In-vitro sperm immobilization activity and biocompatibility study of NVD terpolymer


  • Sachin Gupta Department of Pharmacology, Advance Institute of Biotech and Paramedical Sciences, Kanpur, India
  • Ruchi Yadav Department of Community Medicine, Uttar Pradesh University of Medical Sciences, Saifai, India
  • Mayank Thakur Department of Pharmacology, Advance Institute of Biotech and Paramedical Sciences, Kanpur, India



Boronate polymer, Contraceptive, Sander-crammer assay, Sperm immobilization


Background: Boronate derivatives have been used in affinity chromatography for separation of cells based on their glycoprotein content. Boronate containing polymers when used intravaginally before sexual intercourse could bind to the glycoproteins present on sperm cell surface and render them immobile, which potentially may work as female-controlled contraceptive. To study this hypothesis NVD terpolymer which contains boronic acid was studied on goat sperm and its biocompatibility was accessed on NIH3T3 fibroblast.

Methods: Sperm motility study was carried out on goat sperm cells. The study was divided into two groups, test group (NVD terpolymer in simulated vaginal fluid) and negative control (simulated vaginal fluid only) performed using Sander-crammer assay. In the test group, the study was started from 0.1 % of the polymer solution, until the half of the sperms became non-motile as compared to normal control. The biocompatibility study was performed by culturing the NIH3T3 fibroblast with different concentrations of NVD polymer, followed by cell viability assay by performing 3-(4, 5-dimethylthiazol-2-Yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at 6, 12 and 24 hour spectrophotometrically.

Results: Sander-crammer assay resulted in significant (P-value < 0.05) decrease in motile sperm count in test group when compared to control. At 7.5 % concentration, the half of the sperms rendered immobile, and this was termed as effective concentration 50 (EC50). In-vitro biocompatibility study using NIH3T3 fibroblasts culture and MTT assay with cultured cells at 6, 12 and 24 hour, revealed that the polymer is biologically compatible as there were no significant change (P-value < 0.05) in the absorbance.

Conclusions: Boronate containing polymer, such as NVD terpolymer has in-vitro sperm immobilizing activity in goat sperm model, with further research in this area could yield a potential female control contraceptive agent.

Author Biography

Mayank Thakur, Department of Pharmacology, Advance Institute of Biotech and Paramedical Sciences, Kanpur, India




Minnis AM, Padian NS. Choice of female- controlled barrier methods among young women and their male sexual partners. Fam Plann Perspec. 2001;33:3328-34.

Jain RK, Maikhuri JP, Kumar STVS, Sharma VL, Dwivedi AK, Mitra K et al. Novel disulphide esters of carbotioicacid as potent, non-detergent spermicides with low toxicity to Lactobacillus and HeLa cells in vitro. Human Reprod. 2007;22:708-16.

McMohan S, Hansen L, Mann J, Sevigny C, Wong T, Roache M. Contraception. BMC Women’s Health. 2004;4:S25.

Catalone BJ, Kish-Catalone TM, Neely EB, Budgeon LR, Ferguson ML, Stiller C et al. Comparative safety evaluation of the candidate vaginal microbicide C31G. Antimicro Agent Chemother. 2005;49:1509-20.

Hagga J, Tsui A, Wasser HJ. Reproductive health in developing countries: Expanding dimensions, building solutions. National Academy Press, Washington; 1997.

Darroch JE, Singh S. Trends in contraceptive need and use in developing countries in 2003, 2008, and 2012: an analysis of national surveys. Lancet. 2013;381:1756-62.

Piot D. Nonoxynol-9 not effective microbicide, trial shows. UNAIDS press release, 2000.

Khillare B, Shrivastav TG. Spermicidal Activity of Azadirachta indica (neem) leaf extract. Contracep. 2003;68:225-9.

Khaled H, Zairi A. In vitro spermicidal activity of peptides from amphibian skin: Dermaseptin S4 and derivatives. Bioorg Med Chem. 2008;16:266-75.

Sarwar G, Arif M, Khillare B, Thakur SC. In vitro study of the contraceptive spermicidal activity of Ensete Superbum on human sperm. Int J Pharm Pharmaceu Sci. 2014;8:205-7.

Singh H, Jabba MS, Ray AR, Vasudevan P. Effect of anionic polymeric hydrogels on spermatozoa motility. Biomaterials. 1984;5:307-9.

Srivastava A, Shakya AK, Kumar A. Boronate affinity chromatography of cells and biomolecules using cryogel matrices. Enzyme Microb Tech. 2012;51:373-81.

Angyal S, Greeves D, Pickles V. The stereochemistry of complex formation of polyols with borate and periodate anions, and with metal cations. Carbohydr Res. 1974;35:165-73.

Bergold A, Scouten WH. Boronate Chromatography. Solid Phase Biochem. 1983;34:149-87.

Li YC, Larsson EL, Jungvid H, Galaev IY, Mattiasson B. Separation of neoglycoproteins with different degree of glycosylation by boronate chromatography. Chromatographia. 2001;54:213-7.

Kumar A. Hydrogel scaffolds for tissue engineering. USPTO 2011, Publication number US8883503 B2.

Owen DH, Katz DF. Vaginal fluid stimulant. Contracep. 1999;93:91-5.

Reddy RM, Srivastava A, Kumar A. Monosaccharide-responsive phenylboronate-polyol cell scaffolds for cell sheet and tissue engineering applications. PLoS One. 2013;8(10):e77861.

Sander FV, Cramer SD. A practical method for testing the spermicidal action of chemical contraceptives. Human Fertil. 1941;6:134-53.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunologic Methods. 1983;65:55-63.

Rodolfo V, Ortiz CR, Tapia S, Gonzalez G, Trillio E, Garza KM, Stafford SW et al. In vitro biocompatibility studies of fibroblast cells on Ti-Ta alloys. Materials Transactions. 2002;43:2991-94.




How to Cite

Gupta, S., Yadav, R., & Thakur, M. (2017). In-vitro sperm immobilization activity and biocompatibility study of NVD terpolymer. International Journal of Basic & Clinical Pharmacology, 6(5), 1221–1226.



Original Research Articles