Human Osteoblasts' Response to Biomaterials for Subchondral Bone Regeneration in Standard and Aggressive Environments.

Osteochondral lesions, when not properly treated, may evolve into osteoarthritis (OA), especially in the elderly population, where altered joint function and quality are usual. To date, a collagen/collagen-magnesium-hydroxyapatite (Col/Col-Mg-HAp) scaffold (OC) has demonstrated good clinical results, although suboptimal subchondral bone regeneration still limits its efficacy. This study was aimed at evaluating the in vitro osteogenic potential of this scaffold, functionalized with two different strategies: the addition of Bone Morphogenetic Protein-2 (BMP-2) and the incorporation of strontium (Sr)-ion-enriched amorphous calcium phosphate (Sr-ACP) granules. Human osteoblasts were seeded on the functionalized scaffolds (OC+BMP-2 and OC+Sr-ACP, compared to OC) under stress conditions reproduced with the addition of H2O2 to the culture system, as well as in normal conditions, and evaluated in terms of morphology, metabolic activity, gene expression, and matrix synthesis. The OC+BMP-2 scaffold supported a better osteoblast morphology and stimulated scaffold colonization, cell activity, and extracellular matrix secretion, especially in the stressed culture environment but also in normal culture conditions, with increased expression of genes related to osteoblast differentiation. In conclusion, the incorporation of BMP-2 into the Col/Col-Mg-HAp scaffold also represents an improvement of the osteochondral scaffold in more challenging conditions, supporting further preclinical studies to optimize it for use in clinical practice.

Gender-Specific Differences in Human Vertebral Bone Marrow Clot.

Recently, our group described the application of vertebral bone marrow (vBMA) clot as a cell therapy strategy for spinal fusion. Its beneficial effects were confirmed in aging-associated processes, but the influence of gender is unknown. In this study, we compared the biological properties of vBMA clots and derived vertebral mesenchymal stem cells (MSCs) from female and male patients undergoing spinal fusion procedures and treated with vBMA clot. We analyzed the expression of growth factors (GFs) in vBMA clots and MSCs as well as morphology, viability, doubling time, markers expression, clonogenicity, differentiation ability, senescence factors, Klotho expression, and HOX and TALE gene profiles from female and male donors. Our findings indicate that vBMA clots and derived MSCs from males had higher expression of GFs and greater osteogenic and chondrogenic potential compared to female patients. Additionally, vBMA-clot-derived MSCs from female and male donors exhibited distinct levels of HOX and TALE gene expression. Specifically, HOXA1, HOXB8, HOXD9, HOXA11, and PBX1 genes were upregulated in MSCs derived from clotted vBMA from male donors. These results demonstrate that vBMA clots can be effectively used for spinal fusion procedures; however, gender-related differences should be taken into consideration when utilizing vBMA-clot-based studies to optimize the design and implementation of this cell therapy strategy in clinical trials.

Anti-Oxidant Multi-Functionalized Materials: Strontium-Substituted Monetite and Brushite as Delivery Systems for Curcumin.

Curcumin has numerous biological activities and pharmaceutical applications related to its ability to inhibit reactive oxygen species. Herein, strontium-substituted monetite (SrDCPA) and strontium-substituted brushite (SrDCPD) were synthesized and further functionalized with curcumin with the aim to develop materials that combine the anti-oxidant properties of the polyphenol, the beneficial role of strontium toward bone tissue, and the bioactivity of calcium phosphates. Adsorption from hydroalcoholic solution increases with time and curcumin concentration, up to about 5-6 wt%, without affecting the crystal structure, morphology, and mechanical response of the substrates. The multi-functionalized substrates exhibit a relevant radical scavenging activity and a sustained release in phosphate buffer. Cell viability, morphology, and expression of the most representative genes were tested for osteoclast seeded in direct contact with the materials and for osteoblast/osteoclast co-cultures. The materials at relatively low curcumin content (2-3 wt%) maintain inhibitory effects on osteoclasts and support the colonization and viability of osteoblasts. The expressions of Alkaline Phosphatase (ALPL), collagen type I alpha 1 chain (COL1A1), and osteocalcin (BGLAP) suggest that curcumin reduces the osteoblast differentiation state but yields encouraging osteoprotegerin/receptor activator for the NFkB factor ligand (OPG/RANKL) ratio.

Evaluating Mentors in Violence Prevention: A Longitudinal, Multilevel Assessment of Outcome Changes.

There is a need to increase understanding of the effectiveness of bystander programmes targeting gender-based violence in the United Kingdom. There is also a need to utilise a robust theoretical models of decision-making while doing so. Changes were examined in bystanders' attitudes, beliefs, motivations towards intervening, and intervention behavior in situations of gender-based violence. To achieve this, a quantitative examination of Mentors in Violence Prevention was conducted. There were 1396 participants (50% female, 50% male) who were aged 11 to 14 years old (M = 12.25, SD = 0.84) attending high school at the first time point. Participants were attending 17 schools (53% Mentors in Violence Prevention and 47% control) in Scotland. Outcome variables were assessed approximately one year apart using questionnaires. Multilevel linear regressions revealed that Mentors in Violence Prevention did not change outcomes reflecting bystanders' attitudes, beliefs, motivations towards intervening, or intervention behavior in gender-based violence. Discrepancies between the current findings and those of other evaluations may be due to other studies including small numbers of schools that may be more motivated to implement the program. This study also identified two key issues that need to be addressed at stakeholder level before concluding that Mentors in Violence Prevention is ineffective at targeting gender-based violence. That the program has moved towards a more gender-neutral approach in the United Kingdom could explain the null results of this study. Furthermore, the current findings could be attributed to a failure to adequately address the theoretical model underpinning the program in practice.

RNA Extraction from Cartilage: Issues, Methods, Tips.

The increase in degenerative diseases involving articular cartilage has pushed research to focus on their pathogenesis and treatment, exploiting increasingly complex techniques. Gene expression analyses from tissue are representative of the in vivo situation, but the protocols to be applied to obtain a reliable analysis are not completely cleared through customs. Thus, RNA extraction from fresh samples and specifically from musculoskeletal tissue such as cartilage is still a challenging issue. The aim of the review is to provide an overview of the techniques described in the literature for RNA extraction, highlighting limits and possibilities. The research retrieved 65 papers suitable for the purposes. The results highlighted the great difficulty in comparing the different studies, both for the sources of tissue used and for the techniques employed, as well as the details about protocols. Few papers compared different RNA extraction methods or homogenization techniques; the case study reported by authors about RNA extraction from sheep cartilage has not found an analog in the literature, confirming the existence of a relevant blank on studies about RNA extraction from cartilage tissue. However, the state of the art depicted can be used as a starting point to improve and expand studies on this topic.

Evaluating the Mentors in Violence Prevention Program: A Process Examination of How Implementation Can Affect Gender-Based Violence Outcomes.

Gender-based violence is a global public health issue and major human rights concern. It is also a type of violence that is disproportionately experienced by women and girls. This study is the first to examine multiple implementation process (dosage, fidelity, and adaptation) effects on changes in anticipated outcomes of a school-based bystander program targeting gender-based violence, Mentors in Violence Prevention (MVP). Data were collected from two participant groups: mentees (students receiving MVP) and mentors (students delivering MVP), across nine participating high schools. The mentee sample comprised 698 students (about 48.9% males and 49.7% females), aged 11 to 14 years old (M = 11.86, SD = 0.64). The mentor sample comprised 118 students (17.80% males, 82.20% females), aged 15 to 18 years old (M = 16.42, SD = 0.60). Anticipated outcomes were changes in bystanders' attitudes, social influences, control perceptions, intentions, willingness, and intervention behavior, measured using mentees' self-reports at two time points approximately 1 year apart. Implementation processes were measured using mentors' self-reports. Analyses revealed no effects for any of the implementation variables across changes in any of the outcomes measured. These results highlight important implications for the implementation of the MVP program going forward, given its widespread implementation in the United Kingdom. Possible ways that MVP may be enhanced in future are discussed. For example, furthering understanding into how gender-based violence and bystander intervention are addressed and framed during MVP lessons would give more insight into how the current implementation of the program can be improved to maximize its potential benefits.

Ageing and Osteoarthritis Synergically Affect Human Synoviocyte Cells: An In Vitro Study on Sex Differences.

Osteoarthritis is a chronic inflammatory disease that affects all of the joints, especially those of the elderly. Aging is a natural and irreversible biological process implicated in the pathophysiology of many chronic diseases, such as osteoarthritis. Inflammation and oxidative stress are the main factors involved in osteoarthritis and aging, respectively, with the production of several pro-inflammatory cytokines such as Interleukin 1ß (IL1ß) and reactive oxygen species. The aim of the study was to set-up an in vitro model of osteoarthritis and aging, focusing on the sex differences by culturing male and female fibroblast-like synoviocytes (FLSs) with IL1ß, hydrogen peroxide (H2O2), IL1ß+H2O2 or a growth medium (control). IL1ß+H2O2 reduced the cell viability and microwound healing potential, increased Caspase-3 expression and reactive oxygen species and IL6 production; IL1ß increased IL6 production more than the other conditions did; H2O2 increased Caspase-3 expression and reactive oxygen species production; Klotho expression showed no differences among the treatments. The FLSs from female donors demonstrated a better response capacity in unfavorable conditions of inflammation and oxidative stress than those from the male donors did. This study developed culture conditions to mimic the aging and osteoarthritis microenvironment to evaluate the behavior of the FLSs which play a fundamental role in joint homeostasis, focusing on the sex-related aspects that are relevant in the osteoarthritis pathophysiology.

Monetite vs. Brushite: Different Influences on Bone Cell Response Modulated by Strontium Functionalization.

Monetite and brushite are regarded with increasing interest for the preparation of biomaterials for applications in the musculoskeletal system. Herein, we investigated the influence of strontium substitution in the structures of these two phosphates on bone cell response. To achieve this aim, co-cultures of human primary osteoclasts and human osteoblast-like MG63 cells were tested on strontium-substituted monetite and strontium-substituted brushite, as well as on monetite and brushite, as controls. In both structures, strontium substitution for calcium amounted to about 6 at% and provoked enlargement of the cell parameters and morphologic variations. Cumulative release in physiological solution increased linearly over time and was greater from brushite (up to about 160 and 560 mg/L at 14 days for Sr and Ca, respectively) than from monetite (up to about 90 and 250 mg/L at 14 days for Sr and Ca, respectively). The increasing viability of osteoblast-like cells over time, with the different expression level of some typical bone markers, indicates a more pronounced trigger toward osteoblast differentiation and osteoclast inhibition by brushite materials. In particular, the inhibition of cathepsin K and tartrate-resistant acid phosphatase at the gene and morphological levels suggests strontium-substituted brushite can be applied in diseases characterized by excessive bone resorption.

Bystander intervention among secondary school pupils: Testing an augmented Prototype Willingness Model.

This study augmented the Prototype Willingness Model (PWM) to assess reactive and deliberative decision-making underpinning bystander intervention in gender-based violence contexts. There were 2079 participants (50% male, 49% female, and 1% unreported), aged 11-15 years old (M = 12.32, SD = 0.91), attending 19 secondary schools across Scotland. Participants self-reported the augmented PWM variables, then their intervention behaviour approximately 1 month later. Path analyses mostly supported the predicted relationships between positive and negative bidimensional attitudes, subjective norms, prototype perceptions, perceived behavioural control, and self-efficacy on intentions and willingness. Willingness predicted positive (speaking with a teacher) and negative (doing nothing) intervention in less serious violence. Self-efficacy predicted negative intervention in more serious violence. Subjective norms positively moderated the attitudes-intentions relationship. Overall, the results suggested that reactive (willingness) more so than deliberative (intention) decision-making account for intervention when young people witness gender-based violence. Additionally, the findings highlight the complexity of bystander intervention decision-making, where adding control perceptions, bidimensional attitudes, and moderators have independent contributions. Furthermore, self-comparison to the typical bystander who positively intervenes (prototype perceptions) was the strongest predictor of intentions and willingness, highlighting in a novel way the importance of image and group membership on decision-making.

Gender and Sex Are Key Determinants in Osteoarthritis Not Only Confounding Variables. A Systematic Review of Clinical Data.

Many risk factors for osteoarthritis (OA) have been noted, while gender/sex differences have been understated. The work aimed to systematically review literature investigating as primary aim the relationship between gender/sex related discriminants and OA. The search was performed in PubMed, Science Direct and Web of Knowledge in the last 10 years. Inclusion criteria were limited to clinical studies of patients affected by OA in any joints, analyzing as primary aim gender/sex differences. Exclusion criteria were review articles, in vitro, in vivo and ex vivo studies, case series studies and papers in which gender/sex differences were adjusted as confounding variable. Of the 120 records screened, 42 studies were included. Different clinical outcomes were analyzed: morphometric differences, followed by kinematics, pain, functional outcomes after arthroplasty and health care needs of patients. Women appear to use more health care, have higher OA prevalence, clinical pain and inflammation, decreased cartilage volume, physical difficulty, and smaller joint parameters and dimensions, as compared to men. No in-depth studies or mechanistic studies analyzing biomarker differential expressions, molecular pathways and omic profiles were found that might drive preclinical and clinical research towards sex-/gender-oriented protocols.

Mechanical interaction between additive-manufactured metal lattice structures and bone in compression: implications for stress shielding of orthopaedic implants.

One of the main biomechanical causes for aseptic failure of orthopaedic implants is the stress shielding. This is caused by an uneven load distribution across the bone normally due to a stiff metal prosthesis component, leading to periprosthetic bone resorption and to implant loosening. To reduce the stress shielding and to improve osseointegration, biocompatible porous structures suitable for orthopaedic applications have been developed. Aim of this study was to propose a novel in-vitro model of the mechanical interaction between metal lattice structures and bovine cortical bone in compression. Analysis of the strain distribution between metal structure and bone provides useful information on the potential stress shielding of orthopaedic implants with the same geometry of the porous scaffold. Full density and lattice structures obtained by the repetition of 1.5 mm edge cubic elements via Laser Powder Bed Fusion of CoCrMo powder were characterized for mechanical properties using standard compressive testing. The two porous geometries were characterized by 750 µm and 1000 µm pores resulting in a nominal porosity of 43.5% and 63.2% respectively. Local deformation and strains of metal samples coupled with fresh bovine cortical bone samples were evaluated via Digital Image Correlation analysis up to failure in compression. Visualization and quantification of the local strain gradient across the metal-bone interface was used to assess differences in mechanical behaviour between structures which could be associated to stress-shielding. Overall stiffness and local mechanical properties of lattice and bone were consistent across samples. Full-density metal samples appeared to rigidly transfer the compression force to the bone which was subjected to large deformations (2.2 ± 0.3% at 15 kN). Larger porosity lattice was associated to lower stiffness and compressive modulus, and to a smoother load transfer to the bone. While tested on a limited sample size, the proposed in-vitro model appears robust and repeatable to assess the local mechanical interaction of metal samples suitable for orthopaedic applications with the bone tissue. CoCrMo scaffolds made of 1000 µm pores cubic cells may allow for a smoother load transfer to the bone when used as constitutive material of orthopaedic implants.

Mechanical and in vitro biological properties of uniform and graded Cobalt-chrome lattice structures in orthopedic implants.

Human bones are biological examples of functionally graded lattice capable to withstand large in vivo loading and allowing optimal stress distribution. Disruption of bone integrity may require biocompatible implants capable to restore the original bone structure and properties. This study aimed at comparing mechanical properties and biological behavior in vitro of uniform (POR-FIX) and graded (POR-VAR) Cobalt-chrome alloy lattice structures manufactured via Selective Laser Melting. In compression, the POR-VAR equivalent maximum stress was about 2.5 times lower than that of the POR-FIX. According to the DIC analysis, the graded lattice structures showed a stratified deformation associated to unit cells variation. At each timepoint, osteoblast cells were observed to colonize the surface and the first layer of both scaffolds. Cell activity was always significantly higher in the POR-VAR (p < 0.0005). In terms of gene expression, the OPG/RANKL ratio increased significantly over time (p < 0.0005) whereas IL1ß and COX2 significantly decreased (7 day vs 1 day; p < 0.0005) in both scaffolds. Both uniform- and graded-porosity scaffolds provided a suitable environment for osteoblasts colonization and proliferation, but graded structures seem to represent a better solution to improve stress distribution between implant and bone of orthopedic implants.

A Social-Ecological Approach to Understanding Adolescent Sexting Behavior.

This study examined the extent to which active and passive sexting behaviors are associated with family-, school-, peer-, and romantic-level variables. Young people (N = 3,322; 49.1% female, 48.3% male, 2.6% other) aged 11 to 15 years old (M = 12.84, SD = 0.89) took part, and all attended mainstream secondary schools in Scotland. Participants completed self-report measures of school connectedness, parental love and support, perceived susceptibility to peer- and romantic-pressure (e.g., to display behaviors just to impress others), and their involvement in active and passive sexting. The importance of both school- and family-level factors was evident, though perceived romantic-pressure had the largest effect. However, neither school- nor family-level variables were moderated by either perceived romantic-pressure or perceived peer-pressure. Efforts to reduce sexting or increase its safety should primarily seek to tackle young people's ability to respond effectively to romantic-pressure. It may also be helpful to develop school connectedness and to help families provide support that is constructive and not intrusive.

Nanostructure and biomimetics orchestrate mesenchymal stromal cell differentiation: An in vitro bioactivity study on new coatings for orthopedic applications.

The choice of the appropriate material having suitable compositional and morphological surface characteristics, is a crucial step in the development of orthopedic implants. The purpose of this paper is to elucidate, on this regard, the influence of two important hits, i.e., biogenic apatite with bone-like composition and nanostructured morphology, providing the evidence of the efficacy of nanostructured biogenic apatite coatings in favoring adhesion, growth, proliferation, and in vitro osteogenic differentiation of human mesenchymal stromal cells (hMSCs) isolated from the bone marrow. The specific features of this coating in terms of topographical and biochemical cues, obtained by Ionized Jet Deposition, are perceived by hMSCs, as suggested by changes in different morphologic parameters as Aspect Ratio or Elongation index, suggesting the impact exerted by the nanostructure on early adhesion events, cytoskeleton organization, and cells fate. In addition, the nanostructured CaP coating sustained the metabolic activity of the cells and facilitated the osteogenic differentiation of MSC by supporting the osteogenesis-related gene expression. These findings support the use of a combined approach between technological advancement and instructive surfaces, both from the topographical and the biochemical point of view, in order to manufacture smart biomaterials able to respond to different needs of the orthopedic practice.

Autologous Protein Solution Effect on Chondrogenic Differentiation of Mesenchymal Stem Cells from Adipose Tissue and Bone Marrow in an Osteoarthritic Environment.

OBJECTIVE: Osteoarthritis (OA) is an inflammatory and degenerative disease, and the numerous treatments currently used are not fully effective. Mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) are proposed for OA treatment as biologic therapies. The aim of the study was to observe the role of autologous protein solution (APS), a type of PRP, on chondrogenic differentiation of 2 types of MSCs, from bone marrow (BMSCs) and adipose tissue (ADSCs), in an in vitro osteoarthritic microenvironment. DESIGN: Inflammatory culture conditions, mimicking OA, were obtained by adding interleukin-1ß (IL-1ß) and tumor necrosis factor α (TNFα), or synovial fluid from patient osteoarthritic knees (OSF), to the culture medium. MSCs were then treated with APS. RESULTS: After 1 month of culture, both cell types formed mature micromasses, partially altered in the presence of IL-1ß and TNFα but quite preserved with OSF. Inflammatory conditions hindered differentiation in terms of gene expression, not counterbalanced by APS. APS triggered type I collagen deposition and above all contributed to decrease the expression of metalloproteinases in the most aggressive conditions (IL-1ß and TNFα in the culture medium). ADSCs originated micromasses more mature and less prone toward osteogenic lineage than BMSCs, thus showing to better adapt in an aggressive environment than BMSC. CONCLUSIONS: APS seems to act better on inflammation front and, between cell types, ADSCs respond better to the inflammatory microenvironment of OA and to the treatment with APS than BMSCs.

Strontium substituted hydroxyapatite with ß-lactam integrin agonists to enhance mesenchymal cells adhesion and to promote bone regeneration.

Multi-functionalization of calcium phosphates to get delivery systems of therapeutic agents is gaining increasing relevance for the development of functional biomaterials aimed to solve problems related to disorders of the muscolo-skeletal system. In this regard, we functionalized Strontium substituted hydroxyapatite (SrHA) with some ß-lactam integrin agonists to develop materials with enhanced properties in promoting cell adhesion and activation of intracellular signaling as well as in counteracting abnormal bone resorption. For this purpose, we selected two monocyclic ß-lactams on the basis of their activities towards specific integrins on promoting cell adhesion and signalling. The amount of ß-lactams loaded on SrHA could be modulated on changing the polarity of the loading solution, from 3.5-24 wt% for compound 1 and from 3.2-8.4 wt% for compound 2. Studies on the release of the ß-lactams from the functionalized SrHA in aqueous medium showed an initial burst followed by a steady-release that ensures a small but constant amount of the compounds over time. The new composites were fully characterized. Co-culture of human primary mesenchymal stem cells (hMSC) and human primary osteoclast (OC) demonstrated that the presence of ß-lactams on SrHA favors hMSC adhesion and viability, as well as differentiation towards osteoblastic lineage. Moreover, the ß-lactams were found to enhance the inhibitory role of Strontium on osteoclast viability and differentiation.

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. None of the therapeutic strategies used to manage breast cancer bone metastases are really curative. Here, we set-up a novel and advanced model by using fresh tissue from human vertebral bone metastasis from breast carcinoma patients able to retain the tumor microenvironment. The tissue model is based on an ex-vivo culture for up to 40 days and on a constant monitoring of tissue viability, gene expression profile (IL10, IL1b, MMP1, MMP7, PTH1R, PTH2R, TNF, ACP5, SPI1, VEGFA, CTSK, TGF-ß) and histological and immunohistochemical analyses (CDH1/E-cadherin, CDH2/N-cadherin, KRT8/Cytokeratin 8, KRT18/Cytokeratin 18, Ki67, CASP3/Caspase 3, ESR1/Estrogen Receptor Alpha, CD68 and CD8). Results confirmed the development of a reliable, reproducible and cost-effective advanced model of breast cancer bone metastasis able to preserve and maintain long-term tissue viability, as well as molecular markers, tissue histomorphology, tissue micro-architecture and antigen expression. The study provides for the first time the feasibility and rationale for the use of a human-derived advanced alternative model for cancer research and testing of drugs and innovative strategies, taking into account patient individual characteristics and specific tumor subtypes so predicting patient specific responses.

Osseointegration of additive manufacturing Ti-6Al-4V and Co-Cr-Mo alloys, with and without surface functionalization with hydroxyapatite and type I collagen.

The introduction of additive manufacturing (AM) technologies has profoundly revolutionized the implant manufacturing industry, with a particularly significant impact on the field of orthopedics. Electron Beam Melting (EBM) and Direct Metal Laser Sintering (DMLS) represents AM fabrication techniques with a pivotal role in the realization of complex and innovative structure starting from virtual 3D model data. In this study, Ti-6Al-4V and Co-Cr-Mo materials, developed by EBM (Ti-POR) and DMLS (Co-POR) techniques, respectively, with hydroxyapatite (Ti-POR + HA; Co-POR + HA) and type I collagen (Ti-POR-COLL; Co-POR-COLL) coatings, were implanted into lateral femoral condyles of rabbits. Osseointegration process was investigated by histological, histomorphometrical and microhardness evaluations at 4 and 12 weeks after implantation. Both Ti-6Al-4V and Co-Cr-Mo implants, with or without HA and COLL coatings, demonstrated good biocompatibility. As expected, HA coating hastened bone-to-implant contact (BIC) process, while collagen did not significantly improved the osseointegration process in comparison to controls. Regarding newly trabecular bone formation (B.Ar/T.Ar), Co-POR presented the highest values, significantly different from those of Co-POR-COLL. Over time, an increase of BIC parameter and a decrease of B.Ar/T.Ar were detected. Higher mineral apposition rate was observed for Ti-POR and Co-POR in comparison to Ti-POR-COLL and Co-POR-COLL, respectively, at 12 weeks. The same behavior was found for bone formation rate between Co-POR and Co-POR-COLL at 12 weeks. In conclusion, the AM materials guarantee a good osseointegration and provide a suitable environment for bone regeneration with the peculiarity of allowing personalized and patient-specific needs customization to further improve the long-term clinical outcomes.

Two Hits for Bone Regeneration in Aged Patients: Vertebral Bone Marrow Clot as a Biological Scaffold and Powerful Source of Mesenchymal Stem Cells.

Recently, the use of a new formulation of bone marrow aspirate (BMA), the BMA clot, has been described. This product entails a naturally formed clot from the harvested bone marrow, which retains all the BMA components preserved in a matrix biologically molded by the clot. Even though its beneficial effects were demonstrated by some studies, the impact of aging and aging-associated processes on biological properties and the effect of BMA cell-based therapy are currently unknown. The purpose of our study was to compare selected parameters and properties of clotted BMA and BMA-derived mesenchymal stem cells (MSCs) from younger (<45 years) and older (>65 years) female donors. Clotted BMA growth factors (GFs) expression, MSCs morphology and viability, doubling time, surface marker expression, clonogenic potential, three-lineage differentiation, senescence-associated factors, and Klotho synthesis from younger and older donors were analyzed. Results indicated that donor age does not affect tissue-specific BMA clot regenerative properties such as GFs expression and MSCs morphology, viability, doubling time, surface antigens expression, colony-forming units, osteogenic and adipogenic differentiation, and Klotho and senescence-associated gene expression. Only few differences, i.e., increased platelet-derived growth factor-AB (PDGF-AB) synthesis and MSCs Aggrecan (ACAN) expression, were detected in younger donors in comparison with older ones. However, these differences do not interfere with all the other BMA clot biological properties. These results demonstrated that BMA clot can be applied easily, without any sample processing and avoiding potential contamination risks as well as losing cell viability, proliferation, and differentiation ability, for autologous transplantation in aged patients. The vertebral BMA clot showed two successful hits since it works as a biological scaffold and as a powerful source of mesenchymal stem cells, thus representing a novel and advanced therapeutic alternative for the treatment of orthopedic injuries.