Evaluation of an Engineered Hybrid Matrix for Bone Regeneration via Endochondral Ossification.

Despite its regenerative ability, long and segmental bone defect repair remains a significant orthopedic challenge. Conventional tissue engineering efforts induce bone formation through intramembranous ossification (IO) which limits vascular formation and leads to poor bone regeneration. To overcome this challenge, a novel hybrid matrix comprised of a load-bearing polymer template and a gel phase is designed and assessed for bone regeneration. Our previous studies developed a synthetic ECM, hyaluronan (HA)-fibrin (FB), that is able to mimic cartilage-mediated bone formation in vitro. In this study, the well-characterized HA-FB hydrogel is combined with a biodegradable polymer template to form a hybrid matrix. In vitro evaluation of the matrix showed cartilage template formation, cell recruitment and recruited cell osteogenesis, essential stages in endochondral ossification. A transgenic reporter-mouse critical-defect model was used to evaluate the bone healing potential of the hybrid matrix in vivo. The results demonstrated host cell recruitment into the hybrid matrix that led to new bone formation and subsequent remodeling of the mineralization. Overall, the study developed and evaluated a novel load-bearing graft system for bone regeneration via endochondral ossification.

Hybrid extracellular matrix design for cartilage-mediated bone regeneration.

Recapitulating long bone repair through endochondral ossification (EO) is increasingly becoming a more popular approach. A successful EO Process depends greatly on the establishment of a healthy hypertrophic-cartilage template (HCT). The aim of this work is to design a hydrogel system, which closely mimics the extracellular matrix of HCT. We examined the combinatorial effect of two commonly used hydrogels for bone and cartilage regeneration strategies, hyaluronan (HA) and fibrin (FB), to induce HCT formation. Hydrogel combinations were evaluated using a clinically relevant cell source, human bone marrow mesenchymal stem cells (hBMSCs). The results establish that with increasing HA (50-90%) the chondrogenic and its subsequent hypertrophy trend improved, with 70:30 HA:FB combination showing the highest and most uniform expression of chondrogenic and hypertrophic stage specific markers. This combination also showed superior support for cell micro-aggregation and differentiation. Thus, 70:30 HA-FB matrix demonstrated a healthy formation of chondrogenic and hypertrophic stages with rich stage-specific ECM components. This study demonstrates that with the appropriate hydrogel design it is possible to develop effective tissue engineering therapies for bone defect repair and regeneration through endochondral ossification by establishing a healthy HCT. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 300-309, 2018.

Osteogenic Treatment Initiating a Tissue-Engineered Cartilage Template Hypertrophic Transition.

Hypertrophic chondrocytes play a critical role in endochondral bone formation as well as the progress of osteoarthritis (OA). An in vitro cartilage hypertrophy model can be used as a platform to study complex molecular mechanisms involved in these processes and screen new drugs for OA. To develop an in vitro cartilage hypertrophy model, we treated a tissue-engineered cartilage template, living hyaline cartilaginous graft (LhCG), with osteogenic medium for hypertrophic induction. In addition, endothelial progenitor cells (EPCs) were seeded onto LhCG constructs to mimic vascular invasion. The results showed that osteogenic treatment significantly inhibited the synthesis of endostatin in LhCG constructs and enhanced expression of hypertrophic marker-collagen type X (Col X) and osteogenic markers, as well as calcium deposition in vitro. Upon subcutaneous implantation, osteogenic medium-treated LhCG constructs all stained positive for Col X and showed significant calcium deposition and blood vessel invasion. Col X staining and calcium deposition were most obvious in osteogenic medium-treated only group, while there was no difference between EPC-seeded and non-seeded group. These results demonstrated that osteogenic treatment was of the primary factor to induce hypertrophic transition of LhCG constructs and this model may contribute to the establishment of an in vitro cartilage hypertrophy model.