No abstract

Senescent cells (SnCs) accumulate in many vertebrate tissues with age and contribute to age-related pathologies1–3, presumably through their secretion of factors contributing to the senescence-associated secretory phenotype (SASP)4–6. Removal of SnCs delays several pathologies7–9 and increases healthy lifespan8. Aging and trauma are risk factors for the development of osteoarthritis (OA)10, a chronic disease characterized by degeneration of articular cartilage leading to pain and physical disability. Senescent chondrocytes are found in cartilage tissue isolated from patients undergoing joint replacement surgery11–14, yet their role in disease pathogenesis is unknown. To test the idea that SnCs might play a causative role in OA, we used the p16-3MR transgenic mouse, which harbors a p16INK4a (Cdkn2a) promoter driving the expression of a fusion protein containing synthetic Renilla luciferase and monomeric red fluorescent protein domains, as well as a truncated form of herpes simplex virus 1 thymidine kinase (HSV-TK)15,16. This mouse strain allowed us to selectively follow and remove SnCs after anterior cruciate ligament transection (ACLT). We found that SnCs accumulated in the articular cartilage and synovium after ACLT, and selective elimination of these cells attenuated the development of post-traumatic OA, reduced pain and increased cartilage development. Intra-articular injection of a senolytic molecule that selectively killed SnCs validated these results in transgenic, non-transgenic and aged mice. Selective removal of the SnCs from in vitro cultures of chondrocytes isolated from patients with OA undergoing total knee replacement decreased expression of senescent and inflammatory markers while also increasing expression of cartilage tissue extracellular matrix proteins. Collectively, these findings support the use of SnCs as a therapeutic target for treating degenerative joint disease.

Nanoparticles have great potential as controllable drug delivery vehicles because of their size and modular functionality. Timing and location are important parameters when optimizing nanoparticles for delivery of chemotherapeutics. Here we show that positively- and negatively-charged gold nanoparticles carrying either fluorescein or doxorubicin molecules move and localize differently in an in vitro three dimensional model of tumour tissue. Fluorescence microcopy and mathematical modelling showed that uptake, and not diffusion, is the dominant mechanism in particle delivery. Our results suggest that positive particles may be more effective for drug delivery because they are more significantly taken up by proliferating cells. Negative particles, which diffused faster, may perform better when delivering drugs deep into the tissues. An understanding of how surface charge can control tissue penetration and drug release may overcome some of the current limitations in drug delivery.

Gold colloids functionalized with amino acids provide a scaffold for effective DNA binding with subsequent condensation. Particles with lysine and lysine dendron functionality formed particularly compact complexes and provided highly efficient gene delivery without any observed cytotoxicity. Nanoparticles functionalized with first generation lysine dendrons (NP−LysG1) were ∼28-fold superior to polylysine in reporter gene expression. These amino acid-based nanoparticles were responsive to intracellular glutathione levels, providing a tool for controlled release and concomitant expression of DNA.