Background: Platelet-rich plasma (PRP) has emerged as a promising therapeutic intervention for knee osteoarthritis (OA), attracting substantial clinical and research attention. However, the clinical relevance of the treatment benefit remains controversial. Purpose: To evaluate the effectiveness of PRP compared with placebo in patients with knee OA in terms of minimal clinically important difference (MCID) and to investigate the possible influence of platelet concentration on the clinical outcome. Study Design: Meta-analysis. Level of evidence 1. Methods: The search was conducted on 5 databases (PubMed, Cochrane Library, Scopus, Embase, Web of Science) using the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. Inclusion criteria were randomized controlled trials comparing PRP and placebo injections to treat knee OA, written in the English language, with no time limitation. The effects were quantified at 1-, 3-, 6-, and 12-month follow-up points. Visual analog scale (VAS) for pain and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were used, with subanalyses based on platelet concentration performed using a 1,000,000 ± 20% platelets/µL cutoff. The MCID values (VAS, 1.37; WOMAC, 6.4) were used to interpret clinical improvement. The articles’ quality was assessed using the Revised Tool for Risk of Bias in Randomized Trials and the Grading of Recommendations Assessment, Development and Evaluation guidelines. Results: Among the 5499 articles retrieved, 18 randomized controlled trials (1995 patients) were included. PRP presented statistically superior improvements in VAS and WOMAC scores compared with placebo at all follow-up points, exceeding the MCID at 3- and 6-month follow-up points for VAS and at all follow-up points for WOMAC. The subanalysis based on platelet concentration showed that high-platelet PRP provided clinically significant pain relief with the improvement exceeding the MCID compared with placebo at 3-, 6-, and 12-month follow-up points. In contrast, low-platelet PRP failed to offer a clinically perceivable benefit in terms of VAS score. WOMAC results showed that both products provided a clinically significant improvement at 3 and 6 months of follow-up. This benefit was maintained up to the 12-month follow-up in the high-platelet group but not in the low-platelet group, where the improvement compared with placebo did not reach statistical significance. Conclusion: This meta-analysis showed that PRP offered clinically relevant functional improvement at 1-, 3-, 6-, and 12-month follow-up points and pain relief at 3- and 6-month follow-up points compared with placebo for the treatment of knee OA. Platelet concentration was found to influence treatment efficacy, with high-platelet PRP providing superior pain relief and more durable functional improvement compared with low-platelet PRP.

The characterization of cancer stem cell (CSC) subpopulation, through the comparison of the gene expression signature in respect to the native cancer cells, is particularly important for the identification of novel and more effective anticancer strategies. However, CSC have peculiar characteristics in terms of adhesion, growth, and metabolism that possibly implies a different modulation of the expression of the most commonly used housekeeping genes (HKG), like b-actin (ACTB). Although it is crucial to identify which are the most stable HKG genes to normalize the data derived from quantitative Real-Time PCR analysis to obtain robust and consistent results, an exhaustive validation of reference genes in CSC is still missing. Here, we isolated CSC spheres from different musculoskeletal sarcomas and carcinomas as a model to investigate on the stability of the mRNA expression of 15 commonly used HKG, in respect to the native cells. The selected genes were analysed for the variation coefficient and compared using the popular algorithms NormFinder and geNorm to evaluate stability ranking. As a result, we found that: 1) Tata Binding Protein (TBP), Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (YWHAZ), Peptidylprolyl isomerase A (PPIA), and Hydroxymethylbilane synthase (HMBS) are the most stable HKG for the comparison between CSC and native cells; 2) at least four reference genes should be considered for robust results; 3) the use of ACTB should not be recommended, 4) specific HKG should be considered for studies that are focused only on a specific tumor type, like sarcoma or carcinoma. Our results should be taken in consideration for all the studies of gene expression analysis of CSC, and will substantially contribute for future investigations aimed to identify novel anticancer therapy based on CSC targeting.

Objective: The aim of this study was to document the survival rate in the middle-aged patient group up to 65 years old and to compare it with other age groups of patients undergoing total knee arthroplasty (TKA) for knee osteoarthritis (OA). Design: The Register of Orthopaedic Prosthetic Implants (RIPO) regional registry was used to analyze the results of patients <80 years old affected by primary OA and treated with TKA from 2000 to 2019. The database was investigated according to the age group: younger than 50 years, 50-65 years, or 66-79 years, with the aim to estimate revision surgeries and implant survivorship. Results: A total of 45,488 TKAs for primary OA were included in the analysis (M: 11,388; F: 27,846). The percentage of patients <65 years old increased from 13.5% to 24.8% between 2000 and 2019 ( P < 0.0001). The survival analysis showed an overall influence of age on the implant revision rate ( P < 0.0001), with an estimated survival rate of 78.7%, 89.4%, and 94.8% at 15 years in the 3 groups, respectively. Compared with the older-aged group, the relative risk of failure was 3.1 (95% confidence interval [CI] = 2.2-4.3; P < 0.001) higher in patients <50 years old and 1.8 (95% CI = 1.6-2.0; P < 0.001) higher in patients 50-65 years old. Conclusions: TKA use in the middle-aged patient population up to 65 years old increased significantly over time. These patients present a double risk of failure with respect to older patients. This is particularly important considering the increasing life expectancy and the emergence of new joint preserving strategies, which could postpone the need for TKA to an older age.

Rhabdomyosarcoma is the most frequent soft tissue sarcoma in children and adolescents, with a high rate of relapse that dramatically affects the clinical outcome. Multiagent chemotherapy, in combination with surgery and/or radiation therapy, is the treatment of choice. However, the relapse rate is disappointingly high and identification of new therapeutic tools is urgently needed. Under this respect, the selective block of key features of cancer stem cells (CSC) appears particularly promising. In this study, we isolated rhabdomyosarcoma CSC with stem-like features (high expression of NANOG and OCT3/4, self-renewal ability, multipotency). Rhabdomyosarcoma CSC showed higher invasive ability and a reduced cytotoxicity to doxorubicin in comparison to native cells, through a mechanism unrelated to the classical multidrug resistance process. This was dependent on a high level of lysosome acidity mediated by a high expression of vacuolar ATPase (V-ATPase). Since it was not associated with other paediatric cancers, like Ewing’s sarcoma and neuroblastoma, V-ATPase higher expression in CSC was rhabdomyosarcoma specific. Inhibition of lysosomal acidification by the V-ATPase inhibitor omeprazole, or by specific siRNA silencing, significantly enhanced doxorubicin cytoxicity. Unexpectedly, lysosomal targeting also blocked cell growth and reduced the invasive potential of rhabdomyosarcoma CSC, even at very low doses of omeprazole (10 and 50 µM, respectively). Based on these observations, we propose lysosome acidity as a valuable target to enhance chemosensitivity of rhabdomyosarcoma CSC, and suggest the use of anti-V-ATPase agents in combination with standard regimens as a promising tool for the eradication of minimal residual disease or the prevention of metastatic disease.

Musculoskeletal sarcomas are aggressive malignancies often characterized by an adverse prognosis despite the use of intense multiagent chemotherapy or molecular targeted therapy in combination to surgery and radiotherapy. Stem-like cells identified within solid tumors have been recently implicated in drug resistance, metastasis and local relapse. Here, we report the identification of putative cancer stem cells (CSCs) in sarcomas using a sphere culture system. These sarcospheres, able to grow in anchorage-independent and serum-starved conditions, express the pluripotent embryonic stem cell marker genes OCT3/4, Nanog and SOX2. Expression levels of these genes were greater in sarcospheres than in the parental tumor cultures. Importantly, the isolated tumor spheres transplanted into mice were tumorigenic and capable of recapitulating the human disease. Finally, we demonstrated that low (1%) O2 conditions, reproducing those found within the tumor microenvironment, significantly increase the number and the size of sarcospheres. The sphere formation assay is, therefore, a valuable method for the isolation of putative CSCs from human sarcomas and its efficiency is improved by controlling oxygen availability. This method provides a reliable preclinical model that can be used for future studies aimed at investigating crucial aspects of sarcoma biology, such as resistance to treatments and relapse.

Nanoparticles (NPs) formed from polymers conjugated with bisphosphonates (BPs) allow the bone targeting of loaded drugs, such as doxorubicin, for the treatment of skeletal tumours. The additional a...