ABSTRACT
Periosteum, the thin layer covering adjacent to bone containing specific architecture, is important for functional bone regeneration and remodeling. Synthetic periosteum investigated presently lacks the resemblance of natural periosteum, suffering from poor mechanical strength and cell attachment. Here, we report a newly-developed biomimetic film to function as synthetic periosteum. Based on poly(ε-caprolactone) (PCL), where surface wettability of the synthetic periosteum is enhanced by microtantalum (mTa) particle blending and after a cold drawing process, further obtains topographical anisotropy without any involvement of solvent. This new blend shows mechanical enhancement over pure PCL, with yield stress and elastic strain approaching the natural periosteum. A distinct degradation mechanism is proposed for the blend, and by seeding with mouse calvarial preosteoblasts, cell proliferation is promoted on surface of the drawn PCL but delayed on the mTa-blended PCL. However, cell mineralization is accelerated on the mTa-blended surface. This is less on the drawn PCL. The synergistical integration of cellular proliferation, alignment and osteogenic enhancement suggest that the cold drawn PCL/Ta blend has unique potential for developing into a synthetic periosteum and other tissue-engineering products.
Subject(s)
Periosteum , Polyesters , Animals , Mice , Tissue Engineering , Osteogenesis , Tissue ScaffoldsABSTRACT
Recurrent miscarriage (RM) is one of the common complications of pregnancy, which is closely related to gene mutation. The profiling of non-coding RNAs showed that the expression level of long non-coding RNA LINC01347 (LINC01347) in the serum of patients with recurrent abortion was significantly increased, which could serve as a potential marker for early diagnosis. However, the biological functions of LINC01347 in the miscarriage remain to be elucidated. In this study, LINC01347 expression levels in HTR-8/SVneo cells and placenta samples were measured by RT-qPCR. The migration ability of HTR-8/SVneo cells was detected by wound-healing assay. Western blotting (WB) assay was conducted to measure E-cadherin, Vimentin, N-cadherin, PTEN, phospho-AKT(S473), phospho-AKT(T308) and AKT levels. Dual luciferase reporter assay and RNA pull-down analysis were performed to validate the molecular interactions. The results showed an upregulation of LINC01347 in the placenta samples of RM patients and HTR-8/SVneo cells. LINC01347 overexpression impaired the invasion and migration of trophoblast cells, while LINC01347 silencing promoted cell migration and invasion. LINC01347 level was also negatively correlated with the changes of epithelial-mesenchymal transition (EMT) markers in trophoblasts. We further demonstrated that miR-101-3p/PTEN/AKT axis played an important role in mediating the biological roles of LINC01347 in the invasion and migration of trophoblasts. In conclusion, our results revealed that LINC01347 suppresses the migratory ability and regulates the EMT processes in trophoblasts by regulating miR-101-3p/PTEN/AKT axis, suggesting that targeting LINC01347 may serve as a strategy to ameliorate RM.
Subject(s)
Abortion, Habitual , RNA, Long Noncoding , Trophoblasts , Abortion, Habitual/genetics , Abortion, Habitual/metabolism , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition/genetics , Female , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolism , Pre-Eclampsia/genetics , Pre-Eclampsia/metabolism , Pregnancy , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolism , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Trophoblasts/metabolismABSTRACT
Recurrent spontaneous abortion (RSA) is a common complication during early gestation, which is associated with aberrant DNA methylation. Zinc Finger and BTB Domain Containing 24 (ZBTB24) plays a critical role in facilitating DNA methylation and cell proliferation. However, the regulatory role of ZBTB24 on trophoblast development in RSA remains unclear. In this study, ZBTB24 expression was compared between decidua tissues of RSA patients and induced abortion controls from a published dataset, which was further validated in placental villi tissues by RT-qPCR and Western blot. The roles of ZBTB24 in trophoblast proliferation, differentiation, and migration were investigated by functional assays after ZBTB24 knockdown or overexpression in HTR-8/SVneo cells. Our results showed that ZBTB24 expression was significantly decreased in RSA patients, and ZBTB24 expression level positively regulated cell viability, differentiation, and migration in HTR-8/SVneo cells. We further demonstrated that ZBTB24 modulated the expression of E-cadherin by altering the DNA methylation at the promoter region. Overall, the downregulation of ZBTB24 is implicated in RSA by inhibiting trophoblast proliferation, differentiation, and migration. Therefore, ZBTB24 may serve as a promising therapeutic target and diagnostic marker for RSA.
