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Purification and biocompatibility of fermented hyaluronic acid for its applications to biomaterials.

Choi S, Choi W, Kim S, Lee SY, Noh I, Kim CW - Biomater Res (2014)

Bottom Line: While low molecular weight impurities and insoluble impurities were successfully removed using a ultrafiltration membrane with 50 KDa molecular weight cut, endotoxins, high molecular weight proteins and nucleic acids were removed from the broth by employing adsorbents such as alumina and activated carbons.Alumina showed the best results for the removal of endotoxins, all of the activated carbons were very effective in the removal of high molecular weight proteins and nucleic acids.We obtained high molecular weight HA with excellent biocompatibility as judged by both measurement of cell proliferation and viability, indicating high possibility of its applications to biomaterials.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Biotechnology, Korea University, Seoul, 136-701 Republic of Korea ; Department of Bioplant, Hanmi Pharm. Co, Pyeongtaek, 451-805 Korea.

ABSTRACT

Background: Hyaluronic acid (HA) is of importance due to its diverse applications in pharmaceuticals and medical devices such as dermal filler, adhesion barriers, carrier for cells and bioactive molecules as well as scaffold biomaterials for tissue engineering. Evaluations of purification and biocompatibility of HA are required for its applications to biomaterials.

Results: After synthesizing HA by fermentation of streptococcus zooepidemicus for 25 hr, extensively purification of the fermented broth was performed to remove impurities using a filtration process for insoluble components and cells, and diverse adsorbents for soluble impurities. Its in vitro biocompatibility has been evaluated by measurement of cell counting and assay of cell live and dead. 60% yield of white HA powder was obtained, having 15-17 dL/g intrinsic viscosity with a molecular weight of approximately 1,000 kDa. While low molecular weight impurities and insoluble impurities were successfully removed using a ultrafiltration membrane with 50 KDa molecular weight cut, endotoxins, high molecular weight proteins and nucleic acids were removed from the broth by employing adsorbents such as alumina and activated carbons. Alumina showed the best results for the removal of endotoxins, all of the activated carbons were very effective in the removal of high molecular weight proteins and nucleic acids. The purified HA solution showed excellent cell compatibility with no cell damages as observed by both measurement of cell proliferation and observation of cell viability.

Conclusions: We obtained high molecular weight HA with excellent biocompatibility as judged by both measurement of cell proliferation and viability, indicating high possibility of its applications to biomaterials.

No MeSH data available.


Related in: MedlinePlus

Proliferation of MC3TC on purified HA solution with assays of CCK-8 over 7 days (A) and live and dead assay (B) afterin vitrocell culture for 7 days (B, ×100).
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Fig5: Proliferation of MC3TC on purified HA solution with assays of CCK-8 over 7 days (A) and live and dead assay (B) afterin vitrocell culture for 7 days (B, ×100).

Mentions: Biocompatibility of HA solution were excellent in terms of cell proliferation and viability as observed by measurement of CCK-8 and observation of their cell morphologies (Figure 5). The optical density of the samples increased from 0.15 to 0.19 to 0.58 as measured by CCK-8 (Figure 5A) and all the cells were viable without observation of any cell death (Figure 5B).Figure 5


Purification and biocompatibility of fermented hyaluronic acid for its applications to biomaterials.

Choi S, Choi W, Kim S, Lee SY, Noh I, Kim CW - Biomater Res (2014)

Proliferation of MC3TC on purified HA solution with assays of CCK-8 over 7 days (A) and live and dead assay (B) afterin vitrocell culture for 7 days (B, ×100).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4549129&req=5

Fig5: Proliferation of MC3TC on purified HA solution with assays of CCK-8 over 7 days (A) and live and dead assay (B) afterin vitrocell culture for 7 days (B, ×100).
Mentions: Biocompatibility of HA solution were excellent in terms of cell proliferation and viability as observed by measurement of CCK-8 and observation of their cell morphologies (Figure 5). The optical density of the samples increased from 0.15 to 0.19 to 0.58 as measured by CCK-8 (Figure 5A) and all the cells were viable without observation of any cell death (Figure 5B).Figure 5

Bottom Line: While low molecular weight impurities and insoluble impurities were successfully removed using a ultrafiltration membrane with 50 KDa molecular weight cut, endotoxins, high molecular weight proteins and nucleic acids were removed from the broth by employing adsorbents such as alumina and activated carbons.Alumina showed the best results for the removal of endotoxins, all of the activated carbons were very effective in the removal of high molecular weight proteins and nucleic acids.We obtained high molecular weight HA with excellent biocompatibility as judged by both measurement of cell proliferation and viability, indicating high possibility of its applications to biomaterials.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Biotechnology, Korea University, Seoul, 136-701 Republic of Korea ; Department of Bioplant, Hanmi Pharm. Co, Pyeongtaek, 451-805 Korea.

ABSTRACT

Background: Hyaluronic acid (HA) is of importance due to its diverse applications in pharmaceuticals and medical devices such as dermal filler, adhesion barriers, carrier for cells and bioactive molecules as well as scaffold biomaterials for tissue engineering. Evaluations of purification and biocompatibility of HA are required for its applications to biomaterials.

Results: After synthesizing HA by fermentation of streptococcus zooepidemicus for 25 hr, extensively purification of the fermented broth was performed to remove impurities using a filtration process for insoluble components and cells, and diverse adsorbents for soluble impurities. Its in vitro biocompatibility has been evaluated by measurement of cell counting and assay of cell live and dead. 60% yield of white HA powder was obtained, having 15-17 dL/g intrinsic viscosity with a molecular weight of approximately 1,000 kDa. While low molecular weight impurities and insoluble impurities were successfully removed using a ultrafiltration membrane with 50 KDa molecular weight cut, endotoxins, high molecular weight proteins and nucleic acids were removed from the broth by employing adsorbents such as alumina and activated carbons. Alumina showed the best results for the removal of endotoxins, all of the activated carbons were very effective in the removal of high molecular weight proteins and nucleic acids. The purified HA solution showed excellent cell compatibility with no cell damages as observed by both measurement of cell proliferation and observation of cell viability.

Conclusions: We obtained high molecular weight HA with excellent biocompatibility as judged by both measurement of cell proliferation and viability, indicating high possibility of its applications to biomaterials.

No MeSH data available.


Related in: MedlinePlus