Chondrocyte Survival and Material Properties of Hypothermically Stored Cartilage

Williams, Riley J.; Dreese, James C.; Chen, Christopher T.

doi:10.1177/0095399703258733

Cited by 150 publications

(36 citation statements)

References 42 publications

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“…6,10,15,19,30,33,39,42 In addition, during prolonged 4°C storage, chondrocytes, particularly in the superficial zone of cartilage, succumb with increasing storage duration; chondrocyte viability deteriorates substantially in the superficial zone by 14days, 3,26 and overall by 28days. 3,26,31,32,40,43,44,47 While shorter 4°C storage durations (i.e. 14days) preserve overall chondrocyte viability, acquisition and distribution of these allografts is logistically challenging.…”

Section: Introductionmentioning

confidence: 99%

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: Introductionmentioning

confidence: 99%

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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“…51 During routine 4°C storage of OCA, chondrocytes, especially those at the articular surface with PRG4-secreting function, are susceptible, with increasing storage duration, to cell death. 52-55 While usage of 4°C stored OCA leads to some clinical improvement compared to pre-operative patient function, the long-term efficacy of 4°C stored OCA in animal models suggests that such grafts are not as effective as fresh grafts, and susceptible to deterioration with variable outcomes, 56-58 supporting the importance of maintaining cell viability during OCA storage. Screening stored OCA for PRG4-secretion prior to implantation and/or storing OCA in a manner that preserves this function of cartilage may be useful to improve the long-term biological performance of osteochondral allografts.…”

Section: Discussionmentioning

confidence: 99%

In vivo efficacy of fresh versus frozen osteochondral allografts in the goat at 6 months is associated with PRG4 secretion

Pallante-Kichura

Chen

Temple-Wong

et al. 2013

Journal Orthopaedic Research

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The long-term efficacy of osteochondral allografts is due to the presence of viable chondrocytes within graft cartilage. Chondrocytes in osteochondral allografts, especially those at the articular surface that normally produce the lubricant proteoglycan-4 (PRG4), are susceptible to storage-associated death. The hypothesis of this study was that the loss of chondrocytes within osteochondral grafts leads to decreased PRG4 secretion, after graft storage and subsequent implant. The objectives were to determine the effect of osteochondral allograft treatment (FROZEN vs. FRESH) on secretion of functional PRG4 after (i) storage, and (ii) 6months in vivo in adult goats. FROZEN allograft storage reduced PRG4 secretion from cartilage by ~85% compared to FRESH allograft storage. After 6months in vivo, the PRG4-secreting function of osteochondral allografts was diminished with prior FROZEN storage by ~81% versus FRESH allografts and by ~84% versus non-operated control cartilage. Concomitantly, cellularity at the articular surface in FROZEN allografts was ~96% lower than FRESH allografts and non-operated cartilage. Thus, the PRG4-secreting function of allografts appears to be maintained in vivo based on its state after storage. PRG4 secretion may be not only a useful marker of allograft performance, but also a biological process protecting the articular surface of grafts following cartilage repair.

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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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Chondrocyte Survival and Material Properties of Hypothermically Stored Cartilage

Abstract: This observed decline may have significant ramifications on long-term graft survival following stored-allograft implantation.

Cited by 150 publications

References 42 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

In vivo efficacy of fresh versus frozen osteochondral allografts in the goat at 6 months is associated with PRG4 secretion

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

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