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Human Dental Pulp Stem Cells and Gingival Fibroblasts Seeded into Silk Fibroin Scaffolds Have the Same Ability in Attracting Vessels.

Woloszyk A, Buschmann J, Waschkies C, Stadlinger B, Mitsiadis TA - Front Physiol (2016)

Bottom Line: Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts.Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds.Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration.

View Article: PubMed Central - PubMed

Affiliation: Orofacial Development and Regeneration, Center of Dental Medicine, Institute of Oral Biology, University of Zurich Zurich, Switzerland.

ABSTRACT
Neovascularization is one of the most important processes during tissue repair and regeneration. Current healing approaches based on the use of biomaterials combined with stem cells in critical-size bone defects fail due to the insufficient implant vascularization and integration into the host tissues. Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts. Perfusion capacity was evaluated by non-invasive in vivo Magnetic Resonance Imaging while the number and density of blood vessels were measured by histomorphometry. Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds. Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration.

No MeSH data available.


Related in: MedlinePlus

Experimental setup and MRI analysis. (A,B) The scaffold is placed in the middle of a silicone ring on top of the CAM of a fertilized egg, which is stabilized in a Petri dish and incubated for 1 week. (C) Top and (D) bottom views of the scaffold after 1 week of incubation in ovo. (E) Scheme showing the analyzed parts of the samples. (F) Magnetic resonance image of the vascularized scaffold before and after the injection of the contrast agent Gd-DOTA. (G) Relaxation rate change ΔR1 in the interface, middle, and surface region of the scaffolds. Values are given as mean ± standard deviation. No statistically significant differences were found. (H) Percent change of the mean ΔR1 and slope of regression line between the interface and the surface layers within each group. CAM, chorioallantoic membrane; sc, scaffold; sr, silicone ring; vs, vessel; hDPSCs, human dental pulp stem cells; hGFs, human gingival fibroblasts.
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Figure 1: Experimental setup and MRI analysis. (A,B) The scaffold is placed in the middle of a silicone ring on top of the CAM of a fertilized egg, which is stabilized in a Petri dish and incubated for 1 week. (C) Top and (D) bottom views of the scaffold after 1 week of incubation in ovo. (E) Scheme showing the analyzed parts of the samples. (F) Magnetic resonance image of the vascularized scaffold before and after the injection of the contrast agent Gd-DOTA. (G) Relaxation rate change ΔR1 in the interface, middle, and surface region of the scaffolds. Values are given as mean ± standard deviation. No statistically significant differences were found. (H) Percent change of the mean ΔR1 and slope of regression line between the interface and the surface layers within each group. CAM, chorioallantoic membrane; sc, scaffold; sr, silicone ring; vs, vessel; hDPSCs, human dental pulp stem cells; hGFs, human gingival fibroblasts.

Mentions: No IACUC approval is necessary when performing experiments in chicken embryos until embryonic day 14 according to Swiss animal care guidelines (TSchV, Art. 112). Fertilized Lohman white LSL chicken eggs (Animalco AG, Staufen AG, Switzerland) were pre-incubated for 3 days at 38°C at a rotation speed of 360°/4 h (Bruja 3000, Brutmaschinen-Janeschitz GmbH, Hammelburg, Germany). On embryonic day 3 (ED 3) the eggs were processed for in ovo cultivation, which requires the opening of the shell with a drill (Dremel§, Conrad Electronic AG, Wollerau SZ, Switzerland). 2 mL of albumen was always removed with a syringe to increase the empty space under the top of the egg shell. The eggs were stabilized in 60 mm Petri dishes (Greiner Bio-One GmbH, Frickenhausen, Germany) and the created holes of the shells were covered with another 60 mm Petri dish that was fixed with a tape before the incubation of eggs at 37°C. On ED 7, empty and cell-seeded scaffolds were placed on the CAM (1-2/egg) in the middle of silicone rings that ensure a flat surface (Figures 1A,B) during their incubation period of 7 days.


Human Dental Pulp Stem Cells and Gingival Fibroblasts Seeded into Silk Fibroin Scaffolds Have the Same Ability in Attracting Vessels.

Woloszyk A, Buschmann J, Waschkies C, Stadlinger B, Mitsiadis TA - Front Physiol (2016)

Experimental setup and MRI analysis. (A,B) The scaffold is placed in the middle of a silicone ring on top of the CAM of a fertilized egg, which is stabilized in a Petri dish and incubated for 1 week. (C) Top and (D) bottom views of the scaffold after 1 week of incubation in ovo. (E) Scheme showing the analyzed parts of the samples. (F) Magnetic resonance image of the vascularized scaffold before and after the injection of the contrast agent Gd-DOTA. (G) Relaxation rate change ΔR1 in the interface, middle, and surface region of the scaffolds. Values are given as mean ± standard deviation. No statistically significant differences were found. (H) Percent change of the mean ΔR1 and slope of regression line between the interface and the surface layers within each group. CAM, chorioallantoic membrane; sc, scaffold; sr, silicone ring; vs, vessel; hDPSCs, human dental pulp stem cells; hGFs, human gingival fibroblasts.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835714&req=5

Figure 1: Experimental setup and MRI analysis. (A,B) The scaffold is placed in the middle of a silicone ring on top of the CAM of a fertilized egg, which is stabilized in a Petri dish and incubated for 1 week. (C) Top and (D) bottom views of the scaffold after 1 week of incubation in ovo. (E) Scheme showing the analyzed parts of the samples. (F) Magnetic resonance image of the vascularized scaffold before and after the injection of the contrast agent Gd-DOTA. (G) Relaxation rate change ΔR1 in the interface, middle, and surface region of the scaffolds. Values are given as mean ± standard deviation. No statistically significant differences were found. (H) Percent change of the mean ΔR1 and slope of regression line between the interface and the surface layers within each group. CAM, chorioallantoic membrane; sc, scaffold; sr, silicone ring; vs, vessel; hDPSCs, human dental pulp stem cells; hGFs, human gingival fibroblasts.
Mentions: No IACUC approval is necessary when performing experiments in chicken embryos until embryonic day 14 according to Swiss animal care guidelines (TSchV, Art. 112). Fertilized Lohman white LSL chicken eggs (Animalco AG, Staufen AG, Switzerland) were pre-incubated for 3 days at 38°C at a rotation speed of 360°/4 h (Bruja 3000, Brutmaschinen-Janeschitz GmbH, Hammelburg, Germany). On embryonic day 3 (ED 3) the eggs were processed for in ovo cultivation, which requires the opening of the shell with a drill (Dremel§, Conrad Electronic AG, Wollerau SZ, Switzerland). 2 mL of albumen was always removed with a syringe to increase the empty space under the top of the egg shell. The eggs were stabilized in 60 mm Petri dishes (Greiner Bio-One GmbH, Frickenhausen, Germany) and the created holes of the shells were covered with another 60 mm Petri dish that was fixed with a tape before the incubation of eggs at 37°C. On ED 7, empty and cell-seeded scaffolds were placed on the CAM (1-2/egg) in the middle of silicone rings that ensure a flat surface (Figures 1A,B) during their incubation period of 7 days.

Bottom Line: Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts.Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds.Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration.

View Article: PubMed Central - PubMed

Affiliation: Orofacial Development and Regeneration, Center of Dental Medicine, Institute of Oral Biology, University of Zurich Zurich, Switzerland.

ABSTRACT
Neovascularization is one of the most important processes during tissue repair and regeneration. Current healing approaches based on the use of biomaterials combined with stem cells in critical-size bone defects fail due to the insufficient implant vascularization and integration into the host tissues. Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts. Perfusion capacity was evaluated by non-invasive in vivo Magnetic Resonance Imaging while the number and density of blood vessels were measured by histomorphometry. Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds. Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration.

No MeSH data available.


Related in: MedlinePlus