Barthold, J. E. | bioRxiv Science

bioRxiv2022

Acellular Cartilage-Bone Allografts Restore Structure, Mechanics, and Surface Properties In Vivo, but Limit Recellularization and Integrative Repair

The repair of articular cartilage after damage is challenging, and clinical interventions to promote regeneration remain elusive. The most effective treatment for cartilage defects utilizes viable osteochondral allografts from young donors, but unfortunately suffers from severe source limitations and short storage time. Decellularized tissue offers the potential to utilize native tissue structure and composition while also overcoming source limitations, but the long-term efficacy of acellular allografts is unknown. Here, we show that acellular osteochondral allografts improve functional and integrative cartilage repair in defect regions after 6 months in a preclinical (sheep) animal model. Functional measures of intratissue strain and structure assessed by MRI demonstrate similar biomechanical performance between implants and native cartilage. Compared to native tissue, the structure, composition, and tribology of acellular allografts conserve surface roughness and lubrication, native cartilage material properties under compression and relaxation, and compositional ratios of collagen:glycosaminoglycan and collagen:phosphate. However, while high cellularity was observed in the integration zones between native cartilage and acellular allografts, recellularization throughout the chondral implant was largely lacking, potentially limiting long-term cellular maintenance in the graft and repair success. Our results advance a suite of joint-to-cellular functional assays, demonstrate the biomechanical efficacy of acellular allografts for at least six months in vivo, and suggest that long-term implant success may suffer from a lack of cell migration into the dense decellularized chondral tissue.

bioengineering↗

bioRxiv2021

Dedifferentiation alters chondrocyte nuclear mechanics during in vitro culture and expansion

Dedifferentiation of chondrocytes during in vitro passaging before implantation, and post implantation in vivo, is a critical limitation in cartilage tissue engineering. Several biophysical features define the dedifferentiated state including a flattened cell morphology and increased stress fiber formation. However, how dedifferentiation influences nuclear mechanics, and the possible long-term implications of this state, are unknown. In this study, we investigated how chondrocyte dedifferentiation affects the mechanics of the chromatin architecture inside the cell nucleus and the gene expression of the structural proteins located at the nuclear envelope. Through an experimental model of cell stretching and a detailed spatial intranuclear strain quantification, we identified that strain is amplified and distribution of strain within the chromatin is altered under tensile loading in the dedifferentiated state. Further, using a confocal microscopy image-based finite element model and simulation of cell stretching, we found that the cell shape is the primary determinant of the strain amplification inside the chondrocyte nucleus in the dedifferentiated state. Additionally, we found that nuclear envelope proteins have lower gene expression in the dedifferentiated state suggesting a weaker nuclear envelope which can further intensify the intranuclear strain amplification. Our results indicate that dedifferentiation and altered nuclear strain could promote gene expression changes at the nuclear envelope, thus promoting further deviation from chondrocyte phenotype. This study highlights the role of cell shape on nuclear mechanics and lays the groundwork to design biophysical strategies for the maintenance and enhancement of the chondrocyte phenotype during expansion with a goal of successful cartilage tissue engineering. SIGNIFICANCEChondrocytes dedifferentiate into a fibroblast-like phenotype in a non-native biophysical environment. Using high resolution microscopy, intranuclear strain analysis, finite element method based computational modeling, and molecular biology techniques, we investigated how mechanical force causes abnormal intranuclear strain distribution in chondrocytes during the dedifferentiation process. Overall, our results suggest that the altered cell geometry aided by an altered or weakened nuclear envelope structure are responsible for abnormal intranuclear strain during chondrocyte dedifferentiation that can further deviate chondrocytes to a more dedifferentiated state.

biophysics↗