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. 2022 Mar 10;33(3):31.
doi: 10.1007/s10856-022-06653-8.

Three-dimensional printing of polycaprolactone/hydroxyapatite bone tissue engineering scaffolds mechanical properties and biological behavior

Naghme Rezania  1   2 , Mitra Asadi-Eydivand  3   4 , Nabiollah Abolfathi  2 , Shahin Bonakdar  5 , Morteza Mehrjoo  2   5 , Mehran Solati-Hashjin  2   6
Affiliations

Three-dimensional printing of polycaprolactone/hydroxyapatite bone tissue engineering scaffolds mechanical properties and biological behavior

Naghme Rezania et al. J Mater Sci Mater Med. .

Abstract

Controlled pore size and desirable internal architecture of bone scaffolds play a significant role in bone regeneration efficiency. In addition to choosing appropriate materials, the manufacturing method is another significant factor in fabricating the ideal scaffold. In this study, scaffolds were designed and fabricated by the fused filament fabrication (FFF) technique. Polycaprolactone (PCL) and composites films with various percentages of hydroxyapatite (HA) (up to 20%wt) were used to fabricate filaments. The influence of (HA) addition on the mechanical properties of filaments and scaffolds was investigated. in vitro biological evaluation was examined as well as the apatite formation in simulated body fluid (SBF). The addition of HA particles increased the compressive strength and Young's modulus of filaments and consequently the scaffolds. Compared to PCL, Young's modulus of PCL/HA20% filament and three-dimensional (3D) printed scaffold has increased by 30% and 50%, respectively. Also, Young's modulus for all scaffolds was in the range of 30-70 MPa, which is appropriate to use in spongy bone. Besides, the MTT assay was utilized to evaluate cell viability on the scaffolds. All the samples had qualified cytocompatibility, and it would be anticipated that addition of HA particles raise the biocompatibility in vivo. Alkaline phosphatase (ALP) evaluation shows that the addition of HA caused higher ALP activity in the PCL/HA scaffolds than PCL. Furthermore, calcium deposition in the PCL/HA specimens is higher than control. In conclusion, the addition of HA particles into the PCL matrix, as well as utilizing an inexpensive commercial FFF device, lead to the fabrication of scaffolds with proper mechanical and biological properties for bone tissue engineering applications. Graphical abstract.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Fig. 1
Fig. 1
The schema of the fused filament fabrication (FFF) process, from left to right, shows the computational design of scaffold, filament fabrication, and 3D printing process of scaffolds
Fig. 2
Fig. 2
Design of three-dimensional (3D) scaffold in SolidWorks 2017 software. It shows over-view and cross-section view of scaffolds with diameter and height of 20 mm and 5.2 mm, respectively
Fig. 3
Fig. 3
A XRD pattern of synthesized hydroxyapatite to show the crystalline structure of HA
Fig. 4
Fig. 4
A SEM images of PCL/HA5%, PCL/HA10%, PCL/HA15% and PCL/HA20% composite films. White particle corresponds to the presence of HA. B XRD pattern of PCL/HA10% and PCL/HA20% composite films. The particular peaks of PCL and HA are at the angles of 2θ = 23.67, 21.37°, and 2θ = 32,27°, respectively. C Rheology behavior of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% composites at 180 °C. Error bars show standard deviation (n = 3). The rheology has decreased by increasing of HA percentage. (**p < 0.01, ****p < 0.0001)
Fig. 5
Fig. 5
A Stress-strain curve of fabricated PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% under tensile strength. B Elastic section of stress-strain curves of filaments under tensile strength. C, D Young’s modulus and yield stress of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% filaments under tensile strength which is calculated from the stress-strain curves. Error bar presents the standard deviation. (*p < 0.05, **p < 0.01, and ***p < 0.001, n = 3)
Fig. 6
Fig. 6
SEM images of PCL, PCL/HA10%, and PCL/HA20% scaffolds. The first and second columns show top and cross-section views of scaffolds (The scale bar is 1 mm and 500 µm, respectively). The third column pictures show the presence of HA particles in the polymeric matrix. (The scale bar is 5 µm)
Fig. 7
Fig. 7
A Punched specimens for the mechanical testing. The diameter and height of the samples were 6 mm and 12 mm, respectively. B The stress-strain curves of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% scaffolds under compression test (n = 3). C The Young’s modulus of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% scaffolds under compression test. Error bar indicateس the standard deviation. (*p < 0.05 and **p < 0.01, n = 3)
Fig. 8
Fig. 8
A Cell viability of MG-63 cell line after contact with PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% scaffold’s extraction. The extraction time was performed for 7 and 14 days. B SEM images of cell adhesion on PCL, PCL/HA5%, PCL/HA10%, PCL/HA15% and PCL/HA20% after 3 days. C, D SEM image of MG-63 cell adhesion PCL/HA20% scaffolds after 3 ad 14 days. E Alkaline Phosphatase (ALP) activity of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% after 7 and 14 days. F Microscopic images of calcium deposition on PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% scaffolds after 10 days. The red color shows the amount of deposited calcium. (The error bar shows the standard deviation, n = 3)
Fig. 9
Fig. 9
SEM images of PCL, PCL/HA5%, PCL/HA10%, PCL/HA15%, and PCL/HA20% scaffolds after 21 days of soaking in SBF show the bioactivity of scaffolds after this duration. (The scale bar for all the images is 1 µm)
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