Subacute Toxicity Study Research Articles

Toxicity Studies and Phytochemical Screening of Ficus Racemosa and Kalanchoe Pinnata

Kalanchoe pinnata and Ficus racemosa are naturally occurring plant products that are gaining importance because of their excellent medicinal properties. Determining the toxicity of a plant extract is crucial, based on which the therapeutic dose of extracts can be adjusted. The present study evaluates phytochemical screening and toxicity studies (acute and sub-acute) of the Kalanchoe pinnata and Ficus racemosa plant products. Phytochemical screening showed the presence of alkaloids, tannins, flavonoids, steroids, phenols, and glycosides. The acute toxicity study of both the plant extracts was performed in Wistar albino rats at a single dose of 2000mg/kg body weight in both genders. Sub-acute toxicity study was performed in Wistar albino rats (both male and female) for 28 days at various doses of 500, 1000, and 2000 mg/kg body weight. The evaluation results of the acute toxicity study did not show any signs of toxicity, behavioral changes, mortality, or differences in gross histopathology appearance. The results of the sub-acute toxicity study also showed no changes in body weight, toxic signs, mortality, or behavioral changes. All biochemical and hematological parameters were in normal ranges comparable to the control group's. At the end of the treatment period, all the rats were sacrificed, and the liver and kidneys were sent for histopathological examination. No abnormalities were observed in the treatment groups. The present study concludes that both plant extracts are safe, and the LD50 of both plant extracts was above 2000mg/kg bw.

Evaluation of Safety Profile of Costus Pictus D Don Methanolic Leaf Extract on Albino Wistar Rats

With the emerging trend of preferring naturopathy over modern medicine, common people are in more danger when consuming natural plant products without the knowledge of the toxic agents present in them. Aim and Objective: To evaluate the safety of methanolic leaf extract of Costus pictus D Don on albino Wistar rats. Methodology: The current study was designed strictly based on the organization for economic cooperation and development (OECD) guideline 423 for acute toxicity study to determine LD50 and guideline 407 for sub-acute toxicity study for hazard identification and risk assessment of the test solution. Methanolic leaf extract was prepared by soxhlation. 12 animals (each 12-week-old nulliparous, non-pregnant female Wistar rats with a mean weight of 142 ± 2 g), 3 per step were used for the acute toxicity study. The test was initiated with a single test dose of 300 mg/kg BW on three animals and continued till 2000 mg/kg BW. After ingesting the test dose each animal was observed individually for the first 4 hours and later every day for 2 weeks for signs of toxicity. For the sub-acute toxicity study, 30 adult Wistar rats (each 16-week-old rat weighing 250±12g) were randomized into 3 groups (1 control and 2 study groups) of 10 each consisting of five males and 5 females. Animals in the control group received 1% Carboxymethyl cellulose (CMC) at a dose of 10 ml/kg BW whereas the animals in the study group received 500 and 1000 mg/kg body weight (BW) of the extract respectively for 28 days. Later, all the animals were sacrificed and blood samples were studied for hematological and biochemical changes. Results: The lethal dose of Costus pictus D Don methanolic leaf extract was fixed as more than 2000 mg/kg Body weight. No obvious change was observed in feeding habits, weight, hematology, biochemical parameters, and histopathology. Conclusion: Methanolic leaf extract of Costus pictus D Don was observed to be absolutely safe when given orally in albino Wistar rats.

Assessment of the Safety of Aqueous Extract of Maize (Zea mays) Husks in Wistar Albino Rats

Aims: The possible toxic impact of aqueous extract of maize husks (HA) on Wistar albino rats was investigated.
 Study Design: Experimental design.
 Place and Duration of Study: This study was conducted at the Federal University of Technology, Akure, Nigeria between January 2020 and April 2020.
 Methodology: The extract was prepared adopting the cold extraction procedure. Forty-eight (48) Wistar albino rats were used for this study. Acute toxicity study was carried out by administering HA at a single dose of 2000, 4000 and 8000 mg/kg body weight to the rats by oral gavage. The rats were observed for 14 days for any mortality or signs of toxicity. For sub-acute study, doses of 200, 400 and 800 mg/kg body weight were orally administered daily for 28 days. Biochemical and haematological parameters as well as histopathological studies were carried out after the experiments.
 Results: Acute toxicity results indicated that the median lethal dose (LD50) of HA was greater than 8000 mg/kg. There was remarkable body weight gain (P ≤ 0.05) in both male and female rats in all the sub-acute groups and acute group treated with 8000mg/kg. In the acute and sub-acute toxicity study, slight difference was recorded between the biochemical and haematological parameters of the treated rats dosed with the extract and the control. There was a significant increase in Packed Cell Volume (PCV) of female rats and lymphocytes of male rats treated with 800 mg/mL and 400 mg/mL respectively. The total protein, glucose and urea level of male rats treated with 200 mg/mL reduced while urea level of females treated with 200 mg/mL and 400 mg/mL also reduced remarkably. However, histological data showed no significant difference.
 Conclusion: In general, the extract was found to show no toxic effect on the rats and hence it is safe for potential therapeutic use.

Sub-acute toxicity study of the aqueous extract from leaves and flowers of Acmella caulirhiza on female albino Wistar rats

Sub-acute toxicity study of the aqueous extract from leaves and flowers of Acmella caulirhiza on female albino Wistar rats

Toxicological Study of n-hexane and Chloroform Extracts of Lantana camara L. in Experimental Animals

Abstract: Aim: The aim of the current study is to investigate the acute, sub-acute and sub-chronic toxicity study of extracts (n-hexane and chloroform) of L. camara in Wistar rats. Materials and Methods: HPTLC and GC-MS analysis were carried out to analyze different constituents present in L. camara extract. Acute toxicity study was conducted as per OECD 420 guidelines, while sub-acute and sub-chronic studies were conducted according to OECD 407 and 408 guidelines respectively. Toxicopathological effects of both extracts were tested with the help of hemotoxylin and eosin staining of vital organs (liver, kidney, brain, testes, and ovaries) in experimental animals. Antioxidant and behavioral tests were also conducted on both the extracts, so as to evaluate the possible effects of toxicity. Results: The toxicity investigation of chloroform and n-hexane extracts exhibits no toxicity in terms of hematological (p<0.01) and biochemical parameters (p<0.05). Histopathological sections also confirmed the non-toxic effects on the liver, brain, kidney and on reproductive organs. Conclusion: The n-hexane and chloroform extracts of L. camara can be safely used for therapeutic purposes. Keywords: Lantana camara, GC-MS, HPTLC, Toxicity, Histopathology, Antioxidant, PI uptake.

Acute and Sub-Acute Toxicity Studies of Starch Hyaluronate in Wistar Rats

Starch hyaluronate is used as a new superdisintegrating agent in the development of fastdissolving tablets of poorly soluble drugs because of its improved biocompatibility, and hydrophilicity. The purpose of this study is to assess the acute and subacute toxicity profiles of starch hyaluronate in Wistar rats. The starch hyaluronate was administered to Wistar rats in a single-dose acute study and monitored for seven days. Wistar male and female rats were used in a 28-day sub-acute study that examined the effects of oral doses of 100, 200, and 600 mg/kg body weight/day on body weight, food intake, mortality, biochemical analysis, and histopathologicalevaluation. An acute study revealed that the synthesized starch hyaluronate minimal oral fatal dose for rats was larger than 2000 mg/kg body weight. The subacute toxicity evaluation found no significant changes when compared to the control group, and there was no change in hematological or biochemical parameters. The weights of the liver, kidneys, and pancreas remained constant. Based on the findings, it was concluded that starch hyaluronate at 600mg/kg body weight/day was neither immunogenic nor sensitizing, and was not a reproductive or developmental toxicant, implying that it is relatively safe when taken orally.&nbsp

EVALUATION OF ORAL TOXICOLOGICAL INVESTIGATION OF A HERBAL COMPOSITE (HERBODIL®) IN EXPERIMENTAL ANIMALS

Purpose: The aim of the investigation was to find out the degree of toxicity of the herbal composite for humans, animals, or the environment. Acute and 28 days repeated sub-acute oral toxicity study of herbal composite (Herbodil®) carried out as per the current OECD guidelines.Materials and Methods: 2000mg/kg of the herbal composite was orally administered to the animals to find out the acute toxicity. The treated animals were observed for toxic signs at thirty min, one, two and four hours and thereafter once a day for the next 14 days. In sub-acute study i.e.,28 days repeated dose oral study, the mice were segregated into four groups (two set for each sex) of six mice each. Group-1 mice served as a control (untreated). Group II mice consumed lower dose of herbal composite i.e., 100 mg/kg, Group III mice consumed middle/moderate dose i.e., 200 mg/kg and Group IV mice received high dose of 400 mg/kg (orally) once daily for 28 days respectively.Results: This experimental in vivo study confirm that neither the acute toxicity study of herbal composite at the dose level of 2000mg/kg nor the sub-acute 28 days oral toxicity study developed any toxic signs, behavioural changes, or mortality during the whole study. Haematological and biochemical parameters dose not changes during the sub-acute study. Relative body weight of the mice also not change after the study.Conclusion: Experimental results obtained from the current investigation suggest that LD50 of developed herbal composite was &gt;2000 mg /kg and the herbal composite is completely safe and non-toxic for therapy.

Safety Evaluation of Arsenicum album in Acute and Sub-Acute Toxicity Studies in Rats

Arsenic album is frequently prescribed in homoeopathy for many diseases. However, its safety data is not available. Thus, the study’s purpose is to evaluate the oral toxicity of Arsenic album 6C, 30C, and 200C in rats. Arsenic album (6C, 30C, and 200C) was given at 2000 μl/kg for acute toxicity and observed for up to 14 days. For subacute toxicity, it was given for 28 days and observed for clinical signs, change in body weight and Mortality. Hematological, biochemical, organ weight and histopathological analyses were assessed. Results indicate no mortality of arsenic album in acute toxicity and LD50 is &gt;2000 μl/kg. In the subacute toxicity study, arsenic album (200 μl/kg) did not show any significant changes in above parameters. It may be concluded that the arsenic album (6C, 30C, and 200C) is safe and produces no toxicity when administered orally for a prolonged duration at 200 μl/kg in rats.

In Vivo Toxicity Assessment of the Probiotic Bacillus amyloliquefaciens HTI-19 Isolated from Stingless Bee (Heterotrigona itama) Honey.

This study evaluated the acute and sub-acute toxicity of B. amyloliquefaciens HTI-19 (isolated from stingless bee honey) in female Sprague Dawley rats. In an acute toxicity study, the rats received a low dosage (1 × 109 CFU·mL-1), medium dosage (3 × 109 CFU·mL-1), or high dosage (1 × 1010 CFU·mL-1) of B. amyloliquefaciens HTI-19 daily orally by syringe-feeding for 14 days. For the subacute toxicity study, rats received a low dosage (1 × 109 CFU·mL-1) or a high dosage (1 × 1010 CFU·mL-1) for 28 days. The probiotic feeding in acute and sub-acute toxicity studies showed no mortality or significant abnormalities in rats throughout the experimental period. In week 2 of the acute study, the body weight of the rats showed a significant increase (p < 0.05) compared to the control. By gross and microscopic examination of organs, no evidently significant changes were observed in the morphology of organs. Serum biochemical tests and blood hematology tests also revealed no treatment-related changes. Overall, these data indicated that oral administration of B. amyloliquefaciens HTI-19 up to 1 × 109 CFU·mL-1 for 28 days can be considered safe.

Study of acute and subacute toxicity of the aqueous extract of the rhizomes of Anchomanes difformis Blum Engl. (Araceae) in rats

The aim of the study was to evaluate the acute and subacute toxicity of Anchomanes difformis rhizomes extract. The qualitative and quantitative chemical analyzes of the phenolic compounds from Anchomanes difformis rhizomes aqueous extract. were carried out by CCM and spectrophotometry. The acute toxicity study was conducted on female swiss mice at doses of 2000, 3500 and 5000 mg / kg per os. The subacute toxicity study was carried out in Wistar rats quatidianly treated with 400 mg / kg per os of the aqueous extract for 28 days. Control lots received 10 ml / kg. Chemical analyzes revealed the presence of polyphenols and flavonoids. The acute toxicity study of the aqueous extract did not result in any deaths. The LD50 is greater than 5000 mg / kg. The subacute toxicity study showed weight gain in rats and various organsː uterus, heart, kidneys, liver, and lungs. On biochemical parameters, aqueous extract significantly increase Creat, Asat, Alat levels, but not Gly and Chol. In hematological parameters Anchomanes difformis aqueous extract caused increase in WBCs, IDR-CD, PCT and decrease in PLT, GMV, IDR-DS.

TOXICITY PROFILES OF METHANOL EXTRACT OF CROSSOPTERYX FEBRIFUGA LEAF (AFZEL. EX G. DON) BENTH

Crossopteryx febrifuga is one of the plants used for the local treatment of some disease conditions by the people of Nigeria via its leaf, root and bark without knowing the toxic nature of the plant recipes. This research was designed to ascertain the toxicity profiles of the plant leaf. Acute toxicity was determined in two phases using rats. In sub-acute toxicity studies, haematology, histopathology and evaluation of liver function indices were conducted. In the first phase of the toxicity study, the mice showed no observable toxic effects as high as at 1000 mg/kg. However, it caused sluggishness, respiratory distress at higher doses in the second phase. On haematological indices, white blood cell, lymphocyte, erythrocytes, haemoglobin, haematocrit rose significantly across the doses administered. The liver function test revealed significant increase in serum alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST), albumin, total protein and conjugated bilirubin while 800 mg/kg extract reduced the aspartate transaminase. On histopathology, liver revealed normal hepatocytes but dilated central vein while in kidney, renal corpuscle, glomeruli and tubule were normal. On heart, revealed was active vascular congestion with normal bundle of myocardial fibres and coronary artery while in aorta, it revealed normal architecture. With the nature of the toxicity profile revealed especially at doses less than 500 mg/kg of C. febrifuga extract, the plant could be considered to be relatively safe. This study is significant as it provides information that may suggest some safe doses out of the doses used for clinical trial and application

Acute and Subacute Toxicity Studies of Ethanol, Chloroform extracts and Flower oil of Senecio edgeworthii hook plant

This study investigated the acute and subacute oral toxicity of SE- Oil, SE-Alcohol and SE-Chloro extracts of Senecio edgeworthii (SE) in a rodent for evaluating its safety profile. Senecio edgeworthii (SE) plant for the acute oral toxicity (300 to 5000mg/kg) and subacute oral toxicity SE- Oil (SE-Oil 50, 200, 500mg/kg) and SE-Chloro (SEC 100, 500, 1000mg/kg) and SE-Alcohol (SEA 100, 500, 1000mg/kg) studies were administered orally according to the guidelines 423 and 407 of OECD, respectively. In the acute oral toxicity study, doses are administered by 300mg/kg, 1000mg/kg, 2000mg/kg and 5000mg/kg (OECD guideline no. 423). SEA and SEC do not show toxicity up to 5000mg/kg; SE- Oil shows toxicity at a dose of 2500mg/kg. In the subacute toxicity study, the daily oral treatment with extracts of SEA and SEC 100, 500, 1000mg/kg, SE-Oil 50, 250, 500mg/kg of extracts for 28 days did not produce any death or hazard. Likewise, SEA and SEC 100, 500, 1000mg/kg, SE-Oil 50, 250mg/kg of Senecio edgeworthii hook no significant changes were recorded in food intake, body weight gain, the biochemical and haematological parameter of control and treated rats. SE-Oil 500mg/kg of significant changes were recorded in food intake and body weight gains are decreased compared to the control group and SE-Oil 500mg/kg there was a significant (P &lt; 0.05) decrease in RBC counts in the treated animals compared to that of the control group counts in the treated animals and induction (ALT, AST, and alkaline phosphatase and Total protein). SE- Oil 500mg/kg of extract treaded group saw mild liver pathological findings characterized by abnormal hepatic configuration was observed with treated animals. The present result from the acute and subacute toxicity study Senecio edgeworthii hook alcohol and chloroform extract is considered relatively safe on acute and subacute oral exposure. Senecio edgeworthii hook flower hexane extract shows acute toxicity at dose 2500mg/kg and in subacute toxicity study, higher dose 500mg/kg.

Preclinical In vivo Acute Toxicity Testing of Hyrdoethanolic Extracts of Ficus thonningii Blume (Moraceae) on Wistar Rat Models

Ficus thonningii (Blume) is a medicinal plant whose biological activity has been demonstrated in the management of peptic ulcers. Unfortunately, little scientific data exists on its toxicity. This is why we studied the acute and subacute oral toxicity profile of the hydro-ethanolic extract of the bark of F. thonningii and the systemic exposure of the bioactive components of the plant.Acute toxicity was assessed by administering a single dose of 2000 mg/kg to 5-week-old rats and observed for 14 days. The subacute toxicity study was performed in 6-week-old rats. Animals were orally treated with a daily dose of 125 mg/kg, 250 mg/kg, 500 mg/kg extract for 28 days. A satellite group of animals received 500 mg/kg per day for 28 days and after discontinuation of treatment they were observed for 14 days. Hematological and blood biochemical parameters, as well as kidney and liver histology, were recorded at the end of each experiment. For acute treatment, administration of a dose of 2000 mg/kg did not induce any critical behavioral changes or deaths. Following subacute dosing, biochemical analysis revealed a slight elevation of liver parameters in the 500mg/kg hydroethanolic extract, while no significant increase was observed for kidney parameters. This study showed that daily administration of Ficus thonningii (Blume) for 28 days resulted in the potential to reverse impairment of liver function.

Acute and sub-acute dermal toxicity of meloxicam emulgel: Analysis of biochemical, hematological, histopathological and immunohistochemical expression

Meloxicam, a non-steroidal anti-inflammatory drug (NSAID) for the treatment of osteoarthritis. Despite being more effective against pain mediated by inflammation, it is associated with gastrointestinal, cardiovascular, and renal toxicity. In the current study, acute single-dose (2000 mg/kg) and subacute (500, 1000, and 2000 mg kg−1 for 28 days) dermal toxicity analyses of meloxicam emulgel were conducted in Wistar rats. Various biochemical, hematological, histopathological and immunohistochemical parameters were evaluated. The dermal LD50 (lethal dose) of meloxicam emulgel was found to be > 2000 mg/kg. No significant adverse effects of meloxicam emulgel following topical administration in subacute toxicity studies were noticed. IL-1β was not expressed post treatment with meloxicam emulgel. IL-1β is an influential pro-inflammatory cytokine that is decisive for host-defence consequence to injury and infection. Therefore, using data gleaned from the extant study, topical administration of meloxicam emulgel may be regarded as safe as the “no observed adverse effect level” (NOAEL) was >2000 mg/kg in experimental animals.

Synthesis, cytotoxicity, and pharmacokinetic evaluations of niclosamide analogs for anti-SARS-CoV-2

Niclosamide, an oral anthelmintic drug, could inhibit SARS-CoV-2 virus replication through autophagy induction, but high cytotoxicity and poor oral bioavailability limited its application. Twenty-three niclosamide analogs were designed and synthesized, of which compound 21 was found to exhibit the best anti-SARS-CoV-2 efficacy (EC50 = 1.00 μM for 24 h), lower cytotoxicity (CC50 = 4.73 μM for 48 h), better pharmacokinetic, and it was also well tolerated in the sub-acute toxicity study in mice. To further improve the pharmacokinetics of 21, three prodrugs have been synthesized. The pharmacokinetics of 24 indicates its potential for further research (AUClast was 3-fold of compound 21). Western blot assay indicated that compound 21 could down-regulate SKP2 expression and increase BECN1 levels in Vero-E6 cells, indicating the antiviral mechanism of 21 was related to modulating the autophagy processes in host cells.

Phytochemical component and toxicological evaluation of purple sweet potato leaf extract in male Sprague-Dawley rats.

This study assessed the toxicity of lutein-rich purple sweet potato leaf (PSPL) extract in male Sprague-Dawley rats. Methods and study design: A total of 54 adult male Sprague-Dawley rats were used. For the acute toxicity study, three rats in the acute control group were fed 2,000mg/kg of PSPL for 14days. The subacute toxicity study included six rats each in four groups administered 50, 250, 500, or 1,000mg/kg for 28days and observed for further 14days without treatment in the subacute control and subacute satellite groups. Changes in body weight; blood biochemistry; hematological parameters; relative organ weight; and histological sections of the heart, kidney, liver, pancreas, aorta, and retina were observed for signs of toxicity. Results: The gradual increase in weekly body weight, normal level full blood count, normal liver and kidney profile, relative organ weight, and histological sections of all stained organ tissue in the treated group compared with the acute, subacute, and satellite control groups demonstrated the absence of signs of toxicity. Conclusion: Lutein-rich PSPL extract shows no signs of toxicity up to 2,000mg/kg/day.

Phytochemical composition and toxicity assessment of Ammi majus L.

Objective: To assess the acute and subacute toxicity as well as the phytochemical composition of two extracts and three fractions of Ammi majus L. Methods: The aqueous extracts were prepared separately by maceration for 48 h and by infusion for 1 h, while the fractions were prepared by the Soxhlet extractor, successively employing cyclohexane, ethyl acetate, and ethanol. The acute toxicity study was carried out in accordance with the OECD N°423 guideline at a single dose (2000 mg/kg) in mice for 14 days. The subacute toxicity study was performed by a daily oral administration of 250 mg/kg for 10 days and 100 mg/kg doses for 28 days. Phytochemical screening was performed using staining and precipitation reactions, while the chemical characterization of some analytes was detected by HPLC-MS/MS analysis. Results: In the acute toxicity study, no signs of toxicity such as convulsion, salivation, diarrhea, sleep and coma were observed during 30 minutes and 14 days, so the lethal dose was higher than 2000 mg/kg for each extract and fraction. The subacute toxicity results showed that at a dose of 250 mg/kg, 61.10% of the animals died and the rest developed morbidity. On the other hand, at a dose of 100 mg/kg, all the animals were still alive after 28 days, with no morbidity and the biochemical parameters were normal with no abnormalities in the liver, kidneys and pancreas. Phytochemical screening indicated the presence of flavonoids, tannins, coumarins, and free quinones and the absence of alkaloids and anthocyanins. Conclusions: The extracts and fractions of Ammi majus L. are not toxic in the short and long term with a varied chemical composition. Toxicological tests on animals other than rodents and in the long term (more than 28 days) are needed to further confirm the safety of Ammi majus extracts.

Assessment of the Acute and Subacute Toxicity Profile of Diethylether Extract of Jatropha Tanjorensis leaf in Male Wistar Rats

The aim of study was to evaluate the acute and subacute toxicity of diethylether extract of Jatropha tanjorensis leaves in male Wistar rats. The acute toxicity study was conducted by placing Wistar albino rats into four groups of five rats each. These were sequentially dosed one at a time, using four graded doses of 1250, 2500, 3750 and 5000 mg/kg b.wt of the plant extract. Observations were made and recorded systematically daily for a seven days period for mortality, breathing and other behavioral changes after dose administration. For the sub-acute toxicity study, the animals were divided into ten groups of five rats each. Group 1 served as normal control and received distilled water for a 42 days period. Groups 2,3 and 4 received 167mg/kg b.wt of the plant extract for 14 days, 28 days and 42 days respectively. Groups 5, 6 and 7 received 250mg/kg b.wt of the plant extract for 14 days, 28 days and 42 days period respectively. Groups 8, 9 and 10 received 500mg/kg b.wt of the plant extract for 14, 28 and 42 days respectively. The animals were sacrificed at 14 days interval and the hematological and biochemical parameters were determined. There was a significant (p &lt;0.05) increase in RBC, Hb, HT, MCV and MCH and a decrease in WBC for 14, and 28 days treatment. There was significant increase in the levels of TP, ALB and a decrease in the levels of TBIL, ALP while ALT and AST remained unchanged. The serum markers of lipid profile, total–CHOL, LDL-CHOL, VLDL-CHOL and TG were significantly reduced and HDL-CHOL level was increased. The activities of the stress enzymes GPx, CAT and SOD were increased while the level of MDA was reduced. The K+ level was unchanged, Na+, Cl- and HCO3- levels increased while urea and creatinine levels were decreased. Jatropha tanjorensis diethylether extract elicited non toxic effect in the hematological and biochemical parameters. Thus, the extract can be considered safe for oral administration.

Acute and sub-acute toxicity studies of hydro-alcoholic extract of dried fruits of Piper longum Linn in Wistar rats

Acute and sub-acute toxicity studies of hydro-alcoholic extract of dried fruits of Piper longum Linn in Wistar rats

Reproductive Toxicity Evaluation of Graphene-Based Tumor Cell Nucleus-Targeting Fluorescent Nanoprobes on Reproduction and Offspring Health.

A new graphene-based fluorescent nanoprobe for tumor cell nucleus (GTTNs) was synthesized in our laboratory that penetrates the cell membrane and particularly targets cancer cell nucleus and displays tremendous potential for clinical applications. Although acute and subacute toxicity studies have been conducted on GTTNs, a primary result could be drawn that GTTNs appear to have almost no acute and subacute toxicity. However, as an important part of safety evaluation, the influences on reproductive and offspring developmental toxicity are still absent. In this study, male mice were injected intravenously with GTTNs, and the survival status, histopathology of the testes and epididymides, proliferation and apoptosis of testicular tissue, and sperm motility of mice were measured. To evaluate the short- and long-term fertility in male mice, different male mice resided with untreated female mice on days 1 and 30 after the end of the last treatment, and the offspring health parameters were assessed by measuring pup numbers, body weight, and organ indexes of the pups. The results indicated that GTTNs-exposed male mice retained good fertility, healthy structure of testes and epididymides, and production of healthy sperm. Meanwhile, there were no significant differences between the offspring and the control group. In consideration of GTTNs with broad prospects for biomedical applications, our results contribute a basis for further understanding of its biosafety.