Department of Chemical Engineering, Sharif University of Technology, Tehran, Iran.
Department of Chemical Engineering, Sharif University of Technology, Tehran, Iran; Environmental Applied Science and Management, Ryerson University, Toronto, Canada.
Acta Biomater. 2019 Jan 1;83:233-244. doi: 10.1016/j.actbio.2018.10.031. Epub 2018 Oct 24.
Developing scaffolds that can provide cells and biological cues simultaneously in the defect site is of interest in tissue engineering field. In this study, platelet lysate (PL) as an autologous and inexpensive source of growth factors was incorporated into a cell-laden injectable hyaluronic acid-tyramine (HA-TA) hydrogel. Subsequently, the effect of platelet lysate on cell attachment, viability and differentiation of human mesenchymal stem cell (hMSCs) toward chondrocytes was investigated. HA-TA conjugates having a degree of substitution of 20 TA moieties per 100 disaccharide units were prepared and crosslinked in the presence of horseradish peroxidase and low concentrations of hydrogen peroxide. The storage moduli of the gels ranged from 500 to 2000 Pa and increased with increasing polymer concentration. In contrast to a retained round shape of the cells when using pure HA-TA hydrogel, the hMSCs attached and spread out in PL enriched matrix. The enrichment of hMSCs laden HA-TA hydrogels with PL induced a cartilage like extra cellular matrix deposition in vitro. The hMSCs increasingly deposited collagen type II and proteoglycans over time. The deposition of the new extracellular matrix (ECM) is simultaneous with gel degradation and resulted ultimately in the formation of a tough dense matrix. These findings demonstrate the potential of injectable HA-TA-PL hydrogel as a cell delivery system for cartilage regeneration. STATEMENT OF SIGNIFICANCE: Cartilage tissue has limited ability to self-repair because of its avascular nature. To have an efficient cartilage tissue regeneration, we combined platelet lysate (PL), as an autologous and inexpensive source of growth factors, with an injectable hyaluronic acid tyramine (HA-TA) hydrogel scaffold. Platelet lysate had a vital role in supporting human mesenchymal stem cells (hMSCs) activities, like cell attachment, viability and proliferation in the 3D hydrogel structure. Also, the hMSCs encapsulated HA-TA induced hyaline cartilage generation when placed in chondrogenic differentiation medium. This study introduces a new system for cartilage tissue engineering, which can be injected in a minimally invasive manner and is rich with patient's own growth factors and biological cues.
在组织工程领域,开发能够在缺陷部位同时提供细胞和生物线索的支架是很有意义的。在这项研究中,血小板裂解液 (PL) 作为一种自体且廉价的生长因子来源,被掺入到负载细胞的可注射透明质酸-酪胺 (HA-TA) 水凝胶中。随后,研究了血小板裂解液对人骨髓间充质干细胞 (hMSCs) 向软骨细胞附着、活力和分化的影响。制备了取代度为每 100 个二糖单位 20 TA 部分的 HA-TA 缀合物,并在辣根过氧化物酶和低浓度过氧化氢存在下交联。凝胶的储能模量范围为 500 至 2000 Pa,并随聚合物浓度的增加而增加。与使用纯 HA-TA 水凝胶时细胞保持圆形形状相反,hMSCs 在富含 PL 的基质中附着并展开。PL 丰富的负载 hMSCs 的 HA-TA 水凝胶在体外诱导形成类似于软骨的细胞外基质沉积。随着时间的推移,hMSCs 逐渐沉积胶原蛋白 II 和糖胺聚糖。新细胞外基质 (ECM) 的沉积与凝胶降解同时发生,最终形成坚硬致密的基质。这些发现表明,可注射的 HA-TA-PL 水凝胶作为软骨再生的细胞递送系统具有潜力。 意义声明:由于软骨组织的血管生成特性,其自我修复能力有限。为了实现有效的软骨组织再生,我们将血小板裂解液 (PL) 作为一种自体且廉价的生长因子来源,与可注射的透明质酸-tyramine (HA-TA) 水凝胶支架结合使用。血小板裂解液在支持人类间充质干细胞 (hMSCs) 的活性方面发挥了重要作用,例如在 3D 水凝胶结构中的细胞附着、活力和增殖。此外,当置于软骨分化培养基中时,负载 HA-TA 的 hMSCs 诱导透明软骨生成。这项研究介绍了一种新的软骨组织工程系统,该系统可以微创方式注射,并富含患者自身的生长因子和生物线索。