novel methods to treat tissue damage caused by trauma, infection, or diseases. [1] Nevertheless, tissue engineering therapy using biologically active factors and stem cell-loaded scaffolds are not considered viable strategies for clinical applications due to their high time and cost requirements, as well as the regulatory challenges of ex vivo stem cell manipulation. [2] Endogenous stem cells are tissuespecific adult stem cells that are capable of self-renewal and can differentiate into specific cells. [3][4][5] These cells occur in specific niches in the body and can respond to signals or inflammatory substances emitted by tissue damage, among other processes. Importantly, these cells migrate to the specific regions of the body that require tissue repair. Because endogenous stem cells can be involved in the process of repair and regeneration of damaged or diseased tissues or organs, in situ tissue regeneration using endogenous stem cells could overcome several of the challenges caused by ex vivo stem cell manipulation in conventional tissue engineering. [6,7] Chemoattractants such as substance P (SP), stromal-derived factor, stem cell factor, granulocyte colony-stimulating factor, and monocyte chemotactic proteins are potent endogenous stem cell activating factors and can thus be used to prolong and enhance endogenous stem cell recruitment. [8][9][10] However, the number of chemoattractant-guided endogenous stem cells that migrate during the endogenous healing process is generally too low to achieve full in situ tissue regeneration. [9][10][11][12][13] Thus, novel chemoattractants must be developed to increase the migration of endogenous stem cells to the damaged area.Therefore, our first aim was to increase the migration of endogenous stem cells to the damaged area using a more efficient chemoattractant. SP-based nociceptive signaling plays an important role in the migration of blood-mediated stem cells, as well as in neurotransmission and neuromodulation. Compared to other complex 3D chemoattractants, SP is a relatively simple peptide comprised of only 11 amino acids (RPKPQQFFGLM). [14] The use of chemoattractants to promote endogenous stem cell-based in situ tissue regeneration has recently garnered much attention. This study is the first to assess the endogenous stem cell migration using a newly discovered substance P (SP) analog (SP1) by molecular dynamics simulations as an efficient chemoattractant. Further, a novel strategy based on electrostatic interaction using cationic chitosan (Ch) and anionic hyaluronic acid (HA) to prepare an SP1-loaded injectable C/H formulation without SP1 loss is developed. The formulation quickly forms an SP1-loaded C/H hydrogel in situ through in vivo injection. The newly discovered SP1 is found to possess human mesenchymal stromal cells (hMSCs) migration-inducing ability that is approximately two to three times higher than that of the existing SP. The designed VEGF-mimicking peptide (VP) chemically reacts with the hydrogel (C/H-VP) to sustain the release of VP, thus inducing ...