C. Struijk (Rochester, US)
Mayo ClinicPresenter Of 1 Presentation
24.1.6 - Biological Enhancement of Meniscus Allograft Transplantation: Combining Meniscus and Mesenchymal Stromal Cells
Abstract
Purpose
Meniscal allograft transplantation (MAT) represents established surgical procedures with proven outcomes. Yet, storage as frozen specimens and limited cellular repopulation may impair graft viability and performance, and lead to the most common complications of shrinkage and graft tear. We hypothesize that a cell-based injection therapy combining human mesenchymal stromal cells (MSCs) and meniscus cells (MC) may enhance meniscus allograft cell repopulation, viability and healing after implantation.
Methods and Materials
Both MSCs and MCs were obtained from single donors, during a liposuction procedure and total knee arthroplasty respectively, based on IRB approved protocols. For all experiments, passage 4 MCs and MSCs were suspended with lactated ringers containing cell ratios 100% MSC, 80:20% MSC:MC or 100% MCs. MSCs membranes were labelled with PKH26 Red Fluorescent Cell Linker (Sigma-Aldrich) to investigate cell ratio and migration over time. All allografts, acquired through JRF Ortho®, were manually injected with 50ml solution, containing 0,7*10E6 cells of one of the cell ratios, each accompanied by a mock injected control sample. Injected allografts and medium were collected on multiple timepoints for histological and biochemical analysis. For each experiment, a minimum of n=3 was used per experimental group. Data are presented as a mean-standard deviation and statistical analysis was performed by multiple unpaired t tests.
Results
All groups show living cells (live/dead stain) on day 28 using confocal microscopy with a general repopulation of the graft, especially for grafts treated with cell ratios 80:20% (MSC:MC) and 100% (MC) (Fig.1 and 2). Positive immunofluorescence stain for connexin-43 was achieved for all cell ratios, indicating cell-cell communication through gap junctions. DsDNA analysis revealed a significant difference in DNA content between treated groups and controls.
Conclusion
Biological enhancement of meniscus allograft transplantation through early repopulation of the allograft is feasible with a cell-based injection therapy with monocultures of native MCs or cocultures of MSCs and meniscus cells.