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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

Conductivities and permeance of filtrate through the membrane filters over different cycles of diafiltration.
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Fig2: Conductivities and permeance of filtrate through the membrane filters over different cycles of diafiltration.

Mentions: We employed poly(ether sulphone) membranes with two kinds of pore sizes, i.e. with molecular weight cut-off sizes of both 30 and 50 KD, to measure the conductivity and permeance of the membrane filters (Figure 2). The conductivities of the two membranes were similar to each other at the same number of diafiltration cycles, but the values significantly decreased as the number of diafiltration cycles was increased. More specifically, the conductivities of the membranes with the 30 KD molecular weight cut-off size decreased from 19.6 to 9.01, 4.01, 1.7 and 0.7 ms/cm for diafiltration cycles of 1, 2, 3 and 4, respectively. However, the permeance of the membrane filters were displayed a significant different behavior than the conductivities. As expected, the peremances of the membrane filters for both molecular weight cut-off sizes, i.e. 30 and 50 KDa, increased when the number of diafiltration cycles was increased. The permeance of the membrane filters with a 50 KDa molecular weight cut-off size increased from 3.5 to 5.1, 7.6, 10.5 and 11.7 L/hr/m2, while the permeance of the 30 kDa molecular weight cut-off size increased from 1.7 to 2.6, 5.2, 7.1 and 7.9 L/hr/m2. These results indicated that the membrane filters with higher molecular weight cut-off sizes and samples subject to more diafiltration cycles had higher rates of permeate flow. The reason for the increase in permeances by repeated diafiltrations was previously shown to be due to the removal of air entrapped in the membrane pores [24].Figure 2


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)

Conductivities and permeance of filtrate through the membrane filters over different cycles of diafiltration.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig2: Conductivities and permeance of filtrate through the membrane filters over different cycles of diafiltration.
Mentions: We employed poly(ether sulphone) membranes with two kinds of pore sizes, i.e. with molecular weight cut-off sizes of both 30 and 50 KD, to measure the conductivity and permeance of the membrane filters (Figure 2). The conductivities of the two membranes were similar to each other at the same number of diafiltration cycles, but the values significantly decreased as the number of diafiltration cycles was increased. More specifically, the conductivities of the membranes with the 30 KD molecular weight cut-off size decreased from 19.6 to 9.01, 4.01, 1.7 and 0.7 ms/cm for diafiltration cycles of 1, 2, 3 and 4, respectively. However, the permeance of the membrane filters were displayed a significant different behavior than the conductivities. As expected, the peremances of the membrane filters for both molecular weight cut-off sizes, i.e. 30 and 50 KDa, increased when the number of diafiltration cycles was increased. The permeance of the membrane filters with a 50 KDa molecular weight cut-off size increased from 3.5 to 5.1, 7.6, 10.5 and 11.7 L/hr/m2, while the permeance of the 30 kDa molecular weight cut-off size increased from 1.7 to 2.6, 5.2, 7.1 and 7.9 L/hr/m2. These results indicated that the membrane filters with higher molecular weight cut-off sizes and samples subject to more diafiltration cycles had higher rates of permeate flow. The reason for the increase in permeances by repeated diafiltrations was previously shown to be due to the removal of air entrapped in the membrane pores [24].Figure 2

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