Abstract
Osteoarthritis (OA) is a degenerative joint disease characterized by progressive deterioration and loss of articular cartilage. There is currently no treatment to reverse the onset of OA. Thus, we developed a targeted delivery strategy to transfer genes into primary human chondrocytes as a proof-of-concept study. We displayed a chondrocyte-affinity peptide (CAP) on the pIII minor coat protein of the M13 filamentous bacteriophage (phage)-based particle carrying a mammalian transgene cassette under cytomegalovirus CMV promoter and inverted terminal repeats (ITRs) cis elements of adeno-associated virus serotype 2 (AAV-2). Primary human articular chondrocytes (HACs) were used as an in vitro model, and the selectivity and binding properties of the CAP ligand in relation to the pathogenic conditions of HACs were characterized. We found that the CAP ligand is highly selective toward pathogenic HACs. Furthermore, the stability, cytotoxicity, and gene delivery efficacy of the CAP-displaying phage (CAP.Phage) were evaluated. We found that the phage particle is stable under a wide range of temperatures and pH values, while showing no cytotoxicity to HACs. Importantly, the CAP.Phage particle, carrying a secreted luciferase (Lucia) reporter gene, efficiently and selectively delivered transgene expression to HACs. In summary, it was found that the CAP ligand preferably binds to pathogenic chondrocytes, and the CAP.Phage particle successfully targets and delivers transgene to HACs.
Highlights
The signal of FITC-chondrocyte-affinity peptide (CAP) was found in the cytoplasm of human articular chondrocytes (HACs) but barely detected in primary human synovial fibroblasts (PHSFs) (Figure 1A)
We found that normal chondrocytes bound to the control peptide at flow were incubated with both groups of HACs, and the FITC signals were measured by
As extracellular matrix (ECM) is a major barrier to successful gene therapy, we previously showed that modulation of the ECM by enzymatic digestion enhances targeted gene delivery by phage [38]
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Advances in regenerative medicine technology significantly extend human longevity. Life expectancy has rapidly been extended, resulting in greater numbers of old-age population [1]. Osteoarthritis (OA) is a long-term chronic disease caused by the deterioration of joint cartilage, resulting in pain, impaired movement, and disability. The disease is the most common cause of disability in older adults.
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