Tissue barbs are small-scale structures that may be used for sutureless joining of tissues. In this study, several types of tissue barbs were fabricated using two-photon polymerization/micromolding, including two-pronged tissue barbs, eight-pronged tissue barbs, 10-pronged tissue barbs, and 16-pronged tissue barbs. Tissue barb penetration in porcine tissue was observed using confocal laser scanning microscopy. Constructs containing medical tape and tissue barbs were created by applying tissue barbs in a parallel arrangement to Transpore™ medical tape. These results suggest that two-photon polymerization/micromolding is an indirect rapid prototyping approach that may be used for high-throughput replication of tissue barbs and other microstructured solid wound sealants.