Nano‐Engineered Titanium Implants Loaded With Gingival Fibroblasts‐Derived Microvesicles Enhance Early Osseointegration And Soft Tissue Attachment In Vivo

Titanium dental implants require both reliable osseointegration and peri-implant soft tissue seal formation to ensure long-term success. While osseointegration has been widely studied, strategies to optimize soft tissue healing remain underexplored. In this study, we developed titania nanopore (TNP) implant surfaces functionalized with human primary gingival fibroblasts-derived microvesicles (hGFs-MVs) to investigate early peri-implant soft and hard tissue healing in vivo. hGFs-MVs were enriched, characterized, and loaded onto TNP implants at 10⁹ particles per implant (2 mm × 3 mm) via passive adsorptive loading. Rat first molars were extracted, and implants were placed into healed sockets after 4 weeks. Animals were assigned to TNP or hGFs-MVs/TNP groups (n = 5-6) and assessed at 1 and 4 weeks by micro-CT and histology. Cryo-electron microscopy confirmed the double-membrane structure of hGFs-MVs, and in vitro release studies showed peak vesicle release at day 3. Histology revealed early osteoid formation at week 1 and mature mineralized bone by week 4. hGFs-MVs/TNP implants exhibited significantly higher percentage of bone-to-implant contact (BIC%) at 4 weeks. Importantly, the hGFs-MVs/TNP group showed markedly enhanced soft tissue attachment and mature collagen fibers. These 'proof-of-concept' findings suggest that hGFs-MVs-loaded TNP implants enhance both in vivo osseointegration and soft tissue integration, providing a clinically relevant, cell-free strategy for improving peri-implant healing.