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In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds.

Sun H, Qu Z, Guo Y, Zang G, Yang B - Biomed Eng Online (2007)

Bottom Line: In addition, histochemical analysis with CD31 and von-Willebrand factor staining showed that VEC retained their endothelial characteristics.In vivo implantation of MSC and VEC co-cultures into rat's muscle resulted in pre-vascular network-like structure established by the VEC in the PLGA.These results suggest that the vascular endothelial cells could efficiently stimulate the in vitro proliferation and differentiation of osteoblast-like cells and promote osteogenesis in vivo by the direct contact or interaction with the MSC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oral Pathology, School of Stomatology, JiLin University, ChangChun 130041, PR China. hcsun@jlu.edu.cn

ABSTRACT
It is well established that vascularization is critical for osteogenesis. However, adequate vascularization also remains one of the major challenges in tissue engineering of bone. This problem is further accentuated in regeneration of large volume of tissue. Although a complex process, vascularization involves reciprocal regulation and functional interaction between endothelial and osteoblast-like cells during osteogenesis. This prompted us to investigate the possibility of producing bone tissue both in vitro and ectopically in vivo using vascular endothelial cells because we hypothesized that the direct contact or interaction between vascular endothelial cells and bone marrow mesenchymal stem cells are of benefit to osteogenesis in vitro and in vivo. For that purpose we co-cultured rat bone marrow mesenchymal stem cells (MSC) and kidney vascular endothelial cells (VEC) with polylactide-glycolic acid scaffolds. In vitro experiments using alkaline phosphatase and osteocalcin assays demonstrated the proliferation and differentiation of MSC into osteoblast-like cells, especially the direct contact between VEC and MSC. In addition, histochemical analysis with CD31 and von-Willebrand factor staining showed that VEC retained their endothelial characteristics. In vivo implantation of MSC and VEC co-cultures into rat's muscle resulted in pre-vascular network-like structure established by the VEC in the PLGA. These structures developed into vascularized tissue, and increased the amount and size of the new bone compared to the control group (p < 0.05). These results suggest that the vascular endothelial cells could efficiently stimulate the in vitro proliferation and differentiation of osteoblast-like cells and promote osteogenesis in vivo by the direct contact or interaction with the MSC. This technique for optimal regeneration of bone should be further investigated.

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Hematoxylin and eosin staining of the implanted area from rat thigh and Quantitative measurements of new blood vessel and new bone from HE staining slides. A. Control group at 8 weeks post-implanted. B. Experiment group at 8 weeks post-implanted. C. Control group at 12 weeks post-implanted. D. Experiment group at 12 weeks post-implanted. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. Red arrow indicates PLGA material. Black arrow indicates new bone. Yellow arrow indicates blood vessel. E. New blood vessel. F. New bone. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. The data shown are the means ± SD from six experiments.
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Figure 4: Hematoxylin and eosin staining of the implanted area from rat thigh and Quantitative measurements of new blood vessel and new bone from HE staining slides. A. Control group at 8 weeks post-implanted. B. Experiment group at 8 weeks post-implanted. C. Control group at 12 weeks post-implanted. D. Experiment group at 12 weeks post-implanted. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. Red arrow indicates PLGA material. Black arrow indicates new bone. Yellow arrow indicates blood vessel. E. New blood vessel. F. New bone. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. The data shown are the means ± SD from six experiments.

Mentions: After 8 or 12 weeks following implantation specimens were collected and examined by soft X-rays. The corresponding radiographs are shown in figure 3. Eight weeks post-implantation, diffuse radio-opacity could be seen in the right thigh but not in the left (data not shown). This difference between the right thigh (with the MSC and VEC plated PLGA) and the left thigh (with the MSC plated PLGA) was even more pronounced at 12 weeks after the implantation (Fig. 3A and 3B). Densitometry of the various samples was performed using the NIH-image software and the results are displayed in figure 4. While the left thigh was no different from control, a 2-fold increase in density was seen in the right thigh (Fig. 3C). These results suggest that addition of VEC to cultured MSC induced osteogenesis in vivo.


In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds.

Sun H, Qu Z, Guo Y, Zang G, Yang B - Biomed Eng Online (2007)

Hematoxylin and eosin staining of the implanted area from rat thigh and Quantitative measurements of new blood vessel and new bone from HE staining slides. A. Control group at 8 weeks post-implanted. B. Experiment group at 8 weeks post-implanted. C. Control group at 12 weeks post-implanted. D. Experiment group at 12 weeks post-implanted. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. Red arrow indicates PLGA material. Black arrow indicates new bone. Yellow arrow indicates blood vessel. E. New blood vessel. F. New bone. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. The data shown are the means ± SD from six experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2186340&req=5

Figure 4: Hematoxylin and eosin staining of the implanted area from rat thigh and Quantitative measurements of new blood vessel and new bone from HE staining slides. A. Control group at 8 weeks post-implanted. B. Experiment group at 8 weeks post-implanted. C. Control group at 12 weeks post-implanted. D. Experiment group at 12 weeks post-implanted. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. Red arrow indicates PLGA material. Black arrow indicates new bone. Yellow arrow indicates blood vessel. E. New blood vessel. F. New bone. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. The data shown are the means ± SD from six experiments.
Mentions: After 8 or 12 weeks following implantation specimens were collected and examined by soft X-rays. The corresponding radiographs are shown in figure 3. Eight weeks post-implantation, diffuse radio-opacity could be seen in the right thigh but not in the left (data not shown). This difference between the right thigh (with the MSC and VEC plated PLGA) and the left thigh (with the MSC plated PLGA) was even more pronounced at 12 weeks after the implantation (Fig. 3A and 3B). Densitometry of the various samples was performed using the NIH-image software and the results are displayed in figure 4. While the left thigh was no different from control, a 2-fold increase in density was seen in the right thigh (Fig. 3C). These results suggest that addition of VEC to cultured MSC induced osteogenesis in vivo.

Bottom Line: In addition, histochemical analysis with CD31 and von-Willebrand factor staining showed that VEC retained their endothelial characteristics.In vivo implantation of MSC and VEC co-cultures into rat's muscle resulted in pre-vascular network-like structure established by the VEC in the PLGA.These results suggest that the vascular endothelial cells could efficiently stimulate the in vitro proliferation and differentiation of osteoblast-like cells and promote osteogenesis in vivo by the direct contact or interaction with the MSC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oral Pathology, School of Stomatology, JiLin University, ChangChun 130041, PR China. hcsun@jlu.edu.cn

ABSTRACT
It is well established that vascularization is critical for osteogenesis. However, adequate vascularization also remains one of the major challenges in tissue engineering of bone. This problem is further accentuated in regeneration of large volume of tissue. Although a complex process, vascularization involves reciprocal regulation and functional interaction between endothelial and osteoblast-like cells during osteogenesis. This prompted us to investigate the possibility of producing bone tissue both in vitro and ectopically in vivo using vascular endothelial cells because we hypothesized that the direct contact or interaction between vascular endothelial cells and bone marrow mesenchymal stem cells are of benefit to osteogenesis in vitro and in vivo. For that purpose we co-cultured rat bone marrow mesenchymal stem cells (MSC) and kidney vascular endothelial cells (VEC) with polylactide-glycolic acid scaffolds. In vitro experiments using alkaline phosphatase and osteocalcin assays demonstrated the proliferation and differentiation of MSC into osteoblast-like cells, especially the direct contact between VEC and MSC. In addition, histochemical analysis with CD31 and von-Willebrand factor staining showed that VEC retained their endothelial characteristics. In vivo implantation of MSC and VEC co-cultures into rat's muscle resulted in pre-vascular network-like structure established by the VEC in the PLGA. These structures developed into vascularized tissue, and increased the amount and size of the new bone compared to the control group (p < 0.05). These results suggest that the vascular endothelial cells could efficiently stimulate the in vitro proliferation and differentiation of osteoblast-like cells and promote osteogenesis in vivo by the direct contact or interaction with the MSC. This technique for optimal regeneration of bone should be further investigated.

Show MeSH
Related in: MedlinePlus