A Novel Fibrin Matrix Derived from Platelet-Rich Plasma: Protocol and Characterization.

Although fibrin matrices derived from Platelet-Rich Plasma (PRP) are widely used in regenerative medicine, they have some limitations that can hinder their application. Modifying the composition of the PRP-derived fibrin matrix may improve its properties, making it suitable for certain medical uses. Three types of fibrin matrices were obtained: a PRP-derived fibrin matrix (FM), a PRP-derived fibrin matrix with a high fibrinogen content and platelets (FM-HFP) and a PRP-derived fibrin matrix with a high fibrinogen content (FM-HF). The fibrinogen levels, biomechanical properties and cell behavior were analyzed. The presence of platelets in the FM-HFP generated an inconsistent fibrin matrix that was discarded for the rest of the analysis. The fibrinogen levels in the FM-FH were higher than those in the FM (p < 0.0001), with a concentration factor of 6.86 ± 1.81. The values of clotting and swelling achieved using the FM-HF were higher (p < 0.0001), with less clot shrinkage (p < 0.0001). The FM had a significantly higher stiffness and turned out to be the most adherent composition (p = 0.027). In terms of cell viability, the FM-HF showed less cell proliferation but higher live/dead ratio values (p < 0.01). The increased fibrinogen and platelet removal in the FM-HF improved its adhesion and other biomechanical properties without affecting cell viability.

Method to obtain a plasma rich in platelet- and plasma-growth factors based on water evaporation.

Platelet-Rich Plasma, also known as PRP, is an autologous biologic product used in medicine as a treatment for tissue repair. Nowadays, the majority of PRP obtention methods enrich only platelets, not considering extraplatelet biomolecules, which take part in several cell processes. In the present work, a novel PRP preparation method was developed to obtain a PRP rich in both platelet and plasma extraplatelet molecules. The method is based on the evaporation of the water of the plasma using a rotary evaporator. With this new methodology an increase in plasmatic growth factors and, as a consequence, a better dermal fibroblast cell viability was achieved, compared to a standard PRP formulation. This novel PRP product obtained with this new methodology showed promising results in vitro as an improved PRP treatment in future application.

New Formulation of Platelet-Rich Plasma Enriched in Platelet and Extraplatelet Biomolecules Using Hydrogels.

Platelet-rich plasma (PRP) is an autologous biologic product used in several fields of medicine for tissue repair due to the regenerative capacity of the biomolecules of its formulation. PRP consists of a plasma with a platelet concentration higher than basal levels but with basal levels of any biomolecules present out of the platelets. Plasma contains extraplatelet biomolecules known to enhance its regenerative properties. Therefore, a PRP containing not only a higher concentration of platelets but also a higher concentration of extraplatelet biomolecules that could have a stronger regenerative performance than a standard PRP. Considering this, the aim of this work is to develop a new method to obtain PRP enriched in both platelet and extraplatelet molecules. The method is based on the absorption of the water of the plasma using hydroxyethyl acrylamide (HEAA)-based hydrogels. A plasma fraction obtained from blood, containing the basal levels of platelets and proteins, was placed in contact with the HEAA hydrogel powder to absorb half the volume of the water. The resulting plasma was characterized, and its bioactivity was analyzed in vitro. The novel PRP (nPRP) showed a platelet concentration and platelet derived growth factor (PDGF) levels similar to the standard PRP (sPRP), but the concentration of the extraplatelet growth factors IGF-1 (p < 0.0001) and HGF (p < 0.001) were significantly increased. Additionally, the cells exposed to the nPRP showed increased cell viability than those exposed to a sPRP in human dermal fibroblasts (p < 0.001) and primary chondrocytes (p < 0.01). In conclusion, this novel absorption-based method produces a PRP with novel characteristics compared to the standard PRPs, with promising in vitro results that could potentially trigger improved tissue regeneration capacity.

Method Based on Ultrafiltration to Obtain a Plasma Rich in Platelet and Plasma Growth Factors.

Platelet-Rich Plasma (PRP) is an autologous biological product which, due to its regenerative capacity, is currently used in different fields of medicine. This biological treatment has proven to be effective in numerous research studies due to its high content of growth factors released by platelets. However, the current systems used to obtain PRP do not enrich the growth factors and cytokines outside platelets. Considering this, the present work aims to develop a new technique by which all the biomolecules present in plasma are enriched. Thus, a new method based on ultrafiltration has been developed for the obtaining of the novel PRP. By this method, ultrafiltration of the plasma water is carried out using a 3KDa filtering unit. The results showed that the technique was able to concentrate extraplatelet factors, such as IGF-1 and HGF, in contrast with conventional plasmas. Thus, the cultured cells responded with increased viability to this new PRP. These results could provide a new approach to the treatment of injuries requiring regenerative medicine, potentially improving the outcomes of the conventional PRPs.

Action of Platelet-Rich Plasma on In Vitro Cellular Bioactivity: More than Platelets.

Platelet-rich plasma (PRP) is a biological therapy in which one of the mechanisms of action is the stimulation of biological processes such as cell proliferation. The size of PRP's effect depends on multiple factors, one of the most important being the composition of PRP. The aim of this study was to analyze the relationship between cell proliferation and the levels of certain growth factors (IGF-1, HGF, PDGF, TGF-ß and VEG) in PRP. First, the composition and effect on cell proliferation of PRP versus platelet-poor plasma (PPP) were compared. Subsequently, the correlation between each growth factor of PRP and cell proliferation was evaluated. Cell proliferation was higher in cells incubated with lysates derived from PRP compared to those cultured with lysates derived from PPP. In terms of composition, the levels of PDGF, TGF-ß, and VEGF were significantly higher in PRP. When analyzing the PRP growth factors, IGF-1 was the only factor that correlated significantly with cell proliferation. Of those analyzed, the level of IGF-1 was the only one that did not correlate with platelet levels. The magnitude of PRP's effect depends not only on platelet count but also on other platelet-independent molecules.

Bioactive and degradable hydrogel based on human platelet-rich plasma fibrin matrix combined with oxidized alginate in a diabetic mice wound healing model.

In the present study we developed an injectable, bioactive and degradable hydrogel composed of alginate at 2.5% oxidation degree and calcium-activated platelet rich plasma (PRP) for wound healing applications (PRP-HG-2.5%). The alginate gives mechanical support to the hydrogel while the activated PRP provides growth factors that enhance wound healing and fibrin which creates an adequate microenvironment for cell migration and proliferation. The rheological and mechanical properties of the hydrogel were characterized. Further characterization revealed that PRP-HG-2.5% showed a faster hydrolitic degradation rate than unmodified alginate and a similar platelet derived growth factor (PDGF-BB) release profile. In vitro efficacy studies, carried out in human fibroblasts and keratinocytes, showed that PRP-HG-2.5% was not cytotoxic and that it was able to promote cell adhesion and proliferation. Thereafter, in an in vivo full thickness wound healing study conducted in diabetic mice, no differences were found among PRP-HG-2.5% and its counterpart without PRP, likely due to the xenogeneic origin of the PRP. This hypothesis was validated in vitro, since a cytotoxic effect was observed after human PRP application to mouse fibroblasts. Therefore, PRP-HG-2.5% might be a promising strategy for chronic woundstreatment, although its effectiveness should be evaluated in a more reliable preclinical model.

Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization.

Platelet-Rich Plasma (PRP) is enriched in molecular messengers with restorative effects on altered tissue environments. Upon activation, platelets release a plethora of growth factors and cytokines, either in free form or encapsulated in exosomes, which have been proven to promote tissue repair and regeneration. Translational research on the potential of exosomes as a safe nanosystem for therapeutic cargo delivery requires standardizing exosome isolation methods along with their molecular and morphological characterization. With this aim, we isolated and characterized the exosomes released by human PRP platelets. Western blot analysis revealed that CaCl2-activated platelets (PLT-Exos-Ca2+) released more exosomes than non-activated ones (PLT-Exos). Moreover, PLT-Exos-Ca2+ exhibited a molecular signature that meets the most up-to-date biochemical criteria for platelet-derived exosomes and possessed morphological features typical of exosomes as assessed by transmission electron microscopy. Array analysis of 105 analytes including growth factors and cytokines showed that PLT-Exos-Ca2+ exhibited lower levels of most analytes compared to PLT-Exos, but relatively higher levels of those consistently validated as components of the protein cargo of platelet exosomes. In summary, the present study provides new insights into the molecular composition of human platelet-derived exosomes and validates a method for isolating highly pure platelet exosomes as a basis for future preclinical studies in regenerative medicine and drug delivery.

Platelet Lysate Nebulization Protocol for the Treatment of COVID-19 and Its Sequels: Proof of Concept and Scientific Rationale.

One of the most severe effects of coronavirus disease 2019 (COVID-19) is lung disorders such as acute respiratory distress syndrome. In the absence of effective treatments, it is necessary to search for new therapies and therapeutic targets. Platelets play a fundamental role in respiratory disorders resulting from viral infections, being the first line of defense against viruses and essential in maintaining lung function. The direct application of platelet lysate (PL) obtained from the platelet-rich plasma of healthy donors could help in the improvement of the patient due its anti-inflammatory, immunomodulatory, antifibrotic, and repairing effects. This work evaluates PL nebulization by analyzing its levels of growth factors and its biological activity on lung fibroblast cell cultures, besides describing a scientific basis for its use in this kind of pathology. The data of the work suggest that the molecular levels and biological activity of the PL are maintained after nebulization. Airway administration would allow acting directly on the lung tissue modulating inflammation and stimulating reparative processes on key structures such as the alveolocapillary barrier, improving the disease and sequels. The protocol developed in this work is a first step for the study of nebulized PL both in animal experimentation and in clinical trials.

Effects of Platelet-Rich Plasma on Cellular Populations of the Central Nervous System: The Influence of Donor Age.

Platelet-rich plasma (PRP) is a biologic therapy that promotes healing responses across multiple medical fields, including the central nervous system (CNS). The efficacy of this therapy depends on several factors such as the donor's health status and age. This work aims to prove the effect of PRP on cellular models of the CNS, considering the differences between PRP from young and elderly donors. Two different PRP pools were prepared from donors 65‒85 and 20‒25 years old. The cellular and molecular composition of both PRPs were analyzed. Subsequently, the cellular response was evaluated in CNS in vitro models, studying proliferation, neurogenesis, synaptogenesis, and inflammation. While no differences in the cellular composition of PRPs were found, the molecular composition of the Young PRP showed lower levels of inflammatory molecules such as CCL-11, as well as the presence of other factors not found in Aged PRP (GDF-11). Although both PRPs had effects in terms of reducing neural progenitor cell apoptosis, stabilizing neuronal synapses, and decreasing inflammation in the microglia, the effect of the Young PRP was more pronounced. In conclusion, the molecular composition of the PRP, conditioned by the age of the donors, affects the magnitude of the biological response.

Platelet-rich plasma injections delay the need for knee arthroplasty: a retrospective study and survival analysis.

PURPOSE: The biological action of platelet-rich plasma (PRP) could slow down the osteoarthritis progression, resulting in a delay of joint replacement. This work aims to evaluate the ability of PRP to postpone and even avoid knee replacement in patients with knee osteoarthritis (KOA) analyzing, on the one hand, the time of delay and on the other hand the percentage of patients without undergoing total knee arthroplasty (TKA). METHODS: A retrospective analysis and a survival analysis were conducted. KOA patients who underwent knee replacement between 2014 and 2019 and previously received PRP infiltrations were included in the retrospective analysis. Regarding survival analysis, KOA patients who received PRP treatment during 2014 and with follow-up until 2019 were included. The dates of PRP treatment and TKA, KOA severity, age of the patients, number of PRP cycles, and administration route were analyzed. RESULTS: This work included 1084 patients of which 667 met the inclusion criteria. 74.1% of the patients in the retrospective study achieved a delay in the TKA of more than 1.5 years, with a median delay of 5.3 years. The survival analysis showed that 85.7% of the patients did not undergo TKA during the five year follow-up. The severity degree, age, PRP cycles, and administration route had a statistically significant influence on the efficacy of PRP in delaying surgery. CONCLUSION: These data suggest that the application of PRP in KOA patients is a treatment that could delay TKA, although further studies are needed to understand and improve this therapy.

Intraosseous infiltrations of Platelet-Rich Plasma for severe hip osteoarthritis: A pilot study.

OBJECTIVE: Addressing the subchondral bone through intraosseous infiltrations of Platelet-Rich Plasma (PRP) may improve the effectiveness of this technique for severe hip osteoarthritis (HOA). METHODS: Forty patients with HOA degree 2 and 3 according to the Tönnis scale were recruited for this study. They were susceptible to a total hip arthroplasty, without response to previous treatment based on intraarticular infiltrations of PRP. Patients received a combination of intraosseous injections into the acetabulum and the femoral head, as well as intraarticular PRP infiltrations. The clinical outcome was evaluated at 2, 6 and 12 months using the Hip Osteoarthritis Outcome Score (HOOS) and the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index. RESULTS: At 2, 6 and 12 months, patients had significant pain improvement according to HOOS pain, WOMAC pain, and VAS scores. After the treatment, the percentage of patients with minimal clinically important improvement was 40% (16 over 40 patients) at 2 months, 37.5% (15 over 40) at 6 months, and 40% (16 over 40) at 12 months. Conclusion: The combination of intra-articular and intra-osseous infiltrations of PRP showed a pain reduction and improvement in hip joint functionality up to 12 months in patients with severe HOA, with no severe adverse effects.

Biological and structural effects after intraosseous infiltrations of age-dependent platelet-rich plasma: An in vivo study.

Platelet-rich plasma (PRP) is an increasingly widespread treatment for joint pathologies. Its characteristics and administration route are variables that may influence the clinical outcome. The aim of this in vivo study was to analyze in aged rats the biological and structure effects of intraosseous infiltrations of two different types of PRP obtained from young and old donors. During 6 months intraosseous infiltrations were performed and 4 days after the last infiltration, animals were sacrificed, and bones were extracted for micro-computed tomography (micro-CT) and histological analysis. Molecular composition of the PRP of aged donors presented higher levels of proinflammatory molecules. The histological studies showed a greater cellularity of bone marrow in groups treated with PRP. Concerning micro-CT analysis, young PRP showed a better femoral bone structure according to values of percentage of trabecular bone, trabecular space, trabecular density, and subchondral bone plate volume. In summary, this study has demonstrated that intraosseous infiltrations of PRP from young donors prevent from age-related bone degeneration. This treatment could stimulate the biological processes that maintain homeostasis and bone structure and avoid osteoarticular pathologies.

Platelet-rich plasma combined with allograft to treat osteochondritis dissecans of the knee: a case report.

BACKGROUND: Osteochondritis dissecans of the knee is a prevalent pathology in young, active people that is brought about by either traumatic, developmental, or iatrogenic etiologies. CASE PRESENTATION: A 40-year-old Caucasian man reporting pain, swelling, and functional reduction was evaluated and diagnosed with internal condyle osteochondritis dissecans of the knee. Harnessing the trophic, chondroprotective, anti-inflammatory, and immunomodulatory properties of platelet-rich plasma, we carried out a knee open-sky surgical technique in which we combined autologous therapy with osteochondral allograft to treat the focal, large, and deep traumatic-iatrogenic osteochondritis dissecans of the knee. The axial computed tomographic scan taken 1 year after surgery revealed an area of abnormal signal intensity that was reduced on a computed tomographic scan 2 years later. The computed tomographic scan obtained 2 years later and the magnetic resonance imaging scan 3 years later also showed a clear reattachment and incorporation of the graft. Seven years after the surgery, the patient resumed his daily routine without any recurrent symptoms. CONCLUSION: Platelet-rich plasma application in osteochondral allograft implantation open surgery could enhance the healing process of medial condyle osteochondritis dissecans of the knee.

Treating Severe Knee Osteoarthritis with Combination of Intra-Osseous and Intra-Articular Infiltrations of Platelet-Rich Plasma: An Observational Study.

OBJECTIVE: Assessing the therapeutic effects of a combination of intra-articular and intra-osseous infiltrations of platelet-rich plasma (PRP) to treat severe knee osteoarthritis (KOA) using intra-articular injections of PRP as the control group. DESIGN: In this observational study, 60 patients suffering from severe KOA were treated with intra-articular infiltrations of PRP (IA group) or with a combination of intra-osseous and intra-articular infiltrations of PRP (IO group). Both groups were matched for sex, age, body mass index, and radiographic severity (III and IV degree according to Ahlbäck scale). Clinical outcome was evaluated at 2, 6, and 12 months, using the Knee injury and Osteoarthritis Outcome Score (KOOS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaires. RESULTS: At 2, 6 and 12 months after treatment, IO group had a significant improvement in all KOOS and WOMAC subscales ( P < 0.05). On the contrary, patients of the IA group did not improve in any of the scores. Sixteen out of 30 IO group patients showed minimal clinically important improvement (MCII) whereas 8 out of 30 IA group patients showed this response at 6 months (26.7%; 95% CI -0.4 to 49.9; P = 0.037). At 12 months, 14 patients of IO group and 5 patients of the IA group showed MCII (30%; 95% CI 4.3 to 51.9; P = 0.013). No differences between groups were observed at 2 months. CONCLUSIONS: PRP intra-articular injections in severe KOA were not effective and did not provide any benefit. Combination of intra-articular and intra-osseous infiltrations of PRP was not clinically superior at 2 months, but it showed superior clinical outcomes at 6 and 12 months when compared with intra-articular injections of PRP.

Autologous bioscaffolds based on different concentrations of platelet rich plasma and synovial fluid as a vehicle for mesenchymal stem cells.

In the field of tissue engineering, diverse types of bioscaffolds are being developed currently for osteochondral defect applications. In this work, a novel scaffold based on platelet rich plasma (PRP) and hyaluronic acid with mesenchymal stem cells (MSCs) has been evaluated to observe its effect on immobilized cells. The bioscaffolds were prepared by mixing different volumes of synovial fluid (SF) with PRP from patients obtaining three formulations at PRP-SF ratios of 3:1, 1:1 and 1:3 (v/v). The live/dead staining revealed that although the cell number of each type of bioscaffold was different, these this constructs provide cells with a suitable environment for their viability and proliferation. Moreover, immobilized MSCs showed their ability to secrete fibrinolytic enzymes, which vary depending on the fibrin amount of the scaffold. Immunohistochemical analysis revealed the positive staining for collagen type II in all cases, proving the biologic action of SF derived MSCs together with the suitable characteristics of the bioscaffold for chondrogenic differentiation. Considering all these aspects, this study demonstrates that these cells-based constructs represent an attractive method for cell immobilization, achieving completely autologous and biocompatible scaffolds. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 377-385, 2018.

Application of 3D technology and printing for femoral derotation osteotomy: case and technical report.

In some surgical techniques like femoral derotation osteotomy, accuracy is a key factor that often is not optimal because of the lack of appropriate technology. 3D printing is emerging in many professional areas and its use in the medical field may enhance the results of certain surgeries. This case describes a patient who underwent an intramedullary nail fixation to treat a femoral shaft fracture. After nine months, the patient presented hip pain and "in toe" walking caused by a malrotation produced during the surgery. To address the consequent femoral derotation osteotomy, 3D technology was used throughout the whole process. A 3D model of the patient's femur was created to conduct a real and accuracy assessment of femoral anteversion. Then, a customized surgical guide was designed and printed to ensure the proper alignment during surgery. Given the success of this surgery, 3D printing can be considered a quick and inexpensive tool to improve surgical results.

Cryopreservation of Human Mesenchymal Stem Cells in an Allogeneic Bioscaffold based on Platelet Rich Plasma and Synovial Fluid.

Transplantation of mesenchymal stem cells (MSCs) has emerged as an alternative strategy to treat knee osteoarthritis. In this context, MSCs derived from synovial fluid could provide higher chondrogenic and cartilage regeneration, presenting synovial fluid as an appropriate MSCs source. An allogeneic and biomimetic bioscaffold composed of Platelet Rich Plasma and synovial fluid that preserve and mimics the natural environment of MSCs isolated from knee has also been developed. We have optimized the cryopreservation of knee-isolated MSCs embedded within the aforementioned biomimetic scaffold, in order to create a reserve of young autologous embedded knee MSCs for future clinical applications. We have tested several cryoprotectant solutions combining dimethyl sulfoxide (DMSO), sucrose and human serum and quantifying the viability and functionality of the embedded MSCs after thawing. MSCs embedded in bioscaffolds cryopreserved with DMSO 10% or the combination of DMSO 10% and Sucrose 0,2 M displayed the best cell viabilities maintaining the multilineage differentiation potential of MSCs after thawing. In conclusion, embedded young MSCs within allogeneic biomimetic bioscaffold can be cryopreserved with the cryoprotectant solutions described in this work, allowing their future clinical use in patients with cartilage defects.

Intraosseous Infiltration of Platelet-Rich Plasma for Severe Hip Osteoarthritis.

This work describes a technique of platelet-rich plasma (PRP) infiltration for the treatment of severe hip osteoarthritis (OA). Although the results achieved with intra-articular infiltrations of PRP are promising, they may be insufficient in the long-term for severe hip OA. The technique consists of a combined intra-articular and intraosseous infiltration of PRP to reach all joint tissues, especially the subchondral bone, and hence facilitate a greater distribution of PRP. Diagnosis is based on clinical and radiographic findings, and patients with grade III OA according to the Tönnis scale, as well as patients who have not responded to conventional treatment, are considered candidates for this technique. After an ultrasound-guided intra-articular PRP infiltration is performed, 2 intraosseous infiltrations are conducted with a fluoroscope; the first injection is applied into the acetabulum and the second into the femoral head. However, this technique presents more difficulty than the conventional administration, so it is necessary to consider several aspects described in this work.

Adult Cells Combined With Platelet-Rich Plasma for Tendon Healing: Cell Source Options.

BACKGROUND: The combination of cells with platelet-rich plasma (PRP) may fulfill tendon deficits and help overcome the limited ability of tendons to heal. PURPOSE: To examine the suitability of 3 human cell types in combination with PRP and the potential impact of the tenocyte-conditioned media (CM) to enhance tendon healing. STUDY DESIGN: Controlled laboratory study. METHODS: Tenocytes, bone marrow-derived mesenchymal stem cells, and skin fibroblasts were cultured in 3-dimensional PRP hydrogels supplemented or not with CM, and cell proliferation and migration were examined. The effect of tendon-derived CM on matrix-forming phenotype and secretion of inflammatory proteins was determined through their administration to mesenchymal stem cells, tendon, and skin fibroblasts by reverse transcription quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: Differences were found in the matrix-forming phenotype between each of the cell types. The ratio of collagen I:collagen III was greater in bone marrow-derived mesenchymal stem cells than in skin fibroblasts and tenocytes. The bone marrow-derived mesenchymal stem cells expressed increased levels of cartilage-related genes than tenocytes or skin fibroblasts. The presence of the tenocyte-CM stimulated basic healing mechanisms including proliferation and chemotaxis in all cell types. In addition, the tenocyte-CM modified the matrix-forming phenotype of every cell type when cultured in PRP hydrogels. Each cell type secreted interleukin-6, interleukin-8, and monocyte chemotactic protein-1 in PRP hydrogels, but mesenchymal stem cells secreted less interleukin-8 and monocyte chemotactic protein-1 than tenocytes or skin fibroblasts. CONCLUSION: The tenocyte-CM combined with PRP stimulated tenogenesis in mesenchymal stem cells and in skin fibroblasts and reduced the secretion of inflammatory proteins. CLINICAL RELEVANCE: Modifying the target tissue with PRP prior to cell implantation may optimize the effect of cell therapies. Skin fibroblasts and bone marrow-derived mesenchymal stem cells combined with PRP could be used to regenerate tendons.

Modulation of Synovial Fluid-Derived Mesenchymal Stem Cells by Intra-Articular and Intraosseous Platelet Rich Plasma Administration.

The aim of this study was to evaluate the effect of intra-articular (IA) or a combination of intra-articular and intraosseous (IO) infiltration of Platelet Rich Plasma (PRP) on the cellular content of synovial fluid (SF) of osteoarthritic patients. Thirty-one patients received a single infiltration of PRP either in the IA space (n = 14) or in the IA space together with two IO infiltrations, one in the medial femoral condyle and one in the tibial plateau (n = 17). SF was collected before and after one week of the infiltration. The presence in the SF of mesenchymal stem cells (MSCs), monocytes, and lymphocytes was determined and quantified by flow cytometry. The number and identity of the MSCs were further confirmed by colony-forming and differentiation assays. PRP infiltration into the subchondral bone (SB) and the IA space induced a reduction in the population of MSCs in the SF. This reduction in MSCs was further confirmed by colony-forming (CFU-F) assay. On the contrary, IA infiltration alone did not cause variations in any of the cellular populations by flow cytometry or CFU-F assay. The SF of osteoarthritic patients contains a population of MSCs that can be modulated by PRP infiltration of the SB compartment.