Physico-Chemical, In Vitro, and In Vivo Evaluation of a 3D Unidirectional Porous Hydroxyapatite Scaffold for Bone Regeneration

Abstract

Abstract:The unidirectional porous hydroxyapatite HAp (UDPHAp) is a scaffold with continuous communicated pore structure in the axial direction. We evaluated and compared the ability of the UDPHAp as a three-dimensional (3D) bone tissue engineering scaffold to the interconnected calcium porous HAp ceramic (IP-CHA). To achieve this, we evaluated in vitro the compressive strength, controlled rhBMP-2 release behavior, adherent cell morphology, cell adhesion manner, and cell attachment of UDPHAp. As a further in vivo experiment, UDPHAp and IP-CHA with rhBMP-2 were transplanted into mouse calvarial defects to evaluate their bone-forming ability. The Results demonstrated that the maximum compressive strengths of the UDPHAp was 7.89 ± 1.23 MPa and higher than that of IP-CHA (1.92 ± 0.53 MPa) (p = 0.0039). However, the breaking energies were similar (8.99 ± 2.72 vs. 13.95 ± 5.69 mJ, p = 0.055). The UDPHAp released rhBMP-2 more gradually in vivo. Cells on the UDPHAp adhered tightly to the surface, which had grown deeply into the scaffolds. A significant increase in cell number on the UDPHAp was observed compared to the IP-CHA on day 8 (102,479 ± 34,391 vs. 32,372 ± 29,061 estimated cells per scaffold, p = 0.0495). In a mouse calvarial defect model, the percentages of new bone area (mature bone + trabecular bone) in the 2x field were 2.514% ± 1.224% for the IP-CHA group and 7.045% ± 2.055% for the UDPHAp group, and the percentage was significantly higher in the UDPHAp group (p = 0.0209). While maintaining the same strength as the IP-CHA, the UDPHAp with 84% porosity showed a high cell number, high cell invasiveness, and excellent bone formation. We believe the UDPHAp is an excellent material that can be applied to bone regenerative medicine.

Craniofacial Bone Regeneration using iPS Cell-Derived Neural Crest Like Cells

“…First, we evaluated the engraftment and diff erentiation capacity of iPS-NCLC-MSCs on HA, the major mineral component of bone, and demonstrated that they exhibited the typical morphology of MSCs and could differentiate into osteoblasts by osteogenic induction. These results coincided with a previous study showing that MC3T3-E1 cells grew well on the UDPHAp surface 20) and suggested that iPS-NCLC-MSCs could maintain the differentiation capacity into osteoblasts on HA when transplanted. Second, we transplanted iPS-NCLC-MSCs with β-TCP in a calvarial defect and showed that the cells contributed to bone regeneration with no formation of tumor in vivo.…”

“…After checking the expression of NC marker as previously described 12) , iPS-NCLCs were cultured and maintained in DMEM containing 10% FBS and 50 U/ml and 50 µg/ml PenicillinStreptomycin on CorningTM PrimariaTM tissue culture dishes (Thermo Fisher Scientifi c) to induce MSCs. For iPS-NCLC-MSCs cultured on hydroxyapatite (HA), unidirectional porous HA scaffolds [18][19][20] (Kurarife, Kuraray Medical Inc., Tokyo, Japan) were coated with fi bronectin overnight at 4°C and washed with PBS 3 times. Dissociated iPS-NCLC-MSCs were seeded on the HA scaff olds and cultured in DMEM supplemented with 10% FBS for 2 days before osteoinduction.…”

Applications of Carbon Nanotubes in Bone Regenerative Medicine

“…Recent studies have reported excellent results in new scaffolds with unidirectional interconnection. It has a porosity of 75-84% and a porous network with a diameter of 100-350 µm [46,50].…”

Fabrication of Porous Al2O3 Ceramics with Submicron-Sized Pores Using a Water-Based Gelcasting Method

“…Porous materials (including ceramic, metallic and polymeric types) are being developed and employed in different fields [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 ]. Among these types of porous materials, many researchers are investigating porous ceramics [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 ].…”