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Enhancing osteochondral allograft viability: effects of storage media composition.

ABSTRACT: Osteochondral allograft transplantation is a well-accepted treatment for articular cartilage damage. However, chondrocyte viability declines during graft storage, which may compromise graft performance. We first tested the hypothesis that the composition of commonly used storage media affects the viability of articular chondrocytes over time; we then tested the hypothesis that the addition of insulin growth factor-1 or the apoptosis inhibitor ZVAD-fmk could enhance the storage properties of serum-free media. Bovine osteochondral grafts were stored at 4 degrees C in lactated Ringer's, Dulbecco's modified eagle's media (DMEM), DMEM supplemented with either insulin growth factor-1 or ZVAD-fmk, and a commercial storage media. Chondrocyte viability in lactated Ringer's declined rapidly to 20.4% at 2 weeks. Viability in DMEM declined more slowly to 54.8% at 2 weeks and 31.2% at 3 weeks. Viability in commercial storage media was 83.6% at 3 weeks and 44.8% at 4 weeks. Viability was increased in DMEM + insulin growth factor-1 (56.4%) and DMEM + ZVAD (52.4%) at 3 weeks compared with DMEM alone. These results confirm the hypotheses that media composition greatly influences chondrocyte viability during cold storage and that insulin growth factor-1 and ZVAD improve the storage properties of DMEM.

SUBMITTER: Teng MS

PROVIDER: S-EPMC2584242 | biostudies-other | 2008 Aug

REPOSITORIES: biostudies-other

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Project description:Introduction:We performed a survivorship analysis of patients treated with patellofemoral osteochondral allograft transplantation (OCA) using either a shell or plug technique and identified variables associated with graft failure. Methods:Consecutive patients at two institutions who underwent OCA of the patellofemoral compartment between March 1, 2001 to March 1, 2015, were retrospectively identified at minimum 2 years' follow-up. Demographic information, intraoperative findings, and postoperative data were collected. Patients were divided into two groups on the basis of surgical technique (plug group and shell group). Failure was defined as revision OCA, conversion to arthroplasty, or gross appearance of graft degeneration on second-look arthroscopy. Logistic regression analysis was performed to identify patient- and surgery-specific variables associated with survivorship. Results:Fifty patients were identified (36 women and 14 men; mean age 37.43 ± 8.87 years). Sixteen patients received plug allografts, whereas 34 received shell allografts. Forty percent of patients underwent a concomitant meniscal, ligamentous, malalignment, or chondral procedure. Five patients in the Plug Group (31.3%) underwent reoperation at a mean 1.37 ± 1.34 years, and 28 patients (82.4%) who received Shell OCA underwent reoperation at a mean 1.94 ± 1.92 years. Two patients in the plug group had graft failure at a mean 9.17 ± 0.93 years, whereas 13 patients in the shell group had graft failure at a mean 3.81 ± 2.78 years. Patients with plug allografts demonstrated 100% and 66% survival at 5 and 9.8 years, respectively. For the shell group, survivorship was 65.8% and 37% at 5 and 10.6 years, respectively. Increasing body mass index was associated with failure for the case series overall (odds ratio 1.33, P = .020). Traumatic cause was found to be protective of failure (odds ratio = 0.02, P = .035). Conclusions:Plug OCA of the patellofemoral compartment can be an efficacious procedure with quality mid-term outcomes. Shell OCA led to high failure rates at midterm outcomes. Increasing body mass index may predispose patients to failure, whereas traumatic cause of their lesion was associated with improved outcomes. Level of Evidence:Level IV, Therapeutic case series.

Project description:BACKGROUND:Despite improvements in treatment options and techniques, articular cartilage repair continues to be a challenge for orthopedic surgeons. This study provides data to support that the 2-year Cryopreserved, Thin, Laser-Etched Osteochondral Allograft (T-LE Allograft) embodies the necessary viable cells, protein signaling, and extracellular matrix (ECM) scaffold found in fresh cartilage in order to facilitate a positive clinical outcome for cartilage defect replacement and repair. METHODS:Viability testing was performed by digestion of the graft, and cells were counted using a trypan blue assay. Growth factor and ECM protein content was quantified using biochemical assays. A fixation model was introduced to assess tissue outgrowth capability and cellular metabolic activity in vitro. Histological and immunofluorescence staining were employed to confirm tissue architecture, cellular outgrowth, and presence of ECM. The effects of the T-LE Allograft to signal bone marrow-derived mesenchymal stem cell (BM-MSC) migration and chondrogenic differentiation were evaluated using in vitro co-culture assays. Immunogenicity testing was completed using flow cytometry analysis of cells obtained from digested T-LE Allografts and fresh articular cartilage. RESULTS:Average viability of the T-LE Allograft post-thaw was found to be 94.97 ± 3.38%, compared to 98.83 ± 0.43% for fresh articular cartilage. Explant studies from the in vitro fixation model confirmed the long-term viability and proliferative capacity of these chondrocytes. Growth factor and ECM proteins were quantified for the T-LE Allograft revealing similar profiles to fresh articular cartilage. Cellular signaling of the T-LE Allograft and fresh articular cartilage both exhibited similar outcomes in co-culture for migration and differentiation of BM-MSCs. Flow cytometry testing confirmed the T-LE Allograft is immune-privileged as it is negative for immunogenic markers and positive for chondrogenic markers. CONCLUSIONS:Using our novel, proprietary cryopreservation method, the T-LE Allograft, retains excellent cellular viability, with native-like growth factor and ECM composition of healthy cartilage after 2?years of storage at -?80?°C. The successful cryopreservation of the T-LE Allograft alleviates the limited availably of conventionally used fresh osteochondral allograft (OCA), by providing a readily available and simple to use allograft solution. The results presented in this paper supports clinical data that the T-LE Allograft can be a successful option for repairing chondral defects.

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Enhancing osteochondral allograft viability: effects of storage media composition.

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