Prenatal Administration of Spermine Promotes Maturation of Premature Fetal Gut Epithelial Tight Junction: Experimental Study on Fetal Rabbit

Study Overview
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Brief Summary
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Infections, particularly on the gastrointestinal tract, has been known to be one of the leading causes of death in preterm infants. This is due to the immaturity of the intestinal epithelial cells. Recent studies have shown that polyamines have a role on the development of cells during embryonal phase. By this experimental study, the investigators would like to evaluate the administration of spermine on the maturation of premature fetal gut epithelial tight junction.

Detailed Description
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This experimental study is conducted with minimum 24 subjects which divided into 6 groups; 1) 24-days pregnant and is given prenatal spermine, 2) 26-days pregnant and is given prenatal spermine, 3) 28-days pregnant and is given prenatal spermine, 4) 24-days pregnant and is not given prenatal spermine, 5) 26-days pregnant and is not given prenatal spermine, and 6) 28-days pregnant and is not given prenatal spermine. At the end of desired pregnancy period, hysterectomy is done and fetus are born. From each parent subject, three fetal samples are chosen using a random sampling. Intestinal tissues are taken from each fetal sample to be examined. Several data will be collected i.e. occludin, β-catenin, and β-actin, as well as histological morphologies.

Official Title
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Prenatal Administration of Oral Spermine Promotes Maturation of Premature Fetal Gut Epithelial Tight Junction: Experimental Study on Oryctolagus Cuniculus

Conditions
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Premature Birth, NEC

Intervention / Treatment
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* Biological: Spermine


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Intestinal tissue of fetal rabbit that is prematurely alive Exclusion Criteria: Intestinal tissue of fetal rabbit that is dead before termination Intestinal tissue of fetal rabbit, in which the parent died before termination

Ages Eligible for Study
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Minimum Age: 24 Days
Maximum Age: 28 Days

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: 24-Day Spermine Group<br>Subjects are given prenatal spermine (20 mg per body weight) during 24 days of pregnancy. On day 24 pregnancy is terminated and intestinal tissue sample is taken to be examined. | Biological: Spermine<br>* Spermine is a polyamine. It is an organic molecule that is involved in cellular metabolism and development.<br>|
| Experimental: 26-Day Spermine Group<br>Subjects are given prenatal spermine (20 mg per body weight) during 26 days of pregnancy. On day 26 pregnancy is terminated and intestinal tissue sample is taken to be examined. | Biological: Spermine<br>* Spermine is a polyamine. It is an organic molecule that is involved in cellular metabolism and development.<br>|
| Experimental: 28-Day Spermine Group<br>Subjects are given prenatal spermine (20 mg per body weight) during 28 days of pregnancy. On day 28 pregnancy is terminated and intestinal tissue sample is taken to be examined. | Biological: Spermine<br>* Spermine is a polyamine. It is an organic molecule that is involved in cellular metabolism and development.<br>|
| No Intervention: 24-Day Non Spermine Group<br>Subjects are not given any intervention. On day 24 pregnancy is terminated and intestinal tissue sample is taken to be examined. | |
| No Intervention: 26-Day Non Spermine Group<br>Subjects are not given any intervention. On day 26 pregnancy is terminated and intestinal tissue sample is taken to be examined. | |
| No Intervention: 28-Day Non Spermine Group<br>Subjects are not given any intervention. On day 28 pregnancy is terminated and intestinal tissue sample is taken to be examined. | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Occludin | Measurement of occludin (ng/mg protein) from intestinal tissue sample is assessed using Rabbit Occludin ELISA Kit | 2 weeks | 
| β-catenin | Measurement of β-catenin (pg/mg protein) from intestinal tissue sample is assessed using Rabbit β-Catenin ELISA Kit | 2 weeks | 
| β-actin | Measurement of β-actin (ng/mg protein) from intestinal tissue sample is assessed using β-Actin ELISA Kit | 2 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Histologic Findings | Identifying the development of epithelial tight junction cells based on the morphologic findings of the villi structure, including Goblet cells' distribution, enterocytes, enteroendocrine cells, and crypts. | 2 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Premature Birth, Spermine, Necrotizing Enterocolitis, Tight Junction