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Oct 2013

Glutaraldehyde cross-linking of amniotic membranes affects their nanofibrous structures and limbal epithelial cell culture characteristics

Abstract Given that the cells can sense nanometer dimensions, the chemical cross-linking-mediated alteration in fibrillar structure of collagenous tissue scaffolds is critical to determining their cell culture performances. This article explores, for the first time, the effect of nanofibrous structure of glutaraldehyde (GTA) cross-linked amniotic membrane (AM) on limbal epithelial cell (LEC) cultivation. Results of ninhydrin assays demonstrated that the amount of new cross-links formed between the collagen chains is significantly increased with increasing the cross-linking time from 1 to 24 hours. By transmission electron microscopy, the AM treated with GTA for a longer duration exhibited a greater extent of molecular aggregation, thereby leading to a considerable increase in nanofiber diameter and resistance against collagenase degradation. In vitro biocompatibility studies showed that the samples cross-linked with GTA for 24 hours are not well-tolerated by the human corneal epithelial cell cultures. When the treatment duration is less than 6 hours, the biological tissues cross-linked with GTA for a longer time may cause slight reductions in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt, and anti-inflammatory activities. Nevertheless, significant collagen molecular aggregation also enhances the stemness gene expression, indicating a high ability of these AM matrices to preserve the progenitors of LECs in vitro. It is concluded that GTA cross-linking of collagenous tissue materials may affect their nanofibrous structures and corneal epithelial stem cell culture characteristics. The AM treated with GTA for 6 hours holds promise for use as a niche for the expansion and transplantation of limbal epithelial progenitor cells.


Keywords: glutaraldehyde, amniotic membrane, nanofibrous structure, limbal epithelial cell, stemness

Lai J-Y, Ma DH-K. Glutaraldehyde cross-linking of amniotic membranes affects their nanofibrous structures and limbal epithelial cell culture characteristics. International Journal of Nanomedicine. 2013;8:4157-4168. doi:10.2147/IJN.S52731.
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Aug 2012

Amniotic Membrane: From Structure And Functions To Clinical Applications

Abstract Amniotic membrane (AM) or amnion is a thin membrane on the inner side of the fetal placenta; it completely surrounds the embryo and delimits the amniotic cavity, which is filled by amniotic liquid. In recent years, the structure and function of the amnion have been investigated, particularly the pluripotent properties of AM cells, which are an attractive source for tissue transplantation. AM has anti-inflammatory, anti-bacterial, anti-viral and immunological characteristics, as well as anti-angiogenic and pro-apoptotic features. AM is a promoter of epithelialization and is a non-tumorigenic tissue and its use has no ethical problems. Because of its attractive properties, AM has been applied in several surgical procedures related to ocular surface reconstruction and the genito-urinary tract, skin, head and neck, among others. So far, the best known and most auspicious applications of AM are ocular surface reconstruction, skin applications and tissue engineering. However, AM can also be applied in oncology. In this area, AM can prevent the delivery of nutrients and oxygen to cancer cells and consequently interfere with tumour angiogenesis, growth and metastasis.

Mamede AC, Carvalho MJ, Abrantes AM, Laranjo M, Maia CJ, Botelho MF. Amniotic membrane: from structure and functions to clinical applications. Cell Tissue Res. 2012;349(2):447-458.
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Apr 2008

Properties of the Amniotic Membrane for Potential Use in Tissue Engineering

An important component of tissue engineering (TE) is the supporting matrix upon which cells and tissues grow, also known as the scaffold. Scaffolds must easily integrate with host tissue and provide an excellent environment for cell growth and differentiation. Most scaffold materials are naturally derived from mammalian tissues. The amniotic membrane (AM) is considered an important potential source for scaffolding material. The AM represents the innermost layer of the placenta and is composed of a single epithelial layer, a thick basement membrane and an avascular stroma. The special structure and biological viability of the AM allows it to be an ideal candidate for creating scaffolds used in TE. Epithelial cells derived from the AM have the advantages of stem cells, yet are a more suitable source of cells for TE than stem cells. The extracellular matrix components of the basement membrane of the AM create an almost native scaffold for cell seeding in TE. In addition, the AM has other biological properties important for TE, including anti-inflammatory, anti-microbial, anti-fibrosis, anti-scarring, as well as reasonable mechanical property and low immunogenicity. In this review, the various properties of the AM are discussed in light of their potential use for TE.

Eur Cell Mater. 2008 Apr 29;15:88-99.
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May 2005

Amniotic Fluid: Not Just Fetal Urine Anymore

Amniotic fluid (AF) is a complex substance essential to fetal well-being. This article reviews recent discoveries and the current understanding of the origin and circulation of AF and its nutritive, protective, and diagnostic functions. Future directions for AF research are also discussed.

Amniotic fluid (AF) is a marvelously complex and dynamic milieu that changes as pregnancy progresses. AF contains nutrients and growth factors that facilitate fetal growth, provides mechanical cushioning and antimicrobial effectors that protect the fetus, and allows assessment of fetal maturity and disease. This article will review the development, content, and clinical significance of AF and its essential role in helping the fetus become a newborn.

Journal of Perinatology (2005) 25, 341–348. doi:10.1038/sj.jp.7211290. Mark A Underwood MD1, William M Gilbert MD2 and Michael P Sherman MD1
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Mar 2017

Efficacy of freeze-dried amnion graft following hysteroscopic adhesiolysis of severe intrauterine adhesions

Objective To evaluate the efficacy of freeze-dried amnion graft for prevention of intrauterine adhesion (IUA) reformation after hysteroscopic adhesiolysis.

Methods A prospective randomized controlled trial was conducted among 88 women with severe IUAs who underwent hysteroscopic adhesiolysis at Beijing Obstetrics and Gynecology Hospital between July 15, 2015, and July 1, 2016. All participants had a balloon inserted into the uterine cavity for 1 week. Sterilized freeze-dried amnion graft covered the balloon portion of the Foley catheter among patients allocated to the amnion group (n=44), whereas patients in the control group (n=44) did not receive the graft. Follow-up hysteroscopy was performed 3 months after surgery. Preoperative and postoperative IUA scores, menstruation scores, and pregnancy rates were assessed.

Results Both groups exhibited reductions in IUA scores and improvements in menstruation scores following treatment (P<0.001 for each measure). Compared with the control group, the amnion group had a lower IUA score (P=0.032) and a higher menstruation score (P<0.001) at follow-up. By contrast, the rates of IUA reformation and pregnancy were not significantly different between the two groups.
Conclusion Use of freeze-dried amnion graft was effective in reducing IUA reformation and improving menstruation (according to pictorial blood-loss assessment chart) following hysteroscopic adhesiolysis of severe IUAs.

Gan, L., Duan, H., Sun, F.-Q., Xu, Q., Tang, Y.-Q. and Wang, S. (2017), Efficacy of freeze-dried amnion graft following hysteroscopic adhesiolysis of severe intrauterine adhesions. Int J Gynecol Obstet, 137: 116–122. doi:10.1002/ijgo.12112
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Jun 2016

Amniotic Membrane can be a Valid Source for Wound Healing

Abstract Amniotic membrane (AM) can promote proper epithelialization with suppression of excessive fibrosis by creating a supportive milieu for regeneration of chronic ulcer bed.

Objective The objective of this study is to investigate whether AM scaffold can modulate the healing of a wound by promoting tissue reconstruction rather than promoting scar tissue formation.

Subjects and methods AM was obtained and prepared and then applied to patients with chronic leg ulcers who were randomly divided into two different groups. Group I (control group) included eleven patients in whom ulcers were treated with conventional wound dressings that were changed daily for 8 weeks. Group II (study group) included 14 patients in whom the AM was placed in contact with the ulcer and held in place with a secondary dressing, which was changed daily. Follow-up was done to detect healing rate and detection of ulcer size, assessment of pain, and to take ulcer images (days 0, 7, 14, 21, 30, 45, and 60).


ElHeneidy H, Omran E, Halwagy A, Al-Inany H, Al-Ansary M, Gad A (2016) Amniotic membrane can be a valid source for wound healing. Dove Press (2016:8) 225-231. DOI https://doi.org/10.2147/IJWH.S96636
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Nov 2015

Human Amniotic Membrane–Derived Products in Sports Medicine

Background: Amniotic membrane (AM)–derived products have been successfully used in ophthalmology, plastic surgery, and wound care, but little is known about their potential applications in orthopaedic sports medicine.

Purpose: To provide an updated review of the basic science and preclinical and clinical data supporting the use of AM-derived products and to review their current applications in sports medicine.

Study Design: Systematic review.

Methods: A systematic search of the literature was conducted using the Medline, EMBASE, and Cochrane databases. The search term amniotic membrane was used alone and in conjunction with stem cell, orthopaedic, tissue engineering, scaffold, and sports medicine.

Results: The search identified 6870 articles, 80 of which, after screening of the titles and abstracts, were considered relevant to this study. Fifty-five articles described the anatomy, basic science, and nonorthopaedic applications of AM-derived products. Twenty-five articles described preclinical and clinical trials of AM-derived products for orthopaedic sports medicine. Because the level of evidence obtained from this search was not adequate for systematic review or meta-analysis, a current concepts review on the anatomy, physiology, and clinical uses of AM-derived products is presented.

Conclusion: Amniotic membranes have many promising applications in sports medicine. They are a source of pluripotent cells, highly organized collagen, antifibrotic and anti-inflammatory cytokines, immunomodulators, and matrix proteins. These properties may make it beneficial when applied as tissue engineering scaffolds, improving tissue organization in healing, and treatment of the arthritic joint. The current body of evidence in sports medicine is heavily biased toward in vitro and animal studies, with little to no human clinical data. Nonetheless, 14 companies or distributors offer commercial AM products. The preparation and formulation of these products alter their biological and mechanical properties, and a thorough understanding of these differences will help guide the use of AM-derived products in sports medicine research.

Keywords: amniotic membrane; stem cell; regenerative medicine; scarless healing

Jonathan C. Riboh, MD, Bryan M. Saltzman, MD, Adam B. Yanke, MD, Brian J. Cole, MD, MBA. Human Amniotic Membrane–Derived Products in Sports Medicine. The American Journal of Sports Medicine. Vol 44, Issue 9, pp. 2425 – 2434. First published date: November-19-2015; 10.1177/0363546515612750
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Nov 2014

Safety Analysis Of Cryopreserved Amniotic Membrane/Umbilical Cord Tissue In Foot And Ankle Surgery: A Consecutive Case Series Of 124 Patients

AbstractCryopreserved amnion tissues derived from amniotic membrane/umbilical cord (AM/UC) have been used extensively in ophthalmology for minimizing postoperative inflammation, pain, and adhesion formation following various surgical procedures. There is limited data in the current literature regarding the use of amnion tissue product in foot and ankle surgery. The purpose of this retrospective study is to report the short-term safety profile after in vivo application of cryopreserved AM/UC tissue use in foot and ankle surgery. A retrospective consecutive case series was performed for cases where amnion tissue was used with a minimum follow-up of 120 days between 2011 and 2012. The clinical outcomes of interest were postoperative infections, delayed or nonhealing wounds, adverse surgical site reactions, and repeat surgery for formal irrigation and debridement. One hundred twenty-four patients qualified for inclusion. Cryopreserved AM/UC tissue was introduced into the surgical wound and placed adjacent to the compromised and repaired tendons, most frequently in peroneal and Achilles tendon repairs. In this level IV consecutive case series cohort, there was an overall wound complication rate of 5.64%, with a re-operation rate of 1.6% (2/124). In each of these cases, patients had an irrigation and debridement with ultimate successful wound healing. The results of this study demonstrate that the use of amnion tissue in the foot and ankle setting is safe with a decreasing trend in overall complication rates compared with historically published norms.

DeMill SL, Granata JD, McAlister JE, Berlet GC, Hyer CF. Safety analysis of cryopreserved amniotic membrane/umbilical cord tissue in foot and ankle surgery: a consecutive case series of 124 patients. Surg Technol Int. 2014;25:257-261.
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Jun 2016

Surgical Therapies and Tissue Engineering: At the Intersection Between Innovation and Regulation

Innovations in tissue engineering and regenerative medicine are often realized in the operating room as surgeons restore form and function using biomaterials and grafted tissues. Examples seen every day in operating rooms around the world include the use of decellularized dermal matrix allograft or xenograft for reconstructing the abdominal wall, chest wall, or the pelvic floor…. In essence, surgeons are applying regenerative medicine concepts in real time in the operating room.

Rubin J. Peter, Gurtner Geoffrey C., Liu Wei, March Keith L., Seppänen-Kaijansinkko Riitta, Yaszemski Michael J., and Yoo James J.. Tissue Engineering Part A. March 2016, 22(5-6): 397-400. doi:10.1089/ten.tea.2016.0002.
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Jul 2013

Complements and the Wound Healing Cascade: An Updated Review

Wound healing is a complex pathway of regulated reactions and cellular infiltrates. The mechanisms at play have been thoroughly studied but there is much still to learn. The health care system in the USA alone spends on average 9 billion dollars annually on treating of wounds. To help reduce patient morbidity and mortality related to abnormal or prolonged skin healing, an updated review and understanding of wound healing is essential. Recent works have helped shape the multistep process in wound healing and introduced various growth factors that can augment this process. The complement cascade has been shown to have a role in inflammation and has only recently been shown to augment wound healing. In this review, we have outlined the biology of wound healing and discussed the use of growth factors and the role of complements in this intricate pathway.

Plast Surg Int. 2013; 2013: 146764. Published online 2013 July 24. doi: 10.1155/2013/146764. Hani Sinno and Satya Prakash1. Copyright © 2013 H. Sinno and S. Prakash.
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Jul 2013

The Pharmacology of Regenerative Medicine

Regenerative medicine is a rapidly evolving multidisciplinary, translational research enterprise whose explicit purpose is to advance technologies for the repair and replacement of damaged cells, tissues, and organs. Scientific progress in the field has been steady and expectations for its robust clinical application continue to rise. The major thesis of this review is that the pharmacological sciences will contribute critically to the accelerated translational progress and clinical utility of regenerative medicine technologies. In 2007, we coined the phrase “regenerative pharmacology” to describe the enormous possibilities that could occur at the interface between pharmacology, regenerative medicine, and tissue engineering. The operational definition of regenerative pharmacology is “the application of pharmacological sciences to accelerate, optimize, and characterize (either in vitro or in vivo) the development, maturation, and function of bioengineered and regenerating tissues.” As such, regenerative pharmacology seeks to cure disease through restoration of tissue/organ function. This strategy is distinct from standard pharmacotherapy, which is often limited to the amelioration of symptoms. Our goal here is to get pharmacologists more involved in this field of research by exposing them to the tools, opportunities, challenges, and interdisciplinary expertise that will be required to ensure awareness and galvanize involvement. To this end, we illustrate ways in which the pharmacological sciences can drive future innovations in regenerative medicine and tissue engineering and thus help to revolutionize the discovery of curative therapeutics. Hopefully, the broad foundational knowledge provided herein will spark sustained conversations among experts in diverse fields of scientific research to the benefit of all.


Christ GJ, Saul JM, Furth ME, Andersson K-E. The Pharmacology of Regenerative Medicine. Nader MA, ed. Pharmacological Reviews. 2013;65(3):1091-1133. doi:10.1124/pr.112.007393
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Apr 2008

Properties of the Amniotic Membrane for Potential Use in Tissue Engineering

An important component of tissue engineering (TE) is the supporting matrix upon which cells and tissues grow, also known as the scaffold. Scaffolds must easily integrate with host tissue and provide an excellent environment for cell growth and differentiation. Most scaffold materials are naturally derived from mammalian tissues. The amniotic membrane (AM) is considered an important potential source for scaffolding material. The AM represents the innermost layer of the placenta and is composed of a single epithelial layer, a thick basement membrane and an avascular stroma. The special structure and biological viability of the AM allows it to be an ideal candidate for creating scaffolds used in TE. Epithelial cells derived from the AM have the advantages of stem cells, yet are a more suitable source of cells for TE than stem cells. The extracellular matrix components of the basement membrane of the AM create an almost native scaffold for cell seeding in TE. In addition, the AM has other biological properties important for TE, including anti-inflammatory, anti-microbial, anti-fibrosis, anti-scarring, as well as reasonable mechanical property and low immunogenicity. In this review, the various properties of the AM are discussed in light of their potential use for TE.

Eur Cell Mater. 2008 Apr 29;15:88-99.
Read More
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