MATERIALS AND METHODS: In this study, reprogramming of human dermal fibroblasts (NHDF) into iPSC was carried out using non-integrative Sendai virus for transduction. The iPSC clones were characterised based on the morphological changes, gene expression of pluripotency markers, and spontaneous and directed differentiation abilities into cells of different germ layers.
RESULTS: On day 18-25 post-transduction, colonies with embryonic stem cell-like morphology were obtained. The iPSC generated were free of Sendai genome and transgene after passage 10, as confirmed by RT-PCR. NHDF-derived iPSC expressed multiple pluripotency markers in qRT-PCR and immunofluorescence staining. When cultured in suspension for 8 days, iPSC successfully formed embryoid body-like spheres. NHDF-derived iPSC also demonstrated the ability to undergo directed differentiation into ectoderm and endoderm.
CONCLUSION: NHDF were successfully reprogrammed into iPSC using non-integrating Sendai virus for transduction.
Materials and Methods: Immediate skin tissue expansion in 18 adult female rats was performed using three different sizes (small, medium, and big) of polymethylmethacrylate tissue expanders at the dorsal surface of the metatarsal area of the right limb. The contralateral limb was served as the control. The tissue expanders were surgically implanted and kept for 15 days.
Results: The immediate skin expansion resulted in histological changes such as the increased thickness of the epidermal layer, the reduction of the dermal layer, an elevated number of fibroblast as well as increased vascularity. Furthermore, skin adnexal structures such as hair follicles and sebaceous glands were farther apart.
Conclusion: The rat skin was able to rapidly adjust and compensate against a specific range of immediate mechanical expansion. The histological changes suggest that the tissues were prepared to withstand the increased external forces, in addition to create possibly additional skin in a relatively short-term period.
METHODS: Whole proteins were extracted and digested in-gel with trypsin. Peptides were detected by Orbitrap liquid chromatography mass spectrometry. Mass spectra were identified and quantitated by MaxQuant software. The data were further filtered and analyzed statistically using Perseus software to identify DEPs. Functional annotations of DEPs were performed using Panther Classification System.
RESULTS: A total of 1217 proteins were identified in young and senescent cells, while 1218 proteins in vehicle- and γT3-treated senescent cells. 11 DEPs were found in young and senescent cells which included downregulation of platelet-derived growth factor (PDGF) receptor beta and upregulation of tubulin beta-2A chain protein expressions in senescent cells. 51 DEPs were identified in vehicle- and γT3-treated senescent cells which included upregulation of 70 kDa heat shock protein, triosephosphate isomerase and malate dehydrogenase protein expressions in γT3-treated senescent cells.
CONCLUSIONS: PDGF signaling and cytoskeletal structure may be dysregulated in senescent HDFs. The pro-proliferative effect of γT3 on senescent HDFs may be mediated through the stimulation of cellular response to stress and carbohydrate metabolism. The expressions and roles of these proteins in relation to cellular senescence are worth further investigations. Data are available via ProteomeXchange with identifier PXD009933.
METHODS: 15wt% of zirconia (ZrO2) as well as 30, 35, and 40wt% of beta-tricalcium phosphate (β-TCP) were compounded with PA 12, followed by the fabrication of filament feedstocks using a single screw extruder. The fabricated filament feedstocks were used to print the impact specimens. The melt flow rate, tensile properties of fabricated filament feedstocks, and 3D printed impact properties of the specimens were assessed using melt flow indexer, universal testing machine, and Izod pendulum tester, respectively. The microstructure of selected filament feedstocks and broken impact specimens were analysed using a field emission scanning electron microscope and universal testing machine. Human periodontal ligament fibroblast cells (HPdLF) were used to evaluate the cytotoxicity of the materials by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid) (MTT) assay.
RESULTS: Hybrid ceramics filled PA 12 indicated sufficient flowability for FDM 3D printing. The tensile strength of hybrid ceramics filled PA 12 filament feedstocks slightly reduced as compared to unfilled PA 12. However, the tensile modulus and impact strength of hybrid ceramics filled PA 12 increased by 8%-31% and 98%-181%, respectively. A significant increase was also detected in the cell viability of the developed composites at concentrations of 12.5, 25, 50 and 100mg/ml.
SIGNIFICANCE: The newly developed hybrid ceramics filled PA 12 filament feedstock with improved properties is suitable for an FDM-based 3D printer, which enables the creation of patient-specific craniofacial implant at a lower cost to serve low-income patients.
MATERIALS & METHODS: Podoplanin expression was evaluated immunohistochemically in 153 breast cancers. Tumours with ≥ 10% distinct cytoplasmic podoplanin staining in CAFs were considered as positive.
RESULTS: In 65.3% of analysed tumours, podoplanin expression was found positive in CAFs. According to our results, podoplanin positive CAFs correlated significantly with tumour size (p= 0.012), tumour grade (p= 0.032) and cerbB2 score (p= 0.032).
DISCUSSION: Our results suggest that podoplanin expression by CAFs could predict poor patient outcome in breast carcinoma.