Mohd Isa Isma Liza, Zulkiflee Izzat, Ogaili Raed H, Mohd Yusoff Nurul Huda, Sahruddin Natasya Nadia, Sapri Shaiful Ridzwan, Mohd Ramli Elvy Suhana, Fauzi Mh Busra, Mokhtar Sabarul Afian
Department of Anatomy, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
CÚRAM, SFI Research Centre for Medical Devices, University of Galway, Galway, Ireland.
Front Bioeng Biotechnol. 2023 Dec 12;11:1296531. doi: 10.3389/fbioe.2023.1296531. eCollection 2023.
A regenerative strategy employing extracellular matrix (ECM)-based biomaterials and stem cells provide a better approach to mimicking the three-dimensional (3D) microenvironment of intervertebral disc for endogenous tissue regeneration. However, there is currently limited understanding regarding the human Wharton Jelly derived-mesenchymal stem cells (hWJ-MSCs) towards nucleus pulposus (NP)-like cells. Our study focused on the development of 3D bioengineered hydrogel based on the predominant ECM of native NP, including type II collagen (COLII) and hyaluronic acid (HA), which aims to tailor the needs of the microenvironment in NP. We have fabricated a 3D hydrogel using from COLII enriched with HA by varying the biomacromolecule concentration and characterised it for degradation, stability and swelling properties. The WJ-MSC was then encapsulated in the hydrogel system to guide the cell differentiation into NP-like cells. We successfully fabricated COLII hydrogel (2 mg/ml) and HA 10 mg/ml at a weight ratio of HA and COLII at 1:9 and 4.5:9, and both hydrogels physically maintained their 3D sphere-shaped structure after complete gelation. The higher composition of HA in the hydrogel system indicated a higher water intake capacity in the hydrogel with a higher amount of HA. All hydrogels showed over 60% hydrolytic stability over a month. The hydrogel showed an increase in degradation on day 14. The hWJ-MSCs encapsulated in hydrogel showed a round morphology shape that was homogenously distributed within the hydrogel of both groups. The viability study indicated a higher cell growth of hWJ-MSCs encapsulated in all hydrogel groups until day 14. Overall, our findings demonstrate that HA/COLII hydrogel provides an optimal swelling capacity, stability, degradability, and non-cytotoxic, thus mimics the NP microenvironment in guiding hWJ-MSCs towards NP phenotype, which is potentially used as an advanced cell delivery system for intervertebral disc regeneration.
一种采用基于细胞外基质（ECM）的生物材料和干细胞的再生策略，为模拟椎间盘的三维（3D）微环境以实现内源性组织再生提供了一种更好的方法。然而，目前对于人脐带华通氏胶间充质干细胞（hWJ-MSCs）向髓核（NP）样细胞的了解有限。我们的研究聚焦于基于天然NP的主要ECM（包括II型胶原蛋白（COLII）和透明质酸（HA））开发3D生物工程水凝胶，旨在满足NP微环境的需求。我们通过改变生物大分子浓度，用富含HA的COLII制备了一种3D水凝胶，并对其降解、稳定性和溶胀特性进行了表征。然后将WJ-MSC封装在水凝胶系统中，以引导细胞分化为NP样细胞。我们成功制备了重量比为HA与COLII分别为1:9和4.5:9、浓度为2mg/ml的COLII水凝胶和10mg/ml的HA水凝胶，并且两种水凝胶在完全凝胶化后均物理性地保持其3D球形结构。水凝胶系统中HA的含量越高，表明水凝胶的吸水能力越强，HA的含量也越高。所有水凝胶在一个月内均表现出超过60%的水解稳定性。水凝胶在第14天显示出降解增加。封装在水凝胶中的hWJ-MSCs呈现圆形形态，均匀分布在两组水凝胶中。活力研究表明，直到第14天，封装在所有水凝胶组中的hWJ-MSCs细胞生长更高。总体而言，我们的研究结果表明，HA/COLII水凝胶具有最佳的溶胀能力、稳定性、可降解性且无细胞毒性，因此在引导hWJ-MSCs向NP表型分化方面模拟了NP微环境，这有可能用作椎间盘再生的先进细胞递送系统。
