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a cisplatin-resistant model using OS organoids, finding predict arrhythmia risks before proceeding to in vivo
that cisplatin treatment significantly upregulates excision electrophysiology studies. Conversely, findings from animal
repair cross-complementation 6 (ERCC6) expression, models can guide the optimization of organoid systems,
and that this expression level is correlated with patients’ such as incorporating mechanical cues from load-bearing
clinical pathological characteristics. Subsequently, the team joints into cartilage organoids or immune cells into tumor
used CRISPR-Cas9 to knock down ERCC6, significantly organoids, to better mimic in vivo microenvironments.
restored the organoid’s sensitivity to cisplatin and promoted This iterative synergy enhances research efficiency and
apoptosis, thereby rapidly validating the gene’s potential as a predictive validity.
therapeutic target. CRISPR-Cas9 can also be applied to the Despite their advantages, organoids face challenges in
treatment of genetic diseases. Saito et al. extracted cells scalability, standardization, and functional maturation that
293
from patients with craniosynostosis (cleidocranial dysplasia limit their standalone utility. Vascularization, innervation,
[CCD]) and induced them into iPSCs. CCD is a dominant and longevity remain significant hurdles, particularly
genetic skeletal disorder, and researchers corrected the for modeling chronic diseases or systemic effects. Thus,
mutation in CCD-derived iPSCs using CRISPR/Cas9 while organoids are transforming preclinical research
technology. The edited iPSCs demonstrated the ability to by offering human-reducible reductionist models, they
stimulate bone regeneration in rat cranial defects, thereby are not yet positioned to replace animal studies entirely.
demonstrating the potential therapeutic applications of Instead, their greatest value lies in their integration with
CRISPR/Cas9-reprogrammed iPSCs. Similar strategies can in vivo systems, refining hypotheses, reducing animal use
also be extended to the construction of MSK organoids. through the 3Rs, and accelerating translational research.
For example, CRISPR-Cas9 technology can be used to Future advancements in multi-organoid systems, OoC
overexpress RUNX2 to enhance stem cell osteogenic technologies, and computational integration with animal
differentiation and promote bone matrix mineralization. data will further solidify this complementary partnership,
Alternatively, DNA methylation modifications can be offering a more holistic and human-relevant approach to
employed for epigenetic regulation to precisely coordinate biomedical discovery.
osteogenesis. Although the application of CRISPR-Cas9
in skeletal muscle organoids is now in an early stage of 8. Conclusion
development with limited research, its immense potential Over the past decade, tissue engineering has experienced
in bone regeneration, disease modeling, and clinical a remarkable surge in the development of MSK organoids.
translation is undeniable, and it will undoubtedly play a These organoids have emerged as a powerful alternative
pivotal role in the future. to traditional 2D cultures and animal models, offering

7. Future perspectives a more physiologically relevant representation of native
tissue architecture, cellular composition, and function.
Organoids have emerged as powerful in vitro models By combining the accessibility of in vitro systems with the
that complement, rather than replace, animal studies complexity of in vivo-like environments, MSK organoids
by addressing key limitations while preserving the have become invaluable tools in disease modeling, drug
unique advantages of whole-organism systems. Unlike screening, and the exploration of regenerative repair
traditional cell cultures, organoids recapitulate human strategies. Their adoption has not only accelerated the
tissue architecture and function, making them particularly advances in regenerative medicine but also deepened our
valuable for studying species-specific disease mechanisms understanding of the pathogenesis of MSK disorders.
and drug responses. However, organoids currently lack Despite these advancements, the intricate nature of the
the systemic complexity of animal models, including MSK system means that organoid technologies remain in an
functional immune, vascular, and neural networks, as early stage of development. Looking ahead, the continued
well as organism-level behaviors and pharmacokinetics. evolution of biomaterials and biotechnological innovations
This makes them unsuitable for studying multi-organ is expected to substantially broaden the applications of
interactions, systemic drug effects, or complex phenotypes MSK organoids in tissue engineering.
such as pain, cognition, or motor function, areas where
rodent and other animal models remain indispensable. Acknowledgments
Organoids excel at high-throughput early-stage None.
screening of drug candidates or genetic therapies,
significantly reducing the number of animals needed for Funding
subsequent validation. For example, liver organoids can This research was supported by National Natural
first identify hepatotoxic compounds in vitro, minimizing Science Foundation of China under the project titled
unnecessary animal testing, while cardiac organoids can “Intervertebral disc regeneration based on precise tissue

Volume 1 Issue 3 (2025) 25 doi: 10.36922/OR025280024