As important challenges in burn injuries, infections often lead to delayed and incomplete healing. Wound infections with antimicrobial-resistant bacteria are other challenges in the management of wounds. Hence, it can be critical to synthesize scaffolds that are highly potential for loading and delivering antibiotics over long periods. Methods: Double-shelled hollow mesoporous silica nanoparticles (DSH-MSNs) were synthesized and loaded with cefazolin. Cefazolin-loaded DSH-MSNs (Cef*DSH-MSNs) were incorporated into polycaprolactone (PCL) to prepare a nanofiber-mediated drug release system. Their biological properties were assessed through antibacterial activity, cell viability, and qRT-PCR. The morphology and physicochemical properties of the nanoparticles and nanofibers were also characterized. Results: The double-shelled hollow structure of DSH-MSNs demonstrated a high loading capacity of cefazolin (51%). According to in vitro findings, the Cef*DSH-MSNs embedded in polycaprolactone nanofibers (Cef*DSH-MSNs/PCL) provided a slow release for cefazolin. The release of cefazolin from Cef*DSH-MSNs/PCL nanofibers inhibited the growth of Staphylococcus aureus. The high viability rate of human adipose-derived stem cells (hADSCs) in contact with PCL and DSH-MSNs/PCL was indicative of the biocompatibility of nanofibers. Moreover, gene expression results confirmed changes in keratinocyte-related differentiation genes in hADSCs cultured on the DSH-MSNs/PCL nanofibers with the up-regulation of involucrin. Conclusion: The high drug-loading capacity of DSH-MSNs presents these nanoparticles as suitable vehicles for drug delivery. In addition, the use of Cef*DSH-MSNs/PCL can be an effective strategy for regenerative purposes.