Osteogenesis of 3D-Printed PCL/TCP/bdECM Scaffold Using Adipose-Derived Stem Cells Aggregates; An Experimental Study in the Canine Mandible.

Joon Seok Lee,Jung Bo Huh,Won-Soo Yun,Sung Hwan Moon,Tae Hyun Park,Hyun Mi Kim,Dong Kyu Kim,Jeong Yeop Ryu,Jin-Hyung Shim,Seong Soo Kang,Eun Jung Oh,Ho Yun Chung

doi:10.3390/ijms22115409

Abstract

Three-dimensional (3D) printing is perceived as an innovative tool for change in tissue engineering and regenerative medicine based on research outcomes on the development of artificial organs and tissues. With advances in such technology, research is underway into 3D-printed artificial scaffolds for tissue recovery and regeneration. In this study, we fabricated artificial scaffolds by coating bone demineralized and decellularized extracellular matrix (bdECM) onto existing 3D-printed polycaprolactone/tricalcium phosphate (PCL/TCP) to enhance osteoconductivity and osteoinductivity. After injecting adipose-derived stem cells (ADSCs) in an aggregate form found to be effective in previous studies, we examined the effects of the scaffold on ossification during mandibular reconstruction in beagle dogs. Ten beagles were divided into two groups: group A (PCL/TCP/bdECM + ADSC injection; n = 5) and group B (PCL/TCP/bdECM; n = 5). The results were analyzed four and eight weeks after intervention. Computed tomography (CT) findings showed that group A had more diffuse osteoblast tissue than group B. Evidence of infection or immune rejection was not detected following histological examination. Goldner trichrome (G/T) staining revealed rich ossification in scaffold pores. ColI, Osteocalcin, and Runx2 gene expressions were determined using real-time polymerase chain reaction. Group A showed greater expression of these genes. Through Western blotting, group A showed a greater expression of genes that encode ColI, Osteocalcin, and Runx2 proteins. In conclusion, intervention group A, in which the beagles received the additional ADSC injection together with the 3D-printed PCL/TCP coated with bdECM, showed improved mandibular ossification in and around the pores of the scaffold.

Highlights

The tissue engineering area has seen active research on manufacturing artificial scaffolds using various materials
These findings suggested that 3D-printed PCL/β-tricalcium phosphate (TCP)/bone-demineralized and decellularized extracellular matrix (bdECM) biomaterial and adipose-derived stem cells (ADSCs) aggregates were more effective in reconstructing the mandibular defect of beagle dogs
After exfoliating and exposing the mandible, the mandibular defect was induced according to the size of the model previously produced using the 3D printing technology

Summary

Introduction

The tissue engineering area has seen active research on manufacturing artificial scaffolds using various materials It aims to restore, maintain, and improve the damaged function of the living body. Cells injected into and attached to the pores manufactured according to the characteristics of the implantation site of the scaffold must be able to differentiate and proliferate These internal connective structures through the pores show excellent biocompatibility by enhancing the penetration, differentiation, and proliferation of cells [5,6,7,8]. Manufacturing of scaffold for tissue engineering through the 3D printing technology allows patient-customized scaffolds based on patients’ computed tomography (CT)/magnetic resonance imaging (MRI) data These scaffolds have excellent internal connectivity because of the 3D shape. Their mechanical properties, pore size, and porosity can be controlled [9,10,11,12,13,14]

Methods

Results

Conclusion