|Year : 2018 | Volume
| Issue : 2 | Page : 114-120
The effect of aspartame ingestion in pregnant female albino rats on placental and fetal weights, umbilical cord length, and histology of fetal pancreas
Azza M.M Aboshanady, Manal El-Sayed El-Sawaf, Abdel-Rahman Abu-El-Enain Abdel-Aziz, Mona M.A Attia
Department of Anatomy and Embryology, Faculty of Medicine, Tanta University, Tanta, Egypt
|Date of Submission||20-Jan-2018|
|Date of Acceptance||02-May-2018|
|Date of Web Publication||31-Oct-2018|
Azza M.M Aboshanady
Department of Anatomy and Embryology, Faculty of Medicine, Tanta University, kafr elhema, Tanta, Gharbiya, 31776
Background and aim Sugar is one of the commonest causes of weight gain and diabetes. Aspartame is one of the most commonly used artificial sweeteners worldwide to replace sugar. This study aimed to study the effects of aspartame in pregnant female albino rats on fetal and placental weights, length of umbilical cord, and possible histological changes in fetal pancreas.
Materials and methods Thirty-six adult female albino rats and nine adult male albino rats weighing between 220 and 250 g were used. Each four female rats were housed with one male rat to allow mating. Pregnant rats were divided into three groups (n=12 each): control group (group І), low-dose-treated group (group ІІ), and high-dose-treated group (group ІІІ). The low-dose-treated group received 14 mg/kg aspartame and the high-dose-treated group received 40 mg/kg aspartame daily from the first day of pregnancy to the 20th day of pregnancy.
Results This study showed a highly significant reduction of maternal weight gain, placental weight, fetal weight, and umbilical cord length in both aspartame-treated groups. Fetal pancreas showed histopathological changes in both aspartame-treated groups as evidenced by light and electron microscopy. All the results in this study were dose related.
Conclusion The use of aspartame during pregnancy might reduce fetal and placental weights and umbilical cord length and alter the histology of fetal pancreas.
Keywords: aspartame, pancreas, placenta, umbilical cord
|How to cite this article:|
Aboshanady AM, El-Sawaf ME, Abdel-Aziz ARA, Attia MM. The effect of aspartame ingestion in pregnant female albino rats on placental and fetal weights, umbilical cord length, and histology of fetal pancreas. Tanta Med J 2018;46:114-20
|How to cite this URL:|
Aboshanady AM, El-Sawaf ME, Abdel-Aziz ARA, Attia MM. The effect of aspartame ingestion in pregnant female albino rats on placental and fetal weights, umbilical cord length, and histology of fetal pancreas. Tanta Med J [serial online] 2018 [cited 2019 Oct 21];46:114-20. Available from: http://www.tdj.eg.net/text.asp?2018/46/2/114/244686
| Introduction|| |
Artificial sweetener is a food additive that duplicates the effect of sugar in taste, but has less food energy. The benefits of substituting artificial sweeteners include lower calorie intake, lower incidence of dental caries , better glycemic control , and weight loss . Aspartame is one of the most commonly used artificial sweeteners nowadays. Hundreds of millions of people use aspartame worldwide including children and women of childbearing period ,.
It was approved for use in dry applications by Food and Drug Administration (FDA) in 1981 followed by approval for use in carbonated soft drinks in 1983. Finally, it is approved for its use as a general sweetener in 1996. It is used in many food products such as desserts, yoghurts, vitamins, medicines, chewing gum, and diet beverages .
Several studies on laboratory animals have been made to verify the toxicity of artificial sweeteners. Some researchers have associated artificial sweeteners with health problems such as hepatotoxicity and cancers. A huge controversy concerning artificial sweeteners still exists. Among all artificial sweeteners, aspartame has been the most controversial because of its potential toxicity and carcinogenicity, even at the acceptable daily intake in humans. Chronic aspartame consumption results in progressive accumulation of formaldehyde products, which are responsible for functional alternation of proteins and of DNA mutations. These effects lead to autoimmunity, cell death, or malignant transformation . Despite, numerous toxicological studies of aspartame, consequences of its intake in pregnancy have been minimally addressed . Since the consumption of aspartame is increasing, further investigations and studies are recommended to prove or disapprove the existing fears concerning aspartame especially in pregnancy outcome . Therefore, the aim of this study was to evaluate the effects of aspartame in pregnant female albino rats on fetal and placental weights, length of umbilical cord, and possible histological changes in fetal pancreas.
| Materials and methods|| |
Thirty-six adult female albino rats and nine adult male albino rats weighing between 220 and 250 g were used.
The animals were housed in clean properly ventilated cages with steel wire tops under similar environmental conditions and fed the same laboratory diet. Each four females per one male were housed in a separate cage to allow mating. Vaginal smears were obtained from female rats for the detection of pregnancy and then pregnant rats were randomly divided into three groups, 12 rats each  and were weighted to obtain the initial weight.
Control group (group І)
Each rat received 3 ml distilled water daily at room temperature by a gastric tube from the first day of pregnancy till the 20th day.
Low-dose-treated group (group ІІ)
Each rat received 14 mg/kg aspartame. The dose was adjusted after Portela et al. . The dose was diluted in 3 ml distilled water at room temperature and given to the rats by gastric tube daily from the first day of pregnancy to the 20th day .
High-dose-treated group (group ІII)
Each rat received 40 mg/kg aspartame. The dose was adjusted according to Magnuson et al.  who mentioned that the highest acceptable daily intake of aspartame was determined to be 40 mg/kg body weight according to the European Food Safety Authority. The dose was diluted in 3 ml distilled water at room temperature and administered to the rats by a gastric tube daily during the same period of group II.
On the 20th day of gestation, the animals were weighted to measure the final weight, and then to obtain the weight gain (weight gain=final weight−initial weight). Then, all the rats were anesthetized by a suitable dose of ether and were killed. The uterine horns were opened and placentae, umbilical cords and fetuses were obtained. Each placenta and fetus was freshly weighed and the length of the umbilical cord was measured. Then, the data were assessed statistically.
The abdominal cavity of the fetus was incised longitudinally through median thoracoabdominal incision and the pancreas was collected, fixed in proper fixative, and examined by both light and electron microscopy.
Aspartame was purchased from Sigma Chemical Product Co. (Quesna, Menoufia, Egypt).
Data were analyzed as mean±SD by using SPSS program, version 12 (developed by IBM Corporation) (using the Mann–Whitney U-test). P values were calculated and a P value of less than 0.05 was considered significant and a P value of less than 0.001 was considered highly significant.
| Results|| |
This study showed a statistically highly significant reduction of weight gain of pregnant rats in both aspartame-treated groups compared with the control group. The reduction was more in group III than in group II ([Table 1]).
|Table 1 Weight gain of the pregnant rats in control group and treated groups and P-value|
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From the 12 control pregnant rats 102 viable fetuses were obtained, while 97 viable fetuses were obtained from the 12 pregnant rats treated with low-dose aspartame and 94 viable fetuses were obtained from the 12 pregnant rats treated with high-dose aspartame.
There were also decrease in the weight of viable fetuses and their placentae in both aspartame-treated groups were compared with the control group. The decrease was more in group III than in group II ([Table 2] and [Table 3]). The umbilical cord length of viable fetuses was also lesser in aspartame-treated groups than that found in the control group ([Table 4]).
|Table 2 Fetal weight in different groups at the time of scarification (20th day of pregnancy)|
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|Table 3 Placental weight in different groups at the time of scarification (20th day of pregnancy)|
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|Table 4 Umbilical cord length in different groups at the time of scarification (20th day of pregnancy)|
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Statistically, the decrease in fetal weight, placental weight, and umbilical cord length in both aspartame-treated groups was highly significant as compared with the control group (P<0.001). And also the decrease in these measurements was highly significant between group II compared with group III.
Light microscopic examination of the fetal pancreas
Hematoxylin and eosin-stained sections
Group I: the sections showed normal pancreatic architecture. The pancreatic tissue consisted of pancreatic lobules divided by thin connective tissue septa. The lobules consisted of pancreatic acini and well-demarcated lightly stained Islets of Langerhans More Details. Interlobular pancreatic ducts and blood vessels were seen ([Figure 1]a). The islets appeared as rounded masses of lightly stained acidophilic cells with rounded central nuclei; the cells were arranged in branching and anastomosing cords separated by blood sinusoids. The acini were formed of large pyramidal cells with basal basophilic nucleus and apical densely stained acidophilic cytoplasm ([Figure 2]a).
|Figure 1 Hematoxylin and eosin, ×100. Sections in the fetal pancreas: (a) group І showing that the pancreatic tissue consists of pancreatic lobules divided by connective tissue septa (C). The lobules consist of pancreatic acini (A) and well-demarcated islets of Langerhans (Is). Interlobular pancreatic ducts (Pd) and blood vessels (b) are seen. (b) Group ІІ and (c) group ІІІ showing apparent reduction of the pancreatic tissue which is divided into lobules by connective tissue septa (C). The lobules consist of pancreatic acini (A) and islets of Langerhans (Is) which are hardly demonstrated and densely stained. The reduction of pancreatic tissue is apparent more in group ІІІ. Splenic tissue (P) is seen.|
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|Figure 2 H&E, ×400. Sections of fetal pancreas: (a) group І showing the islets (Is) appearing as rounded masses of lightly stained acidophilic cells (arrow head) separated by blood sinusoids (★). The acini (A) are formed of pyramidal cells with basal nucleus (arrows). (b) Group ІІ show irregular, disorganized, and densely stained islets (Is). Congested blood sinusoids are seen (★). Some acini show wide lumen (arrows). Some acinar cells are multinucleated (double arrows). (c) Group ІІІ show acinar cells are cuboidal (A). Some cells are multinucleated (arrows). A disorganized islet is hardly identified (Is). Blood vessel (b) is seen.|
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Group II: sections from this group revealed apparent reduction of the pancreatic tissue which appeared divided into lobules by connective tissue septa. The pancreatic lobules consisted of pancreatic acini and islets of Langerhans ([Figure 1]b). However, the islets were hardly demonstrated. They appeared with irregular outlines, disorganized, and densely stained. Congested blood sinusoids between islet cells were seen. Some pancreatic acini showed wide lumen and reduced cell height. Some acinar cells were multinucleated ([Figure 2]b).
Group III: sections from this group showed a marked apparent reduction of the pancreatic tissue compared with both control and group II. The little amount of pancreatic lobules was separated by massive invasion of connective tissue septa. The pancreatic lobules consisted of pancreatic acini and a few number of islets of Langerhans which were disorganized and hardly demonstrated ([Figure 1]c and [Figure 2]c). The cells of the pancreatic acini lost their pyramidal appearance. They appeared cuboidal in shape with large basal basophilic nuclei. The lumens of the pancreatic acini were wide and some pancreatic cells were multinucleated ([Figure 2]c).
Mallory trichrome-stained sections
Mallory trichrome-stained sections of group I showed thin connective tissue septa separating the pancreatic lobules ([Figure 3]a), while sections from group II ([Figure 3]b) and group III ([Figure 3]c) showed increased connective tissue infiltration between the pancreatic lobules and around the blood vessels compared with the control group. The infiltration was massive in group III.
|Figure 3 Mallory trichrome ×200. Sections of the fetal pancreas: (a) group І showing thin connective tissue septa (C) separating the pancreatic lobules. Note: the lobules consist of pancreatic acini (A) and well-demarcated islets of Langerhans (Is). (b) Group ІІ & (fc) group ІІІ showing apparent increased connective tissue infiltration (C) between the pancreatic lobules and around the blood vessels (b) compared with the control group. The infiltration is massive in group ІІІ.|
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Immunostained sections for insulin reactivity of fetal pancreas of embryos obtained from group І rats showed a positive reaction in β cells, within the islets’ core. This reaction appeared as dark brown coloration in the cytoplasm of the immunoreactive cells ([Figure 4]a). Apparent reduced intensity of immunoreactivity in the cells in group II was noticed as compared with the control group ([Figure 4]b). Marked reduction in immunoreactive cells in the sections of group III embryos compared with both control and group II were manifested ([Figure 4]c).
|Figure 4 Insulin immunoreactivity ×200. Sections of the fetal pancreas:(a) group І, (b) group ІІ and (c) group ІІІ showing positively stained β cells in the islet core which appear as dark brown coloration (arrow). Notice, apparent reduced intensity of insulin immunoreactivity in β cells in group ІІ and group ІІІ as compared to control group. The reduction of intensity of immunoreactivity is more manifest in group ІІІ.|
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Electron microscopic examination of the fetal pancreas
Electron microscopic examination of the fetal pancreas of the embryos obtained from group I rats showed the normal ultrastructure of the pancreatic acinus. Each acinus cell was pyramidal in shape, contained basal nucleus with prominent nucleolus, apical zymogen granules, and well-developed rough endoplasmic reticulum. Its apical surface faced the lumen of the acinus ([Figure 5]a).
|Figure 5 TEM ×500. Ultrathin sections in the fetal pancreas: (a) group І rats showing a pancreatic acinus. Each acinus cell contains basal nucleus (N) and its cytoplasm shows abundant apical zymogen granules (Z). Its apical surface faces the lumen of the acinus (Lu). (b) Group ІІ and (c) group ІІІ showing loss of pyramidal shape of pancreatic acinar cells.The cytoplasm shows rough endoplasmic reticulum (rER) which becomes markedly dilated in group III, vacuolation (Va) and scanty apical zymogen granules (Z) especially in group ІІІ. (b) One nucleus appears central, shrunken and indented (N1) with multiple vacuoles basal to it.|
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The pancreatic acinus cells of this group lost their pyramidal appearance. Some of them had central, shrunken, and indented nuclei with multiple vacuoles basal to them. The cytoplasm showed rough endoplasmic reticulum, multiple vacuoles, destructed mitochondria, and apical zymogen granules which were apparently less than that seen in the control group ([Figure 5]b).
The pancreatic acinus cells of this group lost their characteristic pyramidal appearance. The cytoplasm showed markedly dilated rough endoplasmic reticulum, multiple vacuoles, and apical zymogen granules which were scanty or depleted ([Figure 5]c).
| Discussion|| |
This study showed that the weight gain during pregnancy was highly significantly reduced in both aspartame-treated groups in comparison to control group. Other studies that were done on rats used 14 mg/kg aspartame body weight orally on the 9th, 10th, and 11th days of pregnancy also observed a significant reduction of maternal weight gain in pregnant rats ,. Abd Elfatah et al.  who used a single daily dose of aspartame solution (50.4 mg) during gestation period found similar results.
Many researchers explained the mechanisms by which aspartame can reduce weight. Rogers et al.  mentioned that aspartame induced satiety in human beings leading to weight loss. Hall et al.  attributed the satiating effect of aspartame to the rising circulating levels of phenylalanine causing suppression of food intake in humans and animals and increased cholecystokinin secretion that delay gastric emptying. On the other hand, Beck et al.  suggested the decreased effects of neuropeptide Y on lipid metabolism. Neuropeptide Y is the peptide that promotes weight gain and fat deposition. On the contrary, some researchers reported that aspartame contributes to weight gain, hunger, and increased appetite .
The current study also showed a highly significant decrease in fetal weight in both aspartame-treated groups compared with the control group. This reduction can indicate that the fetuses cannot get their requirements of substrates, including glucose, which may be due to the possible diminution of substrates in the blood of maternal rats that utilized a sweetener. Similar results were found by Leme and Azoubel , Martins and Azoubel  and Portela et al. . In contrast, other studies claimed that a low dose of aspartame is safe to all individuals including pregnant women and children except people suffering from genetic disorders .
Placental size, architecture, developmental, and pathological processes affect placental–fetal nutrient exchange qualitatively and quantitatively . In humans, there is a positive correlation between placental weight and birth weight in normal and large-for-gestational-age infants . Thus it is believed that placental and fetal weights are directly related . In our study, we observed a highly significant decrease in placental weight of viable fetuses in both aspartame-treated groups. This result agrees with those obtained by Leme and Azoubel , Martins and Azoubel , and Portela et al. .
The length of the umbilical cord has been considered an indicator of fetal movement. It is accepted that umbilical-cord length is influenced by genetic and other factors. In humans, the existence of excessively long or short cords is uncommon . The present study presents a statistically significant decrease in umbilical-cord length of the treated groups. These results differ from those found by Leme and Azoubel  who did not find significant difference in the umbilical-cord length of aspartame-treated groups. However, this result is supported by Martins and Azoubel  and Portela et al. , who observed that the umbilical cords of aspartame-treated groups both at ambient temperature and solution heated to 40°C were diminished.
Suez et al.  suggested that aspartame cause hyperglycemia and an impaired ability to respond to insulin, which could be due to enhanced gluconeogenesis fueled by the production of the short chain fatty acid propionate by the gut microbiota. However, little information is available on the effect of aspartame consumption on the pancreatic structure and function.
In our study, histological examination of fetal pancreas of embryos obtained from both aspartame-treated groups showed apparent reduction of the pancreatic tissue. The pancreatic lobules were separated by massive invasion of connective tissue septa. This result is in agreement with Abd Elfatah et al. . Similar results were found by Mohammed et al.  in effect of aspartame on submandibular salivary gland and Khidr et al.  in effect of aspartame on the liver.
The increased collagen fiber deposition may be due to the effect of the metabolites of aspartame on the cell proteins. Bacon and Britton  attributed the increase in collagen and ground substance formation to elevated lipid peroxidation which has an oxidative damaging effect on proteins and nucleic acids. This opinion is in agreement with Iman  and Khidr et al. , who observed a significant elevation in lipid peroxidase level in rat tissues after 28 days treatment with aspartame.
In the present study, histological examination also showed that some pancreatic acini had a wide lumen which might be due to reduced cell height. Some pancreatic cells were multinucleated. Ultrastructural examination of fetal pancreas of both aspartame-treated groups confirmed our light microscopic findings. It showed loss of pyramidal appearance of pancreatic acinar cells. Dilated rough endoplasmic reticulum, multiple vacuoles, destructed mitochondria, and manifest reduction of zymogen granules were seen in the cytoplasm.
El-Gamal and Ghafeer  have found similar results on adult pancreas when 250 mg/kg/day aspartame was used once daily for 6 months in adults rats. They found that aspartame led to pancreatic stimulation evidenced by the presence of binucleated acinar cells, prominent nucleoli, an apparent decrease in zymogen granules, and cytoplasmic vacuolation. Ultrastructurally, they demonstrated the dilatation of rough endoplasmic reticulum and depletion of secretory granules in acinar cells. On the other hand, Leme and Azoubel  found that minor diameter of pancreatic acinar cell nuclei was the only significant parameter to present for the group treated with aspartame solution at ambient temperature. The results of the present study regarding cytoplasmic vacuolization of pancreatic acinar cells came in accordance with Abdallah , who found a slight hydropic degeneration in liver hepatocytes after long-term administration of 100 mg/kg body weight of aspartame for 14 weeks.
Some researchers found similar results on the architecture of other organs. Portela et al.  found that orogastric administration of aspartame induced significant reduction in kariometric parameters of fetal hepatocytes nuclei compared with the control group. Abd Elfatah et al.  found moderate to marked degeneration of hepatic parenchyma after orogastic administration of aspartame during the gestational period. Martins and Azoubel  found a statistically significant increased cell volume and decreased numerical cell density in the fetal kidneys of rats whose mothers administered aspartame during gestation. Mohammed et al.  found obvious morphological changes of the submandibular gland as it lost its normal acinar architecture.
In this study, the islets were hardly demonstrated. They appeared having irregular outlines and disorganized. The apparent reduced intensity of insulin immunoreactivity in β cells in groups II and III was noticed as compared with the control group. This result is against that was found by El-Gamal and Ghafeer  who mentioned that chronic administration of aspartame leads to increased insulin secretion from β cells.
Thus, our experiment showed histopathological and ultrastructural changes of both exocrine and endocrine fetal pancreas in aspartame-treated rats. These changes were dose related in all cases as the high-dose-treated rats appeared with more aggravated lesions than the low-dose-treated ones.
| Conclusion|| |
The use of aspartame during pregnancy could cause harmful effects to pregnant rats and their offspring as evidenced by the reduction of both fetal and placental weights and diminished umbilical cord length, and histopathological changes in fetal pancreas. It was also concluded that the effects of aspartame are dose related.
- The use of aspartame either as a sweetener or as an ingredient of manufactured food must be prohibited during pregnancy even within the acceptable daily intake.
- Further researches are needed to study the effects of aspartame during different periods of gestation and to study its effects on other fetal organs.
The authors thank all participants who helped with the study and express their deep sorrow for the loss of Professor Dr Abdel-Rahman Abu-El-Enain. He was such a great person who touched so many lives with his knowledge, kindness, and generosity.
The manuscript has been read and approved by all the authors and each author believes that the manuscript represents honest work.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Hayes C. The effect of non-cariogenic sweeteners on the prevention of dental caries: a review of the evidence. J Dent Educ 2001; 65:1106–1109.
Fitch C, Keim KS. Position of the Academy of Nutrition and Dietetics: use of nutritive and nonnutritive sweeteners. J Acad Nutr Diet 2012; 112:739–758.
Miller PE, Perez V. Low-calorie sweeteners and body weight and composition: a meta-analysis of randomized controlled trials and prospective cohort studies. Am J Clin Nutr 2014; 100:765–777.
Anton SD, Martin CK, Han H, Coulon S, Cefalu WT, Geiselman P et al.
Effects of stevia, aspartame and sucrose on food intake, satiety and postprandial glucose and insulin levels. Appetite 2010; 55:37–43.
Soffritti M, Belpoggi F, Manservigi M, Tibaldi E, Lauriola M, Falcioni L et al.
Aspartame administered in feed, beginning prenatally through life span, induces cancers of the liver and lung in male Swiss mice. Am J Ind Med 2010; 53:1197–1206.
Bawazir AE, Bokhary LE. Study the chronic effect of aspartame on some neurotransmitters content and histological structure of cerebellar cortex in male albino rats. Adv Environ Biol 2015; 9:19–25.
Trocho C, Pardo R, Rafecas I, Virgili J, Remesar X, Fernández-Lópes JA et al.
Formaldehyde derived from dietary aspartame binds to tissue components in vivo. Life Sci 1998; 63:337–349.
Haldorsson TI, Strom M, Petersen SB, Olsen SF. Intake of artificially sweetened soft drinks and risk of preterm delivery: a prospective cohort study in 59,334 Damsh pregnant women. Am J Clin Nutr 2010; 92:626–633.
Alkafafy M, Ibrahim Z, Ahmed M, El-Shazly S. Impact of aspartame and saccharin on the rat liver: Biochemical, molecular, and histological approach. Int J Immunopathol Pharmacol 2015; 28:247–255.
Portela GS, Azoubel R, Batigalia F. Effects of aspartame on maternal-fetal and placental weights, length of umbilical cord and fetal liver: A Kariometric experimental study. Int J Morphol 2007; 25:549–554.
Abd Elfatah AA, Ghaly IS, Hanafy SM. Cytotoxic effect of aspartame (diet sweet) on the histological and genetic structures of female albino rats and their offspring. Pak J Biol Sci 2012; 15:904–918.
Magnuson BA, Burdock GA, Doull J, Kroes RM, Marsh GM, Pariza MW et al.
Aspartame: a safety evaluation based on current use levels, regulations, and toxicological and epidemiological studies. Crit Rev Toxicol 2007; 37:629–727.
Martins MRI, Azoubel R. Effects of aspartame on fetal kidney: a morphometry and stereological study. Int J Morphol 2007; 25:689–694.
Rogers PJ, Fleming HC, Blundell JE. Aspartame ingested without tasting inhibits hunger & food intake. Physiol Behav 1990 47:1239–1243.
Hall WL, Millward DJ, Rogers PJ, Morgan LM. Physiology mechanisms mediating aspartame-induced satiety. Physiol Behav 2003; 78:557–562.
Beck B, Burlet A, Max JP, Sricker-Krongrad A. Effects of long-term ingestion of aspartame on hypothalamic neuropeptide Y, plasma leptin and body weight gain and composition. Physiol Behav 2002; 75:41–47.
Yang Q. Gain weight by going diet? Artificial sweeteners and the neurobiology of sugar cravings. Yale J Biol Med 2010; 83:101–108.
Leme LF, Azoubel R. Effects of aspartame on the exocrine pancreas of rat fetuses. Int J Morphol 2006; 24:679–684.
European food safety authority (EFSA). Scientific opinion on the re-evaluation of aspartame (E951) as a food additive. EFSA J 2013; 11:3496.
Hay WW. The Placenta. Not just a conduit for maternal fuels. Diabetes 1991; 40:44–50.
Ruangvutilert P, Titapant V, Kerdphoo V. Placental ratio and fetal growth pattern. J Med Assoc Thai 2002; 85:488–495.
Cetin I, Taricco E. Clinical causes and aspects of placental insufficiency. In: Burton GJ, Barker DJP, Moffett A, Thornburg K, editors. The placenta and human developmental programming. 1st ed. Cambridge: Cambridge University Press 2011. pp. 114–125.
Moore K, Persaud T, Torchia M. Before we are born: essentials of embryology and birth defects [chapter 8]. Ninth ed. Philadelphia,PA: Elsevier/Saunders 2016. pp. 71–89.
Suez J, Korem T, Zeevi D, Zilberman-Schapira G, Thaiss CA, Maza O et al.
Artificial sweeteners induce glucose intolerance by altering the gut microbiota. Nature 2014; 514:181–186.
Mohammed SS, El-Sakhawy MA, Sherif H, Shredah M. Effect of aspartame on submandibular salivary glands of adult male albino rats. Life Sci J 2015; 12:44–50.
Khidr BM, El-Sokkary GH, Saleh SMM. Study on morphological changes induced by aspartame on liver of normal and diabetic male albino rats. J Histol Histopathol 2017; 4:1.
Bacon BR, Britton RS. Hepatic injury in chronic iron overload. Role of lipid peroxidation. Chem Biol Interact 1989; 70:183–226.
Iman MM. Effect of aspartame on some oxidative stress parameters in liver and kidney of rats. Afr J Pharm Pharmacol 2011; 5:678–682.
El −Gamal DA, Ghafeer HH. Histological changes in adult rat pancreas upon chronic administration of aspartame. Egypt J Histol 2012; 35:883–891.
Abdallah IZA. Physiological changes induced by long term administration of saccharin compared with aspartame to male albino rats. Egypt J Hosp Med 2002; 8:70–81.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3], [Table 4]