Effect of Folic Acid on the Presentation of Nuclear Abnormalities in People With a History Drug Abuse

Objectives.

• General Objective. To determine the effect of folic acid supplementation on the presentation of nuclear abnormalities associated with cytogenotoxic damage in healthy subjects and chronic drug users.

• Specific objectives.

  1. Determine the presence of micronuclei and other nuclear abnormalities in baseline samples from the group of chronic drug users and the healthy reference group.

  2. To compare baseline cytogenotoxic damage determinations with subsequent damage determinations, at 15 and 30 days, in chronic drug users supplemented with folic acid.

     Material and methods.

• Study design. Quasi-experimental study of pretest-posttest design with non-equivalent control group in subjects with substance abuse and healthy subjects. The intervention will consist of the administration of 15 mg of folic acid divided into 3 doses per day. Duplicate oral mucosa samples will be taken from each participant to determine the frequency of micronuclei (MN), nuclear sprouting (NBUD), binucleated cells (BN), condensed chromatin (CC), karyorrhexis (CR), pyknosis (PIC) and karyolysis. (CL) in different temporary events: pre-treatment, at 15 days and 30 days.

This project will be carried out in inmates of a rehabilitation center in El Salto, Jalisco, Mexico, in participants over 18 years of age, chronic drug users (More than 6 months of consumption). The foregoing under prior informed consent, in a convenience sample of 45 chronic drug users, as well as an equivalent sample of 45 healthy individuals without such a history.

• Sample size At convenience. 45 chronic drug users, inmates of a rehabilitation center in Salto, Jalisco, as well as 45 healthy individuals with no history of drug use will be considered.
• Data collection techniques and instruments. It will be done by taking oral cytology, as well as by means of a confidential questionnaire, in the questionnaire sociodemographic data will be collected such as: sex, age, profession, schooling, as well as information related to the consumption of psychoactive substances such as: tobacco, marijuana, alcohol, inhalants, cocaine and methamphetamines, significant pathological and non-pathological personal history. In addition to data referring to the consumption of a nutritional supplement or vitamin such as folates, B12, C, E, A, lutein or melatonin, or some other antioxidant, medications for pharmacological treatment of chronic disease and/or severe active periodontal disease. Exposure to fertilizers, radiation, heavy metals, as well as place of residence to rule out that they live in areas with high levels of air pollution (Tlaquepaque, Miravalle, Las Juntas and Las Pintas). Likewise, a physical examination of the oral cavity will be carried out by a dentist. All of the above in order to identify pathologies that have been shown to generate bias in the results obtained.

Folic acid will be used in the intervention stage in 5 mg tablets, box with 20 tablets, under the trade name Colicid from Bruluagsa.

---Cyto-genotoxic analysis by micronuclei in exfoliated cells of oral mucosa. The cytogenotoxic effect generated by the drugs will be evaluated, as well as the effect conferred to the administration of folic acid on the presentation of nuclear abnormalities associated with cytogenotoxic damage in chronic drug users.

In supplemented and non-supplemented individuals, through the oral mucosa cell micronucleus bioassay, duplicate samples will be obtained from each individual, spread on slides and stained with specific staining of nucleic acids (NA), frequency of micronuclei (MN), nuclear buds (NBUD), binucleated cells (BN), condensed chromatin (CC), karyorrhexis (CR), pyknosis (PIC) and karyolysis (CL).

---Data management and analysis The analysis of the data will be carried out in the GraphPad 9.2.0 program for Windows. For the continuous quantitative variables, the data will be presented with measures of central tendency and dispersion: mean and standard deviation. For the ordinal quantitative variables, they will be presented in frequencies and percentages, for the comparison of frequencies between sexes and age groups with the Fisher X2 test considering a p \< 0.05.

The statistical analysis to find differences between the groups will be carried out by means of the T test and for the ordinal variables it will be calculated by means of Chi2.