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Hyaluronic Acid (800 kDa) Supplementation of University of Wisconsin Solution Improves Viability of Osteochondral Grafts and Reduces Matrix Metalloproteinase Expression during Cold Preservation.

Yamada T, Uchida K, Onuma K, Inoue G, Aikawa J, Takano S, Sekiguchi H, Fujimaki H, Miyagi M, Takaso M - ScientificWorldJournal (2015)

Bottom Line: The supplementation of University of Wisconsin (UW) solution with 800 kDa significantly improved the cell viability of OCT after 14 days at 4°C compared to nonsupplemented UW solution.In contrast, UW solution supplemented with either 1900 or 6000 kDa HA did not markedly improve the cell viability of the OCT.Real-time PCR analysis revealed that the levels of matrix metalloproteinases 2, 3, and 9 were significantly decreased in OCT stored in UW solution supplemented with 800 kDa HA.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Engineering and Technology, School of Allied Health Science, Kitasato University, 1-15-1 Minami-ku, Kitasato, Sagamihara, Kanagawa 252-0374, Japan.

ABSTRACT
Osteochondral allografting is a promising option for the treatment of large cartilage defects. However, because the cell viability of osteochondral tissues (OCTs) gradually reduces during storage at 4°C, methods for maintaining the cell viability of fresh OCTs are needed to improve transplantation outcomes. Here, we evaluated whether the supplementation of preservation solution with one of three different molecular weight forms of hyaluronic acid (HA) improved the viability of rat OCTs during long-term cold storage. The supplementation of University of Wisconsin (UW) solution with 800 kDa significantly improved the cell viability of OCT after 14 days at 4°C compared to nonsupplemented UW solution. In contrast, UW solution supplemented with either 1900 or 6000 kDa HA did not markedly improve the cell viability of the OCT. Real-time PCR analysis revealed that the levels of matrix metalloproteinases 2, 3, and 9 were significantly decreased in OCT stored in UW solution supplemented with 800 kDa HA. Although further studies in human OCT are warranted, these findings demonstrate that the use of 800 kDa HA in place of serum may be a suitable approach for the long-term preservation of osteochondral allografts designated for the repair of large cartilage defects in the clinical setting.

No MeSH data available.


Related in: MedlinePlus

Representative hematoxylin and eosin stained tissue sections of OCTs cold preserved in the presence and absence of hyaluronic acids. Histological analysis of OCTs was performed after 2 weeks of preservation in UW (a), UW with HA800 (b), UW with HA1900 (c), and UW with HA6000 (d). Scale bar, 50 μm.
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fig1: Representative hematoxylin and eosin stained tissue sections of OCTs cold preserved in the presence and absence of hyaluronic acids. Histological analysis of OCTs was performed after 2 weeks of preservation in UW (a), UW with HA800 (b), UW with HA1900 (c), and UW with HA6000 (d). Scale bar, 50 μm.

Mentions: To determine the proportion of degenerative chondrocytes in OCTs after long-term cold storage, sections of OCTs prepared after 14 days of cold preservation in UW solution supplemented with and without HAs were hematoxylin and eosin stained and analyzed by light microscopy (Figure 1). After 14 days of storage in UW solution supplemented with HA800 and HA6000 at 4°C, the OCT samples had significantly more normal chondrocytes and fewer degenerated chondrocytes compared with those stored in UW solution without HA800 or HA6000 (P < 0.05; Table 2). The mean proportions of normal chondrocytes in OCTs preserved in UW solution alone and UW solution supplemented with HA1900 were 3.3% and 5.9%, respectively, whereas those in UW solution supplemented with HA800 and HA6000 were 22.4% and 15.8%, respectively. The mean proportions of degenerated chondrocytes in OCTs preserved in UW solution alone and UW solution supplemented with HA1900 were 96.7% and 94.2%, respectively, whereas those in UW solution supplemented with HA800 and HA6000 were only 77.6% and 84.2%, respectively. In addition, Safranin-O staining revealed that only a slight reduction in glycosaminoglycan levels occurred in OCTs preserved UW solution supplemented with HA800, HA1900, and HA6000 compared to the moderate reduction in glycosaminoglycan levels that was observed in OCTs stored in UW solution alone (Figure 2).


Hyaluronic Acid (800 kDa) Supplementation of University of Wisconsin Solution Improves Viability of Osteochondral Grafts and Reduces Matrix Metalloproteinase Expression during Cold Preservation.

Yamada T, Uchida K, Onuma K, Inoue G, Aikawa J, Takano S, Sekiguchi H, Fujimaki H, Miyagi M, Takaso M - ScientificWorldJournal (2015)

Representative hematoxylin and eosin stained tissue sections of OCTs cold preserved in the presence and absence of hyaluronic acids. Histological analysis of OCTs was performed after 2 weeks of preservation in UW (a), UW with HA800 (b), UW with HA1900 (c), and UW with HA6000 (d). Scale bar, 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4496492&req=5

fig1: Representative hematoxylin and eosin stained tissue sections of OCTs cold preserved in the presence and absence of hyaluronic acids. Histological analysis of OCTs was performed after 2 weeks of preservation in UW (a), UW with HA800 (b), UW with HA1900 (c), and UW with HA6000 (d). Scale bar, 50 μm.
Mentions: To determine the proportion of degenerative chondrocytes in OCTs after long-term cold storage, sections of OCTs prepared after 14 days of cold preservation in UW solution supplemented with and without HAs were hematoxylin and eosin stained and analyzed by light microscopy (Figure 1). After 14 days of storage in UW solution supplemented with HA800 and HA6000 at 4°C, the OCT samples had significantly more normal chondrocytes and fewer degenerated chondrocytes compared with those stored in UW solution without HA800 or HA6000 (P < 0.05; Table 2). The mean proportions of normal chondrocytes in OCTs preserved in UW solution alone and UW solution supplemented with HA1900 were 3.3% and 5.9%, respectively, whereas those in UW solution supplemented with HA800 and HA6000 were 22.4% and 15.8%, respectively. The mean proportions of degenerated chondrocytes in OCTs preserved in UW solution alone and UW solution supplemented with HA1900 were 96.7% and 94.2%, respectively, whereas those in UW solution supplemented with HA800 and HA6000 were only 77.6% and 84.2%, respectively. In addition, Safranin-O staining revealed that only a slight reduction in glycosaminoglycan levels occurred in OCTs preserved UW solution supplemented with HA800, HA1900, and HA6000 compared to the moderate reduction in glycosaminoglycan levels that was observed in OCTs stored in UW solution alone (Figure 2).

Bottom Line: The supplementation of University of Wisconsin (UW) solution with 800 kDa significantly improved the cell viability of OCT after 14 days at 4°C compared to nonsupplemented UW solution.In contrast, UW solution supplemented with either 1900 or 6000 kDa HA did not markedly improve the cell viability of the OCT.Real-time PCR analysis revealed that the levels of matrix metalloproteinases 2, 3, and 9 were significantly decreased in OCT stored in UW solution supplemented with 800 kDa HA.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Engineering and Technology, School of Allied Health Science, Kitasato University, 1-15-1 Minami-ku, Kitasato, Sagamihara, Kanagawa 252-0374, Japan.

ABSTRACT
Osteochondral allografting is a promising option for the treatment of large cartilage defects. However, because the cell viability of osteochondral tissues (OCTs) gradually reduces during storage at 4°C, methods for maintaining the cell viability of fresh OCTs are needed to improve transplantation outcomes. Here, we evaluated whether the supplementation of preservation solution with one of three different molecular weight forms of hyaluronic acid (HA) improved the viability of rat OCTs during long-term cold storage. The supplementation of University of Wisconsin (UW) solution with 800 kDa significantly improved the cell viability of OCT after 14 days at 4°C compared to nonsupplemented UW solution. In contrast, UW solution supplemented with either 1900 or 6000 kDa HA did not markedly improve the cell viability of the OCT. Real-time PCR analysis revealed that the levels of matrix metalloproteinases 2, 3, and 9 were significantly decreased in OCT stored in UW solution supplemented with 800 kDa HA. Although further studies in human OCT are warranted, these findings demonstrate that the use of 800 kDa HA in place of serum may be a suitable approach for the long-term preservation of osteochondral allografts designated for the repair of large cartilage defects in the clinical setting.

No MeSH data available.


Related in: MedlinePlus