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

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Title Combination of Human Amniotic Fluid Derived-Mesenchymal Stem Cells and Nano-hydroxyapatite Scaffold Enhances Bone Regeneration
Creator Mohammed, Eman E. A.
Beherei, Hanan
El-Zawahry, Mohamed
Farrag, Abdel Razik
Kholoussi, Naglaa
Helwa, Iman
Gaber, Khaled
Allam, Mousa A.
Mabrouk, Mostafa
Abdel Aleem, Alice K.
Subject Amniotic Fluid Stem Cells
3D Scaffolds
Nano-hydroxyapatite Chitosan
Bone Healing
Description 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 2nd-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 2nd-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 2nd-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.
Publisher Scientific Foundation SPIROSKI, Skopje, Republic of Macedonia; ID Design 2012/DOOEL Skopje, Republic of Macedonia
Date 2019-09-14
Type info:eu-repo/semantics/article
Format application/pdf
Source Open Access Macedonian Journal of Medical Sciences; Vol 7 No 17 (2019): Sep 15 (OAMJMS); 2739-2750
Language eng
Rights Copyright (c) 2019 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 (Author)

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