Transplantation of decellularized and lyophilized amniotic membrane inhibits endometrial fibrosis by regulating connective tissue growth factor and tissue inhibitor of matrix metalloproteinase‑2

Chen,Xing; Zhou,Yan; Sun,Ying; Ji,Tonghui; Dai,Huihua

doi:10.3892/etm.2021.10400

Transplantation of decellularized and lyophilized amniotic membrane inhibits endometrial fibrosis by regulating connective tissue growth factor and tissue inhibitor of matrix metalloproteinase‑2

Authors:
- Xing Chen
- Yan Zhou
- Ying Sun
- Tonghui Ji
- Huihua Dai
View Affiliations

Affiliations: Department of Gynecology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu 210036, P.R. China, Department of Obstetrics, The First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu 210036, P.R. China
Published online on: July 7, 2021 https://doi.org/10.3892/etm.2021.10400
Article Number: 968
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Intrauterine adhesion (IUA) is a disease characterized by endometrial fibrosis caused by injury to the endometrium. In the present study, decellularized and lyophilized human amniotic membrane (DL‑AM) material was transplanted in a rat model to explore the preventive effect against IUA. A total of 24 Sprague Dawley rats were randomly divided into an IUA (n=12) group and an IUA + DL‑AM (n=12) group. To establish the model, the endometrium of the left uterus was scraped, while that of the right uterus was used as a control. In the IUA group, scraped uteri were sutured without any other treatment, whereas DL‑AM was transplanted onto the scraped uteri in the IUA + DL‑AM group. Uteri were resected for histological and immunohistochemical evaluation at 3, 7, 14 and 28 days after surgery. The results confirmed the development of IUA, which was accompanied by an increase in the rate of fibrotic area. Integral optical density (IOD) values of connective tissue growth factor (CTGF) were elevated in the IUA group, while matrix metalloproteinase‑2 (MMP‑2) decreased relative to the control group (P<0.05). After DL‑AM transplantation, the IOD value of CTGF dropped, while MMP‑2 increased compared with the IUA group (P<0.05). However, compared with that in the control group, the IOD value of CTGF was still higher, whereas MMP‑2 was still lower in the IUA + DL‑AM group (P<0.05). Furthermore, no evidence of endometrial regeneration was detected in both the IUA and IUA + DL‑AM groups. Overall, these results indicated that in the rat model of IUA, transplantation of DL‑AM had the potential to prevent the formation of fibrosis to a certain extent and may thus be an alternative strategy for managing the condition.

View Figures

View References

1	Yu D, Wong YM, Cheong Y, Xia E and Li TC: Asherman syndrome-one century later. Fertil Steril. 89:759–779. 2008.PubMed/NCBI View Article : Google Scholar
2	Westendorp IC, Ankum WM, Mol BW and Vonk J: Prevalence of Asherman's syndrome after secondary removal of placental remnants or a repeat curettage for incomplete abortion. Hum Reprod. 13:3347–3350. 1998.PubMed/NCBI View Article : Google Scholar
3	Vancaillie TG and Garad R: Asherman's syndrome. Aust Nurs J. 20:34–36. 2013.PubMed/NCBI
4	Conforti A, Alviggi C, Mollo A, De Placido G and Magos A: The management of Asherman syndrome: A review of literature. Reprod Biol Endocrinol. 11(118)2013.PubMed/NCBI View Article : Google Scholar
5	Fairbairn NG, Randolph MA and Redmond RW: The clinical applications of human amnion in plastic surgery. J Plast Reconstr Aesthet Surg. 67:662–675. 2014.PubMed/NCBI View Article : Google Scholar
6	Dua HS, Gomes JAP, King AJ and Maharajan VS: The amniotic membrane in ophthalmology. Surv Ophthalmol. 49:51–77. 2004.PubMed/NCBI View Article : Google Scholar
7	Fraser JF, Cuttle L, Kempf M, Phillips GE, Hayes MT and Kimble RM: A randomised controlled trial of amniotic membrane in the treatment of a standardised burn injury in the merino lamb. Burns. 35:998–1003. 2009.PubMed/NCBI View Article : Google Scholar
8	Gholipourmalekabadi M, Sameni M, Radenkovic D, Mozafari M, Mossahebi-Mohammadi M and Seifalian A: Decellularized human amniotic membrane: How viable is it as a delivery system for human adipose tissue-derived stromal cells? Cell Prolif. 49:115–121. 2016.PubMed/NCBI View Article : Google Scholar
9	Wilshaw SP, Kearney JN, Fisher J and Ingham E: Production of an acellular amniotic membrane matrix for use in tissue engineering. Tissue Eng. 12:2117–2129. 2006.PubMed/NCBI View Article : Google Scholar
10	Kakabadze Z, Mardaleishvili K, Loladze G, Javakhishvili I, Chakhunasvili K, Karalashvili L, Sukhitashvili N, Chutkerashvili G, Kakabadze A and Chakhunasvili D: Clinical application of decellularized and lyophilized human amnion/chorion membrane grafts for closing post-laryngectomy pharyngocutaneous fistulas. J Surg Oncol. 113:538–543. 2016.PubMed/NCBI View Article : Google Scholar
11	Chen X, Sun J, Li X, Mao L, Cui L and Bai W: Transplantation of oral mucosal epithelial cells seeded on decellularized and lyophilized amniotic membrane for the regeneration of injured endometrium. Stem Cell Res Ther. 10(107)2019.PubMed/NCBI View Article : Google Scholar
12	Chen X, Sun J, Li X, Mao L, Zhou Y, Cui L and Bai W: Antifibrotic effects of decellularized and lyophilized human amniotic membrane transplant on the formation of intrauterine adhesion. Exp Clin Transplant. 17:236–242. 2019.PubMed/NCBI View Article : Google Scholar
13	Chen X and Zhou YF: Preventive effects of transplantation of oral mucosal epithelial cells seeded on a decellularized amniotic membrane in a model of intrauterine adhesion. Int J Clin Exp Pathol. 11:1510–1519. 2018.PubMed/NCBI
14	Xue X, Chen Q, Zhao G, Zhao JY, Duan Z and Zheng PS: The overexpression of TGF-β and CCN2 in intrauterine adhesions involves the NF-κB signaling pathway. PLoS One. 10(e0146159)2015.PubMed/NCBI View Article : Google Scholar
15	Visse R and Nagase H: Matrix metalloproteinases and tissue inhibitors of metalloproteinases: Structure, function, and biochemistry. Circ Res. 92:827–839. 2003.PubMed/NCBI View Article : Google Scholar
16	Percie du Sert N, Hurst V, Ahluwalia A, Alam S, Avey MT, Baker M, Browne WJ, Clark A, Cuthill IC, Dirnagl U, et al: The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. J Physiol. 598:3793–3801. 2020.PubMed/NCBI View Article : Google Scholar
17	Fan B, Jin XH, Shi Y, Zhu H, Zhou W, Tu W and Ding L: Expression and significance of TIMP-3, PACAP and VIP in vaginal wall tissues of patients with stress urinary incontinence. Exp Ther Med. 13:624–628. 2017.PubMed/NCBI View Article : Google Scholar
18	Yu J, Luo Y, Yang XF, Yang MX, Yang J, Yang XS, Zhou J, Gao F, He LT and Xu J: Effects of perinatal exposure to nonylphenol on delivery outcomes of pregnant rats and inflammatory hepatic injury in newborn rats. Braz J Med Biol Res. 49(e5647)2016.PubMed/NCBI View Article : Google Scholar
19	Chávez-García C, Jiménez-Corona A, Graue-Hernández EO, Zaga-Clavellina V, García-Mejía M, Jiménez-Martínez MC and Garfias Y: Ophthalmic indications of amniotic membrane transplantation in Mexico: An eight years amniotic membrane bank experience. Cell Tissue Bank. 17:261–268. 2016.PubMed/NCBI View Article : Google Scholar
20	Koizumi N, Rigby H, Fullwood NJ, Kawasaki S, Tanioka H, Koizumi K, Kociok N, Joussen AM and Kinoshita S: Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells. Graefes Arch Clin Exp Ophthalmol. 245:123–134. 2007.PubMed/NCBI View Article : Google Scholar
21	Francisco JC, Correa Cunha R, Cardoso MA, Baggio Simeoni R, Mogharbel BF, Picharski GL, Silva Moreira Dziedzic D, Guarita-Souza LC and Carvalho KA: Decellularized amniotic membrane scaffold as a pericardial substitute: An in vivo study. Transplant Proc. 48:2845–2849. 2016.PubMed/NCBI View Article : Google Scholar
22	Roy R, Haase T, Ma N, Bader A, Becker M, Seifert M, Choi YH, Falk V and Stamm C: Decellularized amniotic membrane attenuates postinfarct left ventricular remodeling. J Surg Res. 200:409–419. 2016.PubMed/NCBI View Article : Google Scholar
23	Lian N and Li T: Growth factor pathways in hypertrophic scars: Molecular pathoenesis and therapeutic implications. Biomed Pharmacother. 84:42–50. 2016.PubMed/NCBI View Article : Google Scholar
24	He T, Quan T, Shao Y, Voorhees JJ and Fisher GJ: Oxidative exposure impairs TGF-β pathway via reduction of type II receptor and SMAD3 in human skin fibroblasts. Age (Dordr). 36(9623)2014.PubMed/NCBI View Article : Google Scholar
25	Sharma A, Thakur R, Lingaraju MC and Kumar D, Mathesh K, Telang AG, Singh TU and Kumar D: Betulinic acid attenuates renal fibrosis in rat chronic kidney disease model. Biomed Pharmacother. 89:796–804. 2017.PubMed/NCBI View Article : Google Scholar
26	Paradis V, Dargere D, Vidaud M, De Gouville AC, Huet S, Martinez V, Gauthier JM, Ba N, Sobesky R, Ratziu V and Bedossa P: Expression of connective tissue growth factor in experimental rat and human liver fibrosis. Hepatology. 30:968–976. 1999.PubMed/NCBI View Article : Google Scholar
27	Sgalla G, Franciosa C, Simonetti J and Richeldi L: Pamrevlumab for the treatment of idiopathic pulmonary fibrosis. Expert Opin Investig Drugs. 29:771–777. 2020.PubMed/NCBI View Article : Google Scholar
28	Hafez MM, Hamed SS, El-Khadragy MF, Hassan ZK, Al Rejaie SS, Sayed-Ahmed MM, Al-Harbi NO, Al-Hosaini KA, Al-Harbi MM, Alhoshani AR, et al: Effect of ginseng extract on the TGF-β1 signaling pathway in CCl₄-induced liver fibrosis in rats. BMC Complement Altern Med. 17(45)2017.PubMed/NCBI View Article : Google Scholar
29	Knittel T, Mehde M, Grundmann A, Saile B, Scharf JG and Ramadori G: Expression of matrix metalloproteinases and their inhibitors during hepatic tissue repair in the rat. Histochem Cell Biol. 113:443–453. 2000.PubMed/NCBI View Article : Google Scholar
30	Guan S, Liu Q, Han F, Gu W, Song L, Zhang Y, Guo X and Xu W: Ginsenoside Rg1 ameliorates cigarette smoke-induced airway fibrosis by suppressing the TGF-β1/Smad pathway in vivo and in vitro. Biomed Res Int. 2017(6510198)2017.PubMed/NCBI View Article : Google Scholar
31	Zhang M, Hu X, Li S, Lu C, Li J, Zong Y, Qi W and Yang H: Hepatoprotective effects of ethyl pyruvate against CCl4-induced hepatic fibrosis via inhibition of TLR4/NF-κB signaling and up-regulation of MMPs/TIMPs ratio. Clin Res Hepatol Gastroenterol. 42:72–81. 2018.PubMed/NCBI View Article : Google Scholar
32	Zuo WL, Zhao JM, Huang JX, Zhou W, Lei ZH, Huang YM, Huang YF and Li HG: Effect of bosentan is correlated with MMP-9/TIMP-1 ratio in bleomycin-induced pulmonary fibrosis. Biomed Rep. 6:201–205. 2017.PubMed/NCBI View Article : Google Scholar
33	Zhao D, Wang Y, Du C, Shan S, Zhang Y, Du Z and Han D: Honokiol alleviates hypertrophic scar by targeting transforming growth factor-β/Smad2/3 signaling pathway. Front Pharmacol. 8(206)2017.PubMed/NCBI View Article : Google Scholar