Disinfection of Human Amniotic Membrane Using a Hydrodynamic System with Ozonated Water

Urinary tract infections (UTIs) represent a serious global health issue, especially due to emerging multidrug-resistant UTI-causing bacteria. Recently, we showed that the human amniotic membrane (hAM) could be a candidate for treatments and prevention of UPEC and Staphylococcus aureus infections. However, its role against multidrug-resistant bacteria, namely methicillin-resistant S. aureus (MRSA), extended-spectrum beta-lactamases (ESBL) producing Escherichia coli and Klebsiella pneumoniae, vancomycin-resistant Enterococci (VRE), carbapenem-resistant Acinetobacter baumannii, and Pseudomonas aeruginosa has not yet been thoroughly explored. Here, we demonstrate for the first time that the hAM homogenate had antibacterial activity against 7 out of 11 tested multidrug-resistant strains, the greatest effect was on MRSA. Using novel approaches, its activity against MRSA was further evaluated in a complex microenvironment of normal and cancerous urinary bladder urothelia. Even short-term incubation in hAM homogenate significantly decreased the number of bacteria in MRSA-infected urothelial models, while it did not affect the viability, number, and ultrastructure of urothelial cells. The hAM patches had no antibacterial activity against any of the tested strains, which further exposes the importance of the hAM preparation. Our study substantially contributes to basic knowledge on the antibacterial activity of hAM and reveals its potential to be used as an antibacterial agent against multidrug-resistant bacteria.

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A Novel Phenotype of Junctional Epidermolysis Bullosa with Transient Skin Fragility and Predominant Ocular Involvement Responsive to Human Amniotic Membrane Eyedrops

Genes ◽

10.3390/genes12050716 ◽

2021 ◽

Vol 12 (5) ◽

pp. 716

Author(s):

Daniele Castiglia ◽

Paola Fortugno ◽

Angelo Giuseppe Condorelli ◽

Sabina Barresi ◽

Naomi De Luca ◽

...

Keyword(s):

Amniotic Membrane ◽

Epidermolysis Bullosa ◽

Ocular Involvement ◽

Human Amniotic Membrane ◽

Vitro Assay ◽

Junctional Epidermolysis Bullosa ◽

Mucosal Involvement ◽

Skin Fragility ◽

Laminin 332

Junctional epidermolysis bullosa (JEB) is a clinically and genetically heterogeneous skin fragility disorder frequently caused by mutations in genes encoding the epithelial laminin isoform, laminin-332. JEB patients also present mucosal involvement, including painful corneal lesions. Recurrent corneal abrasions may lead to corneal opacities and visual impairment. Current treatments are merely supportive. We report a novel JEB phenotype distinguished by the complete resolution of skin fragility in infancy and persistent ocular involvement with unremitting and painful corneal abrasions. Biallelic LAMB3 mutations c.3052-5C>G and c.3492_3493delCG were identified as the molecular basis for this phenotype, with one mutation being a hypomorphic splice variant that allows residual wild-type laminin-332 production. The reduced laminin-332 level was associated with impaired keratinocyte adhesion. Then, we also investigated the therapeutic power of a human amniotic membrane (AM) eyedrop preparation for corneal lesions. AM were isolated from placenta donors, according to a procedure preserving the AM biological characteristics as a tissue, and confirmed to contain laminin-332. We found that AM eyedrop preparation could restore keratinocyte adhesion in an in vitro assay. Of note, AM eyedrop administration to the patient resulted in long-lasting remission of her ocular manifestations. Our findings suggest that AM eyedrops could represent an effective, non-invasive, simple-to-handle treatment for corneal lesions in patients with JEB and possibly other EB forms.

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Case report of non-healing surgical wound treated with dehydrated human amniotic membrane

Journal of Translational Medicine ◽

10.1186/s12967-015-0608-8 ◽

2015 ◽

Vol 13 (1) ◽

Cited By ~ 5

Author(s):

Neil H Riordan ◽

Ben A George ◽

Troy B Chandler ◽

Randall W McKenna

Keyword(s):

Case Report ◽

Amniotic Membrane ◽

Human Amniotic Membrane ◽

Surgical Wound

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Treatment ofPseudomonas aeruginosakeratitis with combined corneal cross-linking and human amniotic membrane transplantation

Acta Ophthalmologica ◽

10.1111/aos.12115 ◽

2013 ◽

Vol 91 (5) ◽

pp. e410-e411 ◽

Cited By ~ 11

Author(s):

Jaakko S. Mattila ◽

Anna Korsbäck ◽

Kari Krootila ◽

Juha M. Holopainen

Keyword(s):

Amniotic Membrane ◽

Amniotic Membrane Transplantation ◽

Cross Linking ◽

Human Amniotic Membrane

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Ocular surface rehabilitation: Application of human amniotic membrane in high-risk penetrating keratoplasties

Saudi Journal of Ophthalmology ◽

10.1016/j.sjopt.2014.06.010 ◽

2014 ◽

Vol 28 (3) ◽

pp. 198-202 ◽

Cited By ~ 8

Author(s):

Pho Nguyen ◽

Kelly Rue ◽

Martin Heur ◽

Samuel C. Yiu

Keyword(s):

High Risk ◽

Amniotic Membrane ◽

Ocular Surface ◽

Human Amniotic Membrane

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Effect of the Mode of Application of Cryopreserved Human Amniotic Membrane on Adhesion Formation after Abdomino-Pelvic Surgery in a Mouse Model

Frontiers in Medicine ◽

10.3389/fmed.2016.00010 ◽

2016 ◽

Vol 3 ◽

Author(s):

Joseph Nassif ◽

Sehrish A. Abbasi ◽

Mohamad Karim Kechli ◽

Suzan S. Boutary ◽

Labib Ghulmiyyah ◽

...

Keyword(s):

Mouse Model ◽

Amniotic Membrane ◽

Adhesion Formation ◽

Pelvic Surgery ◽

Human Amniotic Membrane ◽

Mode Of Application

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Effects of Preservation Methods in the Composition of the Placental and Reflected Regions of the Human Amniotic Membrane

Cells Tissues Organs ◽

10.1159/000515448 ◽

2021 ◽

pp. 1-11

Author(s):

Jéssica Tereza Guedes de Oliveira Moraes ◽

Maíra Maftoum Costa ◽

Paula Cristina Santos Alves ◽

Luciana Barros Sant’Anna

Keyword(s):

Regenerative Medicine ◽

Amniotic Membrane ◽

Structural Integrity ◽

Storage Period ◽

Soluble Proteins ◽

Control Group ◽

Human Amniotic Membrane ◽

Glycerol Solution ◽

Preservation Methods ◽

Preservation Method

The human amniotic membrane (AM) is emerging as an interesting biomaterial for regenerative medicine due to its biological and mechanical proprieties. The beneficial effects of the AM are probably related to its bioactive factors produced by local cells and stored in the stromal matrix. However, the search for a preservation method capable of preserving AM properties remains a challenge. The aim of this study was to evaluate important features of 2 anatomical regions of the human AM (reflected and placental amnion) after different preservation methods. For this purpose, human placentas were harvested and processed for AM isolation and storage at 2 different conditions: room temperature for 18 h in DMEM (fresh AM) and −80°C in DMEM/glycerol solution for 30 days (cryopreserved AM). After the storage period, the structural integrity of the membrane was assessed by histological and Picrosirius polarization analysis, cellular viability analysis was performed using the MTT assay, and the soluble proteins were quantified with the Qubit Protein Assay Kit. Both preservation protocols reduced the cell viability, mainly in the placental amnion region of the AM, but preserved the morphology of epithelial and stromal layers, as well as the organization and distribution of collagen fibers. There was a reduction in soluble proteins only in fresh AM. Importantly, the cryopreserved AM group presented the same concentration as the control group. In conclusion, the cryopreservation using DMEM/glycerol was ideal for preserving the structural integrity and soluble protein content, indicating the feasibility of this method in preserving AM for its use in regenerative medicine.

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