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Egg-in-cube: design and fabrication of a novel artificial eggshell with functionalized surface.

Huang W, Arai F, Kawahara T - PLoS ONE (2015)

Bottom Line: To test the effectiveness of the design, the cubic eggshells were used to culture chick embryos and survivability was confirmed when PDMS membranes with adequate oxygen permeability were used.Additionally, the chick embryo tissues could be accessed and manipulated from outside the cubic eggshell, by using mechanical tools without breakage of the eggshell.The proposed "Egg-in-Cube" with functionalized surface has great potential to serve as a promising platform for biomedical research.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Functions Engineering, Kyushu Institute of Technology, Wakamatsu-ku, Kitakyushu, Japan.

ABSTRACT
An eggshell is a porous microstructure that regulates the passage of gases to allow respiration. The chick embryo and its circulatory system enclosed by the eggshell has become an important model for biomedical research such as the control of angiogenesis, cancer therapy, and drug delivery test, because the use of embryo is ethically acceptable and it is inexpensive and small. However, chick embryo and extra-embryonic blood vessels cannot be accessed freely and has poor observability because the eggshell is tough and cannot be seen through, which limits its application. In this study, a novel artificial eggshell with functionalized surface is proposed, which allows the total amount of oxygen to pass into the egg for the chick embryo culturing and has high observability and accessibility for embryo manipulation. First, a 40-mm enclosed cubic-shaped eggshell consisting of a membrane structure and a rigid frame structure is designed, and then the threshold of the membrane thickness suitable for the embryo survival is figured out according to the oxygen-permeability of the membrane structure. The designed artificial eggshell was actually fabricated by using polydimethylsiloxane (PDMS) and polycarbonate (PC) in the current study. Using the fabricated eggshell, chick embryo and extra-embryonic blood vessels can be observed from multiple directions. To test the effectiveness of the design, the cubic eggshells were used to culture chick embryos and survivability was confirmed when PDMS membranes with adequate oxygen permeability were used. Since the surface of the eggshell is transparent, chick embryo tissue development could be observed during the culture period. Additionally, the chick embryo tissues could be accessed and manipulated from outside the cubic eggshell, by using mechanical tools without breakage of the eggshell. The proposed "Egg-in-Cube" with functionalized surface has great potential to serve as a promising platform for biomedical research.

No MeSH data available.


Related in: MedlinePlus

Formation of blood vessels on the top and a side membrane of a cubic eggshell.(a) Top views show that the blood vessel network grew gradually, and the embryo became increasingly larger with the development of its circulatory system. (b) Side views show that when the body of a chick embryo became large, the extra-circulatory system spread onto the side membrane to search for a larger volume of oxygen.
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pone.0118624.g005: Formation of blood vessels on the top and a side membrane of a cubic eggshell.(a) Top views show that the blood vessel network grew gradually, and the embryo became increasingly larger with the development of its circulatory system. (b) Side views show that when the body of a chick embryo became large, the extra-circulatory system spread onto the side membrane to search for a larger volume of oxygen.

Mentions: Blood vessel formation on the faces of the cubic eggshell fabricated using 0.3-mm-thick PDMS membranes could be observed simply without requiring a specific apparatus, as shown in Fig. 5. As shown in Fig. 5(A) and (B), from the top face, we found that the chick embryo increased in size gradually following egg cracking, and similarly, the blood vessel network became increasingly complicated and spread rapidly from day 3 to 7. There were no blood vessels observed on the lateral membrane after the egg contents were transferred into the cubic eggshell on day 3. By contrast, after 7-day development, the blood vessels spread almost all around the lateral-side membrane. These results suggest dramatic development of the circulation system of the chick embryo in the cubic eggshell, which indicates that the phenomenon of extensive blood vessel formation on the inner surface of normal eggshells to absorb oxygen passing through the pores of the eggshell could be observed in the cubic eggshell. Furthermore, when we observed the sidewall of the eggshell, not only the distribution of blood vessels, but also the flow of the blood cells was confirmed, as shown in S4 Movie.


Egg-in-cube: design and fabrication of a novel artificial eggshell with functionalized surface.

Huang W, Arai F, Kawahara T - PLoS ONE (2015)

Formation of blood vessels on the top and a side membrane of a cubic eggshell.(a) Top views show that the blood vessel network grew gradually, and the embryo became increasingly larger with the development of its circulatory system. (b) Side views show that when the body of a chick embryo became large, the extra-circulatory system spread onto the side membrane to search for a larger volume of oxygen.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359160&req=5

pone.0118624.g005: Formation of blood vessels on the top and a side membrane of a cubic eggshell.(a) Top views show that the blood vessel network grew gradually, and the embryo became increasingly larger with the development of its circulatory system. (b) Side views show that when the body of a chick embryo became large, the extra-circulatory system spread onto the side membrane to search for a larger volume of oxygen.
Mentions: Blood vessel formation on the faces of the cubic eggshell fabricated using 0.3-mm-thick PDMS membranes could be observed simply without requiring a specific apparatus, as shown in Fig. 5. As shown in Fig. 5(A) and (B), from the top face, we found that the chick embryo increased in size gradually following egg cracking, and similarly, the blood vessel network became increasingly complicated and spread rapidly from day 3 to 7. There were no blood vessels observed on the lateral membrane after the egg contents were transferred into the cubic eggshell on day 3. By contrast, after 7-day development, the blood vessels spread almost all around the lateral-side membrane. These results suggest dramatic development of the circulation system of the chick embryo in the cubic eggshell, which indicates that the phenomenon of extensive blood vessel formation on the inner surface of normal eggshells to absorb oxygen passing through the pores of the eggshell could be observed in the cubic eggshell. Furthermore, when we observed the sidewall of the eggshell, not only the distribution of blood vessels, but also the flow of the blood cells was confirmed, as shown in S4 Movie.

Bottom Line: To test the effectiveness of the design, the cubic eggshells were used to culture chick embryos and survivability was confirmed when PDMS membranes with adequate oxygen permeability were used.Additionally, the chick embryo tissues could be accessed and manipulated from outside the cubic eggshell, by using mechanical tools without breakage of the eggshell.The proposed "Egg-in-Cube" with functionalized surface has great potential to serve as a promising platform for biomedical research.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Functions Engineering, Kyushu Institute of Technology, Wakamatsu-ku, Kitakyushu, Japan.

ABSTRACT
An eggshell is a porous microstructure that regulates the passage of gases to allow respiration. The chick embryo and its circulatory system enclosed by the eggshell has become an important model for biomedical research such as the control of angiogenesis, cancer therapy, and drug delivery test, because the use of embryo is ethically acceptable and it is inexpensive and small. However, chick embryo and extra-embryonic blood vessels cannot be accessed freely and has poor observability because the eggshell is tough and cannot be seen through, which limits its application. In this study, a novel artificial eggshell with functionalized surface is proposed, which allows the total amount of oxygen to pass into the egg for the chick embryo culturing and has high observability and accessibility for embryo manipulation. First, a 40-mm enclosed cubic-shaped eggshell consisting of a membrane structure and a rigid frame structure is designed, and then the threshold of the membrane thickness suitable for the embryo survival is figured out according to the oxygen-permeability of the membrane structure. The designed artificial eggshell was actually fabricated by using polydimethylsiloxane (PDMS) and polycarbonate (PC) in the current study. Using the fabricated eggshell, chick embryo and extra-embryonic blood vessels can be observed from multiple directions. To test the effectiveness of the design, the cubic eggshells were used to culture chick embryos and survivability was confirmed when PDMS membranes with adequate oxygen permeability were used. Since the surface of the eggshell is transparent, chick embryo tissue development could be observed during the culture period. Additionally, the chick embryo tissues could be accessed and manipulated from outside the cubic eggshell, by using mechanical tools without breakage of the eggshell. The proposed "Egg-in-Cube" with functionalized surface has great potential to serve as a promising platform for biomedical research.

No MeSH data available.


Related in: MedlinePlus