Sex Specific Determinants in Osteoarthritis: A Systematic Review of Preclinical Studies

Osteoarthritis (OA) is a highly prevalent joint disease that primarily affects about 10% of the world's population over 60 years old. The purpose of this study is to systematically review the preclinical studies regarding sex differences in OA, with particular attention to the molecular aspect and gene expression, but also to the histopathological aspects. Three databases (PubMed, Scopus, and Web of Knowledge) were screened for eligible studies. In vitro and in vivo papers written in English, published in the last 11 years (2009-2020) were eligible. Participants were preclinical studies, including cell cultures and animal models of OA, evaluating sex differences. Independent extraction of articles and quality assessments were performed by two authors using predefined data fields and specific tools (Animals in Research Reporting In Vivo Experiments (ARRIVE) guideline and Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) tool). Twenty-three studies were included in the review: 4 in vitro studies, 18 in vivo studies, and 1 both in vitro and in vivo study. From in vitro works, sex differences were found in the gene expression of inflammatory molecules, hormonal receptors, and in responsiveness to hormonal stimulation. In vivo research showed a great heterogeneity of animal models mainly focused on the histopathological aspects rather than on the analysis of sex-related molecular mechanisms. This review highlights that many gaps in knowledge still exist; improvementsin the selection and reporting of animal models, the use of advanced in vitro models, and multiomics analyses might contribute to developing a personalized gender-based medicine

Sharing Circulating Micro-RNAs between Osteoporosis and Sarcopenia: A Systematic Review

Background: Osteosarcopenia, a combination of osteopenia/osteoporosis and sarcopenia,is a common condition among older adults. While numerous studies and meta-analyses have beenconducted on osteoporosis biomarkers, biomarker utility in osteosarcopenia still lacks evidence. Here,we carried out a systematic review to explore and analyze the potential clinical of circulating microR-NAs (miRs) shared between osteoporosis/osteopenia and sarcopenia. Methods: We performed asystematic review on PubMed, Scopus, and Embase for differentially expressed miRs (p-value < 0.05)in (i) osteoporosis and (ii) sarcopenia. Following screening for title and abstract and deduplication,83 studies on osteoporosis and 11 on sarcopenia were identified for full-text screening. Full-textscreening identified 54 studies on osteoporosis, 4 on sarcopenia, and 1 on both osteoporosis andsarcopenia. Results: A total of 69 miRs were identified for osteoporosis and 14 for sarcopenia. Therewere 9 shared miRs, with evidence of dysregulation (up- or down-regulation), in both osteoporo-sis and sarcopenia: miR-23a-3p, miR-29a, miR-93, miR-133a and b, miR-155, miR-206, miR-208,miR-222, and miR-328, with functions and targets implicated in the pathogenesis of osteosarcopenia.However, there was little agreement in the results across studies and insufficient data for miRsin sarcopenia, and only three miRs, miR-155, miR-206, and miR-328, showed the same directionof dysregulation (down-regulation) in both osteoporosis and sarcopenia. Additionally, for mostidentified miRs there has been no replication by more than one study, and this is particularly true forall miRs analyzed in sarcopenia. The study quality was typically rated intermediate/high risk of bias.The large heterogeneity of the studies made it impossible to perform a meta-analysis. Conclusions:The findings of this review are particularly novel, as miRs have not yet been explored in the context ofosteosarcopenia. The dysregulation of miRs identified in this review may provide important clues tobetter understand the pathogenesis of osteosarcopenia, while also laying the foundations for furtherstudies to lead to effective screening, monitoring, or treatment strategies (PDF) Sharing Circulating Micro-RNAs between Osteoporosis and Sarcopenia: A Systematic Review. Available from: https://www.researchgate.net/publication/368667300_Sharing_Circulating_Micro-RNAs_between_Osteoporosis_and_Sarcopenia_A_Systematic_Review [accessed Feb 26 2023]

Development and characterization of a novel human 3D model of bone metastasis from breast carcinoma in vitro cultured

Breast cancer frequently metastasizes to the skeleton causing significant morbidity. Here, we set-up a novel and advanced ex vivo model by using fresh tissue from human vertebral bone metastasis from breast carcinoma patients able to retain the tumor microenvironment and tumor cells heterogeneity

SEX DETERMINANT IN AGING AND OSTEOARTHRITIS: IN VITRO MODELS OF HUMAN CHONDROCYTES AND SYNOVIOCYTES AND IN VIVO EVALUATIONS

Osteoarthritis (OA) is one of the most common degenerative musculoskeletal diseases, mainly characterized by progressive destruction of articular cartilage, inflammation of the synovial membrane and bone deformity, leading to pain and disability. OA is a complex disease with an unclear etiology and multiple risk factors, including sex and aging. The aim of the study was to evaluate the behavior of two cell phenotypes present at the joint level, synoviocytes and chondrocytes, and the role of sex differences in the presence of an inflammatory microenvironment (typical of OA) and/or oxidative stress (typical of ageing), using a two-dimensional (2D) in vitro model of monocultures and co-cultures. Human synoviocytes and chondrocytes isolated from male and female knee OA patients were stimulated for 24 hours with interleukin IL-1β and/or hydrogen peroxide H2O2. The sex differences of the cells and the effect of the stimuli were evaluated through cellular and molecular biology techniques, investigating the cellular proliferation rate, the healing percentage of the microwounds (i.e. cell migration), the gene expression of pro-inflammatory molecules (IL-6), proteolytic enzymes (ADAMTS-4, ADAMTS-5, MMP-13), markers of senescence and apoptosis (Klotho, Caspase-3) and components of the extracellular matrix (COL2A1). Furthermore, using an in vivo model of aging and spontaneous OA with C57BL/6 mice, the presence of OA and sex differences in the structure and morphology of the joint components (bone, articular cartilage, synovial membrane) and of the column were evaluated through histological and microtomographic techniques. The results showed that cells from female patients had a worse response capacity under unfavorable conditions of inflammation and oxidative stress than those from male patients, with lower cell viability and migration, and higher expression of inflammatory and apoptotic markers, release of proteolytic enzymes and oxidative stress products. Furthermore, the study showed that the combined effect of the inflammatory microenvironment and oxidative stress reduced the viability and migration of synoviocytes and chondrocytes. Furthermore, it increases the production of IL-6, Caspase-3, ADAMTS-4, ADAMTS-5, MMP-13 and ROS; while it decreases the expression of Klotho and COL2A1. Differently, the in vivo model confirmed the presence of OA in aged mice without showing differences between the sexes. In conclusion, the obtained data indicate the existence of sex-associated differences in cellular behavior in OA, suggesting the relevance of further studies aimed at identifying the mechanisms of OA pathology and potential sex-related differences through adequate preclinical models, to evaluate new hypotheses, concepts and therapeutic treatments able to arrest or reverse the progression of the disease with a personalized approach.L'osteoartrite (OA) è una delle malattie muscoloscheletriche degenerative più comuni, caratterizzata principalmente da progressiva distruzione della cartilagine articolare, infiammazione della membrana sinoviale e deformità ossea, che porta a dolore e disabilità. L'OA è una patologia complessa con un'eziologia poco chiara e molteplici fattori di rischio, tra cui il sesso e l'invecchiamento. Scopo dello studio è stato quello di valutare il comportamento di due fenotipi cellulari presenti a livello articolare, sinoviociti e condrociti, ed il ruolo delle differenze di sesso in presenza di un microambiente infiammatorio (tipico dell'OA) e/o di stress ossidativo (tipico dell'invecchiamento), utilizzando un modello in vitro bidimensionale (2D) di monocolture e co-colture. Sinoviociti e condrociti umani isolati da pazienti di sesso maschile e femminile con OA del ginocchio, sono stati stimolati per 24 ore con l’interleuchina IL-1β e/o con il perossido d’idrogeno H2O2. Le differenze di sesso delle cellule e l’effetto degli stimoli sono stati valutati attraverso tecniche di biologia cellulare e molecolare, investigando il tasso di proliferazione cellulare, la percentuale di guarigione delle microferite (ovvero la migrazione cellulare), l’espressione genica di molecole pro-infiammatorie (IL-6), enzimi proteolitici (ADAMTS-4, ADAMTS-5, MMP-13), marcatori di senescenza e apoptosi (Klotho, Caspase-3) e componenti della matrice extracellulare (COL2A1). Inoltre, utilizzando un modello in vivo di invecchiamento ed OA spontanea con topi C57BL/6, sono state valuate, attraverso tecniche istologiche e microtomografiche, la presenza di OA e le differenze di sesso nella struttura e morfologia delle componenti articolari (osso, cartilagine articolare, membrane sinoviale) e della colonna vertebrale. I risultati hanno mostrato che le cellule di pazienti di sesso femminile avevano una capacità di risposta peggiore in condizioni sfavorevoli di infiammazione e stress ossidativo rispetto a quelli di pazienti di sesso maschile, con una minore vitalità e migrazione cellulare, e una maggiore espressione di marcatori infiammatori e apoptotici, rilascio di enzimi proteolitici e prodotti di stress ossidativo. Inoltre, lo studio ha evidenziato che l’effetto combinato del microambiente infiammatorio e dello stress ossidativo ha ridotto la vitalità e la migrazione cellulare dei sinoviociti e condrociti. Inoltre, aumenta la produzione di IL-6, Caspase-3, ADAMTS-4, ADAMTS-5, MMP-13 e ROS; mentre diminuisce l'espressione di Klotho e COL2A1. Diversamente, il modello in vivo ha confermato la presenza di OA nei topi anziani senza però mostrare differenze tra i sessi. In conclusione, i dati ottenuti indicano l’esistenza di differenze nel comportamento cellulare associate al sesso nell’OA, suggerendo la rilevanza di ulteriori studi volti ad identificare i meccanismi della patologia OA e le potenziali differenze legate al sesso attraverso adeguati modelli preclinici, per valutare nuove ipotesi, concetti e trattamenti terapeutici in grado di arrestare o invertire la progressione della malattia con un approccio personalizzato

Sex Specific Determinants in Osteoarthritis: A Systematic Review of Preclinical Studies

Osteoarthritis (OA) is a highly prevalent joint disease that primarily affects about 10% of the world’s population over 60 years old. The purpose of this study is to systematically review the preclinical studies regarding sex differences in OA, with particular attention to the molecular aspect and gene expression, but also to the histopathological aspects. Three databases (PubMed, Scopus, and Web of Knowledge) were screened for eligible studies. In vitro and in vivo papers written in English, published in the last 11 years (2009–2020) were eligible. Participants were preclinical studies, including cell cultures and animal models of OA, evaluating sex differences. Independent extraction of articles and quality assessments were performed by two authors using predefined data fields and specific tools (Animals in Research Reporting In Vivo Experiments (ARRIVE) guideline and Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) tool). Twenty-three studies were included in the review: 4 in vitro studies, 18 in vivo studies, and 1 both in vitro and in vivo study. From in vitro works, sex differences were found in the gene expression of inflammatory molecules, hormonal receptors, and in responsiveness to hormonal stimulation. In vivo research showed a great heterogeneity of animal models mainly focused on the histopathological aspects rather than on the analysis of sex-related molecular mechanisms. This review highlights that many gaps in knowledge still exist; improvementsin the selection and reporting of animal models, the use of advanced in vitro models, and multiomics analyses might contribute to developing a personalized gender-based medicine

Nano-Based Biomaterials as Drug Delivery Systems Against Osteoporosis: A Systematic Review of Preclinical and Clinical Evidence

Osteoporosis (OP) is one of the most significant causes of morbidity, particularly in post-menopausal women and older men. Despite its remarkable occurrence, the search for an effective treatment is still an open challenge. Here, we systematically reviewed the preclinical and clinical progress in the development of nano-based materials as drug delivery systems against OP, considering the effects on bone healing and regeneration, the more promising composition and manufacturing methods, and the more hopeful drugs and delivery methods. The results showed that almost all the innovative nano-based delivery systems developed in the last ten years have been assessed by preclinical investigations and are still in the preliminary/early research stages. Our search strategy retrieved only one non-randomized controlled trial (RCT) on oligosaccharide nanomedicine of alginate sodium used for degenerative lumbar diseases in OP patients. Further investigations are mandatory for assessing the clinical translation and commercial purposes of these materials. To date, the main limits for the clinical translation of nano-based materials as drug delivery systems against OP are probably due to the low reproducibility of the manufacturing processes, whose specificity and complexity relies on an adequate chemical, structural, and biomechanical characterization, as the necessary prerequisite before assessing the efficacy of a given treatment or process. Finally, an unsatisfactory drug-loading capacity, an uncontrollable release kinetic, and a low delivery efficiency also limit the clinical application

Osteoporosis Preclinical Research: A Systematic Review on Comparative Studies Using Ovariectomized Sheep

Sheep ovariectomy (OVX) alone or associated to steroid therapy, deficient diet, or hypothalamic–pituitary disconnection has proven to be of critical importance for osteoporosis research in orthopedics. However, the impact of specific variables, such as breed, age, diet, time after OVX, and other variables, should be monitored. Thus, the design of comparative studies is mandatory to minimize the impact of these variables or to recognize the presence of unwanted variables as well as to better characterize bone remodeling in this model. Herein, we conducted a systematic review of the last 10 years on PubMed, Scopus, and Web of Knowledge considering only studies on OVX sheep where a control group was present. Of the 123 records screened, 18 studies were included and analyzed. Results showed that (i) Merino sheep are the most exploited breed; (ii) 5–6 years of age is the most used time for inducing OVX; (iii) ventral midline laparotomy is the most common approach to induce OVX; (iv) OVX associated to steroid therapy is the most widely used osteoporosis model; and (v) success of OVX was mostly verified 12 months after surgery. In detail, starting from 12 months after OVX a significant decline in bone mineral density and in microarchitectural bone parameters as well as in biochemical markers were detected in all studies in comparison to control groups. Bone alteration was also site-specific on a pattern as follows: lumbar vertebra, femoral neck, and ribs. Before 12 months from OVX and starting from 3–5 months, microarchitectural bone changes and biochemical marker alterations were present when osteoporosis was induced by OVX associated to steroid therapy. In conclusion, OVX in sheep influence bone metabolism causing pronounced systemic bone loss and structural deterioration comparable to the situation found in osteoporosis patients. Data for treating osteoporosis patients are based not only on good planning and study design but also on a correct animal use that, as suggested by 3Rs principles and by ARRIVE guidelines, includes the use of control groups to be directly contrasted with the experimental group

Vertebral Bone Marrow Clot towards the Routine Clinical Scenario in Spine Surgeries: What about the Antimicrobial Properties?

Exploring innovative techniques and treatments to improve spinal fusion procedures is a global challenge. Here, we provide a scientific opinion on the ability of a vertebral bone marrow (vBM) clot to provide a local combined delivery system not only of stem cells, signaling biomolecules and anti-inflammatory factors but also of molecules and proteins endowed with antimicrobial properties. This opinion is based on the evaluation of the intrinsic basic properties of the vBM, that contains mesenchymal stem cells (MSCs), and on the coagulation process that led to the conversion of fibrinogen into fibrin fibers that enmesh cells, plasma but above all platelets, to form the clot. We emphasize that vBM clot, being a powerful source of MSCs and platelets, would allow the release of antimicrobial proteins and molecules, mainly cathelicidin LL- 37, hepcidin, kinocidins and cationic host defense peptides, that are per se gifted with direct and/or indirect antimicrobial effects. We additionally highlight that further studies are needed to deepen this knowledge and to propose vBM clot as multifunctional bioscaffold able to target all the main key challenges for spinal fusion surgery

Stromal Vascular Fraction and Amniotic Epithelial Cells: Preclinical and Clinical Relevance in Musculoskeletal Regenerative Medicine

Musculoskeletal regenerative medicine is mainly based on the use of cell therapy to heal damaged tissues such as bone, cartilage, and tendons. Throughout the years, different cell types have been employed for the treatment of musculoskeletal diseases, in particular, mesenchymal stem cells (MSCs) derived from bone marrow (BMSCs) and adipose tissue (ADSCs). Though the results of these literature studies have been encouraging, there are some limitations, especially on long-term results. Recently, some interest has shifted towards new cell types such as the stromal vascular fraction (SVF) and amniotic endothelial cells (AECs). The aim of the present literature review is to evaluate preclinical and clinical studies that used SVF and AECs for musculoskeletal tissue regeneration. Forty-eight preclinical and clinical studies, performed in the last 10 years, were identified. Both SVF and AECs, injected or implanted with or without scaffolds, were shown to be valid alternatives, and in some ways superior, to ADSCs and BMSCs, being able to differentiate towards osteogenic, chondrogenic, and tenogenic lineages, and to promote cell and tissue regenerative potential. The use of SVF and AECs could represent a new regenerative treatment in several musculoskeletal pathologies, solving the problem of cell expansion in vitro

What Is the Role of Interleukins in Breast Cancer Bone Metastases? A Systematic Review of Preclinical and Clinical Evidence

Breast cancer cells produce stimulators of bone resorption known as interleukins (ILs). However, data on the functional roles of ILs in the homing of metastatic breast cancer to bone are still fragmented. A systematic search was carried out in three databases (PubMed, Scopus, Web of Science Core Collection) to identify preclinical reports, and in three clinical registers (ClinicalTrials.gov, World Health Organization (WHO) International Clinical Trials Registry Platform, European Union (EU) Clinical Trials Register) to identify clinical trials, from 2008 to 2019. Sixty-seven preclinical studies and 11 clinical trials were recognized as eligible. Although preclinical studies identified specific key ILs which promote breast cancer bone metastases, which have pro-metastatic effects (e.g., IL-6, IL-8, IL-1β, IL-11), and whose inhibition also shows potential preclinical therapeutic effects, the clinical trials focused principally on ILs (IL-2 and IL-12), which have an anti-metastatic effect and a potential to generate a localized and systemic antitumor response. However, these clinical trials are yet to post any results or conclusions. This inconsistency indicates that further studies are necessary to further develop the understanding of cellular and molecular relations, as well as signaling pathways, both up- and downstream of ILs, which could represent a novel strategy to treat tumors that are resistant to standard care therapies for patients affected by breast cancer bone disease