BackgroundDespite　progress　in　developing　wound　care　strategies,　there　is　currently　no　treatment　that　promotes　the　self-tissue　repair　capabilities.　H-2　has　been　shown　to　effectively　protect　cells　and　tissues　from　oxidative　and　inflammatory　damage.　While　comprehensive　effects　and　how　H-2　functions　in　wound　healing　remains　unknown,　especially　for　the　link　between　H-2　and　extracellular　matrix　(ECM)　deposition　and　epidermal　stem　cells　(EpSCs)　activation.MethodsHere,　we　established　a　cutaneous　aseptic　wound　model　and　applied　a　high　concentration　of　H-2　(66%　H-2)　in　a　treatment　chamber.　Molecular　mechanisms　and　the　effects　of　healing　were　evaluated　by　gene　functional　enrichment　analysis,　digital　spatial　profiler　analysis,　blood　perfusion/oxygen　detection　assay,　in　vitro　tube　formation　assay,　enzyme-linked　immunosorbent　assay,　immunofluorescent　staining,　non-targeted　metabonomic　analysis,　flow　cytometry,　transmission　electron　microscope,　and　live-cell　imaging.ResultsWe　revealed　that　a　high　concentration　of　H-2　(66%　H-2)　greatly　increased　the　healing　rate　(3　times　higher　than　the　control　group)　on　day　11　post-wounding.　The　effect　was　not　dependent　on　O-2　or　anti-reactive　oxygen　species　functions.　Histological　and　cellular　experiments　proved　the　fast　re-epithelialization　in　the　H-2　group.　ECM　components　early　(3　days　post-wounding)　deposition　were　found　in　the　H-2　group　of　the　proximal　wound,　especially　for　the　dermal　col-I,　epidermal　col-III,　and　dermis-epidermis-junction　col-XVII.　H-2　accelerated　early　autologous　EpSCs　proliferation　(1-2　days　in　advance)　and　then　differentiation　into　myoepithelial　cells.　These　epidermal　myoepithelial　cells　could　further　contribute　to　ECM　deposition.　Other　beneficial　outcomes　include　sustained　moist　healing,　greater　vascularization,　less　T-helper-1　and　T-helper-17　cell-related　systemic　inflammation,　and　better　tissue　remodelling.ConclusionWe　have　discovered　a　novel　pattern　of　wound　healing　induced　by　molecular　hydrogen　treatment.　This　is　the　first　time　to　reveal　the　direct　link　between　H-2　and　ECM　deposition　and　EpSCs　activation.　These　H-2-induced　multiple　advantages　in　healing　may　be　related　to　the　enhancement　of　cell　viability　in　various　cells　and　the　maintenance　of　mitochondrial　functions　at　a　basic　level　in　the　biological　processes　of　life.