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Viper and cobra venom neutralization by alginate coated multicomponent polyvalent antivenom administered by the oral route.

Bhattacharya S, Chakraborty M, Mukhopadhyay P, Kundu PP, Mishra R - PLoS Negl Trop Dis (2014)

Bottom Line: Snake antivenom, although effective in minimizing mortality in developed countries, is not equally so in developing countries due to its poor availability in remote snake infested areas as, and when, required.Results from in vivo studies indicated that orally delivered ASVS can significantly neutralize venom effects, depicted by protection against lethality, decreased hemotoxicity and renal toxicity caused by russell viper venom.Alginate was effective in entrapping all the structural components of ASVS, which on release and intestinal absorption effectively reconstituted the function of antivenom in neutralizing viper and cobra venom.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, University of Calcutta, Kolkata, India; Centre for Research in Nanoscience and Nanotechnology, University of Calcutta, Kolkata, India.

ABSTRACT

Background: Snake bite causes greater mortality than most of the other neglected tropical diseases. Snake antivenom, although effective in minimizing mortality in developed countries, is not equally so in developing countries due to its poor availability in remote snake infested areas as, and when, required. An alternative approach in this direction could be taken by making orally deliverable polyvalent antivenom formulation, preferably under a globally integrated strategy, for using it as a first aid during transit time from remote trauma sites to hospitals.

Methodology/principal findings: To address this problem, multiple components of polyvalent antivenom were entrapped in alginate. Structural analysis, scanning electron microscopy, entrapment efficiency, loading capacity, swelling study, in vitro pH sensitive release, acid digestion, mucoadhesive property and venom neutralization were studied in in vitro and in vivo models. Results showed that alginate retained its mucoadhesive, acid protective and pH sensitive swelling property after entrapping antivenom. After pH dependent release from alginate beads, antivenom (ASVS) significantly neutralized phospholipaseA2 activity, hemolysis, lactate dehydrogenase activity and lethality of venom. In ex vivo mice intestinal preparation, ASVS was absorbed significantly through the intestine and it inhibited venom lethality which indicated that all the components of antivenom required for neutralization of venom lethality were retained despite absorption across the intestinal layer. Results from in vivo studies indicated that orally delivered ASVS can significantly neutralize venom effects, depicted by protection against lethality, decreased hemotoxicity and renal toxicity caused by russell viper venom.

Conclusions/significance: Alginate was effective in entrapping all the structural components of ASVS, which on release and intestinal absorption effectively reconstituted the function of antivenom in neutralizing viper and cobra venom. Further research in this direction can strategize to counter such dilemma in snake bite management by promoting control release and oral antivenom rendered as a first aid.

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Related in: MedlinePlus

Swelling, in-vitro release, mucoadhesion capacity study of alginate-ASVS beads and characterization of normal, released ASVS.(A) Swelling property of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. (B) Release profile of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. Result showed as mean ± SEM, n = 6. (Ci) Mucin binding study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. (Cii) Ex vivo mucoadhesion study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. Result showed as mean ±SEM, n = 6. *# † p<0.05 was considered significant (*Alginate vs alginate: ASVS :: 2∶1, alginate: ASVS :: 1∶1, and alginate: ASVS :: 1∶2; # Alginate: ASVS :: 2∶1 vs alginate: ASVS :: 1∶1, alginate: ASVS :: 1∶2; † Alginate: ASVS :: 1∶1 vs alginate: ASVS :: 1∶2). (Di) Native polyacrelamide gel electrophoresis of normal ASVS, (Dii) Native polyacrelamide gel electrophoresis of released ASVS. (E) HPLC of normal ASVS solution (C18 column, 4 mm×250 mm, flow rate: 0.5 ml/min, solvent: 60∶40∶0.2:: methanol: water: acetic acid); (F) Overlay of HPLC data of normal ASVS solution and released ASVS solution from alginate beads.
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pntd-0003039-g002: Swelling, in-vitro release, mucoadhesion capacity study of alginate-ASVS beads and characterization of normal, released ASVS.(A) Swelling property of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. (B) Release profile of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. Result showed as mean ± SEM, n = 6. (Ci) Mucin binding study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. (Cii) Ex vivo mucoadhesion study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. Result showed as mean ±SEM, n = 6. *# † p<0.05 was considered significant (*Alginate vs alginate: ASVS :: 2∶1, alginate: ASVS :: 1∶1, and alginate: ASVS :: 1∶2; # Alginate: ASVS :: 2∶1 vs alginate: ASVS :: 1∶1, alginate: ASVS :: 1∶2; † Alginate: ASVS :: 1∶1 vs alginate: ASVS :: 1∶2). (Di) Native polyacrelamide gel electrophoresis of normal ASVS, (Dii) Native polyacrelamide gel electrophoresis of released ASVS. (E) HPLC of normal ASVS solution (C18 column, 4 mm×250 mm, flow rate: 0.5 ml/min, solvent: 60∶40∶0.2:: methanol: water: acetic acid); (F) Overlay of HPLC data of normal ASVS solution and released ASVS solution from alginate beads.

Mentions: The swelling ability of alginate entrapped ASVS beads were studied using different pH solution of 1.2, 6.8, 7.0 and 7.4. Alginate entrapped ASVS beads did not show any significant swelling at the pH 1.2 after 40 minutes of incubation whereas it showed significant gradual increase in swelling after 10, 20, 30 and 40 minutes of incubation respectively at pH 6.8, 7.0 and 7.4 solution which was expressed as weight change of beads, shown in Fig. 2A. Weight of alginate entrapped ASVS beads were increased by 1.4, 3.3, 3.3 times at pH 6.8, 7.0, 7.4 after 10 minutes incubation, Weight of alginate entrapped ASVS beads were increased by 5.4, 9.2, 7.04 times at pH 6.8, 7.0, 7.4 after 20 min incubation, by 8.2, 10.62, 7.24 times at pH 6.8, 7.0, 7.4 after 30 minutes incubation, and by 8.4, 12.48, 7.4 times at pH 6.8, 7.0, 7.4 after 40 minutes incubation respectively.


Viper and cobra venom neutralization by alginate coated multicomponent polyvalent antivenom administered by the oral route.

Bhattacharya S, Chakraborty M, Mukhopadhyay P, Kundu PP, Mishra R - PLoS Negl Trop Dis (2014)

Swelling, in-vitro release, mucoadhesion capacity study of alginate-ASVS beads and characterization of normal, released ASVS.(A) Swelling property of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. (B) Release profile of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. Result showed as mean ± SEM, n = 6. (Ci) Mucin binding study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. (Cii) Ex vivo mucoadhesion study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. Result showed as mean ±SEM, n = 6. *# † p<0.05 was considered significant (*Alginate vs alginate: ASVS :: 2∶1, alginate: ASVS :: 1∶1, and alginate: ASVS :: 1∶2; # Alginate: ASVS :: 2∶1 vs alginate: ASVS :: 1∶1, alginate: ASVS :: 1∶2; † Alginate: ASVS :: 1∶1 vs alginate: ASVS :: 1∶2). (Di) Native polyacrelamide gel electrophoresis of normal ASVS, (Dii) Native polyacrelamide gel electrophoresis of released ASVS. (E) HPLC of normal ASVS solution (C18 column, 4 mm×250 mm, flow rate: 0.5 ml/min, solvent: 60∶40∶0.2:: methanol: water: acetic acid); (F) Overlay of HPLC data of normal ASVS solution and released ASVS solution from alginate beads.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125299&req=5

pntd-0003039-g002: Swelling, in-vitro release, mucoadhesion capacity study of alginate-ASVS beads and characterization of normal, released ASVS.(A) Swelling property of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. (B) Release profile of alginate entrapped ASVS beads (alginate: ASVS :: 1∶1) at different pH solution. Result showed as mean ± SEM, n = 6. (Ci) Mucin binding study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. (Cii) Ex vivo mucoadhesion study of alginate entrapped ASVS beads at different concentration ratio of alginate: ASVS :: 2∶1; alginate: ASVS :: 1∶1; alginate: ASVS :: 1∶2 in 2% calcium chloride solution. Result showed as mean ±SEM, n = 6. *# † p<0.05 was considered significant (*Alginate vs alginate: ASVS :: 2∶1, alginate: ASVS :: 1∶1, and alginate: ASVS :: 1∶2; # Alginate: ASVS :: 2∶1 vs alginate: ASVS :: 1∶1, alginate: ASVS :: 1∶2; † Alginate: ASVS :: 1∶1 vs alginate: ASVS :: 1∶2). (Di) Native polyacrelamide gel electrophoresis of normal ASVS, (Dii) Native polyacrelamide gel electrophoresis of released ASVS. (E) HPLC of normal ASVS solution (C18 column, 4 mm×250 mm, flow rate: 0.5 ml/min, solvent: 60∶40∶0.2:: methanol: water: acetic acid); (F) Overlay of HPLC data of normal ASVS solution and released ASVS solution from alginate beads.
Mentions: The swelling ability of alginate entrapped ASVS beads were studied using different pH solution of 1.2, 6.8, 7.0 and 7.4. Alginate entrapped ASVS beads did not show any significant swelling at the pH 1.2 after 40 minutes of incubation whereas it showed significant gradual increase in swelling after 10, 20, 30 and 40 minutes of incubation respectively at pH 6.8, 7.0 and 7.4 solution which was expressed as weight change of beads, shown in Fig. 2A. Weight of alginate entrapped ASVS beads were increased by 1.4, 3.3, 3.3 times at pH 6.8, 7.0, 7.4 after 10 minutes incubation, Weight of alginate entrapped ASVS beads were increased by 5.4, 9.2, 7.04 times at pH 6.8, 7.0, 7.4 after 20 min incubation, by 8.2, 10.62, 7.24 times at pH 6.8, 7.0, 7.4 after 30 minutes incubation, and by 8.4, 12.48, 7.4 times at pH 6.8, 7.0, 7.4 after 40 minutes incubation respectively.

Bottom Line: Snake antivenom, although effective in minimizing mortality in developed countries, is not equally so in developing countries due to its poor availability in remote snake infested areas as, and when, required.Results from in vivo studies indicated that orally delivered ASVS can significantly neutralize venom effects, depicted by protection against lethality, decreased hemotoxicity and renal toxicity caused by russell viper venom.Alginate was effective in entrapping all the structural components of ASVS, which on release and intestinal absorption effectively reconstituted the function of antivenom in neutralizing viper and cobra venom.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, University of Calcutta, Kolkata, India; Centre for Research in Nanoscience and Nanotechnology, University of Calcutta, Kolkata, India.

ABSTRACT

Background: Snake bite causes greater mortality than most of the other neglected tropical diseases. Snake antivenom, although effective in minimizing mortality in developed countries, is not equally so in developing countries due to its poor availability in remote snake infested areas as, and when, required. An alternative approach in this direction could be taken by making orally deliverable polyvalent antivenom formulation, preferably under a globally integrated strategy, for using it as a first aid during transit time from remote trauma sites to hospitals.

Methodology/principal findings: To address this problem, multiple components of polyvalent antivenom were entrapped in alginate. Structural analysis, scanning electron microscopy, entrapment efficiency, loading capacity, swelling study, in vitro pH sensitive release, acid digestion, mucoadhesive property and venom neutralization were studied in in vitro and in vivo models. Results showed that alginate retained its mucoadhesive, acid protective and pH sensitive swelling property after entrapping antivenom. After pH dependent release from alginate beads, antivenom (ASVS) significantly neutralized phospholipaseA2 activity, hemolysis, lactate dehydrogenase activity and lethality of venom. In ex vivo mice intestinal preparation, ASVS was absorbed significantly through the intestine and it inhibited venom lethality which indicated that all the components of antivenom required for neutralization of venom lethality were retained despite absorption across the intestinal layer. Results from in vivo studies indicated that orally delivered ASVS can significantly neutralize venom effects, depicted by protection against lethality, decreased hemotoxicity and renal toxicity caused by russell viper venom.

Conclusions/significance: Alginate was effective in entrapping all the structural components of ASVS, which on release and intestinal absorption effectively reconstituted the function of antivenom in neutralizing viper and cobra venom. Further research in this direction can strategize to counter such dilemma in snake bite management by promoting control release and oral antivenom rendered as a first aid.

Show MeSH
Related in: MedlinePlus