Nutritional Intervention For Optimizing Mitochondrial Performance
Increasing Mitochondrial Efficiency in Cultured C1C2 Muscle Cells
Shuk Yee Ngan and Frank Merante Seneca College of Applied Arts and Technology
April 2, 2018
Introduction
Mental and physical stress from the demands of life, work, or athletic activity can reduce mental clarity and physical performance. Everyday people and athletes alike seek improvements in stamina, strength, and cognitive performance in order to live fulfilling meaningful lives.
This is a preliminary study designed to assess the utility of a specific nutritional Intervention for optimizing mitochondrial performance. If mitochondrial performance can be achieved through the use of nutrients, the benefits associated with improving mitochondrial function may also be realized. These may include:
- Increased energy.
- Improved cognitive function.
- Better recovery after physical effort.
- Anti-aging and well-being.
We collaborated on an in vitro model to demonstrate biochemically the potential performance enhancement characteristics of the commercially available supplement, Mitronite. To this end, a muscle cell culture system (C2C12 mouse myoblasts) was developed and used to assess the Mitronite formulation.
An in vitro culture simplifies the evaluation by removing confounding variables seen in whole body assessments or studies using whole blood.
An indicator of performance, particularly mitochondrial performance, is reflected in the level of lactic acid produced in a stressed or exercised cell. Commonly, serum lactic acid levels, as a means of monitoring the degree of substrate flux through the mitochondria represents an indirect measure of mitochondrial performance. Progressive exertion and exercise can increase blood lactate levels and metabolic acidosis, which impact exercise performance.
Example of "differentiated C2c12 cells form multinucleated skeletal muscle myotubes in vitro" Image take from the website of Stephen Badylak Laboratory of the McGowan Institute for Regenerative Medicine (http://www.mirm.pitt.edu/badylak/projects/projects.Muscul.asp)
Method
In this study, in vitro enzymatic methods were developed assess mouse myoblast extracellular lactate levels produced during various metabolic interventions. Myoblast cells were washed and depleted of their intracellular glycogen stores. Subsequently they were provided one of two supplements, either a glucose containing solution, or a glucose + Mitronite-formulation . The level of lactic acid was determined after 30 minutes. Lower levels of lactic acid suggest improved metabolism and cellular homeostasis.
Results
Enzymatic assessment of lactic acid suggests that Kreb cycle intermediates present in the Mitronite formulation reduce lactate production, fostering mitochondrial performance even in the presence of fermentable carbon sources, namely glucose. Incidentally, cells provided only the glucose containing solution – a common ingredient in many sports drinks - were irreparably stressed and would dislodge from the surface of the plate indicating cellular damage or death (Figure 1).
Figure 1. (a) Glucose only solution. Cells were irreparably stress and and would dislodge from the surface of the plate. This indicates cellular damage or death.
Figure 1. (b) Mitronite plus Glucose solution. Damaged or death, cells were revivified and resumed normal function.
Cells provided the same level of glucose in the presence of the Mitronite formulation appeared health and also produced reduced levels of lactic acid, indicating improved mitochondrial performance and cellular health (Figure 2).
Figure 2. In cell provided the Mitronite formulation lactate production was significantly lower when compared to the cells that were provided the Mitronite formulation plus glucose. This suggests improved mitochondrial activity due to greater uptake of nutrients when glucose is absent.
Discussion and Implications
Muscle cells stress to the point of damage or death can be revivified and resume normal function when provided the kreb's cycle intermediates in the Mitronite formulation.
The observed reduction in cellular lactate levels suggests the nutrients support improved cellular energy and recovery.
This is preliminary research and the conclusions drawn will be used to evaluate further research projects to validate conclusions.