Biofilm may be a consortium of microbial species where the cells of microbes attach to both life form and inanimate surfaces inside a self-made matrix of extracellular polymeric substance (EPS). Biofilm matrix surrounding the polymicrobial environment makes them highly resistant to harsh conditions and antibacterial treatments. The two significant factors that differentiate planktonic from biofilm resident microbes are EPS containing a variety of macromolecules and a diffusible molecule for transferring signals known as quorum sensing (QS). Against this backdrop of microbial resistance and cell signaling, different approaches have been developed to interfere with the specific mechanisms of intracellular and extracellular targets that include herbal active compounds and synthetic nanoparticles. This chapter outlines the features of biofilm development and the approaches with the evidence that can be incorporated into clinical usage.