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Selective laser sintering fabrication of nano-hydroxyapatite/poly-ε-caprolactone scaffolds for bone tissue engineering applications.

Xia Y, Zhou P, Cheng X, Xie Y, Liang C, Li C, Xu S - Int J Nanomedicine (2013)

Bottom Line: The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility.However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds.Thus, they show large potential for use in orthopedic and reconstructive surgery.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China.

ABSTRACT
The regeneration of functional tissue in osseous defects is a formidable challenge in orthopedic surgery. In the present study, a novel biomimetic composite scaffold, here called nano-hydroxyapatite (HA)/poly-ε-caprolactone (PCL) was fabricated using a selective laser sintering technique. The macrostructure, morphology, and mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the nano-HA/PCL scaffolds exhibited predesigned, well-ordered macropores and interconnected micropores. The scaffolds have a range of porosity from 78.54% to 70.31%, and a corresponding compressive strength of 1.38 MPa to 3.17 MPa. Human bone marrow stromal cells were seeded onto the nano-HA/PCL or PCL scaffolds and cultured for 28 days in vitro. As indicated by the level of cell attachment and proliferation, the nano-HA/PCL showed excellent biocompatibility, comparable to that of PCL scaffolds. The hydrophilicity, mineralization, alkaline phosphatase activity, and Alizarin Red S staining indicated that the nano-HA/PCL scaffolds are more bioactive than the PCL scaffolds in vitro. Measurements of recombinant human bone morphogenetic protein-2 (rhBMP-2) release kinetics showed that after nano-HA was added, the material increased the rate of rhBMP-2 release. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both nano-HA/PCL scaffolds and PCL scaffolds were implanted in rabbit femur defects for 3, 6, and 9 weeks. The wounds were studied radiographically and histologically. The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility. However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds. Thus, they show large potential for use in orthopedic and reconstructive surgery.

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Cell viability of the extracts of the scaffolds at different nano-HA contents.Notes: *P<0.05 versus control; #P<0.05 versus pure PCL scaffolds.Abbreviations: HA, hydroxyapatite; PCL, poly-ε-caprolactone.
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f4-ijn-8-4197: Cell viability of the extracts of the scaffolds at different nano-HA contents.Notes: *P<0.05 versus control; #P<0.05 versus pure PCL scaffolds.Abbreviations: HA, hydroxyapatite; PCL, poly-ε-caprolactone.

Mentions: Cell proliferation was analyzed using a CCK-8 assay (Figure 4). Cell proliferation is expressed as the percentage of viable cells exposed to the scaffolds. Cell proliferation of the control cells on days 1, 4, and 7 was always considered to be 100%. On day 1, cell proliferation on scaffolds made with 0 (pure PCL), 5, 10, and 15 wt% nano-HA was 99.7%±3.83%, 103.41%±4.15%, 107.15%±4.23%, and 109.48%±4.68%, respectively. The increase in cell proliferation relative to the control was significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. The increase in cell proliferation relative to the pure PCL was also significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. Cell proliferation values on day 4 were 101.61%±4.36%, 107.23%±4.57%, 109.76%±4.34%, and 113.83%±4.57% for scaffolds containing 0, 5, 10, and 15 wt% nano-HA, respectively. The increase relative to the control was significant (P<0.05) for the scaffolds at 5, 10, and 15 wt% nano-HA, and the cell proliferation of the scaffolds compared to the pure PCL was significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. The cell proliferation values on day 7 for the scaffolds at 0, 5, 10, and 15 wt% nano-HA were 105.94%±4.95%, 112.31%±4.94%, 113.24%±5.33%, and 117.62%±4.83%, respectively. This increase relative to the control was significant (P<0.05) for scaffolds at 5, 10, and 15 wt% nano-HA, and the cell proliferation of the scaffolds relative to pure PCL was also significant (P<0.05) for the scaffolds made with 15 wt% nano-HA only. The CCK-8 assay clearly implies that the scaffolds made of nano-HA and PCL – in particular those made with 15 wt% nano-HA – provided an excellent microenvironment for hMSC growth. We also cultured HaCaT cells with the scaffold for 7 days, and the SEM showed that, after 7 days, HaCaT cells had grown into the scaffold (Figure S1).


Selective laser sintering fabrication of nano-hydroxyapatite/poly-ε-caprolactone scaffolds for bone tissue engineering applications.

Xia Y, Zhou P, Cheng X, Xie Y, Liang C, Li C, Xu S - Int J Nanomedicine (2013)

Cell viability of the extracts of the scaffolds at different nano-HA contents.Notes: *P<0.05 versus control; #P<0.05 versus pure PCL scaffolds.Abbreviations: HA, hydroxyapatite; PCL, poly-ε-caprolactone.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3818022&req=5

f4-ijn-8-4197: Cell viability of the extracts of the scaffolds at different nano-HA contents.Notes: *P<0.05 versus control; #P<0.05 versus pure PCL scaffolds.Abbreviations: HA, hydroxyapatite; PCL, poly-ε-caprolactone.
Mentions: Cell proliferation was analyzed using a CCK-8 assay (Figure 4). Cell proliferation is expressed as the percentage of viable cells exposed to the scaffolds. Cell proliferation of the control cells on days 1, 4, and 7 was always considered to be 100%. On day 1, cell proliferation on scaffolds made with 0 (pure PCL), 5, 10, and 15 wt% nano-HA was 99.7%±3.83%, 103.41%±4.15%, 107.15%±4.23%, and 109.48%±4.68%, respectively. The increase in cell proliferation relative to the control was significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. The increase in cell proliferation relative to the pure PCL was also significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. Cell proliferation values on day 4 were 101.61%±4.36%, 107.23%±4.57%, 109.76%±4.34%, and 113.83%±4.57% for scaffolds containing 0, 5, 10, and 15 wt% nano-HA, respectively. The increase relative to the control was significant (P<0.05) for the scaffolds at 5, 10, and 15 wt% nano-HA, and the cell proliferation of the scaffolds compared to the pure PCL was significant (P<0.05) for the scaffolds at 10 and 15 wt% nano-HA. The cell proliferation values on day 7 for the scaffolds at 0, 5, 10, and 15 wt% nano-HA were 105.94%±4.95%, 112.31%±4.94%, 113.24%±5.33%, and 117.62%±4.83%, respectively. This increase relative to the control was significant (P<0.05) for scaffolds at 5, 10, and 15 wt% nano-HA, and the cell proliferation of the scaffolds relative to pure PCL was also significant (P<0.05) for the scaffolds made with 15 wt% nano-HA only. The CCK-8 assay clearly implies that the scaffolds made of nano-HA and PCL – in particular those made with 15 wt% nano-HA – provided an excellent microenvironment for hMSC growth. We also cultured HaCaT cells with the scaffold for 7 days, and the SEM showed that, after 7 days, HaCaT cells had grown into the scaffold (Figure S1).

Bottom Line: The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility.However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds.Thus, they show large potential for use in orthopedic and reconstructive surgery.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China.

ABSTRACT
The regeneration of functional tissue in osseous defects is a formidable challenge in orthopedic surgery. In the present study, a novel biomimetic composite scaffold, here called nano-hydroxyapatite (HA)/poly-ε-caprolactone (PCL) was fabricated using a selective laser sintering technique. The macrostructure, morphology, and mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the nano-HA/PCL scaffolds exhibited predesigned, well-ordered macropores and interconnected micropores. The scaffolds have a range of porosity from 78.54% to 70.31%, and a corresponding compressive strength of 1.38 MPa to 3.17 MPa. Human bone marrow stromal cells were seeded onto the nano-HA/PCL or PCL scaffolds and cultured for 28 days in vitro. As indicated by the level of cell attachment and proliferation, the nano-HA/PCL showed excellent biocompatibility, comparable to that of PCL scaffolds. The hydrophilicity, mineralization, alkaline phosphatase activity, and Alizarin Red S staining indicated that the nano-HA/PCL scaffolds are more bioactive than the PCL scaffolds in vitro. Measurements of recombinant human bone morphogenetic protein-2 (rhBMP-2) release kinetics showed that after nano-HA was added, the material increased the rate of rhBMP-2 release. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both nano-HA/PCL scaffolds and PCL scaffolds were implanted in rabbit femur defects for 3, 6, and 9 weeks. The wounds were studied radiographically and histologically. The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility. However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds. Thus, they show large potential for use in orthopedic and reconstructive surgery.

Show MeSH
Related in: MedlinePlus