Covid-19, Variants and Quadramune...

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TSOI_Ry
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Covid-19, Variants and Quadramune...

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As I speak with friends and family about the benefits of Quadramune, I’m finding that those who are vaccinated re CV19 ask a similar version of the same question: “I’m vaccinated, why would I need to take Quadramune?” For those, like myself, who have experienced the benefits of QM (im not vaccinated) what is the most concise/best answer to this question, and why is buttressing our immune systems paramount EVEN (and maybe even particularly) if vaccinated. This is a concern as reports of hundreds if not thousands of variants replicate w increasing speed.
2nd Q: Are present CV19 vaccinations a benefit/false sense of security re: Variants?

3rd Q: What's the relationship between mRNA vaccines and variant replication? If mRNA Vaccines don’t “kill” the virus, what exactly is accomplished?
4th Q: The early introduction of booster shots seems to highlight an antibody sustainability issue. Do flu/virus vaccines prevent or dissuade our immune systems from addressing vira loads as intended?

An argument is often made raising the Polio Vaccine when discussing the efficacy/safety of mRNA vaccines. Maybe you could briefly explain the difference for those of us who find this comparison intellectually impotent.


Thx Tim! 😁

Ryan
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TimGDixon
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Re: Covid-19, Variants and Quadramune...

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Hi Ryan,

Good questions.
1) QuadraMune is a serious immune booster in not only unvaccinated people but vaccinated as well. Take away covid-19 for the moment. Our bodies are under constant assault from the environments we live, work, eat and sleep in. By taking QM you keep your immune system in a state of vigilance rather than wait until something comes along and then try and treat it with QM, which may be just as beneficial but you may have prevented that illness in the first place. This doesn't mean QM is for everyone and each person needs to decide. Maybe they have a really good diet rich with poly-phenols, antioxidants and anti-inflammatories already, but most of us don't. We eat on the go, we run ourselves ragged, we don't drink enough water and we breathe bad air.

2) We have a lot of questions about the longevity of certainly the Pfizer vaccine since we are seeing many breakthrough cases. Currently there is no covid test that will tell you which variant you have, they are either positive to sars/cov2 or not. I personally know 3 people right now with breakthrough infections who were vaccinated with Pfizer.

3) There is some thought that something we call antibody enhanced dependency (AED) is occurring in Pfizer vaccinated people, that is, the vaccine itself is facilitating the new infection. On the other hand it's also quite possible that they made a dosing error that is now locked into the manufacturing and administration, meaning there is not enough of the "S" protein (surface glycoprotein) in the vaccine to stimulate long term immune system memory. This is why they are trying to push boosters but FDA said no under age 65. So for now Ryan we just cant say either way, all the more reason to take QM.

4) is related to 3 but let me continue. The vaccine doesn't kill anything. It exposes the immune system to the foreign protein and the immune says hey you guys this is a bad pathogen send out NK cells, t cells, b cells, make tolerogenic memory and produce neutralizing antibodies (yes some little immune guy in a control room sent that message) - all of that "kills" the virus. No one who has been sick and survived did so without their immune system killing it - some need significant help to achieve that and others, young people for example, had no symptoms but now have self-immunity to sars/cov2.

So in the case of a single person who gets covid-19 and survives that person survived because his/her immune system mounted an attack and killed the virus, recorded the event, and is now vigilant to a future exposure/infection. In some ways the same applies to getting the vaccine, you have some memory to ward off future infection/exposure and if they get sick their immune system will mount a new attack. Jonas Salk knew that if polio infected the human host then it didn't matter if they obtained some future immunity because its too late, polio has done the damage and their is no reversing that, so Salk knew his vaccine had to stoop it dead in its track the first time a human was exposed. You only got one polio shot and it lasts a lifetime - that is the definition of a vaccine to me.

Hope that answer your Q's.
Tim
TSOI_Ry wrote: Sat Sep 18, 2021 4:44 am As I speak with friends and family about the benefits of Quadramune, I’m finding that those who are vaccinated re CV19 ask a similar version of the same question: “I’m vaccinated, why would I need to take Quadramune?” For those, like myself, who have experienced the benefits of QM (im not vaccinated) what is the most concise/best answer to this question, and why is buttressing our immune systems paramount EVEN (and maybe even particularly) if vaccinated. This is a concern as reports of hundreds if not thousands of variants replicate w increasing speed.
2nd Q: Are present CV19 vaccinations a benefit/false sense of security re: Variants?

3rd Q: What's the relationship between mRNA vaccines and variant replication? If mRNA Vaccines don’t “kill” the virus, what exactly is accomplished?
4th Q: The early introduction of booster shots seems to highlight an antibody sustainability issue. Do flu/virus vaccines prevent or dissuade our immune systems from addressing vira loads as intended?

An argument is often made raising the Polio Vaccine when discussing the efficacy/safety of mRNA vaccines. Maybe you could briefly explain the difference for those of us who find this comparison intellectually impotent.


Thx Tim! 😁

Ryan
curncman
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Does vehicle-based delivery of mesenchymal stromal cells give superior results in knee osteoarthritis? Meta-analysis of

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Does vehicle-based delivery of mesenchymal stromal cells give superior results in knee osteoarthritis? Meta-analysis of randomized controlled trials

https://www.journal-cot.com/article/S09 ... X/fulltext

Abstract
Study design
Meta-analysis.
Objectives
We aim to analyze and compare the efficacy and safety of vehicle-based delivery of Mesenchymal Stromal Cells (MSCs) in the management of osteoarthritis of the knee from Randomized Controlled Trials (RCTs) available in the literature.
Materials and methods
We conducted independent and duplicate electronic database searches including PubMed, Embase, Web of Science, and Cochrane Library till August 2021 for RCTs analyzing the efficacy and safety of vehicle-based delivery of MSCs in the management of knee osteoarthritis. Visual Analog Score (VAS) for Pain, Western Ontario McMaster Universities Osteoarthritis Index (WOMAC), Magnetic Resonance Observation of Cartilage Repair Tissue(MOCART) score, and adverse events were the outcomes analyzed. Analysis was performed in R-platform using OpenMeta [Analyst] software.
Results
21 studies involving 936 patients were included for analysis. None of the studies made a direct comparison of the direct and vehicle-based delivery of MSCs, hence we pooled the results of all the included studies of both groups and made a comparative analysis of their outcomes. Although at 6 months, both direct and vehicle-based delivery of MSCs showed significantly better VAS improvement (p = 0.002, p = 0.010), it was not consistent at 1 year for the vehicle delivery (p = 0.973). During 6 months and 12 months, direct delivery of MSCs (p < 0.001, p < 0.001) outperformed vehicle delivery (p = 0.969, p = 0.922) compared to their control based on WOMAC scores respectively. Both direct (p = 0.713) and vehicle-based delivery (p = 0.123) of MSCs did not produce significant adverse events compared to their controls.
Conclusion
Our analysis of literature showed that current clinically employed methods of vehicle-based delivery of MSCs such as platelet-rich plasma, hyaluronic acid did not demonstrate superior results compared to direct delivery, concerning the efficacy of treatment measured by improvement in pain, functional outcomes, and safety. Hence, we urge future clinical trials to be conducted to validate the effectiveness of advanced delivery vehicles such as composite bioscaffolds to establish their practical utility in cartilage regeneration with respect to its encouraging in-vitro evidence.
Keywords
Mesenchymal stem cell
Bone-marrow derived mesenchymal stem cell
Adipose-derived mesenchymal stem cell
Cartilage regeneration
Knee osteoarthritis
Meta-analysis
1. Introduction
With the advancement in regenerative and translational medicine, Cartilage Tissue Engineering (CTE) has revolutionized the area of regenerating cartilage since cartilage has less intrinsic potential for regenerating once it is injured. CTE is defined as either in-vitro or in-vivo regeneration of cartilage tissue to restore the native joint homeostasis.1 The integral part of CTE are mesenchymal stromal cells, biomicromolecules (growth factors, cytokines), and scaffolds. The ease of availability from multiple sources makes mesenchymal stromal cells (MSCs) the versatile cell-based therapy of choice in regenerating the cartilage.2 CTE involves the augmentation of injured cartilage with mesenchymal stromal cells (autologous/allogenic, uncultured/culture-expanded), biocompatible scaffolds, and growth factors & cytokines.
Various researchers have studied various methods of delivery of orthobiologics into knee joint (intra-articular, subchondral, intra-osseous, and biomaterial-based delivery) for regenerating the degenerated cartilaginous tissue in knee osteoarthritis (OA) most effectively.3, 4, 5 Once administered, MSCs undergo radical biological and morphological changes and disappear from the site of injection within a few days of injection.6 To regenerate cartilage, the environment needs a continuous release of biological peptides such as growth factors and cytokines to exert the paracrine signaling of MSCs to differentiate into chondrocytes. To achieve a boosted continuous, pulsatile, and prolonged release of biological peptides from MSCs, vehicles are used along with MSCs to augment their action in the native environment for a longer period. Vehicles such as platelet-rich plasma (PRP) are loaded with growth factors that were considered to lead to assisted cartilage repair and an increased rate of new chondrocytes integration into the surrounding tissues.
Literature evidence demonstrated pros and cons in the usage of biomaterials to deliver MSCs in cartilage regeneration. Numerous preclinical trials have demonstrated the use of biomaterial-based MSC delivery to the injured cartilage resulted in chondrocyte proliferation and differentiation.7, 8, 9, 10 Condello et al. demonstrated the safety and efficacy of type I equine collagen and magnesium-enriched hydroxyapatite scaffold in osteochondral knee lesions in patients with early OA, with high patient satisfaction and low complication rate.11 Kremen et al. in Yucatan mini pigs with post-traumatic OA, made an intraarticular injection of 107 human bone marrow-derived mesenchymal stromal cells (BM-MSCs) with fibrin and found successful chondral regeneration.12 Chitosan hydrogel/3D-printed poly(ε‐caprolactone) construct loaded with synovial MSCs and recruiting tetrahedral framework nucleic acid was injected into the articular cavity and found to result in enhanced chondrogenesis.13 Adipose-derived mesenchymal stromal cells (AD-MSCs) admixed PRP has proved to be beneficial in surgically induced OA knee in Beagle dogs via inhibition of inflammation, induction of extracellular matrix (ECM) synthesis, and chondrocyte proliferation.14 Rajagopal et al. emphasized that MSCs loaded with multi-layered chitosan-gelatin scaffolds result in superior chondrogenesis than single-layered scaffoldings.15
Although the use of biomaterial with a structural matrix to hold the cells at the target site has been considered superior to the use of vehicle-based delivery, the use of vehicles such as PRP, hyaluronic acid (HA), human albumin, human serum remains prevalent in clinical practice. The long-term results in the usage of vehicles in delivering MSCs are still in debate. The success of CTE relies on tissue ergonomics until the cartilaginous tissue gets healed completely. Since most of the scaffolds that are being developed and tested in animal models are yet to come into clinical practice, the PRP and HA remain the commonly used vehicles on which the MSCs are delivered.16,17 Despite its wider usage, clinical efficacy and proven benefits of its use is not yet estimated. To arrive at a consensus on its use and effectiveness in CTE, we aim to analyze the clinical efficacy and patient safety with the currently used vehicles in the delivery of MSCs in the management of osteoarthritis of the knee.
2. Materials & methods
This meta-analysis was conducted following the guidelines of the Back Review Group of Cochrane Collaboration18 and reported based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement.19
2.1 Search strategy
Two reviewers performed an independent electronic literature search for studies evaluating the safety and efficacy of vehicle-based delivery of MSCs in the management of osteoarthritis of the knee. We searched the following databases: PubMed, Embase, Web of Science, and the Cochrane Library up to August 2021. No language or date restrictions were applied. Keywords used for the search were as follows: “Knee Osteoarthritis”, “Knee Degeneration”, “Stem Cell Therapy” and “Mesenchymal Stromal Cells”, “Bone marrow”, “Adipose”. A sample search strategy used in one of the included databases was presented in Supplementary File 1. The reference list of the selected articles was also searched to identify studies not identified in the primary search. As per the inclusion and exclusion criteria, eligible studies were included for meta-analysis. The discrepancy between the authors was resolved through discussion until a consensus was obtained. A detailed study selection flow diagram is given in Fig. 1.
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TimGDixon
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Re: Covid-19, Variants and Quadramune...

Post by TimGDixon »

interesting
Does vehicle-based delivery of mesenchymal stromal cells give superior results in knee osteoarthritis? Meta-analysis of randomized controlled trials
curncman
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Re: Covid-19, Variants and Quadramune...

Post by curncman »

Mesenchymal stem cells after the proprocessing of
tanshinone IIA attenuate cognitive deficits and oxidative
stress injury in an amyloid β-peptide (25–35)-induced rodent
model of Alzheimer’s disease
Attachments
Mesenchymal_stem_cells_after_the_proprocessing_of.6.pdf
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curncman
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A comparative study of mesenchymal stems cells cultured as cell-only aggregates and in encapsulated hydrogels

Post by curncman »

A comparative study of mesenchymal stems cells cultured as cell-only aggregates and in encapsulated hydrogels
https://cris.maastrichtuniversity.nl/en ... -as-cell-o
curncman
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New Imaging Tracking System Could Clarify Stem Cell Role in IPF Therapy

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New Imaging Tracking System Could Clarify Stem Cell Role in IPF Therapy

https://www.rarediseaseadvisor.com/news ... f-therapy/
curncman
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The Current Role of Stem Cell Therapy and iPS Cells

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The Current Role of Stem Cell Therapy and iPS Cells

https://link.springer.com/chapter/10.10 ... 79485-9_15
curncman
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Two Hits for Bone Regeneration in Aged Patients: Vertebral Bone Marrow Clot as a Biological Scaffold and Powerful Source

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

https://www.frontiersin.org/articles/10 ... 07679/full

www.frontiersin.orgFrancesca Salamanna1, www.frontiersin.orgDeyanira Contartese1*, www.frontiersin.orgVeronica Borsari1, www.frontiersin.orgStefania Pagani1, www.frontiersin.orgGiovanni Barbanti Brodano2, www.frontiersin.orgCristiana Griffoni2, www.frontiersin.orgAlessandro Ricci3, www.frontiersin.orgAlessandro Gasbarrini2 and www.frontiersin.orgMilena Fini1
1Complex Structure Surgical Sciences and Technologies, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy
2Department of Oncological and Degenerative Spine Surgery, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy
3Anesthesia-Resuscitation and Intensive Care, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy
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.

Introduction
Bone has well-documented natural healing properties which however change throughout life with aging (Boskey and Coleman, 2010; Curtis et al., 2015). Aging is accompanied by the increased incidence of bone diseases and reduced fracture-healing capacity, which require successful therapies able to enhance bone healing and regeneration (Boskey and Coleman, 2010; Curtis et al., 2015). In this context, the therapeutic potential of adult mesenchymal stem cells (MSCs) for bone repair has been long proposed and investigated (Shang et al., 2021). MSCs from several sources have been employed in the field of bone regeneration including bone marrow (BM), adipose tissue, umbilical cord, and dental-related tissues (Shang et al., 2021). BM-MSCs were the first identified and, to date, represent the most commonly used MSCs source for bone regeneration (Shang et al., 2021). These cells can be implanted after culture expansion or injected as whole bone marrow aspirate (BMA) or BM concentrate (BMC) (Shang et al., 2021). Considering regulation on in vitro cell processing, the use of cultured BM-MSCs in Europe and in the United States is restricted. Conversely, whole BMA or BMC involves minimal cell manipulation and can be used in clinical practice to treat bone diseases in a “one-step” procedure (Hernigou et al., 2005). Although only 0.01–0.001% of MSCs is found among the totality of mononuclear cells in BMA, the concurrent presence of nonadherent osteogenic cells and the establishment of cell–cell interactions suggest that the use of whole BMA, instead of BMC or expanded and purified MSCs, is preferable for bone cell therapy (Hernigou et al., 2005). Recently, our research group also demonstrated that MSCs derived from human clotted BMA have higher growth kinetics in comparison to MSCs derived from human un-clotted BMA (whole and concentrate) as well as higher growth factors expression (transforming growth factor beta, TGF-β; vascular-endothelial growth factor, VEGF-A; fibroblast growth factor 2, FGF2) and higher ability to differentiate toward the osteogenic and chondrogenic phenotype (Salamanna et al., 2020). These data were also confirmed by an in vivo study by Lim and colleagues that underlined that the therapeutic potential of autologous bone graft and BMA clot in the repair of ulnar defects are similar, with the advantage of BMA clot being associated with a lower risk of complications (Lim et al., 2019). A recent literature review further elucidates the characteristics found in BMA clot and its application in the clinical scenario (Santos Duarte Lana et al., 2020). In this context, to confirm and strengthen data on the clinical use of BMA clot for bone regeneration, a pilot clinical study on clotted vertebral BMA is ongoing at our institution (Protocol: CVOD. coagulo_CE-AVEC 587/2020/Sper/IORS). Although BMA clot has key properties and characteristics that make it a promising cell therapy strategy for bone repair and regeneration, aging could be a critical point as aging-associated processes could impact on the biological properties and clinical efficacy of BMA clot (D’Ippolito et al., 1999). During aging, resident MSCs inside the BM are affected by both intrinsic and extrinsic factors, which alter their functions (D’Ippolito et al., 1999). Aging effects on BM-MSC properties, such as telomere length, cell proliferation, differentiation ability, epigenetics, and secretome, have been discussed and demonstrated in several preclinical studies and systematic reviews (Boyette and Tuan, 2014; Baker et al., 2015; Charif et al., 2017; Ganguly et al., 2017). Aging also decreases the expression level of FGF2 which in turn leads to the reduction of BM-MSCs proliferative capacity (Gibon et al., 2016). Additionally, aging reduces the density of MSCs in BM and compromises their osteogenic potential (D’Ippolito et al., 1999). Interestingly, a study by D’Ippolito et al. showed that in human vertebral BM, the number of MSCs with osteogenic potential decreases early during aging, which may be responsible for the age-related reduction in bone formation, mechanical properties, and bone integrity (D’Ippolito et al., 1999). Increasing evidence also suggests that the aging BM-MSCs promote adipogenesis at the expense of osteogenesis, thus resulting in decreased bone formation capacity (Marędziak et al., 2016). The secretion of senescence-associated secretory phenotype (SASP) products from senescent MSCs in BM has also been evidenced during aging (Turinetto et al., 2016; Ma et al., 2018; Malaise et al., 2019). Interleukin-6 (IL-6), one of the most critical SASP factors, has been shown to drive osteoclastogenesis and negatively regulate osteoblast differentiation (Turinetto et al., 2016; Ma et al., 2018). The knockdown of IL-6 significantly enhanced runt-related transcription factor 2 (Runx2) and collagen type I alpha 1 chain (COL1A1) expression in osteoblasts while decreasing the expression of osteoclast-related genes such as tartrate-resistant alkaline phosphatase (TRAP), metalloproteinase (MMP)-9, and cathepsin K (CTSK) (Turinetto et al., 2016; Ma et al., 2018; Malaise et al., 2019). Furthermore, a contribution to MSCs senescence is also given by the Klotho gene, a gene that encodes Klotho antiaging protein (Kuro-o, 2012). Loss-of-function mutation of Klotho in mice leads to a syndrome resembling human premature aging, including defects in skeletal mineralization and osteoporosis (Kuro-o, 2012) scenario. These age-related factors could limit the use of BM stem cell as cell-based therapy in the clinical scenario.

Considering that MSCs from clotted vertebral BMA showed better biological properties in comparison to the whole and concentrated BM, it represents a point-of-care orthobiologic product that uniquely and simultaneously delivers growth factors and MSCs (Salamanna et al., 2020). The goal of this study was to evaluate the influence of donor age on human vertebral clotted BMA, one of the skeletal sites with the highest bone turnover, characterizing and comparing the biological properties of MSCs from younger (<45 years) and older female donors (>65 years).

Materials and Methods
Bone Marrow Clot Collection
The study, which is part of an ongoing pilot clinical study on the efficacy of clotted vertebral BMA in spinal fusion surgery, was approved by the Local Ethics Committee of the Emilia Romagna Region (Comitato Etico Indipendente Area Vasta Emilia Centro, Protocol n. CE-AVEC 587/2020/Sper/IOR S) and was carried out in accordance with relevant guidelines and regulations (IRCCS Istituto Ortopedico Rizzoli has kept the ISO 9001 Quality certification since 2008). Written informed consent was obtained from all subjects involved in the study. Inclusion criteria were patients with degenerative spinal diseases submitted to multilevel instrumented arthrodesis (minimum 2–maximum 6 levels), patients of legal age, and patients able to give their consent and answer self-administered questionnaires. Exclusion criteria were human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), diabetes, pregnancy, bone diseases, drugs active on bone metabolism, primary bone tumors, metastases, minors, and/or patients incapable of giving consent personally. Human BMA was harvested from vertebral pedicles of six female patients, 3 younger (mean age: 44 ± 0.5) and 3 older (mean age: 68.6 ± 2.5) (body mass index and bone mineral index matched), undergoing spinal surgical procedures involving posterolateral arthrodesis. None of the patients had a diagnosis of osteoporosis. Eight milliliters (1 or 2 ml from each vertebra depending on the length of the arthrodesis) of human vertebral BMA were harvested into a 10-ml syringe during the preparation of the pilot hole for pedicle screw fixation. Subsequently, BMA was divided into two equal parts of 4 ml each in a sterile disposable container without any anticoagulant and immediately transferred to the laboratory. BMA without anticoagulant clotted in about 15–30 min and was transferred in culture flasks with Dulbecco’s modified Eagles medium (DMEM, Sigma–Aldrich, St. Louis, MO), containing 10% fetal bovine serum (FBS, Lonza), 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco, Life Technologies, Carlsbad, CA), and 5 μg/ml plasmocin (Invivogen, San Diego, CA) (Figure 1). The cultures were incubated at 37°C in 5% CO2 and under hypoxia (2% O2), to better mimic the microenvironment of the stem cells’ niche. Culture media was changed every three days.
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