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High-Resolution X-Ray Techniques as New Tool to Investigate the 3D Vascularization of Engineered-Bone Tissue.

Bukreeva I, Fratini M, Campi G, Pelliccia D, Spanò R, Tromba G, Brun F, Burghammer M, Grilli M, Cancedda R, Cedola A, Mastrogiacomo M - Front Bioeng Biotechnol (2015)

Bottom Line: We compared samples seeded and not seeded with BMSC, as well as samples differently stained or unstained.Thanks to the high quality of the images, we investigated the 3D distribution of both vessels and collagen matrix and we obtained quantitative information for all different samples.We propose our approach as a tool for quantitative studies of angiogenesis in TE and for any pre-clinical investigation where a quantitative analysis of the vascular network is required.

View Article: PubMed Central - PubMed

Affiliation: Consiglio Nazionale delle Ricerche - Istituto NANOTEC, c/o Dipartimento di Fisica, Università Sapienza , Rome , Italy.

ABSTRACT
The understanding of structure-function relationships in normal and pathologic mammalian tissues is at the basis of a tissue engineering (TE) approach for the development of biological substitutes to restore or improve tissue function. In this framework, it is interesting to investigate engineered bone tissue, formed when porous ceramic constructs are loaded with bone marrow stromal cells (BMSC) and implanted in vivo. To monitor the relation between bone formation and vascularization, it is important to achieve a detailed imaging and a quantitative description of the complete three-dimensional vascular network in such constructs. Here, we used synchrotron X-ray phase-contrast micro-tomography to visualize and analyze the three-dimensional micro-vascular networks in bone-engineered constructs, in an ectopic bone formation mouse-model. We compared samples seeded and not seeded with BMSC, as well as samples differently stained or unstained. Thanks to the high quality of the images, we investigated the 3D distribution of both vessels and collagen matrix and we obtained quantitative information for all different samples. We propose our approach as a tool for quantitative studies of angiogenesis in TE and for any pre-clinical investigation where a quantitative analysis of the vascular network is required.

No MeSH data available.


Related in: MedlinePlus

Quantitative analysis of the A, B, C, and D samples. Plots 1 show the number of vessels crossing the samples at the different depth inside the volume. For each sample, the three pictures above the plot 1, are the tomography images of the upper, central and top parts of the sample. Plots 2 report the spatial distribution of large vessels in dark (red on line) and small vessels in gray (green on line).
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Figure 3: Quantitative analysis of the A, B, C, and D samples. Plots 1 show the number of vessels crossing the samples at the different depth inside the volume. For each sample, the three pictures above the plot 1, are the tomography images of the upper, central and top parts of the sample. Plots 2 report the spatial distribution of large vessels in dark (red on line) and small vessels in gray (green on line).

Mentions: While Figure 2 provides an imaging survey, Figure 3 contains the results of the quantitative analyses. In this figure, we considered the samples A, B, C, and D. We explored a central cube volume of 1.4 mm side, for all the samples. The precision of the selected volume was assured by the precision of the experimental setup, which allowed one to always illuminate equal portions of the samples. Exploiting the 3D character of the tomographic approach, we systematically analyzed the computed sections (each 640 nm thick) of the samples, perpendicular to any chosen direction, studying the number, section, and distribution of the vessels crossing each section. To get insights on the structure of the vascular trees in the recovered implants, we first plotted the number of vessels of the vascular network reaching the system at different depths (Figure 3, plot 1) and then we investigated the depth distribution of vessels with different size (Figure 3, plot 2).


High-Resolution X-Ray Techniques as New Tool to Investigate the 3D Vascularization of Engineered-Bone Tissue.

Bukreeva I, Fratini M, Campi G, Pelliccia D, Spanò R, Tromba G, Brun F, Burghammer M, Grilli M, Cancedda R, Cedola A, Mastrogiacomo M - Front Bioeng Biotechnol (2015)

Quantitative analysis of the A, B, C, and D samples. Plots 1 show the number of vessels crossing the samples at the different depth inside the volume. For each sample, the three pictures above the plot 1, are the tomography images of the upper, central and top parts of the sample. Plots 2 report the spatial distribution of large vessels in dark (red on line) and small vessels in gray (green on line).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561513&req=5

Figure 3: Quantitative analysis of the A, B, C, and D samples. Plots 1 show the number of vessels crossing the samples at the different depth inside the volume. For each sample, the three pictures above the plot 1, are the tomography images of the upper, central and top parts of the sample. Plots 2 report the spatial distribution of large vessels in dark (red on line) and small vessels in gray (green on line).
Mentions: While Figure 2 provides an imaging survey, Figure 3 contains the results of the quantitative analyses. In this figure, we considered the samples A, B, C, and D. We explored a central cube volume of 1.4 mm side, for all the samples. The precision of the selected volume was assured by the precision of the experimental setup, which allowed one to always illuminate equal portions of the samples. Exploiting the 3D character of the tomographic approach, we systematically analyzed the computed sections (each 640 nm thick) of the samples, perpendicular to any chosen direction, studying the number, section, and distribution of the vessels crossing each section. To get insights on the structure of the vascular trees in the recovered implants, we first plotted the number of vessels of the vascular network reaching the system at different depths (Figure 3, plot 1) and then we investigated the depth distribution of vessels with different size (Figure 3, plot 2).

Bottom Line: We compared samples seeded and not seeded with BMSC, as well as samples differently stained or unstained.Thanks to the high quality of the images, we investigated the 3D distribution of both vessels and collagen matrix and we obtained quantitative information for all different samples.We propose our approach as a tool for quantitative studies of angiogenesis in TE and for any pre-clinical investigation where a quantitative analysis of the vascular network is required.

View Article: PubMed Central - PubMed

Affiliation: Consiglio Nazionale delle Ricerche - Istituto NANOTEC, c/o Dipartimento di Fisica, Università Sapienza , Rome , Italy.

ABSTRACT
The understanding of structure-function relationships in normal and pathologic mammalian tissues is at the basis of a tissue engineering (TE) approach for the development of biological substitutes to restore or improve tissue function. In this framework, it is interesting to investigate engineered bone tissue, formed when porous ceramic constructs are loaded with bone marrow stromal cells (BMSC) and implanted in vivo. To monitor the relation between bone formation and vascularization, it is important to achieve a detailed imaging and a quantitative description of the complete three-dimensional vascular network in such constructs. Here, we used synchrotron X-ray phase-contrast micro-tomography to visualize and analyze the three-dimensional micro-vascular networks in bone-engineered constructs, in an ectopic bone formation mouse-model. We compared samples seeded and not seeded with BMSC, as well as samples differently stained or unstained. Thanks to the high quality of the images, we investigated the 3D distribution of both vessels and collagen matrix and we obtained quantitative information for all different samples. We propose our approach as a tool for quantitative studies of angiogenesis in TE and for any pre-clinical investigation where a quantitative analysis of the vascular network is required.

No MeSH data available.


Related in: MedlinePlus