期刊
CONTRACEPTION
卷 93, 期 4, 页码 337-346出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.contraception.2015.11.008
关键词
Human cervicovaginal fluid; Buffering capacity; Simulant design; Cytotoxicity
资金
- National Institutes of Health [U19AI076980]
Background: Reported vaginal and seminal fluid simulants have complex compositions with multiple preparatory steps that contribute to physical instability. We report the design and characterization of stable and simplified buffers that mimic the salient physical/chemical properties of the physiological fluids. Study design/methods: Human cervicovaginal and seminal fluid samples were collected and buffering capacity was determined. The major buffering species were identified from published compositions of reproductive tract fluids. These values were used to compute the composition of vaginal and seminal fluid simulants. Ionic strength, buffering capacities, pH and osmolalities were then calculated or experimentally determined. Finally, cytotoxicity was evaluated in HEC-1-A cells and 3D reconstructed EpiVaginal (TM) tissue (VEC-100-FT) using naive cells/tissue and nonoxynol-9 as controls. Results: The use of calculated amounts of conjugate acid and base for buffer development resulted in compositions that did not require endpoint pH adjustment and could be formulated as stable 10 x concentrates. Furthermore, due to the absence of complex divalent salts, all our proposed simulants were stable at 4 degrees C for 1 month whereas precipitation and pH and osmolality changes were noted in reported buffers. Experimental determination of buffering capacities yielded similar values for undiluted cervicovaginal fluid (beta(4.2-5.2)=35.6 +/- 12.3 mM, N=7) and human seminal fluid (beta(7-6)=37.5 +/- 5 mM, N=3). All neat simulants showed significant cytotoxicity in HEC-1-A cells but were well tolerated by organotypic vaginal tissue. Conclusions: We report revised and improved compositions of buffers mimicking salient properties of vaginal and seminal fluid necessary for in vitro product evaluation. (C) 2016 Elsevier Inc. All rights reserved.
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