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Osteogenic Potential of Human Umbilical Cord Mesenchymal Stem Cells on Coralline Hydroxyapatite/Calci 时间:2018-09-06 20:08 来源:未知作者:admin 点击: 次 Abstract:Coralline hydroxyapatite/calcium carbonate (CHACC) is a biodegradable and osteoconductive bone graft material with promising clinical performance. CHACC has been shown to support proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (MSCs) in vitro and demonstrated to work as a functional scaffold for bone formation in vivo. Umbilical cord matrix is a more accessible and abundant tissue source of MSCs, but its osteogenic capacity in comparison to human bone marrow when cultured on CHACC has not yet been demonstrated. In this study, we assessed the osteogenic differentiation capacity of human MSCs, isolated from bone marrow and umbilical cord matrix and characterised by flow cytometry, when cultured on 200–300 μm CHACC granules. The 3D cultures were characterised by brightfield and scanning electron microscopy (SEM). Osteogenic potential was assessed by immunocytochemistry and qPCR for key markers of bone differentiation (alkaline phosphatase, runx2, type I collagen, and osteocalcin). By day 1, the MSCs had enveloped the surface of the CHACC granules to form organoids, and by day 7, cells had proliferated to bridge nearby organoids. Extracellular matrix deposition and osteogenic differentiation were demonstrated by MSCs from both tissue sources at day 21. However, MSCs from bone marrow demonstrated superior osteogenic differentiation capability compared to those from umbilical cord matrix. In conclusion, it is possible to culture and induce osteogenic differentiation of umbilical cord matrix MSCs on CHACC. Further research is required to optimise the osteogenicity of umbilical cord matrix MSCs to release their full potential as a readily available, accessible, and abundant tissue source for bone tissue engineering.
1. Introduction
Of the diverse range of scaffolds available for use in maxillofacial surgery and dentistry, autografts have been reported to be the “gold standard” with respect to bone grafting procedures [1]. However, harvesting of autografts, usually from the iliac crest, requires surgical intervention, which is associated with additional risks of blood loss, infection, and morbidity, and supply is limited [2, 3]. Other types of grafts include allografts and xenografts, but these can cause an immunological reaction and be rejected by the recipient [3]; so, it is vital to identify suitable alternative materials.
Synthetic biomaterials, such as hydroxyapatite, tricalcium phosphate ceramics and cements, and bioglass, are alternative sources for bone graft substitutes. However, these synthetic biomaterials do not mimic the architecture, porosity, and organic components of the natural bone and are not optimal in regard to biodegradation and host tissue integration or practical to implant or inject. Naturally occurring coral exoskeleton has a porous architecture that is similar to the human trabecular bone [4]. Since its main composition is calcium carbonate, a hydrothermal technique was developed to completely convert the calcium carbonate to be coralline hydroxyapatite (CHA) ceramics for clinical application [5–8].
We have previously reported a coralline hydroxyapatite/calcium carbonate (CHACC) material which shows promising clinical performance when implanted in sizes ranging from 10–100 × 10 × 10 mm3 [9, 10]. This material not only has properties such as porosity, surface structure, and osteoconductivity of coralline hydroxyapatite (CHA) as previously investigated [5–7] but also improves host tissue integration and can be completely biodegraded during bone remodelling [9]. Herein, we are focusing on smaller-sized CHACC, 200–300 μm particles, with the potential to be injected facilitating administration for maxillofacial and dentistry applications.
To increase the functionality of bone biomaterials by hopefully contributing towards remodelling and host integration, stem cells are commonly added [11]. In vitro cellular 3D structures [12] created from stem cells resembling living tissue are known as organoids and have been developed as models for translational medicine [13] and gene therapy [14].