Intrauterine adhesions (IUAs) are characterized by the injury of endometrium due to curettage and/or endometritis. The loss of functional endometrium in uterine cavity usually results in hypomenorrhea, amenorrhea, infertility, and/or recurrent pregnancy loss. Recently, stem cell transplantation has been applied to promote the endometrial regeneration. Human amnion epithelial cells (hAECs) have been shown to have stem cell characteristics. In this study, we found that PKH26-labeled hAECs were mainly distributed in the basal layer of endometrium after transplantation, and hAEC transplantation, including uterine injection and tail vein injection, could increase pregnancy rate and the number of embryos in rat model of IUAs. Moreover, hAEC transplantation was demonstrated to increase the endometrial thickness, promote the proliferation of glands and blood vessels, and decrease fibrotic areas in the endometrium. The immunohistochemical and quantitative polymerase chain reaction analysis showed the upregulated expression of growth factors, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1) after hAEC transplantation; and the downregulated expression of collagen type I alpha 1 (COL1A1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and transforming growth factor-β (TGF-β), all of which are associated with the extracellular matrix (ECM) deposition after hAEC transplantation. The mRNA sequencing indicated that platelet-derived growth factor-C (PDGF-C), thrombospondin-1 (THBS1), connective tissue growth factor (CTGF), Wnt5a, and Snai2 were significantly modulated in treatment groups. These results indicate that hAEC transplantation promotes endometrial regeneration and the restoration of fertility in rat model of IUAs.
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References
1.
DeansR and AbbottJ. (2010). Review of intrauterine adhesions. J Minim Invasive Gynecol, 17:555–569.
2.
DaltonVK, SaundersNA, HarrisLH, WilliamsJA and LebovicDI. (2006). Intrauterine adhesions after manual vacuum aspiration for early pregnancy failure. Fertil Steril, 85:1823.e1–1823.e3.
3.
PanayotidisC, WeyersS, BosteelsJ and van HerendaelB. (2009). Intrauterine adhesions (IUA): has there been progress in understanding and treatment over the last 20 years?. Gynecol Surg, 6:197–211.
4.
SchenkerJG. (1996). Etiology of and therapeutic approach to synechia uteri. Eur J Obstet Gynecol Reprod Biol, 65:109–113.
5.
LiuL, YangH, GuoY, YangG and ChenY. (2019). The impact of chronic endometritis on endometrial fibrosis and reproductive prognosis in patients with moderate and severe intrauterine adhesions: a prospective cohort study. Fertil Steril, 111:1002.e2–1010.e2.
6.
Di SpiezioSA, SpinelliM, BramanteS, ScognamiglioM, GrecoE, GuidaM, CelaV and NappiC. (2011). Efficacy of a polyethylene oxide-sodium carboxymethylcellulose gel in prevention of intrauterine adhesions after hysteroscopic surgery. J Minim Invasive Gynecol, 18:462–469.
7.
MaisV, CirronisMG, PeirettiM, FerrucciG, CossuE and MelisGB. (2012). Efficacy of auto-crosslinked hyaluronan gel for adhesion prevention in laparoscopy and hysteroscopy: a systematic review and meta-analysis of randomized controlled trials. Eur J Obstet Gynecol Reprod Biol, 160:1–5.
8.
AmerMI, Abd-El-MaeboudKHI, AbdelfatahI, SalamaFA and AbdallahAS. (2010). Human amnion as a temporary biologic barrier after hysteroscopic lysis of severe intrauterine adhesions: pilot study. J Minim Invasive Gynecol, 17:605–611.
9.
Di SpiezioSA, CalagnaG, ScognamiglioM, O'DonovanP, CampoR and De WildeRL. (2016). Prevention of intrauterine post-surgical adhesions in hysteroscopy. A systematic review. Eur J Obstet Gynecol Reprod Biol, 203:182–192.
10.
MarchCM. (2011). Management of Asherman's syndrome. Reprod Biomed, 23:63–76.
11.
GargettCE and HealyDL. (2011). Generating receptive endometrium in Asherman's syndrome. J Hum Reprod Sci, 4:49–52.
12.
TaylorHS. (2004). Endometrial cells derived from donor stem cells in bone marrow transplant recipients. JAMA, 292:81–85.
13.
GargettCE and ChanRW. (2006). Endometrial stem/progenitor cells and proliferative disorders of the endometrium. Minerva Ginecol, 58:511–526.
14.
GargettCE. (2007). Uterine stem cells: what is the evidence?. Hum Reprod Update, 13:87–101.
15.
GargettCE and YeL. (2012). Endometrial reconstruction from stem cells. Fertil Steril, 98:11–20.
16.
KilicS, YukselB, PinarliF, AlbayrakA, BoztokB and DelibasiT. (2014). Effect of stem cell application on Asherman syndrome, an experimental rat model. J Assist Reprod Genet, 31:975–982.
17.
GanL, DuanH, XuQ, TangYQ, LiJJ, SunFQ and WangS. (2017). Human amniotic mesenchymal stromal cell transplantation improves endometrial regeneration in rodent models of intrauterine adhesions. Cytotherapy, 19:603–616.
18.
ZhangJ, LiN, ChenS, ChenM, LiB, LiX, et al. (2016). Transplantation induced pluripotent stem cells in situ improves fertility outcome impaired by intrauterine adhesions in mice. Int J Clin Exp Med, 9:10651–10661.
19.
IlancheranS, MichalskaA, PehG, WallaceEM, PeraM and ManuelpillaiU. (2007). Stem cells derived from human fetal membranes display multilineage differentiation potential. Biol Reprod, 77:577–588.
20.
MikiT, LehmannT, CaiH, StolzDB and StromSC. (2005). Stem cell characteristics of amniotic epithelial cells. Stem Cells, 23:1549–1559.
21.
MikiT, MarongiuF, DorkoK, EllisECS and StromSC. (2010). Isolation of amniotic epithelial stem cells. Curr Protoc Stem Cell Biol Unit, 1E.3; DOI: 10.1002/9780470151808.sc01e03s12.
22.
MikiT, MarongiuF, EllisECS, DorkoK, MitamuraK, RanadeA, GramignoliR, DavilaJ and StromSC. (2009). Production of hepatocyte-like cells from human amnion. Methods Mol Biol, 481:155–168.
23.
WeiJP, ZhangTS, KawaS, AizawaT, OtaM, AkaikeT, KatoK, KonishiI and NikaidoT. (2003). Human amnion-isolated cells normalize blood glucose in streptozotocin-induced diabetic mice. Cell Transplant, 12:545–552.
24.
TeradaS, MatsuuraK, EnosawaS, MikiM, HoshikaA, SuzukiS and SakuragawaN. (2000). Inducing proliferation of human amniotic epithelial (HAE) cells for cell therapy. Cell Transplant, 9:701–704.
25.
LefebvreS, AdrianF, MoreauP, GourandL, DaussetJ, Berrih-AkninS, CarosellaED and PaulP. (2000). Modulation of HLA-G expression in human thymic and amniotic epithelial cells. Hum Immunol, 61:1095–1101.
26.
AdinolfiM, AkleCA, McCollI, FensomAH, TansleyL, ConnollyP, HsiBL, FaulkWP, TraversP and BodmerWF. (1982). Expression of HLA antigens, beta 2-microglobulin and enzymes by human amniotic epithelial cells. Nature, 295:325–327.
27.
MikiT. (2016). A rational strategy for the use of amniotic epithelial stem cell therapy for liver diseases. Stem Cells Transl Med, 5:405–409.
28.
HoriJ, WangM, KamiyaK, TakahashiH and SakuragawaN. (2006). Immunological characteristics of amniotic epithelium. Cornea, 25:S53–S58.
29.
CaiH, LiH and HeY. (2016). Interceed and estrogen reduce uterine adhesions and fibrosis and improve endometrial receptivity in a rabbit model of intrauterine adhesions. Reprod Sci, 23:1208–1216.
30.
BaiX, LiuJ, CaoS and WangL. (2019). Mechanisms of endometrial fibrosis and the potential application of stem cell therapy. Discov Med, 27:267–279.
31.
VosdoganesP, HodgesRJ, LimR, WestoverAJ, AcharyaRY, WallaceEM, and MossTJM. (2011). Human amnion epithelial cells as a treatment for inflammation-induced fetal lung injury in sheep. Am J Obstet Gynecol, 205:156.e26–156.e33.
32.
MoodleyY, IlancheranS, SamuelC, VaghjianiV, AtienzaD, WilliamsED, JenkinG, WallaceE, TrounsonA and ManuelpillaiU. (2010). Human amnion epithelial cell transplantation abrogates lung fibrosis and augments repair. Am J Respir Crit Care Med, 182:643–651.
33.
SkvorakKJ, DorkoK, MarongiuF, TahanV, HanselMC, GramignoliR, ArningE, BottiglieriT, GibsonKM and StromSC. (2013). Improved amino acid, bioenergetic metabolite and neurotransmitter profiles following human amnion epithelial cell transplant in intermediate maple syrup urine disease mice. Mol Genet Metab, 109:132–138.
34.
WuQ, FangT, LangH, ChenM, ShiP, PangX and QiG. (2017). Comparison of the proliferation, migration and angiogenic properties of human amniotic epithelial and mesenchymal stem cells and their effects on endothelial cells. Int J Mol Med, 39:918–926.
35.
VáczG, CselenyákA, CserépZ, BenkőR, KovácsE, PankotaiE, LindenmairA, WolbankS and SchwarzCM. (2016). Effects of amniotic epithelial cell transplantation in endothelial injury. Interv Med Appl Sci, 8:164–171.
36.
DingD, MaoD, LiK, WangX, QinW, LiuR, ChiamDS, TomczakN, YangZ, et al. (2014). Precise and long-term tracking of adipose-derived stem cells and their regenerative capacity via superb bright and stable organic nanodots. ACS Nano, 8:12620–12631.
37.
GanL, DuanH, XuQ, TangYQ, LiJJ, SunFQ and WangS. (2017). Human amniotic mesenchymal stromal cell transplantation improves endometrial regeneration in rodent models of intrauterine adhesions. Cytotherapy, 19:603–616.
38.
LinN, LiX, SongT, WangJ, MengK, YangJ, HouX, DaiJ and HuY. (2012). The effect of collagen-binding vascular endothelial growth factor on the remodeling of scarred rat uterus following full-thickness injury. Biomaterials, 33:1801–1807.
39.
WeiP, ChenXL, SongXX, HanCS and LiuYX. (2004). VEGF, bFGF, and their receptors in the endometrium of rhesus monkey during menstrual cycle and early pregnancy. Mol Reprod Dev, 68:456–462.
40.
AbrahamJA, MergiaA, WhangJL, TumoloA, FriedmanJ, HjerrildKA, GospodarowiczD and FiddesJC. (1986). Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor. Science, 233:545–548.
41.
FujitaM, IshiharaM, SimizuM, ObaraK, IshizukaT, SaitoY, YuraH, MorimotoY, TakaseB, et al. (2004). Vascularization in vivo caused by the controlled release of fibroblast growth factor-2 from an injectable chitosan/non-anticoagulant heparin hydrogel. Biomaterials, 25:699–706.
42.
LiX, SunH, LinN, HouX, WangJ, ZhouB, XuP, XiaoZ, ChenB, DaiJ and HuY. (2011). Regeneration of uterine horns in rats by collagen scaffolds loaded with collagen-binding human basic fibroblast growth factor. Biomaterials, 32:8172–8181.
43.
TorryDS, HoltVJ, KeenanJA, HarrisG, CaudleMR and TorryRJ. (1996). Vascular endothelial growth factor expression in cycling human endometrium. Fertil Steril, 66:72–80.
44.
DvorakHF, NagyJA, FengD, BrownLF and DvorakAM. (1999). Vascular permeability factor/vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Microbiol Immunol, 237:97–132.
45.
TammelaT, EnholmB, AlitaloK and PaavonenK. (2005). The biology of vascular endothelial growth factors. Cardiovasc Res, 65:550–563.
46.
FanX, KriegS, KuoCJ, WiegandSJ, RabinovitchM, DruzinML, BrennerRM, GiudiceLC and NayakNR. (2008). VEGF blockade inhibits angiogenesis and reepithelialization of endometrium. FASEB J, 22:3571–3580.
47.
RisauW. (1997). Mechanisms of angiogenesis. Nature, 386:671–674.
48.
HuntTK and PaiMP. (1972). The effect of varying ambient oxygen tensions on wound metabolism and collagen synthesis. Surg Gynecol Obstet, 135:561–567.
49.
JensenJA, HuntTK, ScheuenstuhlH and BandaMJ. (1986). Effect of lactate, pyruvate, and pH on secretion of angiogenesis and mitogenesis factors by macrophages. Lab Invest, 54:574–578.
50.
SuhDY, HuntTK and SpencerEM. (1992). Insulin-like growth factor-I reverses the impairment of wound healing induced by corticosteroids in rats. Endocrinology, 131:2399–2403.
SchultzG, KhawPT, OxfordK, MaCauleyS, Van SettenG and CheginiN. (1994). Growth factors and ocular wound healing. Eye Lond, 8:184–187.
53.
GiudiceLC, DsupinBA, JinIH, VuTH and HoffmanAR. (1993). Differential expression of messenger ribonucleic acids encoding insulin-like growth factors and their receptors in human uterine endometrium and decidua. J Clin Endocrinol Metab, 76:1115–1122.
54.
VancaillieTG and GaradR. (2013). Asherman's syndrome. Aust Nurs J, 20:34–36.
55.
ShanT, ZhangL, ZhaoC, ChenW, ZhangY and LiG. (2014). Angiotensin-(1–7) and angiotensin Ⅱ induce the transdifferentiation of human endometrial epithelial cells in vitro. Mol Med Rep, 9:2180–2186.
56.
SchuppanD, RuehlM, SomasundaramR and HahnEG. (2001). Matrix as a modulator of hepatic fibrogenesis. Semin Liver Dis, 21:351–372.
57.
BornsteinP and SageH. (1989). Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol, 37:67–106.
58.
IredaleJP. (1997). Tissue inhibitors of metalloproteinases in liver fibrosis. Int J Biochem Cell Biol, 29:43–54.
59.
DasSK and VasudevanDM. (2008). Genesis of hepatic fibrosis and its biochemical markers. Scand J Clin Lab Invest, 68:260–269.
60.
YoshijiH, KuriyamaS, MiyamotoY, ThorgeirssonUP, GomezDE, KawataM, YoshiiJ, IkenakaY, NoguchiR, et al. (2000). Tissue inhibitor of metalloproteinases-1 promotes liver fibrosis development in a transgenic mouse model. Hepatology, 32:1248–1254.
61.
DudásJ, KovalszkyI, GallaiM, NagyJO, SchaffZ, KnittelT, MehdeM, NeubauerK, SzalayF and RamadoriG. (2001). Expression of decorin, transforming growth factor-beta 1, tissue inhibitor metalloproteinase 1 and 2, and type IV collagenases in chronic hepatitis. Am J Clin Pathol, 115:725–735.
62.
MurphyFR, IssaR, ZhouX, RatnarajahS, NagaseH, ArthurMJP, BenyonC and IredaleJP. (2002). Inhibition of apoptosis of activated hepatic stellate cells by tissue inhibitor of metalloproteinase-1 is mediated via effects on matrix metalloproteinase inhibition: implications for reversibility of liver fibrosis. J Biol Chem, 277:11069–11076.
63.
NieQH, ChengYQ, XieYM, ZhouYX and CaoYZ. (2001). Inhibiting effect of antisense oligonucleotides phosphorthioate on gene expression of TIMP-1 in rat liver fibrosis. World J Gastroenterol, 7:363–369.
64.
BorderWA, NobleNA, YamamotoT, HarperJR, YamaguchiY, PierschbacherMD and RuoslahtiE. (1992). Natural inhibitor of transforming growth factor-beta protects against scarring in experimental kidney disease. Nature, 360:361–364.
65.
SimePJ, XingZ, GrahamFL, CsakyKG and GauldieJ. (1997). Adenovector-mediated gene transfer of active transforming growth factor-beta1 induces prolonged severe fibrosis in rat lung. J Clin Invest, 100:768–776.
66.
ClouthierDE, ComerfordSA and HammerRE. (1997). Hepatic fibrosis, glomerulosclerosis, and a lipodystrophy-like syndrome in PEPCK-TGF-beta1 transgenic mice. J Clin Invest, 100:2697–2713.
67.
ZeisbergM and KalluriR. (2013). Cellular mechanisms of tissue fibrosis. 1. Common and organ-specific mechanisms associated with tissue fibrosis. Am J Physiol Cell Physiol, 304:C216–C225.
68.
SaedGM and DiamondMP. (2002). Hypoxia-induced irreversible up-regulation of type I collagen and transforming growth factor-beta1 in human peritoneal fibroblasts. Fertil Steril, 78:144–147.
69.
CheginiN. (2008). TGF-beta system: the principal profibrotic mediator of peritoneal adhesion formation. Semin Reprod Med, 26:298–312.
70.
ParkJO, LeeBH, KangYM, KimTH, YoonJY, KimH, KwonUH, LeeKI, LeeHM and MoonSH. (2013). Inflammatory cytokines induce fibrosis and ossification of human ligamentum flavum cells. J Spinal Disord Tech, 26:E6–E12.
71.
SpornMB and RobertsAB. (1990). The Transforming Growth Factors Peptide Growth Factors and Their Receptors Lpp. Springer Verlag, NewYork, pp 419–472.
72.
LiX, PonténA, AaseK, KarlssonL, AbramssonA, UutelaM, BäckströmG, HellströmM, BoströmH, et al. (2000). PDGF-C is a new protease-activated ligand for the PDGF alpha-receptor. Nat Cell Biol, 2:302–309.
73.
CampbellJS, HughesSD, GilbertsonDG, PalmerTE, HoldrenMS, HaranAC, OdellMM, BauerRL, RenHP, et al. (2005). Platelet-derived growth factor C induces liver fibrosis, steatosis, and hepatocellular carcinoma. Proc Natl Acad Sci (USA), 102:3389–3394.
74.
PonténA, LiX, ThorénP, AaseK, SjöblomT, OstmanA and ErikssonU. (2003). Transgenic overexpression of platelet-derived growth factor-C in the mouse heart induces cardiac fibrosis, hypertrophy, and dilated cardiomyopathy. Am J Pathol, 163:673–682.
75.
GilbertsonDG, DuffME, WestJW, KellyJD, SheppardPO, HofstrandPD, GaoZ, ShoemakerK, BukowskiTR, et al. (2001). Platelet-derived growth factor C (PDGF-C), a novel growth factor that binds to PDGF alpha and beta receptor. J Biol Chem, 276:27406–27414.
76.
Schultz-CherryS and Murphy-UllrichJE. (1993). Thrombospondin causes activation of latent transforming growth factor-beta secreted by endothelial cells by a novel mechanism. J Cell Biol, 122:923–932.
77.
DanielC, WiedeJ, KrutzschHC, RibeiroSMF, RobertsDD, Murphy-UllrichJE and HugoC. (2004). Thrombospondin-1 is a major activator of TGF-beta in fibrotic renal disease in the rat in vivo. Kidney Int, 65:459–468.
78.
BornsteinP. (2001). Thrombospondins as matricellular modulators of cell function. J Clin Invest, 107:929–934.
79.
Murphy-UllrichJE and PoczatekM. (2000). Activation of latent TGF-beta by thrombospondin-1: mechanisms and physiology. Cytokine Growth Factor Rev, 11:59–69.
80.
BagavandossP and WilksJW. (1990). Specific inhibition of endothelial cell proliferation by thrombospondin. Biochem Biophys Res Commun, 170:867–872.
81.
DiPietroLA, NebgenDR and PolveriniPJ. (1994). Downregulation of endothelial cell thrombospondin 1 enhances in vitro angiogenesis. J Vasc Res, 31:178–185.
82.
Iruela-ArispeML, BornsteinP and SageH. (1991). Thrombospondin exerts an antiangiogenic effect on cord formation by endothelial cells in vitro. Proc Natl Acad Sci (USA), 88:5026–5030.
83.
Iruela-ArispeML, PorterP, BornsteinP and SageEH. (1996). Thrombospondin-1, an inhibitor of angiogenesis, is regulated by progesterone in the human endometrium. J Clin Invest, 97:403–412.
84.
GressnerOA and GressnerAM. (2008). Connective tissue growth factor: a fibrogenic master switch in fibrotic liver diseases. Liver Int, 28:1065–1079.
85.
LiJ, DuS, ShengX, LiuJ, CenB, HuangF and HeY. (2016). MicroRNA-29b inhibits endometrial fibrosis by regulating the Sp1-TGF-β1/Smad-CTGF axis in a rat model. Reprod Sci, 23:386–394.
86.
HeikkiläM, PeltoketoH and VainioS. (2001). Wnts and the female reproductive system. J Exp Zool, 290:616–623.
87.
YuWZ, ChenXM, NiuWB, WangF, SunB and SunYP. (2015). Role of Wnt5a in the differentiation of human embryonic stem cells into endometrium-like cells. Int J Clin Exp Pathol, 8:5478–5484.
88.
PukropT and BinderC. (2008). The complex pathways of Wnt 5a in cancer progression. J Mol Med, 86:259–266.
89.
ChaJ, BartosA, ParkC, SunX, LiY, ChaSW, AjimaR, HoHYH, YamaguchiTP and DeySK. (2014). Appropriate crypt formation in the uterus for embryo homing and implantation requires Wnt5a-ROR signaling. Cell Rep, 8:382–392.
90.
SavagnerP, KusewittDF, CarverEA, MagninoF, ChoiC, GridleyT and HudsonLG. (2005). Developmental transcription factor slug is required for effective re-epithelialization by adult keratinocytes. J Cell Physiol, 202:858–866.
91.
NewkirkKM, ParentAE, FosseySL, ChoiC, ChandlerHL, Rajala-SchultzPJ and KusewittDF. (2007). Snai2 expression enhances ultraviolet radiation-induced skin carcinogenesis. Am J Pathol, 171:1629–1639.
92.
NewkirkKM, MacKenzieDA, BakaletzAP, HudsonLG and KusewittDF. (2008). Microarray analysis demonstrates a role for Slug in epidermal homeostasis. J Invest Dermatol, 128:361–369.
93.
TurnerFE, BroadS, KhanimFL, JeanesA, TalmaS, HughesS, TselepisC and HotchinNA. (2006). Slug regulates integrin expression and cell proliferation in human epidermal keratinocytes. J Biol Chem, 281:21321–21331.
94.
DuF, YangR, MaHL, WangQY and WeiSL. (2009). Expression of transcriptional repressor Slug gene in mouse endometrium and its effect during embryo implantation. Appl Biochem Biotechnol, 157:346–355.
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