Limits...
3D Printing Bioceramic Porous Scaffolds with Good Mechanical Property and Cell Affinity.

Chang CH, Lin CY, Liu FH, Chen MH, Lin CP, Ho HN, Liao YS - PLoS ONE (2015)

Bottom Line: Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold.The compressive strength was 47 MPa, and the porosity was 34%.The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.

ABSTRACT
Artificial bone grafting is widely used in current orthopedic surgery for bone defect problems. Unfortunately, surgeons remain unsatisfied with the current commercially available products. One of the major complaints is that these products cannot provide sufficient mechanical strength to support the human skeletal structure. In this study, we aimed to develop a bone scaffold with better mechanical property and good cell affinity by 3D printing (3DP) techniques. A self-developed 3D printer with laser-aided gelling (LAG) process was used to fabricate bioceramic scaffolds with inter-porous structures. To improve the mechanical property of the bioceramic parts after heating, CaCO3 was added to the silica ceramic slurry. CaCO3 was blended into a homogenous SiO2-sol dispersion at weight ratios varying from 0/100 to 5/95 to 9/91 (w/w). Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold. The well-mixed biocomposite was used to fabricate the bioceramic green part using the LAG method. The varied scaffolds were sintered at different temperatures ranging from 900 to 1500°C, and the mechanical property was subsequently analyzed. The scaffolds showed good property with the composite ratio of 5:95 CaCO3:SiO2 at a sintering temperature of 1300°C. The compressive strength was 47 MPa, and the porosity was 34%. The topography of the sintered 3DP bioceramic scaffold was examined by SEM, EDS and XRD. The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility. Therefore, the new silica biocomposite is viable for fabricating 3DP bone bioceramics with improved mechanical property and good cell affinity.

Show MeSH
Osteoblast-like MG-63 cells attach on the CS5 scaffold.The high affinity of human bone cells indicated that the fabricated biocomposites act as a biomimic of the human bone scaffold. Live color staining and fluorescence graphs of Nuclear/Actin contact analysis are shown in upper panel. (a) unattached scaffold, (b) live cell staining of formazan on scaffold, and (c) fluorescence on scaffold. The live cells on scaffolds were incubated in a tetrazolium dye bath at 37°C for 4 h. Green signals indicate actin, and blue signals indicate the nuclear site of cells. (d). CS5 scaffolds showing a long-term survival period of 1–6 days. Cell numbers corresponding to the OD570 values represented approximately 836 ± 37 cells for each 0.1 OD value by MTT assay, for normalization with cell counting. ** P<0.01 compared to day 1 group.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664392&req=5

pone.0143713.g010: Osteoblast-like MG-63 cells attach on the CS5 scaffold.The high affinity of human bone cells indicated that the fabricated biocomposites act as a biomimic of the human bone scaffold. Live color staining and fluorescence graphs of Nuclear/Actin contact analysis are shown in upper panel. (a) unattached scaffold, (b) live cell staining of formazan on scaffold, and (c) fluorescence on scaffold. The live cells on scaffolds were incubated in a tetrazolium dye bath at 37°C for 4 h. Green signals indicate actin, and blue signals indicate the nuclear site of cells. (d). CS5 scaffolds showing a long-term survival period of 1–6 days. Cell numbers corresponding to the OD570 values represented approximately 836 ± 37 cells for each 0.1 OD value by MTT assay, for normalization with cell counting. ** P<0.01 compared to day 1 group.

Mentions: To further demonstrate bone cell attachment on the CS5 silica bioceramic bone scaffolds (Fig 10a), we used two methods. First, we attempted to grow cells directly on the scaffold surface for approximately 24 hours and observed the growth using live staining with MTT direct uptake as shown in Fig 10b. Second, the cell morphology of the scaffold is shown by molecular probe fluorescence staining in Fig 10c. We further evaluated MG-63 cells during a long setting period on CS5 scaffold. We incubated the MG-63 cells for 1, 3, and 6 days on CS5 scaffold. As shown in Fig 10d, the cells proliferated twofold and fourfold at day 3 and day 6, respectively. The data showed long-term cell survival on the bioceramic scaffold, meaning that the cells were adapted to the CS5 scaffold environment. Future experiments on real-time cell penetration on 3D scaffolds will allow further evaluation of the long-term cell growth and differentiation states in the scaffolds in vitro and in vivo.


3D Printing Bioceramic Porous Scaffolds with Good Mechanical Property and Cell Affinity.

Chang CH, Lin CY, Liu FH, Chen MH, Lin CP, Ho HN, Liao YS - PLoS ONE (2015)

Osteoblast-like MG-63 cells attach on the CS5 scaffold.The high affinity of human bone cells indicated that the fabricated biocomposites act as a biomimic of the human bone scaffold. Live color staining and fluorescence graphs of Nuclear/Actin contact analysis are shown in upper panel. (a) unattached scaffold, (b) live cell staining of formazan on scaffold, and (c) fluorescence on scaffold. The live cells on scaffolds were incubated in a tetrazolium dye bath at 37°C for 4 h. Green signals indicate actin, and blue signals indicate the nuclear site of cells. (d). CS5 scaffolds showing a long-term survival period of 1–6 days. Cell numbers corresponding to the OD570 values represented approximately 836 ± 37 cells for each 0.1 OD value by MTT assay, for normalization with cell counting. ** P<0.01 compared to day 1 group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664392&req=5

pone.0143713.g010: Osteoblast-like MG-63 cells attach on the CS5 scaffold.The high affinity of human bone cells indicated that the fabricated biocomposites act as a biomimic of the human bone scaffold. Live color staining and fluorescence graphs of Nuclear/Actin contact analysis are shown in upper panel. (a) unattached scaffold, (b) live cell staining of formazan on scaffold, and (c) fluorescence on scaffold. The live cells on scaffolds were incubated in a tetrazolium dye bath at 37°C for 4 h. Green signals indicate actin, and blue signals indicate the nuclear site of cells. (d). CS5 scaffolds showing a long-term survival period of 1–6 days. Cell numbers corresponding to the OD570 values represented approximately 836 ± 37 cells for each 0.1 OD value by MTT assay, for normalization with cell counting. ** P<0.01 compared to day 1 group.
Mentions: To further demonstrate bone cell attachment on the CS5 silica bioceramic bone scaffolds (Fig 10a), we used two methods. First, we attempted to grow cells directly on the scaffold surface for approximately 24 hours and observed the growth using live staining with MTT direct uptake as shown in Fig 10b. Second, the cell morphology of the scaffold is shown by molecular probe fluorescence staining in Fig 10c. We further evaluated MG-63 cells during a long setting period on CS5 scaffold. We incubated the MG-63 cells for 1, 3, and 6 days on CS5 scaffold. As shown in Fig 10d, the cells proliferated twofold and fourfold at day 3 and day 6, respectively. The data showed long-term cell survival on the bioceramic scaffold, meaning that the cells were adapted to the CS5 scaffold environment. Future experiments on real-time cell penetration on 3D scaffolds will allow further evaluation of the long-term cell growth and differentiation states in the scaffolds in vitro and in vivo.

Bottom Line: Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold.The compressive strength was 47 MPa, and the porosity was 34%.The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.

ABSTRACT
Artificial bone grafting is widely used in current orthopedic surgery for bone defect problems. Unfortunately, surgeons remain unsatisfied with the current commercially available products. One of the major complaints is that these products cannot provide sufficient mechanical strength to support the human skeletal structure. In this study, we aimed to develop a bone scaffold with better mechanical property and good cell affinity by 3D printing (3DP) techniques. A self-developed 3D printer with laser-aided gelling (LAG) process was used to fabricate bioceramic scaffolds with inter-porous structures. To improve the mechanical property of the bioceramic parts after heating, CaCO3 was added to the silica ceramic slurry. CaCO3 was blended into a homogenous SiO2-sol dispersion at weight ratios varying from 0/100 to 5/95 to 9/91 (w/w). Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold. The well-mixed biocomposite was used to fabricate the bioceramic green part using the LAG method. The varied scaffolds were sintered at different temperatures ranging from 900 to 1500°C, and the mechanical property was subsequently analyzed. The scaffolds showed good property with the composite ratio of 5:95 CaCO3:SiO2 at a sintering temperature of 1300°C. The compressive strength was 47 MPa, and the porosity was 34%. The topography of the sintered 3DP bioceramic scaffold was examined by SEM, EDS and XRD. The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility. Therefore, the new silica biocomposite is viable for fabricating 3DP bone bioceramics with improved mechanical property and good cell affinity.

Show MeSH