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The influence of emission parameters in low-level-light therapy on cellular responses is not yet fully understood. This study assessed the impact of various light delivery modes on collagen production in human primary fibroblast cultured in monolayers after three treatments with red light-emitting diode illumination (630 nm, 8 J/cm2). Human type I collagen was measured in cell culture supernatants with procollagen type I C-peptide enzyme immunoassay. Results demonstrated that, 72 h post-baseline, specific microsecond pulsing patterns had a more favorable impact on the ability of fibroblasts to produce collagen de novo than comparative conditions of continuous wave, pulsed 50% duty cycle, and millisecond pulsing domains. The cascade of events leading to collagen production by red illumination may be explained by the photodissociation of nitric oxide from cytochrome c oxidase. Short and intermittent light delivery might enhance this cellular event.