Engineering biomaterials to manipulate the immune response to elicit specific therapeutic outcomes is a burgeoning field of research. Mast cells play a distinct and central role in the innate immune response， and are characterized by their rapid release of a myriad of proinflammatory mediators in response to stimulation. These mediators are central to protective actions such as wound healing， angiogenesis， and host defense against pathogens and animal venoms. Considering that mast cells are widely distributed in tissues that interface with the external environment， and are loaded with large amounts of preformed protective compounds， they are ideal targets for novel immunotherapies. Here we report that， by using an engineered nanoscaffold， human mast cells can be contact activated in cell and primary human skin tissue culture using a specific receptor-ligand mechanism. The IgE independent PAMP-12 peptide activates human mast cells through the recently identified Mas-related G-protein coupled receptor member X2 (MRGPRX) receptor. The PAMP-12 motif was conjugated， via a glycine spacer， with the self-assembling peptide (RADA) and mixed with unmodified (RADA) to form a nanofiber matrix; mast cell activation was influenced directly by this ratio. Moreover， conjugating the PAMP-12 motif within the matrix was shown to only activate local， tissue-resident mast cells. The result of ex vivo human skin tissue tests confirmed that the engineered nanoscaffold successfully activated skin-resident mast cells by contact. Thus， this nanoscaffold design may provide a new platform to modulate localized mast cell functions thereby facilitating their protective role in the skin.