Research Article

Anti-Hypothyroidism and Cardiac Effect of Liotrix and PLGA Nanoparticles Loading Liotrix on Induced Hypothyroidism in Rats

Israa Jawad Jaber, Huda F. Hasan*

Department of Physiology, Biochemistry and Pharmacology, College of Veterinary Medicine, University of Baghdad, Iraq.

Received | April 05, 2025; Accepted | May 18, 2025; Published | July 23, 2025

*Correspondence | Huda F. Hasan, Department of Physiology, Biochemistry and Pharmacology, College of Veterinary Medicine, University of Baghdad, Iraq; Email: [email protected]

Citation | Jaber IJ, Hasan HF (2025). Anti-hypothyroidism and cardiac effect of liotrix and plga nanoparticles loading liotrix on induced hypothyroidism in rats. Adv. Anim. Vet. Sci. 13(8): 1746-1755.

DOI | https://dx.doi.org/10.17582/journal.aavs/2025/13.8.1746.1755

ISSN (Online) | 2307-8316; ISSN (Print) | 2309-3331

Copyright: 2025 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

Poly (lactic-co-glycolic acid) (PLGA) is a biodegradable and biocompatible copolymer composed of lactic acid and glycolic acid monomers. The ratio between these monomers can be adjusted, significantly influencing the physicochemical properties of the copolymer, such as degradation rate and mechanical strength. Due to its excellent safety profile, PLGA has been approved by the U.S. Food-Drug Administration (FDA) for a variety of therapeutic applications, including drug delivery systems (Makadia and Siegel, 2022). Recent advancements in nanotechnology have enabled the use of PLGA-based nanoparticles for controlled and targeted drug delivery. These nanoparticles provide significant advantages, including enhanced bioavailability, prolonged drug release, reduced dosing frequency, and minimized systemic side effects (Kumari et al., 2020; Wang et al., 2023). PLGA nanoparticles have been successfully applied for the delivery of anticancer agents, antihypertensives, hormones, vitamins, and biomolecules such as peptides and antibodies. Thyroid disorders, particularly hypothyroidism, represent a major public health concern, affecting nearly all body systems, especially the cardiovascular and renal systems. Hypothyroidism is defined by the insufficient production of thyroid hormones, resulting in decreased metabolic activity and clinical symptoms like fatigue, weight gain, and bradycardia (Chiovato et al., 2019). Liotrix is a synthetic combination of thyroxine (T4) and triiodothyronine (T3) in a 4:1 ratio and is used to restore normal thyroid hormone levels. However, traditional formulations of Liotrix may suffer from limitations including poor bioavailability and side effects due to fluctuations in hormone levels (Al Rubaee et al., 2020). The integration of Liotrix into a PLGA nanoparticle delivery system offers a novel therapeutic approach that could improve treatment efficacy and reduce adverse effects. By enabling sustained and targeted hormone release, this formulation may help stabilize thyroid function more effectively than conventional therapies. This study aimed to prepare and characterize PLGA nanoparticles loaded with Liotrix and to evaluate their therapeutic efficacy in a hypothyroid rat model. Specific objectives include:

• Formulation and physicochemical characterization of PLGA-Liotrix nanoparticles.
• Evaluation of therapeutic outcomes in hypothyroid-induced rats.
• Comparison between conventional Liotrix and PLGA-loaded Liotrix with respect to thyroid function, lipid profile, antioxidant status, and histopathological changes.

MATERIALS AND METHODS

Preparation of PLGA–Liotrix Nanoparticles

PLGA–Liotrix nanoparticles were prepared using the nanoprecipitation method with minor modifications, as described by Ahmed and Hasan (2022) and Crucho and Barros (2017). All procedures involving solvents and polymer handling were performed in a chemical fume hood. Briefly, 40 mg of poly (lactic-co-glycolic acid) (PLGA) was dissolved in 1 mL of dimethyl sulfoxide (DMSO) in a glass test tube, which was then sealed with aluminum foil and parafilm. The solution was incubated overnight, followed by vortexing at high speed (~10 minutes) to ensure complete dissolution. Liotrix (1.6 mg/mL) was then added to the PLGA solution, and the mixture was vortexed at 800 rpm for 5 minutes to achieve homogeneous emulsification. The emulsified solution was transferred to a 20 mL beaker, placed in an ice bath, and sonicated for 9 minutes at 50% amplitude with 15-second on/off pulses. A 0.03% w/v solution of Vitamin E-TPGS was prepared overnight and used as the aqueous phase. The emulsified polymer was then introduced to the aqueous phase under stirring, allowing the solvent to evaporate with the beaker covered in aluminum foil. The resulting suspension was filtered and centrifuged at 14,000 rpm at 4°C. The supernatant was discarded, and the pellet was washed 2–3 times with distilled water. Empty nanoparticles were prepared using the same method, excluding the drug.

Characterization of Liotrix-Loaded PLGA Nanoparticles

Nanoparticles were characterized by measuring particle size and zeta potential, and visualized via transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Drug encapsulation efficiency (%EE) and drug loading (%DL) were determined using UV-Visible spectrophotometry at 243 nm. A standard calibration curve was generated using Liotrix concentrations ranging from 2 to 100 mg/mL, with an R² value of 0.9931. Nanoparticle yield (NY), encapsulation efficiency (EE), and drug loading (DL) were calculated based on methods by Ahmed and Hasan, (2022), Madani et al. (2018) and Jasim et al. (2019).

Induction of Hypothyroidism

Hypothyroidism was induced in rats by administering 1 mg/mL propylthiouracil (PTU) in drinking water for 30 days, following the protocol by Zhang et al. (2022) and Hamed et al. (2024).

Determination of Effective Dose of Liotrix-Loaded Nanoparticles

Thirty-five rats were randomly assigned into seven groups (n=5 per group) to determine the effective dose of PLGA–Liotrix nanoparticles. Doses tested included 1, 1.5, 2.5, 3.0, and 3.5 mg/kg., in addition to positive and negative control groups. After 30 days of treatment, T4 and TSH levels were measured. The dose-response relationship was analyzed using the linear regression equation Y = m + bX to identify the optimal effective dose, as described by Majeed et al. (2022).

Experimental Design for Comparison Between Liotrix and PLGA–Liotrix Treatments

Fifty rats were divided into five groups (n=10 per group) for a 30-day study. One group served as the negative control, while all others were induced with hypothyroidism. Treatments included: Liotrix alone (2.65 mg/kg), PLGA–Liotrix (2.33 mg/kg), and PLGA alone (1.1 mg/kg).

Biochemical Test

Serum lipid profiles (triglycerides, HDL, LDL, oxidized LDL, total cholesterol) were measured using enzymatic kits from Linear Biolaboratory, as per Baggio et al. (1988). CRP concentrations were determined via ELISA. Thyroid hormones (T3, T4, TSH) were measured using commercial RIA kits. Homocysteine and reactive oxygen species (ROS) levels were quantified using SunLong Biotech kits, while total antioxidant capacity was assessed using Cohesion Bioscience kits. Thrombotic factor XII levels were measured using a specific ELISA kit.

Histopathological Study

At the end period of treatment, the animals were1 anesthetized by using a ketamine (80 mg/kg) and xylazine (12 mg/kg) cocktail at a dose of 0.1 ml/100 gm body weight (BW) study on the blood vessels (Heart and Thyroid gland) were prepared on microscopic slides. The samples, putting in a clean container on formalin solution10%. The procedure of (Li et al., 2018).

Statistical Analysis

The statistical analysis on the data from this encounter was conducted utilizing a completely randomized design (CRD), it was the comparison between the averages at the level of probability of (P<0.05) to test the significant differences between the averages of traits and applying the statistical program SAS version 2010 by using one and two ways, T test, and LSD (Wayne and Daniel, 2010).

RESULTS AND DISCUSSION

Maximum Wavelength (λmax) and Liotrix Loading Determination

Liotrix’s maximum absorption wavelength (λmax) was determined by dissolving PLGA-Liotrix in DMSO (1 mg/mL) and scanning from 200–800 nm (Figure 1). The recorded λmax was 243.00 nm with an absorbance of 2.605, which differs slightly from the literature value of 231 nm (Huijun et al., 2014), possibly due to polymer-drug interactions in the formulation. This λmax was used to construct the standard calibration curve.

 

 

 

Morphological and Physicochemical Properties of PLGA-Liotrix Nanoparticles

Transmission electron microscopy (TEM) revealed spherical nanoparticles with monodisperse size distribution ranging from 17 to 100 nm (Figure 2A). Scanning electron microscopy (SEM), performed at a working distance of 13.4 mm and magnification of 110 kx, confirmed this morphology, with a particle size range of 39.38–120.6 nm (Figure 2B). The zeta potential was –28.6 mV, indicating good colloidal stability, with a mobility of –0.000206 cm²/Vs (Figure 4A).

Encapsulation efficiency (EE), drug loading (DL), and nanoparticle yield (NY) were calculated using absorbance values from the standard curve equation (Y = 0.0019X + 0.0232) (Figure 3B) and measured λmax. The results showed (Figure 3C): NY = 6.386%, EE = 37.5%, and DL = 3.25% at λmax = 243.00 nm (Figure 2B). These moderate values can be attributed to the nature of the nanoprecipitation technique, which favors hydrophobic drug encapsulation. Literature supports that using polyvinyl alcohol (PVA) as a stabilizer significantly influences nanoparticle size and zeta potential, with increased PVA reducing both EE and DL (Zhang et al., 2017; Madani et al., 2018).

 

Thyroxine Levels and Dose-Response Relationship

Oral administration of PLGA-Liotrix at 1.5 mg/kg significantly increased serum T3 and T4 levels (P<0.05) compared to the 1 mg/kg dose. A dose dependent increase in T3 and T4 was observed up to 3.5 mg/kg, beyond which the response plateaued (Figure 4). Correspondingly, TSH levels showed a significant decrease (P<0.05) with increasing doses, reaching a saturation point at 2.5–3.5 mg/kg. Additionally, higher doses (20–35 mg/kg) led to a continued increase in T3 and T4 and a notable suppression of TSH compared to the positive control group. These results indicate a clear dose-dependent endocrine response to PLGA-Liotrix, suggesting enhanced thyroid hormone bioavailability via nanoparticle delivery.

Log Dose-Response Curve and Determination of Maximum Effective Dose

A log dose-response curve was constructed for PLGA-Liotrix treatment at doses of 1, 1.5, 2.5, 3, and 3.5 mg/kg, examining serum levels of T3, T4, and TSH (Figure 5A, 5B and 5C). Based on linear regression analysis, the effective doses for T3, T4, and TSH were calculated as 2.23, 1.81, and 2.95 mg/kg, respectively. The mean of these values, 2.33 mg/kg, was selected as the optimal dose for the third experimental phase. This approach helped identify the dose required for maximal therapeutic effect with minimal toxicity, offering insight into the pharmacodynamic profile of PLGA-Liotrix. However, there remains a lack of direct research examining the influence of PLGA itself on thyroid morphology or function in vivo. Therefore, further studies are warranted to explore possible effects of long-term PLGA exposure on thyroid histology. Nanoprecipitation-based Liotrix-PLGA formulations offer several advantages, including smaller particle sizes, higher encapsulation efficiency, and enhanced cellular penetration. These properties, consistent with previous findings (He et al., 2015), contribute to improved drug bioavailability and sustained hormone release. Oliveira et al., (2015) also highlighted PLGA’s potential in drug delivery systems for its ease of production, controlled release properties, and ability to solubilize poorly soluble drugs.

 

Table 1: Effect of Positive group, PLGA+LOTX, LOTX and PLGA alone 30 days’ treatment on Homocysteine levels of induced hypothyroidism in rats.

Parameters groups	Homocysteine (umol/L)
Negative Control	13.27±0.20d
Positive Control	22.57±0.31a
LOTX treated group	17.37±0.13c
LOTX+PLGA group	13.54±0.20d
PLGA treated group	19.19±0.14b
LSD	1.50

 

* Different letters reveal significant difference between groups at level (P˂0.05). Control group: Negative control group, Positive group, LOTX: Liotrix treated group, LOTX + PLGA: combination of Liotrix with poly (lactic-co-glycolic acid treated group, PLGA: Poly (lactic-co-glycolic acid treated group), *n=5.

 

Homocysteine Concentration

As shown in Table 1, hypothyroid rats (positive control) exhibited a significant increase (P<0.05) in plasma homocysteine (Hcy) levels compared to all treated groups. Notably, rats treated with Liotrix-loaded PLGA (LOTX+PLGA) showed a significant reduction (P<0.05) in Hcy levels, approaching values seen in the negative control group. Treatment with PLGA alone also led to reduced Hcy, though still significantly higher than in the negative group. Elevated homocysteine in hypothyroidism may result from reduced activity of key metabolic enzymes (e.g., methionine synthase), impaired folate and B12 metabolism, and oxidative stress—all of which compromise homocysteine clearance (Zhou et al., 2022). The normalization of thyroid hormones with Liotrix treatment was associated with a corresponding decline in Hcy levels, suggesting a cardiovascular benefit, as hyperhomocysteinemia is a known risk factor for atherosclerosis.

 

Table 2: Effect of Liotrix and PLGA loading Liotrix serum lipid profile (cholesterol, triglycerides, LDL, Oxi-LDL and HDL-C) in rats induce hypothyreosis after four weeks of treatment.

Parameters Groups	T.G mg/dl	Cholesterol mg/dl	HDL mg/dl	LDL mg/dl	Oxi-LDL pg/ml
Negative Control	102.60 ±2.45d	104.52 ±1.42d	28.48 ±0.88a	61.32 ±1.52d	55.14 ±1.49d
Positive Control	160.34 ±1.18a	175.51 ±1.92a	11.10 ±1.04d	136.21 ±2.47a	80.33 ±1.76a
LOTX treated group	130.61 ±1.83c	135.68 ±1.88c	19.45 ±0.46b	83.40 ±3.26c	65.68 ±0.97c
LOTX+PLGA group	100.38 ±1.46d	110.58 ±1.44d	27.12 ±0.48a	60.50 ±1.26d	54.44 ±1.05d
PLGA treated group	145.32 ±1.36b	151.45 ±1.43b	15.41 ±0.31c	130.48 ±1.41b	76.11 ±1.53b
LSD	12.40	12.72	3.19	4.35	3.49

 

❖ Different small letters indicate significant (p≤ 0.05) among groups.

 

Table 3: The effect of Positive group, PLGA, LOTX, and PLGA+ LOTX on Reactive oxygen species (ROS) and Total antioxidant capacity concentration (TAC) in serum of rats.

Parameters Groups	Mean±SE
	ROS (pg/ml)	TAC (U/ml)
Negative Control	15.33±1.46d	4.51±0.11a
Positive Control	35.29±1.40a	1.21±0.12d
LOTX treated group	25.11±1.46c	3.31±0.11b
LOTX+PLGA group	15.36±2.34d	4.47±0.12a
PLGA treated group	30.30±1.33b	2.23±0.10c
LSD	4.87	1.01

 

* Different letters reveal significant difference between groups at level (P˂0.05).

 

The sustained release of T3 and T4 from PLGA nanoparticles likely contributed to steady-state hormone levels, optimizing homocysteine metabolism. This mechanism may involve restored function of methionine synthase and cystathionine β-synthase, crucial enzymes in homocysteine clearance, and a reduction in oxidative stress.

Serum Lipid Profile

Hypothyroid rats demonstrated significant dyslipidemia, with increased triglycerides (TG), cholesterol, LDL, and oxidized LDL (Oxi-LDL), and decreased HDL levels (P<0.05), compared to treated groups. However, the group receiving Liotrix-loaded PLGA showed lipid levels comparable to the negative control group (Table 2). In contrast, Liotrix alone only partially corrected lipid disturbances.

 

Table 4: The effect of Positive group, PLGA, LOTX, and PLGA+ LOTX on C-Reactive Protein (CRP) and Thrombotic factor XII concentrations in serum of rats.

Parameters Groups	Mean±SE
	C-Reactive Protein (CRP) (ng/ml)	Thrombotic factor XII pg/ml
Negative Control	4.26±0.21d	1.36±0.08a
Positive Control	8.95±0.23a	1.25±0.11b
LOTX treated group	5.33±0.35c	1.20±0.02c
LOTX+PLGA group	4.14±0.27d	1.32±0.06a
PLGA treated group	6.81±0.18b	1.27±0.08b
LSD	1.01	0.05

 

* Different letters reveal significant difference between groups at level (P˂0.05).

 

These results align with previous findings (Kim et al., 2018) indicating impaired lipid metabolism in hypothyroidism. Increased LDL and Oxi-LDL in untreated animals may be attributed to oxidative stress, promoting cholesterol ester accumulation and atherogenesis (Tall and Yvan-Charvet, 2015). Treatment with Liotrix improved lipid clearance and upregulated hepatic LDL receptor expression, as previously reported (Taylor et al., 2018). The inclusion of PLGA nanoparticles further enhanced lipid profile normalization, potentially due to improved hormone delivery, reduced hepatic inflammation, and modulation of oxidative stress (Kassel et al., 2011; Pashaie et al., 2017). Additionally, the physicochemical properties of PLGA nanoparticles—size, surface charge, and functional groups—may play a role in lipid metabolism regulation.

Ros and Total Antioxidant Capacity Concentration

Reactive oxygen species (ROS) levels in the positive control group increased significantly (P˂0.05) when matched with the PLGA group, LOTX+PLGA group, and control group. The LOTX+PLGA group showed significantly lower ROS levels compared to other treated groups (P<0.05), matching the negative control. This table also showed the antioxidant capacity concentration (TAC) of rats in the LOTX+PLGA group exhibited a significant elevation (P˂0.05) compared to other treated groups.

The increase in ROS in the positive control group may be attributed to overproduction of stresses causing serious cell damage (Tripathi et al., 2016). PTU may lead to stimulation of oxidation of proteins, lipids, carbohydrates, and DNA by accumulation of ROS (Anjum et al., 2015; Demidchik, 2015). However, the activities of antioxidant enzymes are increased by NPs (Giraldo et al., 2014). Furthermore, PLGA-NPs loaded with liotrix have been reported to enhance stress tolerance by activation of antioxidant enzymes, enhancing uptake processes within Liotrix (Tripathi, 2017) (Table 3).

PLGA in the PLGA+ Liotrix group may enhance drug delivery, improving bioavailability, and ensuring targeted therapy. The role of PLGA in Liotrix treatment may be to enhance absorption and bioavailability as nanoparticles improve the solubility of Liotrix, leading to better absorption in the gastrointestinal tract (Ye et al., 2022). Liotrix, a synthetic combination of thyroxine (T₄) and triiodothyronine (T₃), is used to treat hypothyroidism by restoring normal thyroid hormone levels (Majd et al., 2024). Thyroid hormones significantly influence oxidative stress and the body’s antioxidant defense mechanisms, including mitochondrial respiration, which can lead to ROS production (Kochman et al., 2021).

Serum C-Reactive Protein (CRP) and Thrombotic Factor XII Concentrations

The C-Reactive Protein (CRP) concentration of rats in the positive group exhibited a momentous increase (P˂0.05) compared to the PLGA group, LOTX+PLGA group, and control group. The LOTX+PLGA group showed a statistical decrease (P˂0.05) compared to the positive group and all other treated groups with no significant difference compared to the negative control group. The LOTX group exhibited a statistical decrease (P˂0.05) compared to the control group.

The Thrombotic factor XII (Hageman factor) concentration in the positive group exhibited a significant elevation (P˂0.05) compared to other treated and control groups. The LOTX+PLGA group exhibited a significant decrease (P˂0.05) compared to all other treated groups with no significant difference from the negative control group. The LOTX group showed a significant decline (P˂0.05) compared to the positive group but a significant increase (P˂0.05) compared to the LOTX+PLGA group and control group (Table 4).

The increased concentration of CRP in the positive group may be due to PTU-induced hypothyroidism, which increases CRP levels used as prognostic indicators for inflammation and cardiovascular disorders (Shrivastava et al., 2015; Aniss et al., 2020). Hypothyroidism is associated with elevated levels of inflammatory markers including CRP, TNF-α, and IL-6 (Hajje et al., 2014).

In the PLGA+LOTX group, the reduction in CRP concentration may be due to the use of PLGA, a biodegradable polymer for sustained drug release (Yang et al., 2024). Liotrix may help reduce systemic inflammation, although responses can vary (Vudu et al., 2023). The elevated Thrombotic factor XII in the positive control group may be attributed to PTU’s influence on coagulation, as hypothyroidism is linked to fibrinolytic abnormalities. The LOTX treatment might have an anticoagulant effect by reducing activation of the intrinsic coagulation pathway (Marder et al., 2015), and PLGA + LOTX may reduce FXII activation (Renne et al., 2012).

 

Histopathological Section

Sections of thyroid tissues were shown in Figure 6, the section (A) showed thyroid tissue after 4 weeks of induction (Control positive treated group) appeared hyperplasia of follicles with obliterated lumina, interstitum revealed vascular congestion as in. while section (B) revealed thyroid tissue with hypothyroid treated with PLGA Liotrix showed mild hyperplasia and follicle cells had cuboidal with clear cytoplasm and round nuclei. While in (C), the Section of thyroid tissue treated with Liotrix showed hyperplasia of follicles with obliterated lumina and scanty colloid, follicular cells had low columnar to cuboidal shape clear cytoplasm and round nuclei, interstitum revealed vascular congestion. whereas in (D) the section of thyroid tissue treated with PLGA showed sever hyperplasia of follicles.

The result of positive control group may be attributed to increase TSH lead to accumulation of colloid inside the thyroid follicles, hyperplasia, neovascularize, and change morphologically when exposed to TSH, the results in thyroid hormone imbalance and dysregulation, which can cause emotional and behavioral issues, in addition, TSH increases blood flow to the thyroid gland and other organs and tissues, which can lead to blood vessel destruction and thyroid tissue hemorrhage (Liu et al., 2018; Zhou et al., 2022). Follicular cells in the Liotrix-treated group would become more active and release thyroid hormones throughout the bloodstream. This appeared to have been regulated by TSH and coincided with the findings of other (Ahmed et al., 2022; Sahin et al., 2019). The result of PLGA + Liotrix may be attributed to that PLGA-based delivery system may be induced minor alterations in thyroid histology, which could be indicative of a localized tissue response to the nanoparticle formulation. Similar observations have been reported in studies investigating the biocompatibility of PLGA-based drug delivery systems. For instance, a study on the in vivo biocompatibility of PLGA-polyhexylthiophene nanofiber scaffolds in a rat model demonstrated a moderate tissue response characterized by the presence of lymphocytes, macrophages, fibroblasts, and occasional giant cells, with no significant acute inflammation or tissue necrosis observed (Deng et al., 2020).

 

The histopathological sections of cardiac cells were shown in Figure 7, Control positive treated group) showed severe hemorrhagic myocarditis with multiple hemorrhagic foci.as in section (A). myocardium treated with Liotrix showed moderate hemorrhagic myocarditis, infarction of myofibers with tissue depletion as in section (B). Liotrix showed moderate hemorrhagic myocarditis, infarction of myofibers with tissue depletion as in section (C), while, Section of myocardium treated with PLGA showed long, cylindrical muscle fibers arranged as in section (D).

The histopathological sections of Aorta tissues were shown in Figure 8. And all changes were appeared in all treated group as in different sections (A, B, C and D).

 

Thyroid stimulating hormones also caused heart enlargement. Cardiovascular dysfunctions associated with hypothyroidism include elevated total peripheral vascular resistance and decreased cardiac output, heart rate, and ventricular compliance. The onset of inflammation and heart fibrosis is linked to hypothyroidism (Digeronimo 2020). The Cardiac histopathological effect of liotrix treated group may be attributed to Liotrix treatment may help reestablish equilibrium between ROS production and antioxidant capacity, reducing oxidative stress associated with hypothyroidism lead to attenuate the effect of PTU on cardiac tissue, this result agreed with Macvanin et al., 2023. In other wise, other study recorded by Darda et al. (2024) who they mentioned Liotrix increase myocardial inotropy and heart rate and dilate peripheral arteries lead to increase cardiac output, for this reason, the PLGA –Liotrix treated group showed an improvement in cardiac and aorta tissue and this result may be due to characterization of nanoparticles in decreasing the dose and minimized side effect of drugs.

CONCLUSIONS AND RECOMMENDATIONS

PLGA-Liotrix demonstrated superior therapeutic effects over Liotrix alone, improving thyroid hormones, lipid profiles, antioxidant capacity, and tissue histology. These findings suggest that PLGA-Liotrix may represent a promising therapeutic option for hypothyroidism at a reduced dose. However, several limitations must be acknowledged. The study did not evaluate the bioavailability of PLGA-Liotrix, which is essential for understanding its in vivo performance. Moreover, toxicity and immunogenicity assessments were not performed, and the short duration of exposure limits conclusions regarding long-term safety. Therefore, future research should include pharmacokinetic studies, chronic toxicity evaluations, and immunological assessments to comprehensively establish the safety and efficacy of PLGA-Liotrix.

ACKNOWLEDGEMENTS

The authors are thankful all staff in department Physiology, Biochemistry and Pharmacology in college of Veterinary Medicine / University of Baghdad contributed in assistance a during the period of experiment.

NOVELTY STATEMENT

The current study novelty is focus on the role of PLGA nanoparticles in improving the activity of liotrix drug by decreasing the dose and minimizing side effect of drug, enhancing drug delivery, improving bioavailability, and ensuring targeted therapy.

AUTHOR’S CONTRIBUTIONS

All authors were contributed equally

Conflict of Interest

None.

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