Folia Medica Indonesiana
ISSN 0303-7932
Vol. 39 / No. 1 / Published : 2003-01
Order : 4, and page :10 - 17
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Original Article :
An immunohistochemical analysis of tgf-ß1, as a predictor of delayed wound healing in episiotomy surgery of sprague dawley rats
Author :
- St. Mulyata*1
- Department of Anesthesiology Sebelas Maret University School of Medicine Dr Moewardi Hospital, Surakarta
Abstract :
This animal experiment was carried out in the Laboratory of Inter University Center (PAU), Food and Nutrition Department, Gadjah Mada University, Yogyakarta, to prove an assumption that stress in pregnant women inhibited the delivery-wound healing process after birth. A number of 20 adult female Sprague-Dawley rats, averagely 3 months old, weighing about 300 g, were used for the experiment. Before experiment was started, all experimental animals were housed and maintained for adaptation in 2 weeks, with standard food and water ad libitum in accordance with the guidelines from Gadjah Mada University School of Veterinary Medicine, Yogyakarta. To make those rats pregnant, a male sprague-Dawley rat was put in each stable at a male - female ratio of 1 : 5. After pregnancy occurred, those rats were randomly assigned in two groups of 10 rats each. Group I was control without any treatment given. Group II were subjected to stress by putting them one by one into a box with external dimensions of 30 x 30 x 51 cm, provided a sound abating enclosure, and a transparent access door that permitted observation during treatment. They were subsequently treated by an ultrasound wave produced by an ultrasonic processor with the dose of 6.6 KHz and duration of 60 seconds. Nearby the delivery phase, stress was induced to all samples of group II, one by one according to the protocol. After those rats were stressed, they were subjected to episiotomy incision under general anesthesia on their perineal region from vulva to anus about 1 cm in length and its depth was limited to subcutis layer. The episiotomy wound was then closed by 3 simple suture with cotton no. 000. General anesthesia was performed by combination of ketamine/xylazine, intramuscularly. The dose of ketamine was 90 mg/kg BW, and xylazine was 10 mg/kg BW. Samples of control group were subjected only to an episiotomy incision under general anesthesia with the same procedures as that for group II. On day 3 after episiotomy, 10 rats of both groups, 5 rats each, a biopsy was performed on the site of episiotomy wound (vulva), then those rats were killed. From the biopsy tissues, stained with hematoxylin and eosin, histopathologic preparations were made to count for fibroblast cells. From the same biopsy tissues, but without staining, immunohistochemistry preparations were also made to examine TGF-ß1 cytokines found in the rat tissue. In the rest of samples, i.e., 10 rats, biopsy was also performed with the same procedures as that done on day 3. Then, histopathologic preparations were made for fibroblast count, and also immunohistochemistry preparations for TGF-ß1 examination. Results: Statistical analysis using multivariate test showed that there was statistical difference (p < 0.05) in fibroblasts count between two groups, and there were also TGF-ß1 in both groups but were statistically different between control and treated group. TGF-ß1 in treated group was lesser both in day 3 and day 5 of examination. Conclusions: From statistical analysis of results, it could be concluded that stressed Sprague- Dawley's rats may experience delayed episiotomy healing, as it was shown by less TGF-ß1 in rat's tissues of treated group. It could also be concluded that TGF-ß1 cytokine act as a predictor for wound healing.
Keyword :
Stress, episiotomy, ketamine, immunochemistry, fibroblast and TGF-ß1,
References :
Cotran, Kumar RS, V and Robbins SL,(1996) Robbins Pathologic Basis of Disease. 5 ed Philadelphia : WB Saunders Company, Harcourt Brace, Jovanovich Inc.
Goleman, D,(1995) Emotional Intelligence New York : Batam Books
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Volume : 39 / No. : 1 / Pub. : 2003-01 |
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