Revision as of 01:37, 16 January 2026

Abstract

This study aims to investigate the effects of GlyNAC on planarian tissue regeneration. Planarian and organism regeneration are needed to maintain a healthy body. However, as one ages, their regeneration declines. There has been a focus on anti-aging in the older population due to the adverse effects that come with it such as muscle fatigue, a decrease in tissue regeneration, etc. As a result, there was an increase in research to find an effective supplement to combat health issues and aging. GlyNAC, a combination of Glycine and N-acetylcysteine, has been shown to reverse and improve health by increasing glutathione levels. Glutathione is an antioxidant with multiple health benefits such as protecting against oxidative stress, a main factor in causing cell death. To conduct this experiment, planarians are going to be used because they have a similar central nervous system, tissue regeneration, and glutathione production like humans. There will be 1 control group and 3 experimental groups. The control group will be fed regular ground beef while the experimental group will get ground beef with GlyNAC mixed into it. For tissue regeneration, each group would be weighed and measured before and after the ground beef or GlyNAC-mixed ground beef. The behaviors that are going to be observed are the positive behavior, negative behavior, and the planarians’ motility by recording them for 30 minutes with grid paper underneath their petri dishes.

Introduction GlyNAC

   There has been an interest in anti-aging in the modern day due to one’s fear of aging and the many symptoms that come with it, such as muscle fatigue, inflammation, a decrease in tissue regeneration, and oxidative damage(Kumar, et al. 2023).  Oxidative stress occurs when there's an imbalance between the production of reactive oxygen species (ROS) and the body's ability to detoxify them (Pizzino et al,.2017). Excessive oxidative stress leads to aging and cell death, while low oxidative stress supports cell regeneration (Lee et al.,2018).  
   There is limited research on why older people face these symptoms and the lack of effective treatments to treat these symptoms. Despite this, many try to preserve their health and prevent these symptoms from happening through various means such as dieting, supplements, exercise, and antioxidants. Previous experiments have shown that both glycine and N-acetylcysteine, are antioxidants that can protect against oxidative stress (Ruiz-Ramírez, 2014; Kerksick, et al. 2005). These components are precursors to glutathione(GSH), an important antioxidant that protects against oxidative stress, increased energy, cell proliferation, etc. Therefore, GlyNAC is expected to boost glutathione production by combining Glycine and N-acetylcysteine (Kumar, et al. 2023).
  As one age, there is a deficiency of GSH due to the lack of glycine and cysteine needed to synthesize the GSH. With this deficiency of GSH, fewer antioxidant defenses are protecting against oxidative stress that can damage cells that promote a healthy body. Therefore, GlyNAC demonstrates that it has the potential and ability to benefit those who are aging.

Planarian

  Planaria, specifically Dugesia tigrina, are flatworms known for their remarkable tissue regeneration abilities and nervous system. This makes them ideal for studying tissue regeneration and the nervous system, which helps provide insights into human tissue regeneration and the nervous system since both planaria and humans use stem cells for this process and have similar nervous systems. Planaria are famously used to research tissue regeneration, toxicology, and other pharmacology studies. Planarians can regenerate their tissue by using stem cells which are unspecialized cells that can develop into various types of cells, such as skin and nerve cells (Reddien, et al 2018). Additionally, planaria have glutathione in their redox system and glutathione is an essential component in balancing the redox system (Bijnens, et al.2021). With this planaria can create glutathione in balancing their redox system.

Materials and Method:

Preparation of GlyNAC treatment

  Glycine and N-acetylcysteine) were purchased on Amazon (Nutricost). Glycine and N-acetylcysteine were mixed with a 1:1 ratio to make GlyNAC. The GlyNAC was blended into the ground beef with their desired mass concentrations of 0%, 1%, 5%, and 10%  (Table 1.)  

★ The treatments were changed since in trial 1 majority of group D’s planarians had died. So, Group D’s treatment was changed to 5% of GlyNAC.

Preparation of Planarians

  40 planarians have been ordered from the website, Carolina.com, and the catalog number is 132954. The planarians were kept in loosely sealed jars of Poland spring water. They were then separated into 4 groups of 10. Which were each used for different concentrations of GlyNAC. Afterward, the planarian jars were stored in the cabinet with a gap. The planarians were fed ground beef for a week.

Preparation of Treatment Solutions

 All preparation procedures were conducted under sterile conditions. Work surfaces were disinfected using a 10% bleach solution, and hands were washed with soap and water prior to beginning. Standard personal protective equipment (PPE), including laboratory coats, gloves, and safety goggles, was worn throughout the process. Materials required for treatment preparation included beakers, graduated cylinders, pipettes, an analytical balance, weighing boats, and Poland Spring® water. The volume of Poland Spring® water specified in Table 1 was measured using a graduated cylinder. The appropriate volume of Trichosanthes root extract, also specified in Table 1, was then added to the water to create a stock solution, which was mixed thoroughly in a beaker. To prepare the working concentrations, a measured volume of the stock solution (e.g., 10 mL for Group B) was combined with Poland Spring® water (e.g., 90 mL for Group B) to reach a total volume of 100 mL. The solution was mixed by gently swirling the beaker. This procedure was repeated for each treatment group (A–D), based on the concentrations indicated in Table 1. Upon completion, all materials were cleaned, disinfected, and returned to their designated storage locations.

Treating Planarian Before the planarians are used, they must be nurtured. When working with them, safety equipment was worn and materials used in the experiment, as well as the workspace, were disinfected beforehand. The groups were treated with 20g of correspondent food according to Table 1. This was done a maximum of 3 times a week. After feeding, leftover beef was removed from the container using a pipette. Additionally, dirtied water was replaced with clean Poland spring water using a pipette, avoiding taking any planarians along with the dirtied water. The planarians were treated for 7 days before beginning the experiment.

Mobility Measurement Before adding the treatment, the behaviors of planarians were measured. To do this, safety gear was worn, and all materials, gear, and workspace were disinfected. Next, 30 mL of Poland spring water was poured into 4 petri dishes. Then, the planarians were put into the petri dishes based on their respective groups. Afterwards, graph paper “(Figure 1) was placed below the petri dishes, and a camera was used to record the planarian groups for 30 minutes.

Figure 1. Experimental setup for movement