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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

Real-time PCR analysis of OCTs cold preserved in the presence and absence of hyaluronic acid. Expression of MMP-2 (a), MMP-3 (b), MMP-9 (c), and MMP-13 mRNAs (d) in OCT stored for two weeks in UW solution (UW) and UW solution supplemented with 800 kDa HA (HA800). ∗Indicating a statistically significant difference between the UW and UW with HA800 groups. All data are shown as the mean ± SE (n = 6).
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fig4: Real-time PCR analysis of OCTs cold preserved in the presence and absence of hyaluronic acid. Expression of MMP-2 (a), MMP-3 (b), MMP-9 (c), and MMP-13 mRNAs (d) in OCT stored for two weeks in UW solution (UW) and UW solution supplemented with 800 kDa HA (HA800). ∗Indicating a statistically significant difference between the UW and UW with HA800 groups. All data are shown as the mean ± SE (n = 6).

Mentions: To investigate the mechanism by which HA800 increased the cell viability of OCT after cold preservation, OCTs stored in UW supplemented with and without HA800 for 14 days were analyzed by real-time PCR for the expression of several matrix metalloproteinase (MMP) genes (Figure 4). MMP genes were selected because their expression levels are increased in degenerative chondrocytes [13–15] and in livers in response to cold stress [16]. The real-time PCR analysis showed that expression of the genes encoding MMP-2, -3, and -9 was significantly decreased in OCTs stored in UW supplemented with HA800 compared to those in UW alone (Figures 4(a)–4(c)). Expression of the MMP-13 gene was also markedly decreased in OCTs stored in HA800-supplemented UW solution compared to those stored in UW alone, although the difference was not statistically significant (Figure 4(d)).


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)

Real-time PCR analysis of OCTs cold preserved in the presence and absence of hyaluronic acid. Expression of MMP-2 (a), MMP-3 (b), MMP-9 (c), and MMP-13 mRNAs (d) in OCT stored for two weeks in UW solution (UW) and UW solution supplemented with 800 kDa HA (HA800). ∗Indicating a statistically significant difference between the UW and UW with HA800 groups. All data are shown as the mean ± SE (n = 6).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Real-time PCR analysis of OCTs cold preserved in the presence and absence of hyaluronic acid. Expression of MMP-2 (a), MMP-3 (b), MMP-9 (c), and MMP-13 mRNAs (d) in OCT stored for two weeks in UW solution (UW) and UW solution supplemented with 800 kDa HA (HA800). ∗Indicating a statistically significant difference between the UW and UW with HA800 groups. All data are shown as the mean ± SE (n = 6).
Mentions: To investigate the mechanism by which HA800 increased the cell viability of OCT after cold preservation, OCTs stored in UW supplemented with and without HA800 for 14 days were analyzed by real-time PCR for the expression of several matrix metalloproteinase (MMP) genes (Figure 4). MMP genes were selected because their expression levels are increased in degenerative chondrocytes [13–15] and in livers in response to cold stress [16]. The real-time PCR analysis showed that expression of the genes encoding MMP-2, -3, and -9 was significantly decreased in OCTs stored in UW supplemented with HA800 compared to those in UW alone (Figures 4(a)–4(c)). Expression of the MMP-13 gene was also markedly decreased in OCTs stored in HA800-supplemented UW solution compared to those stored in UW alone, although the difference was not statistically significant (Figure 4(d)).

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