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Biocompatibility of Poly-epsilon-caprolactone-hydroxyapatite composite on mouse bone marrow-derived osteoblasts and endothelial cells.

Yu H, Wooley PH, Yang SY - J Orthop Surg Res (2009)

Bottom Line: The results indicated that HA led to a positive stimulation of osteoblasts viability and ALP activity, while HA showed less influence on endothelial cells viability.Supplement of HA into PCL improved biocompatible for bone marrow-derived osteoblasts and endothelial cells.The PCL-HA composite integrating with two types of cells may provide a useful system for tissue-engineered bone grafts with vascularization.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, Wayne State University, Detroit, Michigan, USA. shang-you.yang@wichita.edu.

ABSTRACT

Background: Tissue-engineered bone may be developed by seeding the cells capable of both osteogenesis and vascularization on biocompatible composite scaffolds. The current study investigated the performance of mice bone marrow-derived osteogenic cells and endothelial cells as seeded on hydroxyapatite (HA) and poly-epsilon-caprolactone (PCL) composite scaffolds.

Methods: Mononuclear cells were induced to osteoblasts and endothelial cells respectively, which were defined by the expression of osteocalcin, alkaline phosphatase (ALP), and deposits of calcium-containing crystal for osteoblasts, or by the expression of vascular endothelial growth factor receptor-2 (VEGFR-2) and von Willebrand factor (vWF), and the formation of a capillary network in Matrigel for endothelial cells. Both types of cell were seeded respectively on PCL-HA scaffolds at HA to PCL weight ratio of 1:1, 1:4, or 0:1 and were evaluated using scanning electron microscopy, ALP activity (of osteoblasts) and nitric oxide production (of endothelial cells) plus the assessment of cell viability.

Results: The results indicated that HA led to a positive stimulation of osteoblasts viability and ALP activity, while HA showed less influence on endothelial cells viability. An elevated nitric oxide production of endothelial cells was observed in HA-containing group.

Conclusion: Supplement of HA into PCL improved biocompatible for bone marrow-derived osteoblasts and endothelial cells. The PCL-HA composite integrating with two types of cells may provide a useful system for tissue-engineered bone grafts with vascularization.

No MeSH data available.


Related in: MedlinePlus

Characterizations of osteoblasts. Over 95% of induced osteoblasts expressed osteocalcin visualized by immunocytochemistry stains (Panel A, 200×). Alkaline phosphatase (ALP) activity of osteoblasts was assayed using an ALP kit and visualized as the pink color (Panel B, 200×). In addition, von Kossa staining was performed to reveal ossification nodules in the culture dishes of induced cells, as an indicator of osteoblasts function (Panel C, 200×).
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Figure 2: Characterizations of osteoblasts. Over 95% of induced osteoblasts expressed osteocalcin visualized by immunocytochemistry stains (Panel A, 200×). Alkaline phosphatase (ALP) activity of osteoblasts was assayed using an ALP kit and visualized as the pink color (Panel B, 200×). In addition, von Kossa staining was performed to reveal ossification nodules in the culture dishes of induced cells, as an indicator of osteoblasts function (Panel C, 200×).

Mentions: The capacity of the induced osteoblasts to express osteocalcin was examined by immunocytochemistry (IHC) (Figure 2A). The expression of osteocalcin was detected in over 95% osteogenic-wise induced cells. These cells were also identified by positive staining for ALP (Figure 2B), which indicated that the induced cells possessed distinguishable osteoblastic phenotype. To demonstrate the ability of cells to mineralize matrix, cells cultured on Petri dishes were subjected to von Kossa stain to reveal calcium deposition (Figure 2C) where the darkly stained mineralized nodule were visualized by silver nitrate, indicating normal osteoblasts function in conditioned culture. Differentiation ability of ECs induced from bone marrow was determined by the expression of endothelial markers, VEGFR-2 and vWF, using immunocytofluorescence. The 95% endothelial-wise-induced cells expressed VEGFR-2 and vWF at 2nd passage of culture (Figure 3A, B), indicating the induced cells having normal endothelial phenotype. Matrigel™ culture was performed to monitor the capability of capillary formation. Three-dimensional capillary-like networks from EPCs-derived endothelial cells were clearly established at one-week incubation (Figure 3C).


Biocompatibility of Poly-epsilon-caprolactone-hydroxyapatite composite on mouse bone marrow-derived osteoblasts and endothelial cells.

Yu H, Wooley PH, Yang SY - J Orthop Surg Res (2009)

Characterizations of osteoblasts. Over 95% of induced osteoblasts expressed osteocalcin visualized by immunocytochemistry stains (Panel A, 200×). Alkaline phosphatase (ALP) activity of osteoblasts was assayed using an ALP kit and visualized as the pink color (Panel B, 200×). In addition, von Kossa staining was performed to reveal ossification nodules in the culture dishes of induced cells, as an indicator of osteoblasts function (Panel C, 200×).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654429&req=5

Figure 2: Characterizations of osteoblasts. Over 95% of induced osteoblasts expressed osteocalcin visualized by immunocytochemistry stains (Panel A, 200×). Alkaline phosphatase (ALP) activity of osteoblasts was assayed using an ALP kit and visualized as the pink color (Panel B, 200×). In addition, von Kossa staining was performed to reveal ossification nodules in the culture dishes of induced cells, as an indicator of osteoblasts function (Panel C, 200×).
Mentions: The capacity of the induced osteoblasts to express osteocalcin was examined by immunocytochemistry (IHC) (Figure 2A). The expression of osteocalcin was detected in over 95% osteogenic-wise induced cells. These cells were also identified by positive staining for ALP (Figure 2B), which indicated that the induced cells possessed distinguishable osteoblastic phenotype. To demonstrate the ability of cells to mineralize matrix, cells cultured on Petri dishes were subjected to von Kossa stain to reveal calcium deposition (Figure 2C) where the darkly stained mineralized nodule were visualized by silver nitrate, indicating normal osteoblasts function in conditioned culture. Differentiation ability of ECs induced from bone marrow was determined by the expression of endothelial markers, VEGFR-2 and vWF, using immunocytofluorescence. The 95% endothelial-wise-induced cells expressed VEGFR-2 and vWF at 2nd passage of culture (Figure 3A, B), indicating the induced cells having normal endothelial phenotype. Matrigel™ culture was performed to monitor the capability of capillary formation. Three-dimensional capillary-like networks from EPCs-derived endothelial cells were clearly established at one-week incubation (Figure 3C).

Bottom Line: The results indicated that HA led to a positive stimulation of osteoblasts viability and ALP activity, while HA showed less influence on endothelial cells viability.Supplement of HA into PCL improved biocompatible for bone marrow-derived osteoblasts and endothelial cells.The PCL-HA composite integrating with two types of cells may provide a useful system for tissue-engineered bone grafts with vascularization.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, Wayne State University, Detroit, Michigan, USA. shang-you.yang@wichita.edu.

ABSTRACT

Background: Tissue-engineered bone may be developed by seeding the cells capable of both osteogenesis and vascularization on biocompatible composite scaffolds. The current study investigated the performance of mice bone marrow-derived osteogenic cells and endothelial cells as seeded on hydroxyapatite (HA) and poly-epsilon-caprolactone (PCL) composite scaffolds.

Methods: Mononuclear cells were induced to osteoblasts and endothelial cells respectively, which were defined by the expression of osteocalcin, alkaline phosphatase (ALP), and deposits of calcium-containing crystal for osteoblasts, or by the expression of vascular endothelial growth factor receptor-2 (VEGFR-2) and von Willebrand factor (vWF), and the formation of a capillary network in Matrigel for endothelial cells. Both types of cell were seeded respectively on PCL-HA scaffolds at HA to PCL weight ratio of 1:1, 1:4, or 0:1 and were evaluated using scanning electron microscopy, ALP activity (of osteoblasts) and nitric oxide production (of endothelial cells) plus the assessment of cell viability.

Results: The results indicated that HA led to a positive stimulation of osteoblasts viability and ALP activity, while HA showed less influence on endothelial cells viability. An elevated nitric oxide production of endothelial cells was observed in HA-containing group.

Conclusion: Supplement of HA into PCL improved biocompatible for bone marrow-derived osteoblasts and endothelial cells. The PCL-HA composite integrating with two types of cells may provide a useful system for tissue-engineered bone grafts with vascularization.

No MeSH data available.


Related in: MedlinePlus