We have conducted several experiments on the effect of chloramphenicol and rifampicin.

Chloramphenicol stops the growth of the microorganisms through inhibition of their protein synthesis. In the case of K562 cells chloramphenicol significantly reduced cellgrowth, the activity of the citocrom c oxidase , and the synthesis of ATP and ferritin.

The mechanism of rifampicin is based on the inhibiton of the polymerisation of RNA. It's bactericidal effect is prevalent on intracellular and extracellular pathogens.

Chloramphenicol and rifampicin are the two most commonly used antibiotcs in opthalmology. This is the reason why we got interested in their effects on wound healing.

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What is the limbus?

The limbus is between  the see-through cornea and the opaque sclera. From outside the limbus is composed of the corneal and the conjuctival epithelium. The limbus isn’t rich in cells, but full of high collagen-containing connective tissue.

The study:

We processed the corneal discs according to protocol, then we cultured the recovered limbal cells.

We scratched the completly continuous cell layer with a sterile needle, imitating the injury on the surface of the cornea - this is  the scratch model.

We applied the following concentrations (based on the clinical doses) on the scratch model :

• Control, we did not use any antibiotics
• Rifampicin first concentration 0.1mg/ml
• Rifampicin second concentration 0.2mg/ml
• Chloramphenicol first concentraion 0.5mg/ml
• Chloramphenicol second concentration 1mg/ml

The samples were examined using Long Term Scan(LTS) microscopy and fluorescent microscopy.
The time-lapse videos of the cell cultures showed, that compared to the control, the treated cell cultures recovered slower, or the healing of the scratch did not happen.

The investigation by LTS system we observed that compared to the control in the treated cell cultures the healing of the scratch’s surface was slower or did not happen.

The control cells showed all of the phases of the chromatin condenstaion. On the contrary, we observed the following changes on the treated cultures:

•  As a result of the chloramphenicol treatment, chromatin condenstaion stopped at the praechromosomal phase.
• At higher rifampicin concentration (0.2 mg/ml) chromatin toxicity  manifested  primarily  as  supercoiled  chromatin  ribbons  without  the  appearance  of higher order of chromatin folding.