Morphometric parameters of keratinocyte proliferation and apoptosis following ascorbic acid administration in radiation-induced skin injury

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Abstract

BACKGROUND: Available publications describe the features of radiation-induced skin injury and fibrosis caused by various types of ionizing radiation. Compared with other forms of ionizing radiation, electrons exhibit relatively low cytotoxicity to healthy organs; however, their side effects remain insufficiently explored. One of the key objectives is to develop protective strategies for the epidermis and dermis against electron-induced cytotoxicity during the treatment of malignant tumors and percutaneous tumor exposure.

AIM: The work aimed to perform a morphometric assessment of keratinocyte proliferation and apoptosis following ascorbic acid administration in an experimental model of radiation-induced skin injury.

METHODS: A single-center, prospective, controlled study was conducted. The study object was skin fragments from the outer surface of the thigh of male Wistar rats (aged 9–10 weeks, weighing 220 ± 20 g). Animals (n = 50) were randomly divided into four experimental groups: group 1, controls (n = 20); group 2, local electron irradiation at a dose of 40 Gy (n = 10); group 3, administration of ascorbic acid (intraperitoneally, 50 mg/kg) prior to local electron irradiation at a dose of 40 Gy (n = 10); group 4, administration of ascorbic acid without irradiation (n = 10). After 10 days, skin samples from the irradiated area were harvested for histological and immunohistochemical analysis (using Ki-67 and caspase-3 antibodies).

RESULTS: Ten days after local electron irradiation with the NOVAC-11 linear accelerator (Italy) at a dose of 40 Gy, the exposed skin areas showed signs of radiation-induced injury: moist desquamation, edema, partial desquamation of the basal epidermal layer, formation of microcavities at the dermoepidermal junction, damage to most sebaceous glands, and imbalance in malondialdehyde content and superoxide dismutase activity. Results of Ki-67 and caspase-3 expression analysis revealed reduced keratinocyte proliferative activity and increased apoptosis. However, in animals pretreated with ascorbic acid, the levels of keratinocyte proliferation and apoptosis, as well as epidermal thickness, were comparable to those in the control group.

CONCLUSION: The findings demonstrate the high radioprotective efficacy of ascorbic acid for the epidermis under local electron irradiation at a dose of 40 Gy. Ascorbic acid prevents radiation-induced apoptotic death of keratinocytes by reducing oxidative damage caused by free radicals and by inducing superoxide dismutase expression.

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About the authors

Grigory A. Demyashkin

National Medical Research Radiological Center of the Ministry of Health of the Russian Federation; The First Sechenov Moscow State Medical University

Author for correspondence.
Email: dr.dga@mail.ru
ORCID iD: 0000-0001-8447-2600
SPIN-code: 5157-0177

Dr. Sci. (Medicine)

Russian Federation, Moscow; Moscow

Matvey A. Vadyukhin

The First Sechenov Moscow State Medical University

Email: vma20@mail.ru
ORCID iD: 0000-0002-6235-1020
SPIN-code: 9485-7722
Russian Federation, Moscow

Anna K. Marukyan

The First Sechenov Moscow State Medical University

Email: Marukyan87@mail.ru
ORCID iD: 0000-0002-4619-7385
SPIN-code: 4320-6507
Russian Federation, Moscow

Susanna V. Saakyan

The First Sechenov Moscow State Medical University

Email: drsaakyan@icloud.com
ORCID iD: 0000-0001-8606-8716
SPIN-code: 7742-1420
Russian Federation, Moscow

Elza B. Karakaeva

The First Sechenov Moscow State Medical University

Email: kchr09@mail.ru
ORCID iD: 0000-0001-9833-3433
SPIN-code: 8221-3003
Russian Federation, Moscow

Sergey N. Koryakin

National Medical Research Radiological Center of the Ministry of Health of the Russian Federation

Email: korsernic@mail.ru
ORCID iD: 0000-0003-0128-4538
SPIN-code: 8153-5789

Cand. Sci. (Biology)

Russian Federation, Moscow

Elena Y. Shapovalova

V.I. Vernadsky Crimean Federal University

Email: shapovalova_l@mail.ru
ORCID iD: 0000-0003-2544-7696
SPIN-code: 5321-1246

Dr. Sci. (Medicine), Professor

Russian Federation, Simferopol

Alexey A. Kantorovich

The First Sechenov Moscow State Medical University

Email: w.q.989@mail.ru
ORCID iD: 0009-0007-9370-3600
Russian Federation, Moscow

Anastasiia S. Andrievskikh

The First Sechenov Moscow State Medical University

Email: Andrievskikh2002@mail.ru
ORCID iD: 0009-0007-1787-5910
Russian Federation, Moscow

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2. Fig. 1. Skin fragments from rats on day 10 of the experiment: a, control group (group 1); b, after a single electron irradiation at a dose of 40 Gy (group 2); c, after ascorbic acid administration prior to irradiation (group 3). Hematoxylin and eosin staining; magnification ×100; scale bar 50 µm.

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3. Fig. 2. Skin fragments on day 10 of the experiment, immunohistochemical analysis with Mayer’s hematoxylin counterstaining: a–c, Ki-67 antibodies; d–f, caspase-3 antibodies; a, d, control group (1); b, e, after a single electron irradiation at a dose of 40 Gy (group 2); c, f, after ascorbic acid administration prior to irradiation (group 3). Magnification ×400; scale bar 50 µm.

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