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 (P4) 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 P4 may also affect AM properties in a short-term culture. Results confirm that P4 preserves AM integrity and architecture with respect to untreated AM, which showed alterations in morphology. Transmission electron microscopy (TEM) analyses demonstrate that P4 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.
Highlights
Since the beginning of the twentieth century, researchers paid growing attention to the study of amniotic membrane (AM), even though evidences of the use of AM as a medication have been reported in traditional Chinese medicine (Tyszkiewicz et al, 1999)
The P4-treated AM compared with the fresh AM (Figure 2A) showed an overall higher integrity with a preserved microarchitecture persisting during the culture (Figure 3A)
No structural alterations were observed in the P4 AM until day 3, when only rare membrane breaks occurred on the epithelial layer (Figure 3A)
Summary
Since the beginning of the twentieth century, researchers paid growing attention to the study of amniotic membrane (AM), even though evidences of the use of AM as a medication have been reported in traditional Chinese medicine (Tyszkiewicz et al, 1999). The AM biological properties are mainly ascribable to the secretory abilities of its cellular components, especially of amniotic epithelial cells (AECs) that form the AM’s innermost layer (Litwiniuk and Grzela, 2014) and amniotic mesenchymal stromal cells (AMSCs) that are found disperse into the extracellular matrix underneath the basement membrane (Niknejad et al, 2013). TGF-β acts as extracellular matrix remodeling by stimulating fibroblasts to increase collagen production (Uchide et al, 2012) All these cytokines act in concert to orchestrate the healing process by creating the proper milieu for tissue regeneration
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