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. 2024 Jul 29:15:1399719.
doi: 10.3389/fphar.2024.1399719. eCollection 2024.

Melatonin supplementation counteracts fiber loss in knee ligaments of diabetes-induced rats

Olga Adamska  1 , Artur Wnuk  2 , Agnieszka Kamińska  1 , Małgorzata Poniatowska  3 , Bartosz Maciąg  4 , Maciej Kamiński  5 , Artur Stolarczyk  4 , Maima Matin  6 , Atanas G Atanasov  6   7   8 , Marcin Łapiński  4 , Artur Jóźwik  6
Affiliations

Melatonin supplementation counteracts fiber loss in knee ligaments of diabetes-induced rats

Olga Adamska et al. Front Pharmacol. .

Abstract

Diabetes mellitus (DM) is a prevalent metabolic disease. The clinical impact of sustained hyperglycemia on ligament healing has not been well characterized. Diabetes is a known cause of macro-, microvascular, and diabetic ulcer healing difficulties among tissues. Therefore, we aimed to investigate the healing potential occurring in injured and healthy ligaments among diabetic and healthy individuals using a rat model. We hypothesize that DM may contribute to altering the knee medial collateral ligament (MCL), thus its morphology, biochemical fitness, and functionality. The study cohort consisted of 40 rats. The animals were randomized into four equal groups. Groups I and II (20 rats) received saline subcutaneously and served as controls. Groups III and IV (20 rats) were injected with a single dose of streptozotocin (STZ). All animals underwent surgery to cut the left tibial collateral ligament in the hind limb and suture it. The access site was sutured to create inflammation and study the regenerative capacities of animals with normal carbohydrate metabolism and pharmacologically induced diabetes. Each animal then underwent sham surgery to access and suture the right tibial collateral ligament in the hind limb without ligament intervention. After the animals had undergone surgeries, groups II and IV were given melatonin supplementation for 4 weeks. Rats with DM presented with more fibrosis and calcification of the MCL and decreased healing potential. Treatment with melatonin in diabetic rats mitigated alterations and improved the antioxidant status of ligaments from the diabetic group.

Keywords: diabetes ligaments; diabetes mellitus; ligaments alteration; ligaments alteration in diabetes mellitus; orthopedic complications.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
The right image (Leong et al., 2020) shows the animal model knee joint surgical site during the ligament preparation procedure.
FIGURE 2
FIGURE 2
The left image (Longo et al., 2009) shows a sutured surgical wound in the experimental animal model after the intervention in the knee joint ligament.
FIGURE 3
FIGURE 3
Schematic chronologic representation of the study design.
FIGURE 4
FIGURE 4
Histology-stained sections (longitudinal dissection) from the four groups: normoglycemia and diabetes mellitus, with and without melatonin supplementation. Each column contains light microscopy images obtained from a single representative model and from the same section. A scale bar is shown on the left bottom corner (100 µm). Each row is indicated for one model from which the tissue was retrieved. Sample imaging was used to assess the status of the tissue. The right MCL comes from the extremity on which the sham procedure was performed, and the left MCL comes from the extremity on which a ligament incision was performed. Quantitative results are presented in Tables 3, 4.
FIGURE 5
FIGURE 5
Ligament homogenate TAS, TOS, ROS levels, and OSI rates of all groups. Abbreviations: Control, control group; Control + M, control group + melatonin supplementation; DM, diabetic group; DM + M, diabetic group + melatonin supplementation; OSI, oxidative stress index; ROS, reactive oxygen species; TAS, total antioxidant status; TOS, total oxidant status. Abundance of groups: Control: n = 10; Control + M: n = 10; DM: n = 10; DM + M: n = 10. Data are expressed as mean ± SD. a p < 0.05 versus the control group; b p < 0.05 versus the diabetic group.
FIGURE 6
FIGURE 6
Levels of MDA and GSH in the various ligament homogenates. Abbreviations: Control, control group; Control + M, control group + melatonin supplementation; DM, diabetic group; DM + M, diabetic group + melatonin supplementation; MDA, malondialdehyde; GSH, glutathione. Data are expressed as mean ± SD. a p < 0.05 versus the control group; b p < 0.05 versus the diabetic group.
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Grants and funding

The authors declare that financial support was received for the research, authorship, and/or publication of this article. The authors are thankful for the support from the Project: POPUL/SP/0120/2023/01.

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