Delivery of TEM β-lactamase by gene-transformed Lactococcus lactis subsp. lactis through cervical cell monolayer

Lactococcus lactis subsp. lactis transformed with Plasmid ss80 (encoding the production and secretion of TEM β-lactamase) was used for the delivery of β-lactamase through the C-33A (cervix cell) monolayer. The viability of the cell monolayers co-cultured with L. lactis was examined by the trypan blu...

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Veröffentlicht in:International journal of pharmaceutics 2006-04, Vol.313 (1), p.29-35
Hauptverfasser: Kaushal, Gagan, Trombetta, Louis, Ochs, Raymond S., Shao, Jun
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Trombetta, Louis
Ochs, Raymond S.
Shao, Jun
description Lactococcus lactis subsp. lactis transformed with Plasmid ss80 (encoding the production and secretion of TEM β-lactamase) was used for the delivery of β-lactamase through the C-33A (cervix cell) monolayer. The viability of the cell monolayers co-cultured with L. lactis was examined by the trypan blue exclusion method. The integrity of the monolayers was monitored by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance. The transport rate of β-lactamase through C-33A monolayer was increased by four- and nine-folds ( p < 0.05) at the first hour by the transformed L. lactis compared to the free solution with or without presence of the untransformed L. lactis, respectively. This increase was gradually diminished after the 1st hour: it became 30 and 50% ( p < 0.05) at 10 h. The presence of the untransformed L. lactis with free solution delivery also increased the transport rate by 100% at 1 h ( p < 0.05) and 15% at 10 h ( p > 0.05). The increase in transport rate by the transformed L. lactis is most probably due to the concentrate of β-lactamase on C-33A monolayer. When co-cultured with the L. lactis, the C-33A cell viability and the monolayer TEER remained steady for 10 h. The presence of L. lactis did not change the transport of propranolol and mannitol through the monolayers. In conclusion, the transformed L. lactis significantly ( p < 0.05) increased the transport of β-lactamase through the cervical monolayers, indicating probiotic bacteria delivery may be a promising approach for protein delivery through the vagina.
doi_str_mv 10.1016/j.ijpharm.2006.01.013
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The viability of the cell monolayers co-cultured with L. lactis was examined by the trypan blue exclusion method. The integrity of the monolayers was monitored by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance. The transport rate of β-lactamase through C-33A monolayer was increased by four- and nine-folds ( p &lt; 0.05) at the first hour by the transformed L. lactis compared to the free solution with or without presence of the untransformed L. lactis, respectively. This increase was gradually diminished after the 1st hour: it became 30 and 50% ( p &lt; 0.05) at 10 h. The presence of the untransformed L. lactis with free solution delivery also increased the transport rate by 100% at 1 h ( p &lt; 0.05) and 15% at 10 h ( p &gt; 0.05). The increase in transport rate by the transformed L. lactis is most probably due to the concentrate of β-lactamase on C-33A monolayer. 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When co-cultured with the L. lactis, the C-33A cell viability and the monolayer TEER remained steady for 10 h. The presence of L. lactis did not change the transport of propranolol and mannitol through the monolayers. 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Pharmaceutical industry</topic><topic>Pharmacology. Drug treatments</topic><topic>Protein delivery</topic><topic>TEM β-lactamase</topic><topic>Time Factors</topic><topic>Transformation, Bacterial</topic><topic>Vaginal delivery</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kaushal, Gagan</creatorcontrib><creatorcontrib>Trombetta, Louis</creatorcontrib><creatorcontrib>Ochs, Raymond S.</creatorcontrib><creatorcontrib>Shao, Jun</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>International journal of pharmaceutics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kaushal, Gagan</au><au>Trombetta, Louis</au><au>Ochs, Raymond S.</au><au>Shao, Jun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Delivery of TEM β-lactamase by gene-transformed Lactococcus lactis subsp. lactis through cervical cell monolayer</atitle><jtitle>International journal of pharmaceutics</jtitle><addtitle>Int J Pharm</addtitle><date>2006-04-26</date><risdate>2006</risdate><volume>313</volume><issue>1</issue><spage>29</spage><epage>35</epage><pages>29-35</pages><issn>0378-5173</issn><eissn>1873-3476</eissn><coden>IJPHDE</coden><abstract>Lactococcus lactis subsp. lactis transformed with Plasmid ss80 (encoding the production and secretion of TEM β-lactamase) was used for the delivery of β-lactamase through the C-33A (cervix cell) monolayer. The viability of the cell monolayers co-cultured with L. lactis was examined by the trypan blue exclusion method. The integrity of the monolayers was monitored by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance. The transport rate of β-lactamase through C-33A monolayer was increased by four- and nine-folds ( p &lt; 0.05) at the first hour by the transformed L. lactis compared to the free solution with or without presence of the untransformed L. lactis, respectively. This increase was gradually diminished after the 1st hour: it became 30 and 50% ( p &lt; 0.05) at 10 h. The presence of the untransformed L. lactis with free solution delivery also increased the transport rate by 100% at 1 h ( p &lt; 0.05) and 15% at 10 h ( p &gt; 0.05). The increase in transport rate by the transformed L. lactis is most probably due to the concentrate of β-lactamase on C-33A monolayer. When co-cultured with the L. lactis, the C-33A cell viability and the monolayer TEER remained steady for 10 h. The presence of L. lactis did not change the transport of propranolol and mannitol through the monolayers. In conclusion, the transformed L. lactis significantly ( p &lt; 0.05) increased the transport of β-lactamase through the cervical monolayers, indicating probiotic bacteria delivery may be a promising approach for protein delivery through the vagina.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>16488095</pmid><doi>10.1016/j.ijpharm.2006.01.013</doi><tpages>7</tpages></addata></record>
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subjects Bacterial Adhesion
beta-Lactamases - genetics
beta-Lactamases - metabolism
Bioavailability
Biological and medical sciences
C-33A
Cell Line, Tumor
Cervix Uteri - cytology
Cervix Uteri - metabolism
Epithelial Cells - metabolism
Female
General pharmacology
Humans
Lactococcus lactis
Lactococcus lactis - genetics
Lactococcus lactis - metabolism
Medical sciences
Normal flora
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Protein delivery
TEM β-lactamase
Time Factors
Transformation, Bacterial
Vaginal delivery
title Delivery of TEM β-lactamase by gene-transformed Lactococcus lactis subsp. lactis through cervical cell monolayer
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