Cartilage, an essential connective tissue primarily found in joints, plays a critical role in providing cushioning and facilitating smooth movement. However, human cartilage has limited inherent regenerative capabilities, making it susceptible to damage from trauma, degenerative diseases, and the natural aging process. This vulnerability often leads to conditions such as osteoarthritis, characterized by cartilage degeneration, pain, and joint stiffness.

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Artificial human cartilage holds the potential for improved joint function, enhanced cushioning, and restored mobility. In the realm of regenerative medicine, it plays a vital role in tissue regeneration, employing biomaterials and stem cells to facilitate the repair and restoration of damaged cartilage. In our research, we utilized Auto-QRS to conduct a comprehensive multicomponent quantitative analysis of artificial cartilage samples. Through three-dimensional quantitative assessments, we gained valuable insights into the behavior of various biomolecules at different growth stages, providing crucial guidance for the cultivation process and ultimately improving the survival rate of artificial cartilage. We explored the levels of eight biomolecular components—aggrecan, collagen, elastin, fibronectin, glycogen, laminin, unsaturated lipid, and saturated lipid，in three-week and nine-week human tissue-engineered cartilage (hTEC) samples. Our analysis uncovered intriguing findings regarding the maturation of hTEC towards human elastic cartilage.

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Comparing the two time points, we observed an increase in the levels of aggrecan, total collagen, elastin, glycogen, laminin, and saturated lipid in the nine-week hTEC samples, indicating a maturation trend towards human auricular cartilage. Interestingly, we detected the presence of fibronectin in the nine-week hTEC samples, a component not typically found in human auricular cartilage. Conversely, the levels of unsaturated lipid, total collagen, and elastin were comparatively lower in the nine-week hTEC samples compared to human auricular cartilage. Furthermore, the distribution of both aggrecan and total collagen appeared to be more homogeneous throughout human auricular cartilage compared to the nine-week hTEC samples.

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These findings suggest that the cartilage may not have fully matured at the nine-week time point, indicating the potential need for longer culturing periods before considering transplantation. Further investigations are warranted to gain a deeper understanding of the maturation process of hTEC and optimize culturing conditions to achieve more mature and functional cartilage constructs for transplantation purposes.

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Notably, our research revealed an unexpected observation: a near-complete depletion of glycogen in artificial cartilage samples following a 6-week implantation period. This depletion of glycogen represents a previously unreported phenomenon in conventional bio-analytical methods and holds significant implications for the success of artificial cartilage culture.

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These unexpected results lead us to speculate that the consumption rate of glycogen may play a decisive role in influencing the overall success of artificial cartilage culture. By shedding light on this critical factor, our research underscores the importance of monitoring and understanding the intricate interplay between biomolecules and their impact on the long-term viability and functionality of artificial cartilage constructs.

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Our findings open up avenues for further investigations into the role of glycogen depletion in the cultivation and survival of artificial cartilage. Leveraging advanced analytical tools such as Auto-QRS, we aim to unravel the underlying mechanisms and develop strategies to enhance the success and longevity of artificial cartilage culture. Ultimately, this research has the potential to revolutionize the field of tissue engineering, enabling the development of more robust and clinically viable artificial cartilage constructs for the benefit of patients worldwide.