RESUMO
OBJECTIVE: To compare the ability of three culture strategies of static culture, intermittent centrifugal culture and dynamic bioreactor culture in promoting the infiltration of bone marrow mesenchymal stem cells (BMSCs) throughout electrospun nanoporous aligned nanoyarn scaffold (AYS). METHODS: AYS was constructed by the method of conjugated electrospinning, using the blended solution of poly (L-lactide-co-caprolactone) (P (LLA-CL)) and gelatin. Then the bone marrow mesenchymal stem cells (BMSCs) were transplanted on the scaffolds. Culture the scaffold-cells using three methods of static culture, intermittent centrifugal culture and dynamic bioreactor culture. After 7 and 14 days in culture, the infiltration depth of the cells were observed and measured by hematoxylin and eosin (HE) or 4', 6-diamidino-2-phenylindole (DAPI) staining. RESULT: In the current study, on the 7th day, the BMSCs in the scaffolds of static culture group, intermittent centrifugal culture group, and dynamic bioreactor culture group infiltrated to an average depth of 11.88 ± 1.82 µm, 21.17 ± 13.17 µm, and 26.27 ± 7.42 µm, respectively. There were differences between the bioreactor culture group with the static culture group and the intermittent centrifugal culture group. On the time point of 14 days, the depth of infiltration of BMSCs in dynamic bioreactor culture was the most (115.13 ± 25.44 µm, P < 0.05), and the infiltration of the cells in the intermittent centrifugal culture group was 42.53 ± 13.07 µm, deeper than that of the static culture group (24.53 ± 6.06, P < 0.05). CONCLUSION: Dynamic bioreactor culture may be a preferred method for tissue engineering approaches involving scaffolds with a low porosity, such as those needed for repair of the annulus fibrosus (AF).
Assuntos
Anel Fibroso/citologia , Reatores Biológicos , Células da Medula Óssea/citologia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Proliferação de Células , Células Cultivadas , Nanofibras , Ratos Sprague-DawleyRESUMO
Human endogenous retroviruses (HERVs) play pivotal roles in the development of breast cancer. However, the detailed mechanisms of noncoding HERVs remain elusive. Here, our genome-wide transcriptome analysis of HERVs revealed that a primate long noncoding RNA, which we dubbed TROJAN, was highly expressed in human triple-negative breast cancer (TNBC). TROJAN promoted TNBC proliferation and invasion and indicated poor patient outcomes. We further confirmed that TROJAN could bind to ZMYND8, a metastasis-repressing factor, and increase its degradation through the ubiquitin-proteasome pathway by repelling ZNF592. TROJAN also epigenetically up-regulated metastasis-related genes in multiple cell lines. Correlations between TROJAN and ZMYND8 were subsequently confirmed in clinical samples. Furthermore, our study verified that antisense oligonucleotide therapy targeting TROJAN substantially suppressed TNBC progression in vivo. In conclusion, the long noncoding RNA TROJAN promotes TNBC progression and serves as a potential therapeutic target.