研究目的
To compare the quality of bone mineral formed in osteogenic cell cultures against calvarial bone from postnatal mice and examine the feasibility of extracting information on cell activity and bone matrix quality.
研究成果
Bone formed in osteogenic cell culture exhibited progressive matrix maturation and mineralization but could not fully replicate the high degree of collagen fibril order found in native bone. Raman spectroscopic measures of bone quality used in this study could equally be used to evaluate ex vivo specimens obtained from preclinical models that are used in tissue engineering research.
研究不足
The high initial mineral/matrix ratios observed at Day 8 requires further investigation. The study would have benefited from additional replicate experiments to determine whether the high initial mineral/matrix ratios observed at Day 8 was an outlier or related to the normal biological variation inherent to Sca‐1 stem/progenitor cells isolated fetal mouse calvarial tissues.
1：Experimental Design and Method Selection:
Stem/progenitor cells isolated from fetal mouse calvaria were cultured onto fused‐quartz slides under osteogenic differentiation conditions for 28 days. Raman microscopy and mineral staining techniques were used to analyze the slides at specific time intervals.
2：Sample Selection and Data Sources:
Stem/progenitor cells isolated from fetal mouse calvaria and calvarial tissues from postnatal mice.
3：List of Experimental Equipment and Materials:
Fused‐quartz slides (S1‐UV grade, 12.7‐mm circulars, Esco Products), Raman microscope equipped with a 785 nm laser, 50 × 0.80 numerical aperture (NA) objective (Nikon Instruments, Inc.), MATLAB? software (The MathWorks Inc.), GRAMS/AI? software (ThermoGalactic Inc.), SPSS Statistics Software for Windows (IBM SPSS Statistics Version 24, IBM Corp.).
4：7‐mm circulars, Esco Products), Raman microscope equipped with a 785 nm laser, 50 × 80 numerical aperture (NA) objective (Nikon Instruments, Inc.), MATLAB? software (The MathWorks Inc.), GRAMS/AI? software (ThermoGalactic Inc.), SPSS Statistics Software for Windows (IBM SPSS Statistics Version 24, IBM Corp.).
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Cells were cultured under osteogenic differentiation conditions, slides were removed at specific time intervals, fixed with ice‐cold 70% ethanol, and analyzed by Raman microscopy and mineral staining techniques.
5：Data Analysis Methods:
Spectra were processed in MATLAB? and GRAMS/AI? software, statistical analyses were performed in SPSS Statistics Software.
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