The biological activity of particles is largely dependent on their size in biological systems. Dispersion in the aqueous phase has been both a critical impediment to and a prerequisite for particle studies. Carbon black has been used as a surrogate to investigate the biological effects of carbonaceo...
The biological activity of particles is largely dependent on their size in biological systems. Dispersion in the aqueous phase has been both a critical impediment to and a prerequisite for particle studies. Carbon black has been used as a surrogate to investigate the biological effects of carbonaceous particles. Here, biocompatible methods were established to disperse carbon black into ultrafine and fine particles which are generally distinguished by the small size of 100 nm. Carbon black with a distinct particle size, N330 and N990 were suspended in blood plasma, cell culture media, Krebs-Ringer's solution (KR), or physiological salt solution (PSS). Large clumps were observed in all dispersion preparations; however, sonication improved dispersion - averaged particle sizes for N330 and N990 were $85.0{\pm}42.9$ and $112.4{\pm}67.9$ nm, respectively, in plasma; the corresponding sizes in culture media were $84.8{\pm}38.4$ and $164.1{\pm}77.8$ nm. However, sonication was not enough to disperse N330 less than 100 nm in either KR or PSS. Application of Tween 80 along with sonication reduced the size of N330 to less than 100 nm, and dispersed N990 larger than 100 nm ($73.6{\pm}28.8$ and $80.1{\pm}30.0$ nm for N330 and $349.5{\pm}161.8$ and $399.8{\pm}181.1$ nm for N990 in KR and PSS, respectively). In contrast, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) exhibited little effect. Electron microscopy confirmed the typical aciniform structure of the carbon arrays; however, zeta potential measurement failed to explain the dispersibility of carbon black. The methods established in this study could disperse carbon black into ultrafine and fine particles, and may serve as a useful model for the study of particle toxicity, particularly size-related effects.
The biological activity of particles is largely dependent on their size in biological systems. Dispersion in the aqueous phase has been both a critical impediment to and a prerequisite for particle studies. Carbon black has been used as a surrogate to investigate the biological effects of carbonaceous particles. Here, biocompatible methods were established to disperse carbon black into ultrafine and fine particles which are generally distinguished by the small size of 100 nm. Carbon black with a distinct particle size, N330 and N990 were suspended in blood plasma, cell culture media, Krebs-Ringer's solution (KR), or physiological salt solution (PSS). Large clumps were observed in all dispersion preparations; however, sonication improved dispersion - averaged particle sizes for N330 and N990 were $85.0{\pm}42.9$ and $112.4{\pm}67.9$ nm, respectively, in plasma; the corresponding sizes in culture media were $84.8{\pm}38.4$ and $164.1{\pm}77.8$ nm. However, sonication was not enough to disperse N330 less than 100 nm in either KR or PSS. Application of Tween 80 along with sonication reduced the size of N330 to less than 100 nm, and dispersed N990 larger than 100 nm ($73.6{\pm}28.8$ and $80.1{\pm}30.0$ nm for N330 and $349.5{\pm}161.8$ and $399.8{\pm}181.1$ nm for N990 in KR and PSS, respectively). In contrast, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) exhibited little effect. Electron microscopy confirmed the typical aciniform structure of the carbon arrays; however, zeta potential measurement failed to explain the dispersibility of carbon black. The methods established in this study could disperse carbon black into ultrafine and fine particles, and may serve as a useful model for the study of particle toxicity, particularly size-related effects.
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문제 정의
This study was designed and performed to establish the dispersion methods for carbon black with an emphasis on biocompatibility. With regard to biocompatibility, dispersion in aqueous solutions is of great importance in testing and understanding its biological effects, otherwise it cannot defines the exposure, absorption, delivery, and the consequent biological activity of carbon black in in vivo as well as in vitro.
대상 데이터
All animal experiments were conducted in accordance with protocols approved by the Ethics Committee of Animal Service Center at Dongguk University. Male Sprague-Dawley rats at 6-weeks of age were purchased from Daehan Biolink (Eumseong, Korea), and acclimated for one week before the experiments. The laboratory animal facility was maintained at constant temperature and humidity with a 12-hr light/dark cycle.
Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS) and penicillin/streptomycin were from Invitrogen (Carlsbad, CA, USA). Polyoxyethylene (20) sorbitan monooleate (polysorbate 80, Tween 80), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals used were of the highest purity available and purchased from standard suppliers.
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