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Abstract: tunneling wounds create passageways underneath the skin surface with varying sizes and shapes and can have twists and turns, making their treatment extremely difficult. Available wound care solutions only cater to superficial wounds, and untreated tunneling wounds pose major health concerns. This study aims to fulfill this challenge by fabricating tunnel wound fillers (TWFs) made of natural polymers that mimic the dermal extracellular matrix. In this study, cellulose microfibers (CMFs) derived from banana stem and fish skin-derived collagen were used to formulate bio-inks with varying CMF contents (25, 50, and 75 mg). Trilayered (CMFs, primary and secondary collagen coatings), drug-eluting (Baneocin), and cell-laden (human mesenchymal stem cells) TWFs were three-dimensional (3D)-printed and extensively characterized. CMFs showed the most suitable rheological properties for 3D printing at 50 mg concentration. The Alamar blue data showed significantly increased cell proliferation from Day 1 to Day 7, and scratch tests used to evaluate in vitro wound healing revealed that the best coverage of the wound area was achieved using CMFs in combination with collagen and alginate. Finally, the TWF showed promising capability and tunability in terms of wound shape and size upon testing on a chicken tissue model. The results demonstrate the tremendous potential of TWFs in treating deep tunneling wounds with unique advantages, such as patient-specific customization, good wound exudate absorption capability while releasing wound healing drugs, and the inclusion of stem cells for accelerated healing and tissue regeneration.