Combination of Human Amniotic Fluid Derived-Mesenchymal Stem Cells and Nano-hydroxyapatite Scaffold Enhances Bone Regeneration

Eman E. A.  Mohammed; Hanan  Beherei; Mohamed  El-Zawahry; Abdel Razik  Farrag; Naglaa  Kholoussi; Iman  Helwa; Khaled  Gaber; Mousa A. Allam; Mostafa  Mabrouk; Alice K. Abdel Aleem

doi:10.3889/oamjms.2019.730

Authors

Eman E. A. Mohammed Medical Molecular Genetics Department, National Research Center, Cairo, Egypt; Stem Cell Research Group, Medical Research Centre of Excellence, National Research Center, Cairo, Egypt
Hanan H. Beherei Ceramic Department (Biomaterials), National Research Center, Cairo, Egypt
Mohamed El-Zawahry Fixed and Removable Prosthodontics Department, National Research Center, Cairo, Egypt
Abdel Razik H. Farrag Pathology Departments, National Research Center, Cairo, Egypt
Naglaa Kholoussi Immunity Genetics Department, National Research Center, Cairo, Egypt
Iman Helwa Immunity Genetics Department, National Research Center, Cairo, Egypt
Khaled Gaber Prenatal and Fetal medicine Department, National Research Center, Cairo, Egypt
Mousa A. Allam Spectroscopy Department, National Research Center, Cairo, Egypt
Mostafa Mabrouk Ceramic Department (Biomaterials), National Research Center, Cairo, Egypt
Alice K. Abdel Aleem Medical Molecular Genetics Department, National Research Center, Cairo, Egypt; Stem Cell Research Group, Medical Research Centre of Excellence, National Research Center, Cairo, Egypt; Neurology and Neuroscience Department, Weill Cornell Medical College, Doha, Qatar

DOI:

https://doi.org/10.3889/oamjms.2019.730

Keywords:

Amniotic Fluid Stem Cells, 3D Scaffolds, Nano-hydroxyapatite Chitosan, Bone Healing, Regeneration

Abstract

BACKGROUND: Human amniotic fluid-derived stem cells (hAF-MSCs) have a high proliferative capacity and osteogenic differentiation potential in vitro. The combination of hAF-MSCs with three-dimensional (3D) scaffold has a promising therapeutic potential in bone tissue engineering and regenerative medicine. Selection of an appropriate scaffold material has a crucial role in a cell supporting and osteoinductivity to induce new bone formation in vivo.

AIM: This study aimed to investigate and evaluate the osteogenic potential of the 2^nd-trimester hAF-MSCs in combination with the 3D scaffold, 30% Nano-hydroxyapatite chitosan, as a therapeutic application for bone healing in the induced tibia defect in the rabbit.

SUBJECT AND METHODS: hAF-MSCs proliferation and culture expansion was done in vitro, and osteogenic differentiation characterisation was performed by Alizarin Red staining after 14 & 28 days. Expression of the surface markers of hAF-MSCs was assessed using Flow Cytometer with the following fluorescein-labelled antibodies: CD34-PE, CD73-APC, CD90-FITC, and HLA-DR-FITC. Ten rabbits were used as an animal model with an induced defect in the tibia to evaluate the therapeutic potential of osteogenic differentiation of hAF-MSCs seeded on 3D scaffold, 30% Nano-hydroxyapatite chitosan. The osteogenic differentiated hAF-MSCs/scaffold composite system applied and fitted in the defect region and non-seeded scaffold was used as control. The histopathological investigation was performed at 2, 3, & 4 weak post-transplantation and scanning electron microscope (SEM) was assessed at 2 & 4 weeks post-transplantation to evaluate the bone healing potential in the rabbit tibia defect.

RESULTS: Culture and expansion of 2^nd-trimester hAF-MSCs presented high proliferative and osteogenic potential in vitro. Histopathological examination for the transplanted hAF-MSCs seeded on the 3D scaffold, 30% Nano-hydroxyapatite chitosan, demonstrated new bone formation in the defect site at 2 & 3 weeks post-transplantation as compared to the control (non-seeded scaffold). Interestingly, the scaffold accelerated the osteogenic differentiation of AF-MSCs and showed complete bone healing of the defect site as compared to the control (non-seeded scaffold) at 4 weeks post-transplantation. Furthermore, the SEM analysis confirmed these findings.

CONCLUSION: The combination of the 2^nd-trimester hAF-MSCs and 3D scaffold, 30% Nano-hydroxyapatite chitosan, have a therapeutic perspective for large bone defect and could be used effectively in bone tissue engineering and regenerative medicine.