Effects of Amniotic Membrane Extract Eye Drops on Persistent Epithelial Defects of the Cornea

Purpose: To evaluate the clinical efficacy of amniotic membrane extract eye drops (AMEEDs) in patients with persistent epithelial defects (PEDs) of the cornea.Methods: Sixteen patients with PEDs refractory to the conventional treatment were further treated with AMEEDs six times a day. Visual acuity, visual analog scale (VAS), esthesiometer score, and areas of the epithelial defects before and after 1 and 2 months of treatment were evaluated. After 2 months, AMEEDs were considered effective if all epithelial defects were healed, partially effective if the epithelial defects decreased in size compared with the baseline, and ineffective if the epithelial defects increased in size and required additional treatment.Results: After 2 months of treatment with AMEEDs, there was a reduction in the area of epithelial defects (5.2 ± 3.1 mm2 vs. 0.1 ± 0.1 mm2, respectively, p = 0.01), as well as a significant improvement in best-corrected visual acuity (0.8 ± 0.5 logarithm of minimal angle of resolution [LogMAR] vs. 0.6 ± 0.3 LogMAR, respectively, p = 0.03), and VAS scores (4.3 ± 1.0 vs. 2.8 ± 0.7, respectively, p = 0.04) compared with the baseline values. Treatment with AMEEDs was effective in 13 (81.3%) patients and partially effective in three (18.8%) patients.Conclusions: AMEEDs could stimulate epithelial wound healing and improve ocular symptoms in patients with refractory PED. Therefore, AMEEDs could be considered an effective treatment option for refractory PEDs.

Extracellular Matrix Components Regulate Bone Sialoprotein Expression in MDA-MB-231 Breast Cancer Cells

Bone sialoprotein (BSP) has become a target in breast cancer research as it is associated with tumor progression and metastasis. The mechanisms underlying the regulation of BSP expression have been largely elusive. Given that BSP is involved in the homing of cancer cells in bone metastatic niches, we addressed regulatory effects of proteolytic cleavage and extracellular matrix components on BSP expression and distribution in cell culture models. Therefore, MDA-MB-231 human breast cancer cells were kept in 2D and 3D spheroid cultures and exposed to basement membrane extract in the presence or absence of matrix metalloproteinase 9 or the non-polar protease, dispase. Confocal imaging of immunofluorescence samples stained with different antibodies against human BSP demonstrated a strong inducing effect of basement membrane extract on anti-BSP immunofluorescence. Similarly, protease incubation led to acute upregulation of anti-BSP immunofluorescence signals, which was blocked by cycloheximide, suggesting de novo formation of BSP. In summary, our data show that extracellular matrix components play an important function in regulating BSP expression and hint at mechanisms for the formation of bone-associated metastasis in breast cancer that might involve local control of BSP levels by extracellular matrix degradation and release of growth factors.

Collagen polarization provides a structural memory for the elongation of epithelial anlage

Branched epithelial networks are fundamental features of many organs in the body. The biogenesis of these networks involves distinct processes where multicellular aggregates elongate and branch. In this report we focus on understanding how the extracellular matrix contributes to the process of elongation. Using mammary epithelial organotypic cultures we found that collagen 1, but not a basement membrane extract, induces the formation of elongated multicellular aggregates. Indeed, isotropic aggregates, used as models of epithelial anlage, broke symmetry and elongated when transplanted into collagen 1 gels; this was accompanied by reorganization of collagen fibrils into bundles that were polarized around the elongating aggregates. By applying external stretch as a cell-independent way to reorganize the ECM gels, we found that collagen polarization itself can induce and guide the direction of aggregate elongation. This critically involves cell proliferation, which is selectively enhanced in the regions of anlage that elongate, and requires β1-integrin and ERK signaling. We propose that collagen polarization promotes anlage elongation by providing a structural memory of the initial axis that is generated when aggregates break symmetry.

Amniotic Membrane Extract Protects Islets From Serum-Deprivation Induced Impairments and Improves Islet Transplantation Outcome

Islet culture prior to transplantation is a standard practice in many transplantation centers. Nevertheless, the abundant islet mass loss and function impairment during this serum-deprivation culture period restrain the success of islet transplantation. In the present study, we used a natural biomaterial derived product, amniotic membrane extract (AME), as medium supplementation of islet pretransplant cultivation to investigate its protective effect on islet survival and function and its underlying mechanisms, as well as the engraftment outcome of islets following AME treatment. Results showed that AME supplementation improved islet viability and function, and decreased islet apoptosis and islet loss during serum-deprived culture. This was associated with the increased phosphorylation of PI3K/Akt and MAPK/ERK signaling pathway. Moreover, transplantation of serum-deprivation stressed islets that were pre-treated with AME into diabetic mice revealed better blood glucose control and improved islet graft survival. In conclusion, AME could improve islet survival and function in vivo and in vitro, and was at least partially through increasing phosphorylation of PI3K/Akt and MAPK/ERK signaling pathway.

Effects of Amniotic Membrane Extract on the Hyperplastic Response of the Middle Ear Mucosa in a Bacterially-Induced Otitis Media Rat Model: A Preliminary Study

Objectives. Human amniotic membrane extract (AME) is known to contain numerous bioactive factors and anti-inflammatory substances. However, the anti-inflammatory effects of AME on the middle ear (ME) mucosa are unclear. This study assessed the effects of AME on the growth of the ME mucosa in response to bacterially-induced otitis media (OM).Methods. OM was induced by inoculating nontypeable <i>Haemophilus influenzae</i> (NTHi) into the ME cavity of rats. ME mucosal explants were cultured in AME concentrations of 0, 5, 10, or 50 μg/mL. The area of explant outgrowth was measured in culture and analyzed at 1, 3, 5, and 7 days after explantation. The expression of Ki-67, mucin 5AC (MUC5AC), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) in the explants was also evaluated using quantitative polymerase chain reaction (PCR) and immunocytochemistry (ICC).Results. The NTHi-induced ME mucosa growth increased gradually over the 7-day culture period in all explants at different AME concentrations. There was a trend for mucosal growth inhibition at higher concentrations of AME, although the growth was not significantly different among the groups until day 5. The ME mucosal explants treated with the 50 μg/mL concentration of AME showed significantly suppressed growth on postexplantation day 7 compared with other explants on the same day. PCR and ICC staining revealed that the expression of Ki-67, MUC5AC, TNF-α, and IL-10 further decreased in the explants with higher concentrations of AME than in those with lower concentrations of AME.Conclusion. Our results showed that higher concentrations of AME reduced the mucosal proliferative response in bacterial OM in rats. These findings provide evidence that AME has an influence on the inflammatory and proliferative responses to NTHi infection in ME mucosa.