Transplantation of Osteochondral Allografts After Cold Storage

Malinin, Theodore I.; Temple, H. Thomas; Buck, Billy E.

doi:10.2106/jbjs.d.02991

Cited by 57 publications

(33 citation statements)

References 38 publications

(52 reference statements)

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“…Early signs of tissue structural deterioration in 4°C-stored grafts, including reduced proteoglycan content, increased surface irregularity, and poor graft-host cartilage integration, suggest inadequate biological remodeling capacity of the remaining chondrocytes to sustain long-term function. Although variable gross scores (range 1-9) and qualitatively diminished surface cellularity for 4°C-stored allografts were consistent with previous animal studies, 21,34,37 the significant relationship between surface cellularity and repair outcomes reported here suggests that chondrocytes at the articular surface are vitally important to maintain tissue structure, composition, and function. Since surface chondrocyte viability during allograft storage appears to be a strong determinant of tissue properties, storage conditions that preserve surface viability is essential for long-term repair efficacy.…”

Section: Discussionsupporting

confidence: 90%

The In Vivo Performance of Osteochondral Allografts in the Goat Is Diminished With Extended Storage and Decreased Cartilage Cellularity

et al. 2012

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Background Currently, osteochondral allografts (OCA) are typically used after 4°C storage for prolonged durations (15-43days), which compromises chondrocyte viability, especially at the articular surface. The long-term in vivo performance of these fresh-stored allografts, in association with variable cellularity, is unknown. Hypothesis/Purpose Determine the effect of 4°C storage duration (14, 28days) versus the best (fresh) and worst (frozen) conditions of chondrocyte viability on structure, composition, and function of cartilage in the goat, and the association of retrieved chondrocyte cellularity with those tissue properties. Study Design Controlled Laboratory Study Methods The effect of allograft storage on in vivo repair outcomes was determined for OCA transplanted into fifteen recipient goats and analyzed at 12months. Repair outcomes were assessed by examining cartilage structure (gross, histopathology), composition (cellularity by depth, matrix fixed charge), and biomechanical function (stiffness). Relationships between cellularity and structural scores, matrix fixed charge, and stiffness were assessed by linear regression. Results Repair outcomes in 4°C-stored OCA were inferior to fresh OCA, and were accompanied by diminished cellularity at the surface, matrix fixed charge, and histopathological structure. Overall, cellularity by depth and matrix fixed charge in cartilage of fresh OCA were similar to non-operated controls. However, cellularity at the articular surface and matrix fixed charge in 4°C-stored OCA were lower than fresh, by ~55% (95%CI, 32-76%) and ~20% (95%CI, 9-30%), respectively. In frozen OCA, cellularity and matrix fixed charge were lower than 4°C-stored OCA, by ~93% (95%CI, 88-99%) and ~22% (95%CI: 10-35%), respectively. Cellularity correlated negatively with cartilage health indices, including structural scores, and positively with matrix fixed charge and stiffness. Conclusion Reduced cellularity at the articular surface, resulting from 4°C storage, was associated with variable long-term outcomes, versus consistently good repair by fresh allografts. Cellularity at the articular surface was an important index of biological performance. Clinical Relevance Normal chondrocyte density in vivo, especially in the superficial region of cartilage, is important for maintaining long-term cartilage function and matrix content. In human cartilage, containing cells at ~3-5× lower density than goat, repair outcomes may be related to absolute minimum number of cells rather than density.

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Section: Discussionsupporting

confidence: 90%

The In Vivo Performance of Osteochondral Allografts in the Goat Is Diminished With Extended Storage and Decreased Cartilage Cellularity

et al. 2012

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“…However, the issue with graft storage at 37 °C is that microorganism growth can occur, which increases the infection risk and contributes to high storage costs (Stoker et al 2012). Storage at 4 °C in a tissue bank has been the standard approach for many years, with the advantages of low risk of microbial infection, low storage cost, and an approach that is more widely accepted (Bae et al 2009; Bian et al 2010; Linn et al 2011; Malinin 2006; Onuma et al 2012; Williams et al 2004). With respect to culture medium replacement, some researchers have shown that periodic medium replacement is beneficial for tissue viability during storage (Bian et al 2010; Linn et al 2011; Williams et al 2004), whilst others do not agree with medium replacement (Bae et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro

Song

et al. 2016

Cell Tissue Bank

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Maintenance of articular cartilage allografts in culture media is a common method of tissue storage; however, the technical parameters of graft storage remain controversial. In this study, we examined the optimal temperature and culture medium exchange rate for the storage of osteochondral allografts in vitro. Cylindrical osteochondral grafts (n = 120), harvested from the talar joint surface of ten Boer goats, were randomly classified into four groups and stored under the following conditions: Group A1 was maintained at 4°C in culture medium that was refreshed every 2 days; Group A2 was maintained at 4°C in the same culture medium, without refreshing; Group B1, was maintained at 37°C in culture medium that was refreshed every 2 days; Group B2, was maintained at 37°C in the same culture medium, without refreshing. Chondrocyte viability in the grafts was determined by ethidium bromide/fluorescein diacetate staining on days 7, 21, and 35. Proteoglycan content was measured by Safranin-O staining. Group A1 exhibited the highest chondrocyte survival rates of 90.88 %, 88.31 % and 78.69 % on days 7, 21, and 35, respectively. Safranin O staining revealed no significant differences between groups on days 21 and 35. These results suggest that storage of osteochondral grafts at 4°C with regular culture medium replacement should be highly suitable for clinical application.

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“…Although the cells in heart valve leaflets persist for weeks during hypothermic refrigeration in culture medium [Brockbank et al, 1992; Taylor and Brockbank, 2003], most tissues are refrigerated for only hours before significant loss of cell viability and tissue function occur. There are, however, reports supporting chondrocyte survival for days or weeks of hypothermic storage in their natural extracellular matrix in humans [Williams et al, 2003; Ball et al, 2004; Allen et al, 2005; Malinin et al, 2006] and several animal species [Black et al, 1979; Rodrigo et al, 1980; Wayne et al, 1990; Oates et al, 1995; Kim et al, 1996; Rohde et al, 2004; Williams et al, 2004; Malinin et al, 2006; Teng et al, 2008; Onuma et al, 2009]. Because such survival was considered unusual, compared with other tissue types, we previously initiated research to assess the impact of 4°C storage in DMEM culture medium on cartilage cell viability and establish whether or not cartilage plugs were an acceptable model compared with bisected femoral heads with both cartilage and bone tissue present [Brockbank et al, 2011b].…”

Section: Introductionmentioning

confidence: 99%

Impact of Storage Solution Formulation during Refrigerated Storage upon Chondrocyte Viability and Cartilage Matrix

Wright

Brockbank²,

Rahn³

et al. 2014

Cells Tissues Organs

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Various preservation solutions have been evaluated for longer hypothermic cartilage storage for tissue transplantation; however, the results are mixed. This research was carried out to determine whether phosphate-buffered saline (PBS) or organ preservation solutions would preserve both the extracellular matrix and chondrocytes of articular cartilage better than culture medium during refrigerated storage in the time frame that cartilage is stored for clinical use. Porcine cartilage plugs were stored, without the underlying bone, in culture medium with and without fetal bovine serum (FBS), PBS, Belzer's and Unisol solutions for 1 month at 4°C. Metabolic activity was tested using a resazurin reduction method, and matrix permeability was evaluated by measuring electrical conductivity. Storage in culture medium with 10% FBS was shown to provide good cartilage metabolic function for 7 days, decreasing to about 36% after 1 month of storage. There was no significant difference between samples stored in culture medium with and without FBS after 1 month of storage (p = 0.5005). Refrigerated storage of cartilage in PBS and two different solutions (Belzer's and Unisol) designed for optimal refrigerated tissue and organ storage results in loss of chondrocyte function and retention of matrix permeability. In contrast, the opposite, namely significantly better retention of chondrocyte function and loss of matrix permeability, was observed with culture medium. Future research should be focused on combining retention of chondrocyte function and matrix permeability by storage solution formulation.

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Transplantation of Osteochondral Allografts After Cold Storage

Cited by 57 publications

References 38 publications

The In Vivo Performance of Osteochondral Allografts in the Goat Is Diminished With Extended Storage and Decreased Cartilage Cellularity

The In Vivo Performance of Osteochondral Allografts in the Goat Is Diminished With Extended Storage and Decreased Cartilage Cellularity

Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro

Impact of Storage Solution Formulation during Refrigerated Storage upon Chondrocyte Viability and Cartilage Matrix

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