Insertion of Inorganic-Biomolecular Nanohybrid into Eucaryotic Cell
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Insertion of Inorganic-Biomolecular Nanohybrid into Eucaryotic Cell Seo-Young Kwak1, Sung-Ho Hwang, Yong-Joo Jeong, Jong-Sang Park and Jin-Ho Choy* School of Chemistry & Molecular Engineering, Seoul National University, Seoul 151-747, KOREA 1 Present address: Materials Science & Engineering, University of Illinois at Urbana-Champaign, IL 61801, U.S.A. ABSTRACT It has been clearly demonstrated that ATP could be intercalated into inorganic layered double hydroxide (LDH), giving rise to a biomolecular-inorganic nanohybrid with preserving its physico-chemical and biological integrity. It shows a remarkable transfer efficiency of ATP into target cells by alleviating an electrical repulsion at the cell walls due to the neutralization of negative charge of phosphates by positive hydroxide layers. From cellular uptake experiment with laser scanning confocal fluorescence microscopy, it is revealed that the FITC-LDH hybrid is effectively transferred into 293 cells. Such an unique feature of biomolecule-LDH hybrid will open a new field of reserving and delivering genes, drugs and other functional biomolecules. INTRODUCTION Drug delivery system is gaining attention for life threatening diseases. For the efficient introduction of foreign gene-based therapeutics or drugs into target organ or cells, a carrier system is required. Both viral and non-viral vectors are presently under investigation. However, the conventional approaches have many problems, such as cytotoxicity, immunoreaction, low transfection, etc. Thus, there remains a need for less toxic and more efficient delivery vehicles for drug and other gene-based therapeutics. Generally, the layered double hydroxide (LDH) consisting of magnesium and aluminum has already been used as medicine due to its applicability as an agent for treatment of peptic ulcers. An effective method for treating gastric ulcer is to inhibit the action of hydrochloric acid and pepsin in the gastric juice. Mg2Al(OH)6(CO32-)-LDH meets various requirements imposed on an antiacid, and its excellence may be attributed to its structural features. The rate of reaction of Mg2Al(OH)6(CO32-)-LDH with gastric acid is similar to that of Mg(OH)2, while the buffering pH is rate-controlled by dissolution of Al(OH)3 monomer, and reaches a pH value around 4, which is slightly higher than the one at which Al(OH)3 is dissolved. The high antipeptic activity of LDH can be attributed both to the adsorption of negatively charged pepsin onto positively charged surface of LDH, and to buffering of the pH at about 4 for a long time. Therefore, we believe that the present LDH is quite biocompatible. In the present study, adenosine 5-triphosphate (ATP) is selected as a guest molecule to be inserted into LDH, since it has been well known as energy carrier in biological system. But negatively charged ATP cannot be incorporated effectively into cell due to the negative cell wall. It is, therefore, thought that the encapsulated and charge neutralized ATP with LDH could enter the cell more effectively. In this work, we were able to
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