Impact of Diagnostic X-Ray Exposure on Blood Vessel Integrity in Albino Rats: An Electron Microscopy Analysis

Sameh Fawzy Elsonbaty¹, Mohammad Chand Jamali²*, Wadah M.A Khogali¹, Hend Hamed¹,

Mohamed Abdelfatah Abdelmounim Mohamed², Maxime Merheb³, Adham Elsonbaty⁴,

Rehab Mohd Jamali⁵

¹Department of Health & Laboratory Sciences, College of Medical and Health Sciences,

Liwa University, Abu Dhabi, United Arab Emirates.

²Department of Health & Laboratory Sciences, College of Medical and Health Sciences,

Liwa University, Al Ain, Abu Dhabi, United Arab Emirates.

³College of Health Sciences, Dubai Medical University, Dubai, United Arab Emirates

⁴Faculty of Medicine, October 6 University, Egypt.

⁵College of Medicine, University of Sharjah, Sharjah, United Arab Emirates.

*Corresponding Author E-mail: sameh.elsonbaty@lu.ac.ae, mjamali68@gmail.com,

maxime.merheb@dmu.ae,wadah.khogali@lu.ac.ae,hend.ahmad@lu.ac.ae,

mohammed.abdelfatah@lu.ac.ae,samsonbaty@gmail.com,rehab.jamali@gmail.com

ABSTRACT:

The growing reliance on diagnostic X-ray imaging has raised concerns regarding its potential effects on cellular and tissue health. This research explores the impact of diagnostic X-ray exposure on the structural integrity of blood vessels, with a special focus on the aortic wall in rats. We analyzed the aorta's ultrastructural changes using electron microscopy by comparing patients exposed to X-rays to a control group. The findings revealed significant alterations in the aortic wall, including changes in the smooth muscle cells, elastic fibres, basal lamina, collagen deposition, and intracellular structures. These findings suggest that diagnostic X-ray exposure may disrupt the structural components of the aorta, potentially contributing to long-term vascular damage.

KEYWORDS: X-ray, Blood Vessel Integrity, Microscopic Analysis, Smooth Muscle Cells, Elastic Fibres.

INTRODUCTION:

Diagnostic X-rays are extensively utilized in modern medicine as a reliable imaging modality. While their utility in diagnosing and monitoring various diseases is unquestionable, concerns have been raised regarding the potential long-term effects of X-ray exposure on biological tissues. One area that remains insufficiently explored is the impact of X-ray exposure on vascular integrity¹.

As the body’s largest artery, the aorta plays a pivotal role in the systemic circulation. Thus, any compromise in its structural integrity could significantly affect cardiovascular function². This study aims to evaluate the effects of diagnostic X-ray exposure on the structural integrity of the aortic wall using electron microscopy.

The discovery and early use of ionizing radiation for medical purposes in the late 19th and early 20th centuries revealed its therapeutic potential and significant risks, particularly to the vascular system. Early clinical and pathological observations identified radiation-induced damage to capillaries, characterized by endothelial cell swelling, haemorrhage, and vessel dilation. Endothelial cells were identified as highly radiosensitive, especially in capillaries, compared to larger vessels. Over time, it was recognized that radiation could cause sustained and progressive vascular damage, with late effects such as endothelial loss, vessel thickening, inflammation, and fibrosis contributing to tissue necrosis and ischemia³.

Epidemiological studies, such as those on atomic bomb survivors, linked radiation exposure to increased risks of cardiovascular diseases⁴. Radiation-induced vascular injury is mediated through direct and indirect mechanisms (e.g., oxidative stress and DNA damage) (e.g., immune activation and iron release). These disruptions lead to long-term inflammation and tissue structural damage⁵.

Experimental models, including in vivo dye leakage assays and electron microscopy, provide evidence of multi-phase changes in vascular permeability following radiation exposure⁸. In vitro studies revealed key molecular pathways regulating endothelial barrier integrity, including roles of tight and adherens junction proteins (e.g., VE-cadherin, occludin, claudin), the actin cytoskeleton, and GTPases like RhoA and Rac-1. Transcytosis via caveolae also plays a role in endothelial transport, although its interplay with paracellular pathways remains unclear⁶.

Advanced 3D culture systems and co-culture models enhance the physiological relevance of in vitro vascular models, allowing a more accurate understanding of radiation’s impact on endothelial cells^7,9.

MATERIALS AND METHODS:

Animal Model:

In this experiment, 30 adult rats were used, selecting the samples randomly in two groups; control (n=15) and experimental group (n=15) that was subjected to X-ray radiation. All the animals were kept under usual laboratory conditions where both food and water were available freely. The experimental population was subjected to diagnostic X-ray imaging with a normal clinical dose, how a control was not subjected to the action¹⁰.

Typical Diagnostic X-ray parameters for Soft Tissue Imaging in Small Animals were performed with the following parameters. Typical diagnostic X-ray parameters for soft tissue imaging in small animals are well established. The kilovoltage peak (kVp) is 60 kVp, offering sufficient penetration while maintaining soft tissue contrast¹¹.

The tube current (mA) is generally 200 mA, which allows for reduced exposure time and minimizes motion blur during imaging¹².

Exposure times are 0.05 seconds, which is especially important for small animals with high respiration rates¹³.

The milliampere-seconds (mAs) used in imaging is 10 mAs, balancing image clarity and radiation safety¹⁴.

Electron Microscopy Analysis:

All rat subjects were then killed after being exposed to X-ray, and their aorta extracted to undergo further studies. 2.5 percent glutaraldehyde fixed tissue specimens were processed to make them electron microscopic15. Ultrathin slices (70nm) of the aorta were sectioned with a Reichert Ultracut E ultramicrotome, counterstained with uranyl acetate polyvinyl alcohol and lead citrate and viewed on a transmission electron microscope (TEM) at different magnifications.

RESULTS:

The control group exhibited typical ultrastructural features of the aortic wall. The aortic wall comprised smooth muscle cells interspersed with elastic fibres (Figure 1(B)). The basal lamina beneath the endothelial cells was thin and well-defined (Figure 1(C)). Myofilaments were seen within smooth muscle cells, contributing to the contractile properties of the vessel wall (Figure 1(A)). Smooth muscle cells, elastic fibres, and the basal lamina appeared well-maintained in the control group.

Figure 2: We have gotten an electron microscopic picture of the wall of the aorta in the Rat experimental group, and there were elastic fibers (Orange arrow) and increase collagen fibers (Blue Arrow) shown in picture

A. Original Magnification X3000; the picture B shows that the basal lamina under Endothelial cells have been thickened and folded (White line). Original Magnification X8000. In picture C, the smooth muscles have been shown to have numerous and dilated vesicles in their cytoplasm and have a lot of X5000 Original Magnification (Experimental group, had an increase in the number of elastic fibers and collagen fibers between the cells of smooth muscles); (E) Electron Microscopic image on the wall of the aorta of rat experimental group shows that the cytoplasmic fibrils and collagen fibers (Blue Arrow) are increased between the smooth muscle cells. original magnification 5000 (experimental group, with the presence of fibrils and collagen fibers between smooth-muscle cells, showing more abundance).

Table 1: It is showing the Histological differences shown by the electron microscope between the control and the experimental groups.

Parameter	Control Group	Experimental Group (X-ray Exposure)
Aortic Wall Structure	Well-maintained, normal structure (Figure. 1(A))	Structural alterations observed, including thickened areas (Figure. 2(A), 2(B))
Smooth Muscle Cells	Typical morphology with myofilaments (Figure. 1(A))	Dilated cytoplasmic vesicles, increased myofilaments (Figure. 2(C))
Elastic Fibers	Present between smooth muscle cells (Figure. 1(B))	Increased collagen fibers and elastic fibers between smooth muscle (Figure. 2(A), 2(D))
Collagen Fibers	Sparse and evenly distributed (Figure. 1(B))	Increased collagen deposition between smooth muscle cells (Figure. 2(A), 2(E))
Basal Lamina	Thin and well-defined beneath endothelial cells (Figure. 1(C))	Thickened and folded beneath endothelial cells (Figure. 2(B))
Cytoplasmic Vesicles	Few or none observed	Numerous dilated cytoplasmic vesicles containing glycogen particles (Figure. 2(C))
Rough Endoplasmic Reticulum (RER)	Normal, consistent presence	Increased rough endoplasmic reticulum (RER) in smooth muscle cells (Figure. 2(C))
Glycogen Particles	Normal	Increased glycogen particles within dilated cytoplasmic vesicles (Figure. 2(C))
Myofilament Density	Normal myofilament presence in smooth muscle cells (Figure. 1(A))	Increased number of myofilaments in smooth muscle cells (Figure. 2(C))
General Tissue Integrity	Normal, no signs of damage	Signs of stress response, indicating potential for long-term damage (Figure. 2(A), 2(B))

This table highlights the major structural differences observed between the control and experimental groups based on the electron microscopy analysis. It demonstrates that rats exposed to diagnostic x-rays exhibit several alterations in the aortic wall, smooth muscle cells, collagen fibers, and other intracellular components, indicating a cellular response to radiation-induced stress

DISCUSSION:

Electron microscopy conducted in the control group depicted on the aortic wall characteristic ultrastructural characteristics with all important structural components being normal and being in good status. The structure of the aorta wall was a mixture of smooth muscle cells containing elastic fibers that were instrumental in the flexibility and strength of the vessel (Figure. 1(B)). These fibers give structural strength to aorta to support the pulse driven circulation of blood and the possibility of smooth distribution of blood pressure to the whole body. The smooth muscle cells were arranged in layers, which makes its contribution towards the contractile characteristics important to regulate vascular tone as well as blood flow. The basement membrane below the endothelial cells was thin and well-continuous (Figure. 1(C)), which is an indication of good vascular barrier and functions by maintaining nutrition exchange and endothelial functionality. Myofilaments in the smooth muscle cell were found (Figure. 1(A)) which means the aorta has a contractility. The collection of these myofilaments is very key in the vasoconstriction and vasodilation of the blood vessels in reaction of change of blood pressure. The integrity and organization of the smooth muscle cells as well as the elastic fibres, and the basal lamina were all well-maintained in the control group thus showing evidence of a healthy and functional vascular system. Moreover, no indication of any damages or structural impairment has been observed on the control aorta, indicating the tissue has not been exposed to foreign stressor or damages¹⁷.

Conversely, there were clear changes in the aortic walls of rats that were exposed to X-ray that was used in taking diagnostic tests on the rat. There was enhancement of collagen fibre between the smooth muscle cells as viewed in electron microscopy (Figure. 2(D)). Collagen is a major extracellular matrix protein that furnishes the sturdiness of tissues, and in the case that there is excess collagen deposit it can lead to tissue fibrosis which may subsequently impair the vascular performance. The augmented collagen between the smooth muscle cells shows a possibility of the X-ray exposure causing repair-type reaction. Nevertheless, this is a maladaptive response that may result in stiffness of the vessels in the setting of time. This variability in the quantity of deposited collagen could compromise the normal elasticity of the aorta, which in turn could cause a decreased compliance of the vessel, as well as predispose to the appearance of cardiovascular diseases including, hypertension or atherosclerosis.

The experimental group featured the profound thickening and folding of the basal lamina (Figure. 2(B)), which implies that the X-ray exposure might also trigger the adaptive process in the aortic wall. Nevertheless, this change may affect normal cellular permeability and signalling processes of the vessel wall, which may result in the endothelial dysfunction and increase vulnerability of the aorta to damage. The basal lamina thickening may also tend to decrease the capacity of endothelial cells to interact efficiently with the smooth muscle cells, to disrupt homeostasis of the blood vessel and worsen its capability to control the blood flow and pressure.

Moreover, in the experimental group, smooth muscle cells contain many dilated, cytoplasmic vesicles that were associated with material glycogen particles (Figure. 2(C)). The existence of these vesicles shows that there has been an imbalance in the homeostasis of the cells because due to metabolic stress, the smooth muscle cells may be facing. The cells may also store excess glucose which in turn may lead to their injury through glycogen build-up in the cells in bid to result in energy production in response to stress. Nevertheless, the presence of too much glycogen shows that there is a defect in the metabolic processes, and this may lead to the disturbances in the normal functioning of cells. The foresence of dilated vesicles can also indicate the disruption of an ordinary trafficking and storage of cellular constituents which in turn can impair overall functionality of the smooth muscle cells. Glycogen is normally stored in the form of small regulated quantities in healthy smooth muscle cells; the overloading with glycogen seen in the experimented group thus portrays an indicator that the cells are experiencing great stress due to the damage inflicted to them by radiation. Also, there was a pronounced increase in myofilaments in smooth muscle cells, a factor that resulted in a proportional increase in rough endoplasmic reticulum (RER) (Figure. 2(C)). The RER manufactures proteins especially those that relate to cell structure and functions. RER is benchmarked to elevated rates of protein synthesis in most cases which in turn may be an adaptation to damaged or stressed cells. Given the more abundance of myofilaments in the smooth muscle cells, it is clear that the cells could be trying to hex or remodel their contracting apparatus following the destructive effects of the X-ray exposure. Nevertheless, this compensatory reaction can prove to be long term maladaptive because, as shown in the myofilament replenishment, excessive buildup of these myofilaments might interfere with the normal smooth muscle cell functioning thereby leading to the loss of elasticity of the aortic wall and the resultant stiffness and rigidity.

The noticed increase in amount of both myofilaments and rough endoplasmic reticulum in the experimental group allows applying all the comments on the fact that smooth muscle cells actively reacted to the stress provided by radiation effects through the reasons of raising the production of proteins and rearranging its inner structure. This increased protein production can suggest an adaptive stress response, which however, might not be enough to recover fully the damage produced by the X-ray exposure. The change in composition of the cells (smooth muscle cells) and storage of intracellular contents like glycogen particles and myofilaments in the smooth muscle cell could eventually lead to pathology of the aorta which will fail in its regular capacity to ensure a normal flow and pressure of the blood.

These studies suggest that X-ray related to diagnostic radiation exposure has the potential of producing profound structural changes in the aortic wall especially in smooth muscle cells and extra cellular components of the matrix. The enlargement of collagen fibres, thickening of basal lamina, and the change in the morphology of smooth muscle cells underlines the possibility of the long-term deterioration of the aortic wall that would influence vascular health. The changes can weaken the aorta elasticity and compliance, which can result in the occurrence of a vascular disease like hypertension, atherosclerosis, or aneurysms. The results are indicative of the fact that the X-ray exposure triggers cell stress, which is manifested by means of glycogen storage intensification, accumulation of myofilaments, and rough endoplasmic proliferation of smooth muscle cells. Though these responses can serve to protect against tissue damage they can also reflect maladaptive repair processes that might reduce vascular functioning further. Other research is therefore suggested to be done in the future to understand the functional impact of such structural changes and the measures that can be adopted to eliminate the adverse impact of diagnostic X-rays on vascular tissues.

CONCLUSION:

The exposure of diagnostic X-rays causes a great structural change in the aortic wall such as deposition of collagens, thickening of basal lamina, and deposition of glycogen in smooth muscle cells. The changes would indicate a pathological healing process, which could result in decreased elasticity and compliance of the aorta that can present vascular undertakings like high blood pressure or hormonal failures like atherosclerosis. The article demonstrates the dangers of X-ray to the health of the blood vessels. It gives importance to the necessity of additional studies to examine the long-term functional consequences and prevention of the harm.

REFERENCE:

1. Yoon CW, Park HK, Rha JH. Yield of Screening Tests for Systemic Vasculitis in Young Adults with Ischemic Stroke. Eur Neurol. 2018; 80(5-6): 245-248. doi: 10.1159/000496373.

2. Berman M, et al. Co-culture systems and radiation biology: bridging in vitro and in vivo. Front Physiol. 2022; 13: 900235. doi:10.3389/fphys.2022.900235.

3. Ping Z, Peng Y, Lang H, Xinyong C, Zhiyi Z, Xiaocheng W, Hong Z, Liang S. Oxidative Stress in Radiation-Induced Cardiotoxicity. Oxid Med Cell Longev. 2020 Mar 1; 2020: 3579143. doi: 10.1155/2020/3579143.

4. Jahng JWS, Little MP, No HJ, Loo BW Jr, Wu JC. Consequences of ionizing radiation exposure to the cardiovascular system. Nat Rev Cardiol. 2024 Dec; 21(12): 880-898. doi: 10.1038/s41569-024-01056-4.

5. Harrison D, Lowe S, Patel H. In vivo models for studying radiation-induced vascular permeability and endothelial dysfunction. Radiat Res. 2020; 194(2): 168-176. doi:10.1667/RR2018.2.

6. Johnson LA, et al. Diagnostic X-rays and their impact on vascular integrity. Clin Imaging. 2018;52:12–19..

7. Ponce SB, Seldon C, Yanagihara TK, Horowitz D, Yu JB, Kachnic L, et al. Crazy busy: The blurring of personal and professional boundaries as a diversity, equity, and inclusion issue. Int J Radiat Oncol Biol Phys. 2023; 116(2): 276–9. http://dx.doi.org/10.1016/j.ijrobp.2023.01.010

8. Lavin LM. Radiography in veterinary technology. 5th ed. Elsevier; 2013

9. Lopez CL, Zhang Z, Shen C. Advanced 3D in vitro models to study radiation effects on endothelial cells: innovations and challenges. J Biomed Sci. 2021; 28: 75. doi:10.1186/s12929-021-00091-2.

10. Marin J, et al. Molecular mechanisms of endothelial barrier regulation after radiation. Cell Rep. 2019 Oct 22; 29(4): 1027-1037.

11. Huda W, Abrahams RB. Radiographic techniques, contrast, and noise in x-ray imaging. AJR Am J Roentgenol [Internet]. 2015; 204(2): W126-31. Available from: http://dx.doi.org/10.2214/AJR.14.13116

12. Potts PJ. X-RAY FLUORESCENCE AND EMISSION | wavelength dispersive X-ray fluorescence. In: Encyclopedia of Analytical Science. Elsevier; 2005. p. 419–29.

13. Fritz S, Chaitow L, Hymel GM. Review of pertinent anatomy and physiology. In: Clinical Massage in the Healthcare Setting. Elsevier; 2008. p. 140–95.

14. Watanabe M, et al. Protocol for electron microscopy of rat aortic tissue. Microscopy Techniques. 2015; 22(2): 115–121.

15. Wek RC, Jiang HY, Anthony TG. Coping with stress: eIF2 kinases and translational control. Biochem Soc Trans. 2006 Feb;34(Pt 1):7-11. doi: 10.1042/BST20060007.

16. Yang X, Zhang H, Wu L. The role of immune response in radiation-induced vascular damage: implications for therapy. J Clin Invest. 2023; 133(1): e154230. doi:10.1172/JCI154230.

17. Zhao W, Li Y, Sun H. Epidemiological evidence on radiation exposure and cardiovascular disease risk among atomic bomb survivors. Circ Res. 2022; 130(4): 498-510. doi:10.1161/CIRCRESAHA.121.318013.

Received on 20.04.2025 Revised on 18.08.2025

Accepted on 28.11.2025 Published on 16.03.2026

Available online from March 18, 2026

Research J. Pharmacy and Technology. 2026;19(3):1230-1234.

DOI: 10.52711/0974-360X.2026.00175

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.