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Comparison of Tissue Preservation Using Formalin and Ethanol As Preservative Formula

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Comparison of Tissue Preservation Using Formalin and Ethanol As Preservative Formula View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Althea Medical Journal 359 Comparison of Tissue Preservation using Formalin and Ethanol as Preservative Formula See Woan Shian,1 Arifin Soenggono,2 Sawkar Vijay Pramod3 1Faculty of Medicine Universitas Padjadjaran, 2Department of Anatomy and Cell Biology, 3Department of Surgery Faculty of Medicine Universitas Padjadjaran/Dr. Hasan Sadikin General Hospital Bandung Abstract Background: Tissue preservation can be performed through embalming, by providing the chemical embalming fluid to the human remains. Formalin’s preservative formula is the foundation for modern methodsstudy was of to embalming. compare the Unfortunately, tissue preservation this preservative using formalin formula and has ethanol several as disadvantages.preservative formula. While Ethanol’s Methods:preservative This formula study wasis a carriedconsiderable out from agent September–October to replace formalin’s 2014 preservative in the Laboratory formula. of theThe Department aim of this of Anatomy, Faculty of Medicine, Universitas Padjadjaran. The study used the laboratory experimental method with consecutive sampling of 16 Wistar Rats. Thirty two soleus muscles and thirty two colons were collected and divided into two groups. Each group consisted of 16 soleus muscles and 16 colons. Group 1 wasof bacteria preserved were with determined formalin’s before preservative and after formula treatment. and Group 2 was preserved with ethanol’s preservative Results:formula. The two groups were preserved for six weeks. The tissue’s color, consistency, odor and the growth growth of Tissues bacteria preserved in tissues with but failedethanol’s to retain preservative the consistency. formula hadAll the better data tissue were preservation analyzed with in Chi-square the aspect oftest. color and odor, compared with formalin’s preservative formula. Both preservative formulas showed no Conclusions: preservative formula. [AMJ.2016;3(3):359–63] Ethanol’s preservative formula preserves better quality of tissue compared to formalin’s Keywords: Ethanol, formalin, tissue preservation Introduction infection on contact with dead body, prevent over-hardening and retention of color of tissues and organs, prevent desiccation, inhibit In the framework of undergraduate medical fungal or bacterial growth and has lesser risk education, cadavers are main educational of being a potential environmental chemical tools which are intended for dissection and hazards and biohazards.3,4 to demonstrate prosected specimen through 1,2 Formalin, which is composed of a visual, auditory and tactile pathways. Hence, saturated water solution containing 39–40% tissue preservation plays a pivotal role which of formaldehyde, is discovered in the year is to preserve cadavers, maintaining its life- 1869.3 After formalin was determined to be an like physical characteristics and prevents its 3 excellent preservative, it became the foundation decomposition. This can be done through for modern methods of embalming.3,4 However, embalming, which is an art and science in formalin as preservative formula has several disadvantages for embalming purposes. which is composed of chemical to the human 3 modernremains. culture The aims by of giving embalming the embalming for anatomical fluid health problems, causes over hardening of purposes are to prevent putrefaction progress tissues, coagulates blood, convert tissues to on the cadavers, ensure that there is no risk of Formalin’s preservative formula will lead to Correspondence: See Woan Shian, Faculty of Medicine, Universitasa grey Padjadjaran, hue when Jalan itRaya mixes Bandung-Sumedang with blood, Km.21,fixes Jatinangor, Sumedang, Indonesia, Phone: +6287822004420 Email: [email protected] Althea Medical Journal. 2016;3(3) 360 AMJ September 2016 discolorations, dehydrates tissues, constricts of glycerin, 50ml of phenol, 200g of sodium capillaries and has a suffocating odor.2,3,5,6 In chloride and 600ml of water whereas the addition, ethanol has been phased out for Product Type 22 ‘Embalming and taxidermist 700ml of ethanol, 200ml of phenol, 40ml of 3 Based on several ethanol’sglycerin, 10g preservative of sodium chloride,formula 30mlconsisted of water of researches, ethanol has several advantages and 30ml of formalin. asfluids’ preservative by 1 September formula 2006. and has less risk to Then, the Wistar rats were dissected to health problems.7 Therefore, ethanol can be collect 32 soleus muscles and 32 colons. considered to replace formalin as preservative Firstly, a Wistar rat was put into an inverted formula.3 This study was conducted to compare beaker for anesthesia. The inverted beaker the tissue preservation using formalin and consisted of cotton that was soaked with the ethanol as preservative formula. lethal volume of ether. Next, the Wistar rat was placed on a dissecting tray with needles Methods to secure it. Then, the dissection started by cutting down from the neck to the lower abdomen. Another two lines were cut towards This study was carried out from September– left and right from the end of the center line. October 2014 in the Laboratory of Department The visceral organs were removed and the of Anatomy, Faculty of Medicine, Universitas blood was washed with NaCl 0.9%. Afterward, Padjadjaran. All experiments performed on the the soleus muscles and colons were collected. laboratory animals in this study were approved The dissection procedure was repeated for all by the Health Research Ethics Committee, the Wistar rats. Faculty of Medicine, Universitas Padjadjaran. After all the tissue samples were collected, Formalin and ethanol preservative formula the soleus muscles and colons were divided were obtained from the Laboratory of into two groups. Group 1 was preserved with Department of Anatomy, Faculty of Medicine, Universitas Padjadjaran. The study used the laboratory experimental formula. Each tissue was preserved with 6ml method with consecutive sampling of sixteen formalin’s preservative formula and Group 2 was preserved with ethanol’s preservative healthy male Wistar Rats as study subjects. The two groups were preserved for six The inclusion criteria for the study subjects 0 ofweeks preservative in a temperature fluid in one of 10plastic container. were healthy Wistar rats which were 8 color, consistency, odor and the growth of weeks old male and weighing between 250g, bacteria were determined beforeC. andThe after tissue’s the whereby the exclusion criteria was the Wistar preservation. rats which did not move actively. The colors of the tissues were accessed The preservative chemicals were prepared visually, the odors of the tissues were accessed one week before the dissection by measuring by smelling and the consistencies of the the preservative chemicals according to the tissues were accessed by tactile sensation. volume using a measurement beaker and The growths of bacteria of the tissues were determined by the results on blood agar. The formula consisted of 150ml of formalin, 200ml beam balance. The formalin’s preservative Table 1 Color of Tissue Before and After Preservation Formalin’s Ethanol’s p-value Color Preservative Formula Preservative Formula (Chi-square Test) Before 0.599 Pink 16 16 Pale Red 16 16 After 0.000 Grayish Chocolate 16 0 Reddish Pink 0 16 Grayish White 16 0 Yellowish White 0 16 Althea Medical Journal. 2016;3(3) See Woan Shian, Arifin Soenggono, Sawkar Vijay Pramod: Comparison of Tissue Preservation using Formalin 361 and Ethanol as Preservative Formula Figure 1 Color of Soleus Muscle: (a) soleus muscle before preservation showed pink color. (b)soleus muscle preserved by formalin’s preservative formula preservation showed grayish chocolate color. (c)soleus muscle preserved by ethanol’s preservative formula preservation showed reddish pink color. Figure 2 Color of Colon: (a) colon before preservation showed pale red color. (b)colon preserved by formalin’s preservative formula preservation showed grayish white color.(c)soleus muscle preserved by ethanol’s preservative formula preservation showed yellowish white color procedure of detection of the growth of bacteria the p-value was more than 0.05. began by putting the tissue samples into test However, for the comparison of color of tubes which contained brain-heart infusion the tissues after using formalin and ethanol media. After the samples were incubated at a temperature of 370C for 24 hours, each sample difference because the p value was less than was inoculated on blood agar. Then, results as0.05. a preservative The color of theformula, tissue itthat had was a significantpreserved were obtained after the incubation of blood agar for 24 hours at a temperature of 370C. more similar to the color of the tissue before Furthermore, data of color, consistency, by the ethanol’s preservative formula was odor and growth of bacteria of the tissues preservative formula. Thus, the color of the before and after preservation were statistically preservation rather than the formalin’s analyzed using the Chi-square test. Statistically formula was better than the color of the tissue tissue preserved by the ethanol’s preservative Analysis was performed by comparing the formula (Table 1). significant was considered when p<0.05. preservedThe comparison by the formalin’sof odor preservativeof tissue preservation after using formalin and ethanol tissuepreservative preservation formula group.between the formalin’s preservative formula group and the ethanol’s difference because the p-value was less
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