Budharaju Harshavardhan, Sundaramurthi Dhakshinamoorthy, Sethuraman Swaminathan
Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), ABCDE Innovation Centre, School of Chemical & Biotechnology (SCBT), SASTRA Deemed University, Thanjavur, 613401, India.
J Mater Sci Mater Med. 2025 Jan 13;36(1):8. doi: 10.1007/s10856-024-06855-2.
Preservation and long-term storage of readily available cell-laden tissue-engineered products are major challenges in expanding their applications in healthcare. In recent years, there has been increasing interest in the development of off-the-shelf tissue-engineered products using the cryobioprinting approach. Here, bioinks are incorporated with cryoprotective agents (CPAs) to allow the fabrication of cryopreservable tissue constructs. Although this method has shown potential in the fabrication of cryopreservable tissue-engineered products, the impact of the CPAs on the viscoelastic behavior and printability of the bioinks at cryo conditions remains unexplored. In this study, we have evaluated the influence of CPAs such as glycerol and dimethyl sulfoxide (DMSO) on the rheological properties of pre-crosslinked alginate bioinks for cryoprinting applications. DMSO-incorporated bioinks showed a reduction in viscosity and yield stress, while the addition of glycerol improved both the properties due to interactions with the calcium chloride used for pre-crosslinking. Further, tube inversion and printability experiments were performed to identify suitable concentrations and cryobioprinting conditions for bioinks containing CPAs & pre-crosslinked with CaCl. Finally, based on the printability analysis & cell recovery results, 10% glycerol was used for cryobioprinting and preservation of cell-laden constructs at -80 °C and the viability of cells within the printed structures were evaluated after recovery. Cell viability results indicate that the addition of 10% glycerol to the pre-crosslinked bioink significantly improved cell viability compared to bioinks without CPAs, confirming the suitability of the developed bioink combination to fabricate tissue constructs for on-demand applications.
保存和长期储存随时可用的载细胞组织工程产品是扩大其在医疗保健领域应用的主要挑战。近年来，人们对使用低温生物打印方法开发即用型组织工程产品的兴趣日益浓厚。在这里，生物墨水与冷冻保护剂（CPA）混合，以制造可冷冻保存的组织构建体。虽然这种方法在制造可冷冻保存的组织工程产品方面已显示出潜力，但CPA对低温条件下生物墨水的粘弹性行为和可打印性的影响仍未得到探索。在本研究中，我们评估了甘油和二甲基亚砜（DMSO）等CPA对用于低温打印应用的预交联藻酸盐生物墨水流变学特性的影响。掺入DMSO的生物墨水粘度和屈服应力降低，而添加甘油由于与用于预交联的氯化钙相互作用而改善了这两种特性。此外，进行了试管倒置和可打印性实验，以确定含有CPA并与CaCl预交联的生物墨水的合适浓度和低温生物打印条件。最后，基于可打印性分析和细胞回收结果，使用10%甘油在-80°C下对载细胞构建体进行低温打印和保存，并在回收后评估打印结构内细胞的活力。细胞活力结果表明，与不含CPA的生物墨水相比，向预交联生物墨水中添加10%甘油显著提高了细胞活力，证实了所开发的生物墨水组合适用于制造按需应用的组织构建体。
