Glutamic acid-based dendritic peptides for scaffold-free cartilage tissue engineering

Current treatment modalities for cartilage regeneration often result in the production of fibrous-type cartilage tissue at the defect site, which has inferior mechanical properties as compared to native hyaline cartilage. Further, effective treatments are not available at present, for preventing age-related as well as disease-related hypertrophic development of chondrocytes. In the present study, we designed and synthesized three sets of glutamic acid-based dendritic peptides, differing in degree of lipidation as well as branching. Each set constitutes of N-terminal protected as well as corresponding N-deprotected peptides. Altogether, six peptides [BE12, E12, BE3(12)4, E3(12)4, BE3OMe, E3OMe] were tested for their chondrogenesis enhancing potential in vitro, using rabbit adipose derived mesenchymal stem cells (ADMSCs). Immunohistochemical and gene expression studies as well as biochemical analyses revealed that the lipopeptides [E12 and BE3(12)4] are able to enhance chondrogenic differentiation of ADMSCs significantly (p < 0.001) as compared to control group (chondrogenic medium alone). Glycosaminoglycan content, and the chondrogenic marker genes like Aggrecan (Acan), Type II collagen (Col2a1), Hyaluronan synthase 2 (Has2), and SRY-box 9 (Sox9) expressions were found to be significantly increased in E12 and BE3(12)4 treated groups. Most importantly, the BE3(12)4 treated group showed significantly lower Type I collagen (Col1a2) and Type X collagen (Col10a1) transcript levels (p < 0.001), indicating its potential for hyaline cartilage formation and also to prevent hypertrophic development. Thus, the lipopeptides E12 and BE3(12)4 may be useful for preventing chondrocyte hypertrophy and realizing the hyaline nature of regenerated cartilage tissue in tissue engineering. STATEMENT OF SIGNIFICANCE: The current treatment modalities for degenerative cartilage diseases are unsatisfactory as the resultant regenerated cartilage is often fibrous in nature with inferior mechanical properties. Further, there is no proper treatment available for age-related development of chondrocyte hypertrophy at present. In this study we synthesized glutamic acid-based lipopeptides, which differ in the degree of lipidation as well as branching. We used a combinatorial approach of scaffold-free tissue engineering and dendritic lipopeptides to achieve hyaline-like cartilage tissue from adipose derived mesenchymal stem cells in vitro. Gene expression analysis revealed the down regulation of fibrous cartilage marker Col1a2 and hypertrophic marker Col10a1, suggesting that these lipopeptides may be useful for achieving mechanically superior hyaline cartilage regeneration in future.