研究目的
To fabricate a new biocompatible and conductive nanofibrous scaffold from polycaprolactone/polypyrrole/multiwall carbon nanotube (PCL/PPY/MWCNT) and evaluate its properties, and to assess the effects of the scaffold together with electrical stimulation on photoreceptor differentiation of trabecular meshwork mesenchymal stem cells (TM‐MSCs).
研究成果
The PCL/PPY/MWCNTs scaffold combined with electrical stimulation effectively promoted photoreceptor differentiation of TM‐MSCs, as evidenced by increased expression of rhodopsin and peripherin genes and proteins. This approach shows promise for retinal tissue engineering and regenerative medicine, suggesting further research to optimize and validate in vivo applications.
研究不足
The study did not address long-term stability or in vivo performance of the scaffold. Cell viability on the PCL/PPY/MWCNTs scaffold was lower than on tissue culture plastic, indicating potential biocompatibility issues. Optimization of electrical stimulation parameters and scaffold composition may be needed for enhanced efficacy.
1:Experimental Design and Method Selection:
The study involved fabricating a conductive PCL/PPY/MWCNTs scaffold using electrospinning under specific conditions (high-voltage: 24 kV, flow rate:
2:4 ml/hr, distance:
120 cm, rotating disk speed: 2,500 rpm). Electrical stimulation was applied to TM‐MSCs seeded on the scaffold at 115 V/m, 100 Hz frequency for 10 min/day over 7 days. Methods included SEM, TEM, FT‐IR for scaffold characterization, and immunofluorescence and RT‐qPCR for differentiation analysis.
3:Sample Selection and Data Sources:
TM‐MSCs were isolated and characterized based on a modified protocol. Scaffolds were fabricated from PCL, PPY, and MWCNTs. Data were derived from in vitro cell culture experiments.
4:List of Experimental Equipment and Materials:
Electrospinning machine, SEM (AIS2100, Seron Technology), TEM, FT‐IR (Vector 22 Bruker), potentiostat-galvanostat (Zahner/Zennium, Gamry), electricity generating device (GW, INSTEK AFG‐2005), fluorescence microscopy (Olympus), spectrophometer (NanoDrop 2000), microplate reader (ELX800, BioTeK). Materials included PCL, PPY, MWCNTs (Sigma-Aldrich), solvents (dimethylformamide, chloroform), cell culture media (DMEM with FBS), antibodies (Santa Cruz Biotechnology), RNA extraction kit (SinaClon), cDNA synthesis kit (Takara), RT‐qPCR master mix (AMPLIQON).
5:Experimental Procedures and Operational Workflow:
Scaffold fabrication via electrospinning; characterization using SEM, TEM, FT‐IR, and electrochemical studies; cell seeding on scaffolds in a bioreactor; electrical stimulation application; cell fixation and staining for SEM and immunofluorescence; RNA extraction, cDNA synthesis, and RT‐qPCR for gene expression analysis; MTT assay for cell viability.
6:Data Analysis Methods:
Gene expression analyzed using REST? software with RT‐qPCR data; statistical significance set at p ≤ 0.05; immunofluorescence images analyzed by fluorescence microscopy; electrochemical data from CV and impedance spectra.
独家科研数据包,助您复现前沿成果,加速创新突破
获取完整内容-
Fourier Transform Infrared Spectrometer
Vector 22
Bruker
Investigate chemical properties of the scaffold
-
Electrospinning Machine
Fabrication of fibrous scaffold using high-voltage current
暂无现货
预约到货通知
-
Scanning Electron Microscope
AIS2100
Seron Technology
Determine morphology and diameter of nanofibers; observe cellular adhesion and morphology
暂无现货
预约到货通知
-
Transmission Electron Microscope
Evaluate PPY and MWCNTs dispersion within the fibrous scaffold
暂无现货
预约到货通知
-
Potentiostat-Galvanostat
Zahner/Zennium
Gamry
Record cyclic voltammetry and electrochemical impedance spectroscopy measurements
暂无现货
预约到货通知
-
Electricity Generating Device
AFG-2005
GW, INSTEK
Apply electrical stimulation to cells
暂无现货
预约到货通知
-
Fluorescence Microscope
Olympus
Analyze protein expression in differentiated cells
暂无现货
预约到货通知
-
Spectrophotometer
NanoDrop 2000
Measure purity and quantity of RNA
暂无现货
预约到货通知
-
Microplate Reader
ELX800
BioTeK
Measure MTT absorbance for cell viability assay
暂无现货
预约到货通知
-
Polycaprolactone
Sigma-Aldrich
Component of the conductive scaffold
暂无现货
预约到货通知
-
Polypyrrole
Sigma-Aldrich
Component of the conductive scaffold to enhance electrical properties
暂无现货
预约到货通知
-
Multiwall Carbon Nanotubes
Sigma-Aldrich
Component of the conductive scaffold to increase electrical conductivity
暂无现货
预约到货通知
-
RNA Extraction Kit
RNX-Plus
SinaClon
Extract total RNA from cells
暂无现货
预约到货通知
-
cDNA Synthesis Kit
RevertAid First Strand cDNA Synthesis Kit
Takara
Synthesize cDNA from RNA
暂无现货
预约到货通知
-
RT-qPCR Master Mix
RealQ Plus 2x Master Mix Green/High ROX
AMPLIQON
Perform quantitative real-time polymerase chain reaction
暂无现货
预约到货通知
-
Antibody to Rhodopsin
H-100: sc-15382
Santa Cruz Biotechnology
Detect rhodopsin protein expression in immunofluorescence
暂无现货
预约到货通知
-
Antibody to Recoverin
H-100: sc-28849
Santa Cruz Biotechnology
Detect recoverin protein expression in immunofluorescence
暂无现货
预约到货通知
-
FITC-anti-rabbit IgG
Abcam
Secondary antibody for immunofluorescence detection
暂无现货
预约到货通知
-
登录查看剩余16件设备及参数对照表
查看全部