| Summary: | (1) Background: In hospitals, medical and dental clinics, antiseptics or disinfectants play an essential role in the control of nosocomial infections. This study aimed to evaluate <i>R. officinalis</i> and <i>P. paniculata</i> glycolic extracts regarding: (I) their antimicrobial action on planktonic and biofilm (monotypic and cutaneous biofilm model—<i>S. aureus</i>, <i>S. epidermidis</i> and <i>C. acnes</i>); and (II) their cytotoxicity on human keratinocytes (HaCaT). (2) Methods: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were performed (CLSI protocol M7-A6 and M11-A8). MTT analysis was used to evaluate the antibiofilm activity of the extracts on biofilms and their cytotoxicity on human keratinocytes. (3) Results: The combined glycolic extracts MIX A (75% <i>P. paniculata</i> + 25% <i>R. officinalis)</i>; MIX B (50% <i>P. paniculata</i> + 50% <i>R. officinalis</i>); and MIX C (25% <i>P. paniculata</i> + 75% <i>R. officinalis</i>) promoted MBC values by 50 mg/mL on <i>S. aureus</i>, absent on <i>S. epidermidis</i>, and ranged 6.25–50 mg/mL for <i>C. acnes.</i> The cutaneous biofilm model was reduced more than 90%. In addition, it showed biocompatibility with human keratinocytes, resulting in percentages of viability greater than 50%. (4) Conclusions: The combination of extracts promoted antimicrobial action on planktonic cultures, and monotypic and heterotypic biofilms of skin pathogens. Additionally, these extracts are biocompatible against human keratinocytes.
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