Human Amniotic Mesenchymal Stem Cells and Fibroblasts Accelerate Wound Repair of Cystic Fibrosis Epithelium

Abstract

Cystic fibrosis (CF) airways are affected by a deranged repair of the damaged epithelium resulting in altered regeneration and differentiation. Previously, we showed that human amniotic mesenchymal stem cells (hAMSCs) corrected base defects of CF airway epithelial cells via connexin (CX)43-intercellular gap junction formation. In this scenario, it is unknown whether hAMSCs, or fibroblasts sharing some common characteristics with MSCs, can operate a faster repair of a damaged airway epithelium. A tip-based scratch assay was employed to study wound repair in monolayers of CFBE14o- cells (CFBE, homozygous for the F508del mutation). hAMSCs were either co-cultured with CFBE cells before the wound or added to the wounded monolayers. NIH-3T3 fibroblasts (CX43+) were added to wounded cells. HeLa cells (CX43-) were used as controls. γ-irradiation was optimized to block CFBE cell proliferation. A specific siRNA was employed to downregulate CX43 expression in CFBE cells. CFBE cells showed a delayed repair as compared with wt-CFTR cells (16HBE41o-). hAMSCs enhanced the wound repair rate of wounded CFBE cell monolayers, especially when added post wounding. hAMSCs and NIH-3T3 fibroblasts, but not HeLa cells, increased wound closure of irradiated CFBE monolayers. CX43 downregulation accelerated CFBE wound repair rate without affecting cell proliferation. We conclude that hAMSCs and fibroblasts enhance the repair of a wounded CF airway epithelium, likely through a CX43-mediated mechanism mainly involving cell migration.

Elexacaftor/Tezacaftor/Ivacaftor Accelerates Wound Repair in Cystic Fibrosis Airway Epithelium

“…We have previously shown that CFBE cell monolayers (null for CFTR expression) show a delay in the wound closure. CFBE cells closed the wound only after 48 h, while non-CF 16HBE cells closed the wound in 24 h [ 22 ]. Thus, we investigated the effect of ETI on wound closure.…”

“…The defect in wound repair presented by us [ 22 ] and others in various cell models of CF primary and immortalized cell lines [ 19 , 21 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ] strongly suggest that the lack/dysfunction of the CFTR protein may play an essential role. Orkambi ® (lumacaftor/ivacaftor) has been previously shown to significantly increase wound repair rates over a period of 6 h with CF primary airway epithelial cells [ 19 ].…”

“…Of note, we observed that ETI treatment was able to increase the wound healing rate between 0 and 6 h, with a slowing of wound closure at further times, although still significant when the overall time interval 0–24 h was considered. These data suggested that rescuing F508del-CFTR by CFTR modulators increased wound repair approaching that of a non-CF cell line, i.e., 16HBE [ 22 ]. Since the triple combination of small molecules (VX-661, VX-445 and VX-770) directly interacts with CFTR and rescues the dysfunctional F508del-CFTR protein [ 37 , 38 , 39 , 40 , 41 , 42 ], the current study demonstrated that functional CFTR plays an important role for wound repair.…”

“…Digital images of 10x fields were obtained with a Leica DM IRB inverted microscope equipped with a Leica DFC450 C camera (Leica, Wetzlar, Germany). The percentage of wound closure and the wound healing rate, presented in μm/h, were calculated with ImageJ software (National Institutes of Health, Bethesda, MD, USA), as previously done [ 22 ].…”

Solanum lycopersicum (Tomato)-Derived Nanovesicles Accelerate Wound Healing by Eliciting the Migration of Keratinocytes and Fibroblasts

“…NIH mouse fibroblast cell line (NIH-3T3, a kind gift of Dr. Giuseppe Procino, University of Bari “A. Moro”, Bari, Italy) was grown in Dulbecco’s modified Eagle medium (DMEM) with 4.5 g/L glucose and sodium pyruvate (Corning), containing 10% fetal bovine serum (FBS), 1% L-glutamine, and 1% penicillin/streptomycin [ 64 ].…”