. We used a linear regression model to estimate the change in weight and fat mass over the course of a 6 month period. To determine the effect of dietary protein on body size, a multivariate linear model was used to calculate the relative change of body weights and changes of fat masses over a period (6 months). We also calculated the percentage change (percentage change) in fat and lean mass. Because the fat-free mass (FFM) is a measure of total body energy expenditure, it was included as a covariate in our model. This covariance was adjusted for age
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The best way to get the best results from your collagen is to use it in your skin care routine.
, and the collagen in collagen-rich products is the most effective way of getting the benefits of collagen. The best collagen products are the ones that contain at least 10% collagen, which is a very high percentage of the total amount of your body’s collagen that you can get from the skin. This means that the more collagen you have in the body, the better your results will be.The most important thing to remember is that collagen does not have to be in a product to have the benefit of it. It can be added to your diet, in supplements, or even in skin creams. If you are looking for a collagen product that is high in protein, try the following:
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1/2 cup (60g) unsweetened cocoa powder
, or to taste
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(Note: I used a 1/4 cup of cocoa, but you can use any amount you like.)
The first step is to make the chocolate. In a medium bowl, whisk together the cocoa and sugar until smooth. Add the egg yolks and whisk until combined. Pour the mixture into a piping bag fitted with a small tip and pipe the filling into the bottom of a 9 x 13 inch pan. (I used my 9×13 pan, which is about the size of my hand.) Bake at 350 degrees for about 30 minutes, until the top is golden brown. Let cool completely before cutting into squares. The chocolate will keep for up to a week in the refrigerator.