Quek Jolene, Vizetto-Duarte Catarina, Ng Kee Woei, Teoh Swee Hin, Choo Yen
Developmental Biology and Regenerative Medicine Programme, Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 308232, Singapore.
School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798, Singapore.
J Tissue Eng Regen Med. 2025 Jul 3;2025:7287217. doi: 10.1155/term/7287217. eCollection 2025.
Despite ongoing efforts, the regeneration of critical-sized bone defects remains a significant challenge for clinicians due to the absence of a standard clinically compliant bone tissue engineering protocol. These challenges are mostly attributed to the inadequacies of current methods, characterized by their high variability and the reliance on animal-derived components, such as fetal bovine serum (FBS) in cell culture. To address these shortcomings, our approach diverges from conventional practices by prioritizing consistency and reproducibility, and the complete elimination of animal derivatives throughout the entire process. We have developed a novel method that utilizes a peptide-functionalized photocrosslinkable methacrylated hyaluronic acid (MeHA-RGD) hydrogel as a cell sealant for loading human adipose-derived stem cells (hASCs) into a 3D porous polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold. Additionally, we created a 3D-printed vacuum-assisted cell loading device to facilitate this process and ensure efficiency and consistency during cell loading. Our findings indicate that the MeHA-RGD hydrogel supports both stem cell viability and osteogenic differentiation, demonstrating outcomes comparable to those achieved with fibrin glue, a conventional cell sealant widely used in BTE from autologous or xenogeneic sources, even under serum- and xeno-free conditions. In the pursuit of clinical translation, it is vital that biomaterials exhibit low variability, are easily accessible, readily available, and completely free of animal derivatives. To our knowledge, this is the first study to employ a 3D-printed vacuum-assisted cell loading device system within a fully defined hybrid 3D system under complete serum- and xeno-free conditions. These findings unravel and encourage alternative approaches in addressing the existing challenges in BTE, thereby facilitating and accelerating clinical translation in the future.
尽管一直在努力，但由于缺乏标准的临床适用骨组织工程方案，临界尺寸骨缺损的再生对临床医生来说仍然是一项重大挑战。这些挑战主要归因于当前方法的不足，其特点是高度可变且依赖动物源性成分，如细胞培养中的胎牛血清（FBS）。为了解决这些缺点，我们的方法与传统做法不同，优先考虑一致性和可重复性，并在整个过程中完全消除动物衍生物。我们开发了一种新方法，利用肽功能化的可光交联甲基丙烯酸化透明质酸（MeHA-RGD）水凝胶作为细胞密封剂，将人脂肪来源干细胞（hASC）加载到三维多孔聚己内酯-磷酸三钙（PCL-TCP）支架中。此外，我们创建了一个3D打印的真空辅助细胞加载装置，以促进这一过程，并确保细胞加载过程中的效率和一致性。我们的研究结果表明，MeHA-RGD水凝胶支持干细胞活力和成骨分化，即使在无血清和无动物源的条件下，其结果与纤维蛋白胶相当，纤维蛋白胶是一种广泛用于自体或异种来源的骨组织工程的传统细胞密封剂。在追求临床转化的过程中，生物材料具有低变异性、易于获取、随时可用且完全不含动物衍生物至关重要。据我们所知，这是第一项在完全无血清和无动物源的条件下，在完全定义的混合三维系统中使用3D打印真空辅助细胞加载装置系统的研究。这些发现揭示并鼓励采用替代方法来应对骨组织工程中现有的挑战，从而在未来促进和加速临床转化。
