Using engineering models to shorten cryoprotectant loading time for the vitrification of articular cartilage

Shardt, Nadia; Chen, Zhirong; Yuan, Shuying Claire; Wu, Kezhou; Laouar, Leila; Jomha, Nadr M.; Elliott, Janet A.W.

doi:10.1016/j.cryobiol.2020.01.008

Cited by 15 publications

(27 citation statements)

References 53 publications

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“…These results indicate that the 7-hour CPA permeation strategy 18 is applicable to the vitrification of full condyle articular cartilage tissue. Importantly, we found no statistically significant differences in the chondrocyte viability of 10-mm diameter osteochondral dowels between the storage in a Falcon plastic tube with surrounding CPAs and storage in a vacuumed bag without surrounding CPAs (Fig.…”

Section: Discussionmentioning

confidence: 74%

“…6E ). These results indicate that the 7-hour CPA permeation strategy 18 is applicable to the vitrification of full condyle articular cartilage tissue.…”

Section: Discussionmentioning

confidence: 74%

“… 12 , 14 , 16 , 17 The development of CPA permeation protocols can be optimized using mathematical modelling. Our group developed an optimized 7-hour stepwise CPA permeation protocol for articular cartilage with 2-mm thickness, 18 and the experimental results confirmed the efficiency of this protocol for 10-mm diameter porcine osteochondral dowels with a promising chondrocyte survival of approximately 75%. 13 Since our CPA permeation approach leads to a vitrifiable concentration of CPAs throughout the cartilage tissue and this has been successful in smaller osteochondral tissue fragments, 18 it is essential to document its effectiveness on a larger scale such as full femoral condyles.…”

Section: Introductionmentioning

confidence: 78%

“…CPA permeation is a main factor that determines whether the vitrification of articular cartilage tissue will be successful. Shardt et al 18 proposed an optimized 7-hour CPA permeation protocol based on an engineering model incorporating predictions of CPA concentration, freezing point, and tissue vitrifiability. In the current study, we found that after applying the optimized 7-hour CPA permeation protocol, the chondrocyte viability after vitrification in small osteochondral dowels (10-mm diameter on a bone base) showed approximately 70% chondrocyte viability after vitrification for both storage in a Falcon plastic tube surrounded with CPAs and storage in a vacuumed bag without CPAs ( Fig.…”

Section: Discussionmentioning

confidence: 99%

“…During the vitrification experiments, samples of either 10-mm diameter osteochondral dowels or full femoral condyles were exposed to the multi-CPA loading solution (by immersion in 50 mL for dowels or 200 mL for full femoral condyles) prepared with DMEM-F12 medium following the 7-hour 3-step loading protocol developed previously 13 , 18 ; see Figure 3 for the CPA concentrations, durations, and temperatures of each step indicated in the orange box. Once the CPA loading procedure was completed, the cartilage samples were transferred to the containers (15 mL Falcon tube with 5 mL vitrification solution for the dowels, 250 mL plastic cup with 100 mL vitrification solution for the femoral condyles, VS+ in Fig.…”

Section: Methodsmentioning

confidence: 99%

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Vitrification of Intact Porcine Femoral Condyle Allografts Using an Optimized Approach

Laouar

Elliott

et al. 2020

CARTILAGE

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Objective Successful preservation of articular cartilage will increase the availability of osteochondral allografts to treat articular cartilage defects. We compared the effects of 2 methods for storing cartilage tissues using 10-mm diameter osteochondral dowels or femoral condyles at −196°C: (a) storage with a surrounding vitrification solution versus (b) storage without a surrounding vitrification solution. We investigated the effects of 2 additives (chondroitin sulfate and ascorbic acid) for vitrification of articular cartilage. Design Healthy porcine stifle joints ( n = 11) from sexually mature pigs were collected from a slaughterhouse within 6 hours after slaughtering. Dimethyl sulfoxide, ethylene glycol, and propylene glycol were permeated into porcine articular cartilage using an optimized 7-hour 3-step cryoprotectant permeation protocol. Chondrocyte viability was assessed by a cell membrane integrity stain and chondrocyte metabolic function was assessed by alamarBlue assay. Femoral condyles after vitrification were assessed by gross morphology for cartilage fractures. Results There were no differences in the chondrocyte viability (~70%) of 10-mm osteochondral dowels after vitrification with or without the surrounding vitrification solution. Chondrocyte viability in porcine femoral condyles was significantly higher after vitrification without the surrounding vitrification solution (~70%) compared to those with the surrounding vitrification solution (8% to 36%). Moreover, articular cartilage fractures were not seen in femoral condyles vitrified without surrounding vitrification solution compared to fractures seen in condyles with surrounding vitrification solution. Conclusions Vitrification of femoral condyle allografts can be achieved by our optimized approach. Removing the surrounding vitrification solution is advantageous for vitrification outcomes of large size osteochondral allografts.

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Section: Discussionmentioning

confidence: 74%

“…6E ). These results indicate that the 7-hour CPA permeation strategy 18 is applicable to the vitrification of full condyle articular cartilage tissue.…”

Section: Discussionmentioning

confidence: 74%