Progesterone Prolongs Viability and Anti-inflammatory Functions of Explanted Preterm Ovine Amniotic Membrane

Abstract

Amniotic membrane (AM) is considered an important medical device with many applications in regenerative medicine. The therapeutic properties of AM are due to its resistant extracellular matrix and to the large number of bioactive molecules released by its cells. An important goal that still remains to be achieved is the identification of cultural and preservation protocols able to maintain in time the membrane morphology and the biological properties of its cells. Recently, our research group demonstrated that progesterone (P 4) is crucial in preventing the loss of the epithelial phenotype of amniotic epithelial cells in vitro. Followed by this premise, it has been evaluated whether P 4 may also affect AM properties in a short-term culture. Results confirm that P 4 preserves AM integrity and architecture with respect to untreated AM, which showed alterations in morphology. Transmission electron microscopy (TEM) analyses demonstrate that P 4 also maintains unaltered cell-cell junctions, nuclear status, and intracellular organelles. On the contrary, an untreated AM experienced an extensive cell death and a strong reduction of immunomodulatory properties, measured in terms of anti-inflammatory cytokine expression and secretion. Overall, these results could open to new strategies to ameliorate the protocols for cryopreservation and tissue culture, which represent preliminary stages of AM application in regenerative medicine.

Amniotic Epithelial Stem Cells Counteract Acidic Degradation By-Products of Electrospun PLGA Scaffold by Improving Their Immunomodulatory Profile In Vitro

“…oAECs are able to secrete cytokines and growth factors with antiapoptotic, proangiogenic, and immune-modulatory molecules, which can alter the pro-inflammatory signals and enhance the anti-inflammatory ones, hence improving tissue regeneration [84][85][86][87][88]. Among them, it was recently demonstrated in the amniotic epithelium that a highly conserved physiological mechanism amongst species controlling the expression and releasing balance of potent anti-inflammatory and proinflammatory cytokines, such as interleukin (IL)-10, IL-4, and IL-12, under both basal and LPS-inductive conditions [54,80,89,90].…”

“…The absorbance (Abs) was measured at 595 nm by using EnSpire ® Multimode Plate Reader (PerkinElmer, Waltham, MA, USA). The percentage of viability was estimated according to Equation (3) [54]:…”

Unveiling the immunomodulatory shift: Epithelial-mesenchymal transition Alters immune mechanisms of amniotic epithelial cells

“… 16 In this regard, amniotic membrane (AM) has been historically considered an important medical device with many applications in regenerative medicine for its structural and biological properties. 17 In particular, the widespread use of AM depends on its anti-inflammatory, anti-bacterial, anti-microbial, anti-fibrotic, and anti-cancer properties and, nevertheless, for its huge immunomodulatory potential. The latter effect is further important in the field of cell-based therapy and mostly depends on the immunological advantages of both cell populations that form AM: amniotic epithelial cells (AECs) and amniotic mesenchymal cells (AMCs).…”

Bio-Fabrication of Human Amniotic Membrane Zinc Oxide Nanoparticles and the Wet/Dry HAM Dressing Membrane for Wound Healing

“…Treatment with EDTA and/or proteolysis may also result in HAM damage ( Saghizadeh et al, 2013 ; Sripriya and Kumar, 2016 ). Canciello et al (2020) acknowledged that “An important goal that still remains to be achieved is the identification of cultural and preservation protocols able to maintain in time the membrane morphology and the biological properties of its cells.” It is critical to develop improved methods of preparation and application in order to address difficulties and limitations in the manufacturing, preservation, sterilization, and packaging methodologies, to maximize HAM protection and long-term storage, and to take advantage of the amniotic membrane’s potential benefits in a variety of applications. Therefore, the main object of the present investigation was to develop a novel process for preparing a sterile, stable, inexpensive human amniotic membrane product, free from the cellular elements of the chorion, for use as a wound dressing in dermal and ocular applications.…”