Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation of CHA, F-CHA, and CPC + CHA Composites
2.3. Strength, Injection, and Dispersion Tests
2.4. In Vitro Measurements
2.4.1. Cytotoxicity toward L929 Cells
2.4.2. Attachment, Proliferation, and Mineralization of D1 Osteoprogenitor Cell Cultures on Sample Surfaces
Cell Attachment and Morphological Observation
Semi-Quantitative Detection of Alkaline Phosphatase (ALP) Activity
2.5. Statistical Analysis
3. Results and Discussion
3.1. Identification of CHA
3.1.1. XRD Phase Identification
3.1.2. FTIR Spectral Analysis
3.1.3. TEM Images and CHA Nanorod Aspect Ratios
3.1.4. Cytotoxicity of CHA
3.2. Identification of F-CHA and the Corresponding Templating Mechanism
3.3. Release Characteristics, Strength, Phase, Morphology, and Injectability of CPC + F-CHA
3.3.1. Individual Time and Cumulative Release Characteristics of Ferulic Acid
3.3.2. Compressive and Diametral Tensile Strengths of CPC + F-CHA Composites
3.3.3. SEM Images of the Fracture Surfaces and XRD Phase Identification of CPC + F-CHA Composites after Immersion for 1 Day
3.3.4. Injection and Disintegration Tests of the CPC + F-CHA Composites in ddH2O
3.4. Cytotoxicity toward L929, Cell Attachment, Proliferation, and Differentiation of D1 Cultured with CPC + F-CHA
3.4.1. Cell Viability and Morphologies of Sample Extract Culture with L929 Cells
3.4.2. Morphology and ALP Activity of D1 Progenitor Bone Cells on Various CPC + F-CHA Composite Surfaces
4. Conclusions
- L25K is a suitable mediator for impregnating bone, promoting ferulic acid, and compositing with CPC bone cement.
- The release process of CPC + F-CHA composites shows three stages, including an early burst release from the composite surface, followed by the diffusion release of ferulic acid within the composites, and, finally, the slow release of the encapsulated drug. These findings prove that F-CHA may be an effective drug carrier.
- The addition of F-CHA did not affect apatite formation. Increases in the addition of F-CHA resulted in a decrease in the strength of the product obtained. The CS of the CPC + 2.5, 5.0% F-CHA 2.5%, and 5% additive groups were approximately 50 MPa; thus, these materials may be used as load-bearing devices. The group added with CPC + 10%F-CHA achieved a CS of only 30 MPa; while this strength is relatively low, the obtained material may still find applications in non-load bearing orthopedic restoration.
- After 1 h of culture, D1 cells cultured in the CPC-only group retained their spherical shape, but the cell morphology of cells in all other experimental CPC + F-CHA groups was flat. These results demonstrate that ferulic acid can promote progenitor bone cell attachment. On the 10th day of D1 cell culture, the amount of ALP production in the CPC + 10.0%F-CHA group was significantly higher than that in other treatment groups. This finding indicates that the addition of F-CHA in CPC increases the ability of a single cell to secrete ALP.
- It can be seen that the controlled amount of heparin can prepare uniform nanorod-shaped CHA with good biocompatibility. The grafting of ferulic acid to CPC yielded a material that could be used as a slow-release drug carrier, thereby increasing the applicability of CPC in orthopedic tissue engineering.
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Designated Groups | Nomenclature |
---|---|
Comparative processes without heparin | Control CHA nanorods |
Templated by heparin LEO INJ 25,000 unit/mL | L25K |
Templated by heparin LEO INJ 50,000 unit/mL | L50K |
Templated by heparin LEO INJ 100,000 unit/mL | L100K |
Using L25K templated CHA for ferulic acid impregnation and composite CPC with a ratio of 2.5 wt% L25K/CPC | CPC + 2.5% F-CHA |
Using L25K templated CHA for ferulic acid impregnation and composite CPC with a ratio of 5.0 wt% L25K/CPC | CPC + 5.0% F-CHA |
Using L25K templated CHA for ferulic acid impregnation and composite CPC with a ratio of 10.0 wt% L25K/CPC | CPC + 10.0% F-CHA |
Dimensions and Aspect Ratio | Control CHA Nanorods Mean (S.D.) a | L25K Mean (S.D.) a | L50K Mean (S.D.) a | L100K Mean (S.D.) a |
---|---|---|---|---|
Length (nm) | 70.08 (28.36) | 70.32 (20.94) | 60.62 (21.04) | 52.76 (8.71) |
Width (nm) | 24.35 (6.16) | 19.84 (5.20) | 17.51 (3.69) | 14.93 (6.16) |
Aspect length-to-width ratio | 2.88 | 3.54 | 3.46 | 3.53 |
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Chang, K.-C.; Chen, J.-C.; Cheng, I.-T.; Haung, S.-M.; Liu, S.-M.; Ko, C.-L.; Sun, Y.-S.; Shih, C.-J.; Chen, W.-C. Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid. Polymers 2021, 13, 2219. https://doi.org/10.3390/polym13132219
Chang K-C, Chen J-C, Cheng I-T, Haung S-M, Liu S-M, Ko C-L, Sun Y-S, Shih C-J, Chen W-C. Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid. Polymers. 2021; 13(13):2219. https://doi.org/10.3390/polym13132219
Chicago/Turabian StyleChang, Kai-Chi, Jian-Chih Chen, I-Tse Cheng, Ssu-Meng Haung, Shih-Ming Liu, Chia-Ling Ko, Ying-Sui Sun, Chi-Jen Shih, and Wen-Cheng Chen. 2021. "Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid" Polymers 13, no. 13: 2219. https://doi.org/10.3390/polym13132219