Page 71 - OR-1-3

Research on intercellular interactions necessitates then replace the cartilage to form bone tissue, promoting
reliance on multicellular and multi-tissue coculture skeletal growth. Inspired by endochondral ossification,
191
systems, even transcending the limitations of single- Dong et al. combined endochondral ossification with
organoid models. For instance, Tong et al. revealed endogenous enzyme-induced mineralization to simulate
183
cellular crosstalk between muscle and bone under IH the mineralization process in natural bone development,
using an MSK OoC platform. The results demonstrate that creating bone organoids based on the endochondral
136
mitochondrial damage in muscle tissue triggers CXCL5 ossification model. In addition, Xie et al. utilized 3D
release, which suppresses expression of osteogenic markers printing technology and hydrogel microspheres loaded
(e.g., Runt-related transcription factor 2 [Runx2, Osterix) with human BMSCs to construct osteo-callus organoids.
while enhancing osteoclast activity. Furthermore, Park They constructed osteo-callus organoids that recapitulate
et al. engineered a trabecular bone organoid model developmental processes, exhibiting cellular composition
132
using DBP. This trabecular bone organoid was employed similar to that of developing endochondral ossification. In
192
to investigate intercellular crosstalk among osteocytes, another study, Kesharwani et al. utilized ESCs-derived
osteoblasts, and osteoclasts during local bone remodeling organoids to model vascular dynamics on a microfluidic
regulation. For multi-organoids construction, Yin et al. chip at the initial stage of endochondral ossification. These
121
investigated the interactions among neuronal, skeletal, and studies provide strong support for a deeper understanding
muscular cells by generating hNMSOs from hPSCs. These of the endochondral ossification process and its
hNMSOs demonstrated that: motor neurons can control mechanisms. Intramembranous ossification is a process
skeletal muscle contraction through the NMJ; skeletal where MSCs directly commit to osteoblasts and mineralize
support fosters the development and maturation of human to form bone tissue, without transitioning through a
muscle; and pathological skeletal degeneration directly cartilaginous intermediate stage. The majority of organoids
precipitates NMJ dysfunction. are constructed by mimicking the developmental process
of intramembranous ossification. For instance, Zhu et al.
193
4.2. Organoids for the developmental/physiological developed a GelMA/DNA double-network hydrogel that
process significantly enhances osteogenic mineralization of BMSCs,
Organoids have significant advantages in the study of while concurrently exhibiting anti-inflammatory properties
development and physiological processes. First, they can and pro-angiogenic functionality. As for modeling
132
highly simulate the development of organs, providing a 3D physiological process, Park et al. constructed trabecular
structure that closely resembles the in vivo environment, bone organoids to investigate molecular mechanisms and
allowing researchers to observe the dynamic processes cellular activities during localized bone remodeling.
of cell differentiation, tissue formation, and organ The development of cartilage is a highly regulated
development in vitro. Second, organoids can be used biological event involving multiple aspects, including cell
187
for personalized research using patient-derived cells, differentiation, matrix synthesis, and remodeling. During
revealing individual developmental differences and cartilage development, yes-associated protein (YAP), a key
disease-related mechanisms. In addition, organoids effector of the Hippo signaling pathway, directly influences
188
are highly manipulable, enabling the study of the effects the chondrogenic differentiation of MSCs through its
of specific genes or signaling pathways on development subcellular localization and activity. Zhu et al. utilized
194
194
through various methods, such as gene editing and drug MSCs in combination with Verteporfin to modulate the
intervention. This model not only serves as a powerful YAP signaling pathway, constructing hyaline cartilage
189
complement to animal models but also enhances research organoids on decellularized cartilage matrix scaffolds to
efficiency and precision, providing a robust tool for mimic the physiological and developmental processes
developmental studies. of cartilage. As for modeling physiological processes,
144
Bone development involves two modes: endochondral Liu et al. constructed a condylar cartilage organoid to
ossification and intramembranous ossification. Both explore the primary cilia’s functions. At present, there are
190
processes work together to drive skeletal development relatively few studies using cartilage organoids to simulate
and growth. A deep understanding of the mechanisms the development of cartilage. A deeper understanding of
underlying bone development is of great significance for this process is of great significance for investigating the
bone fracture repair, regeneration, and the occurrence pathogenesis of cartilage-related diseases and developing
and development of diseases. Endochondral ossification new therapeutic approaches.
begins with the differentiation of mesenchymal cells into Organoids also play an important role in simulating the
chondrocytes, forming a cartilage template. Subsequently, development of skeletal muscle. For example, Shahriyari
chondrocytes undergo hypertrophy and calcification, et al. used hPSCs to construct a new skeletal muscle
195
followed by the integration of blood vessels. Osteoblasts organoid model, successfully reproducing key stages of

Volume 1 Issue 3 (2025) 13 doi: 10.36922/OR025280024