. 2023 May 12;11:1183592. doi: 10.3389/fbioe.2023.1183592

TABLE 2.

Studies of nano-TiO2 on the male reproductive system.

Authors	Year	Organ	Characteristic of nano-TiO₂	Materials	Method	Results				References
Authors	Year	Organ	Characteristic of nano-TiO₂	Materials	Method	Weight	Histopathologic change and other results	Hormone	Mechanism	References
Guo et al.	2009	Testis	Anatase	45 male ICR mice aged 6 weeks	Intraperitoneally injected, 200 and 500 mg/kg	-	Sperm density and motility↓, Sperm malformation↑	-	Germ cell apoptosis	Guo et al. (2009)
Gao et al.	2013	Testis	-	115 CD-1 male mice aged 5 weeks	Intragastric, 2.5, 5, and 10 mg/kg BW for 90 days	Testicular weight↓, relative testicular weight↓, BW↓	Sperm concentration↓ Motility rate↓	FSH, LH, T↓	Spermatogenesis: upregulated: Ly6e, downregulated: Adam3, Tdrd6, Spata19, Tnp2, Prm1	Gao et al. (2013)
Gao et al.	2013	Testis	-	115 CD-1 male mice aged 5 weeks	Intragastric, 2.5, 5, and 10 mg/kg BW for 90 days	Testicular weight↓, relative testicular weight↓, BW↓	Sperm concentration↓ Motility rate↓	FSH, LH, T↓	Apoptosis and oxidative stress: upregulated: Axud1, Cyp1b1, Cyp2e1, Gpx5, Th	Gao et al. (2013)
Jia et al.	2014	Testis	Anatase; 25 nm	60 Kunming male mice aged 3 weeks	Intragastric, 10, 50 or 250 mg/kg BW for 42 days	BW↓	Vacuoles in seminiferous tubules (50, 250 mg/kg); Decreased layers of spermatogenic cells (250 mg/kg)	T↓(250 mg/kg)	Downregulated: P450-17α, 17β-HSD; Upregulated: Cyp19; No effect: P450scc, 3β-HSD, AR	Jia et al. (2014)
Orazizadeh et al.	2014	Testis	-	32 NMRI mice aged 6–8 weeks	Intragastric, 0.2, 10, 300 mg/kg BW for 35 days	Testicular weight↓, relative testicular weight↓	Sperm count, motility↓Vacuolization of the seminiferous tubules, decreased layers of spermatogenic cells	T↓	-	Orazizadeh et al. (2014)
Orazizadeh et al.	2014	Testis	-	32 NMRI mice aged 6–8 weeks	Intragastric, 0.2, 10, 300 mg/kg BW for 35 days	Testicular weight↓, relative testicular weight↓	Johnsen’s score↓	T↓	-	Orazizadeh et al. (2014)
Meena et al.	2015	Testis	-	24 Wistar rats aged 8 weeks	Intravenous injected (through caudal vein), 5, 25, and 50 mg/kg BW for 30 days	Average coefficient of testis↓	Apoptotic cell population increased, Inflammation in testicular cells	T↓ (25 mg/kg)	Oxidative stress: SOD, GPx↓, CAT, lipid peroxidase ↑	Meena et al. (2015)
Hong et al.	2015	Testis	Anatase; 5.5 nm	160 ICR mice aged 4 weeks	Intragastric, 1.25, 2.5, or 5 mg/kg BW for 6 months	BW, testis and epididymis indices↓	Sperm count and motility↓	T↓	Expression of Cdc2, Cyclin B1, Dmcl, TERT, Tesmin, TESP-1, XPD, and XRCCI mRNA↓Gsk3-β mRNA↑ in the testicular tissues.	Hong et al. (2015b)
Hong et al.	2015	Testis	Anatase; 5.5 nm	160 ICR mice aged 4 weeks	Intragastric, 1.25, 2.5, or 5 mg/kg BW for 6 months	BW, testis and epididymis indices↓	Spano-sperm, sperm breakages, rarefaction of Sertoli cells and Leydig cells	T↓	Cdc2, DMC1, TERT, Tesmin, XRCC1 and XPD expressions↓, Gsk3-β and PGAM4↑	Hong et al. (2015b)
Hong et al.	2015	Testis	Anatase; 5–6 nm	160 ICR mice aged 4 weeks	Intragastric, 2.5, 5 or 10 mg/kg BW for 60 days	BW, testicular weight, relative testicular weight↓	Sperm count and motility↓	-	LDH, SODH, SDH, G6PD, Na⁺/K⁺ -ATPase, Ca²⁺-ATPase, and Ca²⁺/Mg²⁺-ATPase ↓	Hong et al. (2015a)
							Sperm malformation↑		ACP, AKP, and NOS↑
							Spano-sperm, sperm breakages, rarefaction of Sertoli cells and Leydig cells		Oxidative stress: ROS↑
							Decreased layers of spermatogenic cells
Asare et al.	2016	Testis	21 nm	6 Ogg1 ^−/− KO mice aged 8–12 weeks, 6 Isogenic Ogg1 ^+/+ mice (WT)	Intravenous injected (tail vein),	-	-	-	DNA damage	Asare et al. (2016)
Asare et al.	2016	Testis	21 nm		5 mg/kg BW for 1 or 7 days	-	-	-	Oxidative stress in testis, Atr and Rad51, Ddb2, Sod1 and Fos expression↑	Asare et al. (2016)
Hong et al.	2016	Testis	Anatase; 5–6 nm	160 ICR mice aged 5 weeks	Intragastric, 1.25, 2.5, or 5 mg/kg BW for 9 months	-	Mating rate, pregnancy rate and number giving birth/foetus↓	-	The expression of Tyro3, Axl, Mer, IκB, SOCS1, and SOCS3 genes and proteins↓	Hong et al. (2016a)
Hong et al.	2016	Testis	Anatase; 5–6 nm	160 ICR mice aged 5 weeks	Intragastric, 1.25, 2.5, or 5 mg/kg BW for 9 months	-	Infiltration of inflammatory cells, Sperm breakages, rarefaction, apoptosis or necrosis of spermatogenic cells and Sertoli cells, and vacuolation of seminiferous tubule	-		Hong et al. (2016a)
Hong et al.	2016	SCs	Anatase; 5–6 nm	Primary cultured Sertoli cells of ICR mice	Intragastric, 5, 15, or 30 ug/mL for 24 h	-	Cell viability↑, LDH activity↑, Apoptosis of Sertoli cells↑	-	Oxidative stress: MMP↓, ROS and lipid peroxidation level↑, SOD, CAT, and GPx↓	Hong et al. (2016b)
Hong et al.	2016	SCs	Anatase; 5–6 nm	Primary cultured Sertoli cells of ICR mice	Intragastric, 5, 15, or 30 ug/mL for 24 h	-	Vacuolization of the seminiferous tubules, nuclear shrinkage, chromatin marginalization, endoplasmic reticulum expansion, and mitochondrial swelling	-	Cells apoptosis: cytochrome c, caspase-3, caspase-12, Bax, GRP78, and CHOP↓, Bcl-2↑	Hong et al. (2016b)
Khorsandi et al.	2017	Testis	-	32 NMRI mice aged 6–8 weeks	Intragastric, 300 mg/kg BW for 42 days	-	Sperm count and motility↓	T↓	Oxidative stress: MDA↑SOD and CAT↓	Khorsandi et al. (2017)
							Sperm malformation↑
							Apoptosis index↓
Morgan et al.	2017	Testis	-	80 albino rats (180–200 g)	Intragastric, 100 mg/kg BW for 8 weeks	-	Sperm viability↓	T↓	Oxidative stress: MDA↑GSH↓ CAT-	Morgan et al. (2017)
							Sperm malformation↑		Apoptosis↑
							Interstitial oedema and sloughing of its germinal epithelium		Testin gene expression↑
Mao et al.	2017	GC-2, TM4 cells	21 nm	GC-2 cells, TM4 cells	0.1, 1, 10, 100 ug/mL for 4 h	-	Microtubule network of GC-2 Cells, microtubule dynamic of GC-2 Cells, microfilamen networks of TM4 Cells, migration ability of GC-2 Cells, phagocytic activity of TM4 Cells changed	-	-	Mao et al. (2017)
Miura et al.	2017	Testis	-	C57BL/6J mice aged 8 weeks	i.v. Injection, 0.1, 1, 2, 10 mg/kg BW for 4 weeks	BW, testicular weight↓	Sperm viability↓	T (−)	-	Miura et al. (2017)
Miura et al.	2017	Testis	-	C57BL/6J mice aged 8 weeks	i.v. Injection, 0.1, 1, 2, 10 mg/kg BW for 4 weeks	BW, testicular weight↓	Sperm malformation↑	T (−)	-	Miura et al. (2017)
Shahin et al.	2017	Testis	Anatase; <25 nm	48 Wistar rats aged 6–7 weeks	Intragastric, 50 mg/kg BW for 1,2,3 weeks	BW↓ prostate gland weight↓	Sperm count, motility↓	T↓, E₂, LH, FSH↑	Apoptotic changes: anti-apoptotic factor Bcl-2↓ pro-apoptotic factor Bax/Fas/Caspase-3↑	Shahin and Mohamed (2017)
							Sperm malformation↑
							Decreased layers of spermatogenic cells
							Johnsen’s score↓
Ye et al.	2017	Sertoli cells	Anatase; 5–6 nm	Primary cultured SD rat Sertoli cells (aged 18 days)	5, 15, or 30 μg/mL TiO₂NPs for 24 h	-	Nano-TiO₂ entered the cytoplasm and cell nuclei	-	Inflammatory cytokines TNF-α, NF-κB, I-κB and IL-1β protein expression↑	Ye et al. (2017)
Ye et al.	2017	Sertoli cells	Anatase; 5–6 nm	Primary cultured SD rat Sertoli cells (aged 18 days)	5, 15, or 30 μg/mL TiO₂NPs for 24 h	-	Nano-TiO₂ entered the cytoplasm and cell nuclei	-	p-PKC and p-p38 MAPK proteins in SCs↑	Ye et al. (2017)
Li et al.	2018	Leydig cells	Anatase	Primary cultured SD rat Leydig cells	0, 10, 20,40 μg/mL for 24 h	-	Nano-TiO₂ entered the cytoplasm and cell nuclei	T↓(20,40 ug/mL)	Mitochondrial damage: MMP↓	Li et al. (2018)
Li et al.	2018	Leydig cells	Anatase	Primary cultured SD rat Leydig cells	0, 10, 20,40 μg/mL for 24 h	-	Nano-TiO₂ entered the cytoplasm and cell nuclei	T↓(20,40 ug/mL)	P-ERK1/2, PKA,PKC↓3βHSD, StAR, P450scc, SR-BI, and DAX1 proteins↑	Li et al. (2018)
Miura et al.	2019	Testis	-	C57BL/6J mice aged 8 weeks	IT injection, 10,50 mg/kg for 1,3,9 days	-	Sperm motility↓		[³H]-thymidine incorporation, and ATP level↓	Miura et al. (2019)
Santonastaso et al.	2019	Human sperm	Mixture of the rutile and anatase, 21 nm	Human sperm	1,10 μg/L for 15,30,45,90 min	-	Sperm motility, sperm genomic stability↓	-	Intracellular ROS in sperm cells↑	Santonastaso et al. (2019)
Santonastaso et al.	2019	Human sperm	Mixture of the rutile and anatase, 21 nm	Human sperm	1,10 μg/L for 15,30,45,90 min	-	Loss of sperm DNA integrity, sperm DNA fragmentation↑	-	Intracellular ROS in sperm cells↑	Santonastaso et al. (2019)
Hussein et al.	2019	Testis	<50 nm	70 SD rats aged 6–8 weeks	Intragastric,	-	Sperm abnormalities↑	LH, FSH,T↓	Oxidative stress: L-MDA↑SOD, CAT,GSH↓	Hussein et al. (2019)
Hussein et al.	2019	Testis	<50 nm	70 SD rats aged 6–8 weeks	300 mg/kg BW for 30 days	-	Decreased layers of spermatogenic cells	LH, FSH,T↓	17β-HSD gene expression↓Bax gene expression↑	Hussein et al. (2019)
Lauvås et al.	2019	Testis	Rutile, 17 nm	47 C57BL/6J BomTac mice aged 9 weeks	Intratracheally instilled, 63 µg for 7 weeks	-	Sperm counts (−)	T (−)	-	Lauvas et al. (2019)
Ogunsuyi et al.	2020	Testis	Anatase; <25 nm	25 Swiss mice aged 11–15 weeks	Intraperitoneally injection, 9.38, 18.75, 37.5, 75 mg/kg BW for 35 days	BW, testicular weight (−)	Sperm count, motility↓	T, LH↓FSH↑	Oxidative stress: SOD↓, CAT↓, GSH↓	Ogunsuyi et al. (2020)
							Sperm malformation↑
							Vacuolization of the seminiferous tubules, decreased layers of spermatogenic cells
Liu et al.	2021	Testis	Anatase and rutile, 40 nm ± 5 nm	15 SD rats aged 8 weeks	Intragastric, 500 mg/kg BW for 3 and 7 days	-	Sperm counts↓	T↓	Inhibit testosterone synthesis, which is related to the reactive oxygen species (ROS)-MAPK (ERK1/2)-StAR signal pathway	Liu et al. (2021)
Liu et al.	2021	Testis	Anatase and rutile, 40 nm ± 5 nm	15 SD rats aged 8 weeks	Intragastric, 500 mg/kg BW for 3 and 7 days	-	Anatase is more toxic than rutile.	T↓		Liu et al. (2021)
Mancuso et al.	2022	Sertoli cells	-	Porcine prepubertal Sertoli cells (SCs)	5,100 μg/mL for 24 h and 1,3 weeks	-	Cell count↓	-	Apoptosis in SCs, intracellular ROS Production and DNA Damage, Stimulated proinflammatory and immunomodulatory responses, activated MAPK and NF-κB signalling pathway	Mancuso et al. (2021)
							Deeply invaginated and shrunk nuclei, disorder of chromatin components, missing endoplasmic reticulum membranes, mitochondria number↓
							Apoptotic mitochondria, enlarged endoplasmic reticulum, increased frequency of lipid droplets
Meng et al.	2022	Testis	Anatase; 5–10 nm	96 ICR mice aged 6–8 weeks	Intragastric, 50 mg/kg BW for 30 days	BW↓	Sperm count and motility↓	-	Oxidative stress: SOD↓, MDA↑	Meng et al. (2022)
							Sperm malformation↑		Germ cell apoptosis by inhibiting mitochondrial apoptotic pathway, expression of Bcl-2↑, Bax, Cleaved Caspase 3, and Cleaved Caspase 9↓
							Vacuolization of the seminiferous tubules, decreased layers of spermatogenic cells
Danafar et al.	2022	Testis	-	32 NMRI mice aged 8–12 weeks	Intragastric, 2.5,5,10 mg/kg BW TiO₂NPs for 40 days	-	Sperm malformation↑, spermatogenesis index and lumen parameters↓	T↓,	Malondyaldehyde in the seminal fluid↑ MDA↑	Danafar et al. (2021)
							Leydig cell count↓	FSH, LH, E2 (−)
							decreased layers of spermatogenic cells
Li et al.	2023	Testis	-	32SD rats aged 8 weeks	Intragastric, 50 mg/kg BW for 90 days		Sperm count, mobility↓		Oxidative stress: GSH-Px, CAT↓, MDA, LDH↑	Li et al. (2023)
Li et al.	2023	Testis	-	32SD rats aged 8 weeks	Intragastric, 50 mg/kg BW for 90 days		Sperm malformation↑		Testicular cell apoptosis, DNA damage in sperm	Li et al. (2023)
Halawa et al.	2022	Testis	50–55 nm	20 Albino rats aged 3–4 months	150 mg/kg BW for 14 days	Testicular weight (−)	Decreased layers of spermatogenic cells, vacuolization of the seminiferous tubules, infiltration of inflammatory cells	T↓	Oxidative stress: GSH,GST,SOD,CAT,GPx, GSH-Px↓MDA↑	Halawa et al. (2022)