Zamorano Mauricio, Aguilar-Gallardo Cristobal, Lugo Aloyma, Jimenez Luis, Farias Jorge G, Mantalaris Athanasios
Chemical Engineering Department, Universidad de La Frontera, Temuco 4811230, Chile.
Biological Systems Engineering Laboratory, Department of Chemical Engineering, Imperial College London, London SW7 2AZ, UK.
Int J Mol Sci. 2025 May 3;26(9):4348. doi: 10.3390/ijms26094348.
Bone tissue engineering (BTE) emerged as a practical approach to tackle prosthetic industry limitations. We merge aspects from developmental biology, engineering and medicine with the aim to produce fully functional bone tissue. Mesenchymal stem cells have the capability of self-renewal and specific lineage differentiation. Herein lies their potential for BTE. Among MSCs, human dental pulp stem cells have a higher proliferation rate, shorter doubling times, lower cellular senescence, and enhanced osteogenesis than hBM-SCs under specific conditions. In addition, these cells are readily accessible and can be extracted through a subtle extraction procedure. Thus, they garner fewer moral concerns than most MSCs available and embody a promising cell source for BTE therapies able to replace hBM-MSCs. Interestingly, their study has been limited. Conversely, there is a need for their further study to harness their true value in BTE, with special emphasis in the design of bioprocesses able to produce viable, homogenous bone constructs in a clinical scale. Here, we study the osteogenic differentiation of hDPSCs encapsulated in alginate hydrogels under suspended culture in a novel perfusion bioreactor. The system is compared with traditional 3D static and fed-batch culture methodologies. The novel system performed better, producing higher alkaline phosphatase activity, and more homogeneous, dense and functional bone constructs. Additionally, cell constructs produced by the in-house-designed system were richer in mature osteoblast-like and mineralizing osteocyte-like cells. In conclusion, this study reports the development of a novel bioprocess able to produce hDPSC-based bone-like constructs, providing new insights into hDPSCs' therapeutic potential and a system able to be transferred from the laboratory bench into medical facilities.
骨组织工程（BTE）作为一种解决假肢行业局限性的实用方法应运而生。我们将发育生物学、工程学和医学的各个方面结合起来，旨在制造出功能完备的骨组织。间充质干细胞具有自我更新和特定谱系分化的能力。这正是它们在骨组织工程中的潜力所在。在间充质干细胞中，人牙髓干细胞在特定条件下比人骨髓间充质干细胞具有更高的增殖率、更短的倍增时间、更低的细胞衰老率以及更强的成骨能力。此外，这些细胞易于获取，可通过精细的提取程序获得。因此，与大多数可用的间充质干细胞相比，它们引发的道德问题更少，是骨组织工程治疗中有望替代人骨髓间充质干细胞的细胞来源。有趣的是，对它们的研究一直很有限。相反，有必要对其进行进一步研究，以挖掘它们在骨组织工程中的真正价值，特别要强调能够在临床规模上生产有活力、均匀的骨构建体的生物工艺设计。在此，我们研究了在新型灌注生物反应器中悬浮培养的藻酸盐水凝胶包裹的人牙髓干细胞的成骨分化。该系统与传统的三维静态和分批补料培养方法进行了比较。新型系统表现更佳，产生了更高的碱性磷酸酶活性，以及更均匀、致密且功能更强的骨构建体。此外，内部设计系统产生的细胞构建体中成熟的成骨细胞样细胞和矿化骨细胞样细胞更为丰富。总之，本研究报告了一种能够生产基于人牙髓干细胞的骨样构建体的新型生物工艺的开发，为深入了解人牙髓干细胞的治疗潜力以及一个能够从实验室工作台转移到医疗设施中的系统提供了新的见解。
