Autologous Bone Marrow Cells Research Articles

Regeneration of Osteochondral Lesion of the Talus with Retrograde Drilling Technique: An In Vitro Pilot Study.

Background: Retrograde Drilling (RD) is a surgical technique employed for osteochondral lesions of the talus (OCLTs) to reach the subchondral bone lesion from behind, thus preserving cartilage integrity. The aim of the present pilot study was to set up an in vitro model of OCLTs to evaluate the regenerative potential of biological approaches that could be associated with the RD technique. Methods: For this purpose, an OCLT was created in human osteochondral specimens, to try to mimic the RD technique, and to compare the regenerative potential of two biological treatments. For this purpose, three groups of treatments were performed in vitro: (1) no treatment (empty defect); (2) autologous bone graft (ABG); (3) hyaluronic membrane enriched with autologous bone marrow cells. Tissue viability; production of Collagen I and II, Vascular Endothelial Growth Factor, and Aggrecan; and histological and microCT evaluations were performed after 30 days of culture in normal culture conditions. Results: It was observed that Group 3 showed the highest viability, and Group 2 showed the highest protein production. From a histological and microtomographic point of view, it was possible to appreciate the structure of the morcellized bone with which the defect of Group 2 was filled, while it was not yet possible to observe the deposition of mineralized tissue in Group 3. Conclusions: To conclude, this pilot study shows the feasibility of an alternative in vitro model to evaluate and compare the regenerative potential of two biological scaffolds, trying to mimic the RD technique as much as possible. The tissues remained vital for up to 4 weeks and both ABG and hyaluronic acid-based scaffolds stimulated the release of proteins linked to regenerative processes in comparison to the empty defect group.

B-ultrasound-guided Intrahepatic Infusion of Autologous Bone Marrow Cells for Decompensated Cirrhosis

Objective: To study the therapeutic effect of B-ultrasound-guided intrahepatic infusion of autologous bone marrow nucleated cells on decompensated cirrhosis. Methods: To observe the clinical treatment of 75 cases of decompensated cirrhosis. Among them, 30 cases received routine liver protection and diuretic treatment. 45 cases were treated by percutaneous transhepatic infusion of autologous bone marrow nucleated cells under the guidance of B ultrasound. There were no significant differences in liver function and blood routine indexes between the two groups before treatment (p > 0.05). Results: The indexes of liver function and blood routine at different time periods of 1 month, 3 months, 6 months, and 12 months in the conventional treatment group did not change significantly. 6 cases died of liver failure within 1 year, the fatality rate was 20%. The indexes of liver function and blood routine of percutaneous liver transhepatic infusion of autologous bone marrow nucleated cells at 1 month, 3 months, 6 months, and 12 months under the guidance of B-ultrasound were significantly better than those of the conventional treatment group (p < 0.05). One case died of gastrointestinal bleeding in the group of percutaneous transhepatic infusion of autologous bone marrow nucleated cells guided by B ultrasound, with a fatality rate of 2.5%. Compared with the conventional treatment group, there were significant differences (p < 0.05). Conclusion: Conventional drug therapy has no obvious effect on decompensated cirrhosis. Intrahepatic infusion of bone marrow nucleated cells can significantly promote liver function reconstruction in decompensated cirrhosis.

Atorvastatin Treatment Significantly Increased the Concentration of Bone Marrow-Derived Mononuclear Cells and Transcutaneous Oxygen Pressure and Lowered the Pain Scale after Bone Marrow Cells Treatment in Patients with "No-Option" Critical Limb Ischaemia.

The present study investigated the outcomes and possible predictive factors of autologous bone marrow cells (BMCs) therapy in patients with "no-option" critical limb ischaemia (CLI). It was focused on exploring the clinical background and prior statin and renin-angiotensin system (RAS)-acting agents pharmacotherapy related to the therapeutic efficacy of BMCs treatment. In the present study, we reviewed thirty-three patients (mean age 64.9 ± 10 years; 31 males) with advanced CLI after failed or impossible revascularisation, who were treated with 40 mL of autologous BMCs by local intramuscular application. Patients with limb salvage and wound healing (N = 22) were considered as responders to BMCs therapy, and patients with limb salvage and complete ischemic wound healing (N = 13) were defined as super-responders. Logistic regression models were used to screen and identify the prognostic factors, and a receiver operating characteristics (ROC) curve, a linear regression, and a survival curve were drawn to determine the predictive accuracy, the correlation between the candidate predictors, and the risk of major amputation. Based on the univariate regression analysis, baseline C-reactive protein (CRP) and transcutaneous oxygen pressure (TcPO2) values were identified as prognostic factors of the responders, while CRP value, ankle-brachial index (ABI), and bone marrow-derived mononuclear cells (BM-MNCs) concentration were identified as prognostic factors of the super-responders. An area under the ROC curve of 0.768 indicated good discrimination for CRP > 8.1 mg/L before transplantation as a predictive factor for negative clinical response. Linear regression analysis revealed a significant dependence between the levels of baseline CRP and the concentration of BM-MNCs in transplanted bone marrow. Patients taking atorvastatin before BMCs treatment (N = 22) had significantly improved TcPO2 and reduced pain scale after BMCs transplant, compared to the non-atorvastatin group. Statin treatment was associated with reduced risk for major amputation. However, the difference was not statistically significant. Statin use was also associated with a significantly higher concentration of BM-MNCs in the transplanted bone marrow compared to patients without statin treatment. Patients treated with RAS-acting agents (N = 20) had significantly reduced pain scale after BMCs transplant, compared to the non-RAS-acting agents group. Similar results, reduced pain scale and improved TcPO2, were achieved in patients treated with atorvastatin and RAS-acting agents (N = 17) before BMCs treatment. Results of the Spearman correlation showed a significant positive correlation between CLI regression, responders, and previous therapy before BMCs transplant with RAS-acting agents alone or with atorvastatin. CRP and TcPO2 were prognostic factors of the responders, while CRP value, ABI, and BM-MNCs concentration were identified as predictive factors of the super-responders. Atorvastatin treatment was associated with a significantly increased concentration of BM-MNCs in bone marrow concentrate and higher TcPO2 and lower pain scale after BMCs treatment in CLI patients. Similarly, reduced pain scales and improved TcPO2 were achieved in patients treated with atorvastatin and RAS-acting agents before BMCs treatment. Positive correlations between responders and previous treatment before BMCs transplant with RAS-acting agents alone or with atorvastatin were significant.

An updated systematic review of neuroprotective agents in the treatment of spinal cord injury.

This systematic review aims to summarize the findings from all clinical randomized trials assessing the efficacy of potential neuroprotective agents in influencing the outcomes of acute spinal cord injuries (SCI). Following the PRISMA guidelines, we conducted comprehensive searches in four electronic databases (PubMed, Scopus, Cochrane Library, and Web of Science) up to September 5th, 2023. Our analysis included a total of 30 studies. We examined the effects of 15 substances/drugs: methylprednisolone, tirilazad mesylate, erythropoietin, nimodipine, naloxone, Sygen, Rho protein antagonist, granulocyte colony-stimulating factor, autologous macrophages, autologous bone marrow cells, vitamin D, progesterone, riluzole, minocycline, and blood alcohol concentration. Notable improvements in neurological outcomes were observed with progesterone plus vitamin D and granulocyte colony-stimulating factor. In contrast, results for methylprednisolone, erythropoietin, Sygen, Rho Protein, and Riluzole were inconclusive, primarily due to insufficient sample size or outdated evidence. No significant differences were found in the remaining evaluated drugs. Progesterone plus vitamin D, granulocyte colony-stimulating factor, methylprednisolone, Sygen, Rho Protein, and Riluzole may enhance neurological outcomes in acute SCI cases. It is worth noting that different endpoints or additional subgroup analyses may potentially alter the conclusions of individual trials. Therefore, certain SCI grades may benefit more from these treatments than others, while the overall results may remain inconclusive.

Autological bone marrow concentrate for the treatment of early stages of femoral head avascular necrosis (literature review)

Relevance. In the treatment of the early stages of the femoral head osteonecrosis core decompression is widely used, which reduces intraosseous pressure and increases blood flow to the femoral head. To increase the efficiency of the core decompression the use of various bioadjuvants, including concentrate of autologous bone marrow cells is needed, the appropriateness of which is actively discussed.Purpose of the systematic review: evaluation of the effectiveness of autologous bone marrow cells in core decompression in the treatment of the early stages of the femoral head osteonecrosis according to the literature.Material and methods. Articles with clinical data in full text in English or Russian available in the Internet (PubMed and elibrary databases) were included in the study. Articles containing actual data on the stage of osteonecrosis, the number of femoral heads operated on, the use of autologous bone marrow cells as an adjunct to the core decompression of the femoral head, the number of ineffective surgical interventions after which total hip arthroplasty was required were included.Results. Sixteen original articles (2011–2022) were analyzed, and the results of 1135 femoral heads treatment (892 with autologous bone marrow cells, 243 without autologous bone marrow cells, control group) were evaluated. Patient follow-up periods ranged from 9 to 120 months, averaging 45 months. When using autologous bone marrow cells, the need for total hip arthroplasty was observed in 168 of 892 patients (18.8 %); when core decompression was used without autologous bone marrow cells, the rate of total hip arthroplasty was 27.2 % (p > 0.05). Only 2 articles out of 16 authors did not consider autologous bone marrow cells to be reasonable in the core decompression of femoral head.Conclusion. The use of autologous bone marrow cells concentrate as a bioadjuvant in the core decompression slows the progression of osteonecrosis. According to the overwhelming number of authors, the core decompression using autologous bone marrow cells is highly effective, significantly reducing the intensity of pain syndrome and improving the function of the hip joint. The results will be more encouraging with the further development of regenerative medicine, which will provide new samples of cellular preparations for bone tissue structure restoration.

Therapy with bone marrow mesenchymal stem cells in bone regeneration in children with osteonecrosis secondary to sickle cell disease.

This study aimed to describe the evolution of bone regeneration in children with hip osteonecrosis associated with sickle cell disease, treated with bone marrow-derived mesenchymal stem cell implants at the Professor Edgar Santos University Hospital Complex. A non-randomized clinical trial was conducted with 48 patients of both sexes, aged between 11 and 18years, diagnosed with femoral head osteonecrosis secondary to sickle cell disease. Patient selection was based on strict criteria, including confirmed diagnosis of sickle cell anemia and a stage of osteonecrosis compatible with the proposed treatment. Bone regeneration assessment was performed through radiographic examinations and magnetic resonance imaging, following the Ficat & Arlet criteria and the Salter-Thompson classification. Statistical analysis revealed a significant association between the patients' age and positive treatment outcomes, suggesting that autologous bone marrow cell implantation is a safe and effective approach in the early stages of osteonecrosis. The majority of patients (87.5%) reported complete pain relief, while 10.42% experienced significant symptom improvement. Only one patient (2.08%) did not observe improvement. The results indicate that cell therapy can regenerate or slow the progression of bone necrosis, reducing the need for more invasive surgical procedures. The study demonstrates the potential of bone marrow-derived mesenchymal stem cell implantation in treating hip osteonecrosis in children with sickle cell disease, emphasizing the importance of long-term monitoring of bone structure stability.

P-785 Promising Reproductive Outcomes In Patients With Refractory Thin Endometrium After Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) – A Case Series

Abstract Study question Does Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) improve endometrial thickness and receptivity and restoration of fertility in refractory cases of thin endometrium? Summary answer Selective Autologous Bone Marrow Regenerative Cell Therapy from Seragen (ABM-RCT) improves endometrial thickness, pregnancy, and live birth rate in refractory cases of thin endometrium. What is known already The impaired endometrial function can limit implantation due to insufficient tissue regeneration. Clinical investigation has proved that stem cells existing in the endometrium might originate from bone marrow and provides evidence that bone marrow-derived cells have a non-hematopoietic physiologic contribution to decidual stroma and play a vital role in implantation and pregnancy maintenance and non-hematopoietic BMSCs are able to impact decidual molecular milieu and overcome implantation defects. ABM-RCT is the most commonly used cell therapy and its safety and efficacy are well-established and documented. This forms the basis for exploring and standardizing ABM-RCT in the management of refractory thin endometrium. Study design, size, duration Eleven patients (from age 29 to 43 with refractory thin endometrium resistant to conventional treatment modalities were recruited and obtained informed consent with detailed explanation that the therapy is still experimental and the risk of failure was given to the couple as part of gynec-oncology treatment. Five patients had asherman’s syndrome, two patients had genital tuberculosis, two patients had thin endometrium hypo-responsive/unresponsive to estrogens, with RIF and two patients underwent chemo and radiotherapy Participants/materials, setting, methods BM aspiration was performed under local anesthesia, from the iliac crest using a disposable BM aspiration needle (Jamshidi, 11 G) and collected in heparinized syringes. Progenitor cell enrichment was done by Seragen’s Selective Enrichment Protocol. Peripheral blood was collected to enrich growth factor concentrate. A 2.9mm Hysteroscope was used with an operating channel and egg pickup needle attached to it and cells were implanted in subendometrial zone in all four walls of the uterine cavity. Main results and the role of chance All patients presented to us with refractory thin endometrium in HRT cycle despite giving all possible medications. Hence the decision was taken for Autologous Bone Marrow Cell Concentrate Therapy.Post ABM-RCT, endometrial thickness showed increase 100% (11/11) of more than 8 mm in all cases on day of embryo transfer with average endometrial thickness improvement was 1.8 mm than previous cycles, with uniform triple layer pattern.10 out of 11 (91%) patients conceived after autologous stem cell injection. 9 Out of 10 (90%) patients conceived had delivered healthy babies and 1 patient (9%) had a miscarriage at 12th week of pregnancy. 1 patient did not conceive will be assessed for immunoprofiling and one more rejuvenation cycle. Limitations, reasons for caution Our study outcomes are consistent with several previous studies and the feasibility of treatment at an IVF setup and encouraging results. Well-planned study with more patients is warranted to evaluate safety, effectiveness, and cost of this modality before it becomes integrated in treatment of this frustrating condition during IVF procedures Wider implications of the findings Our results demonstrates selectively enriched ABM-RCT enhances endometrial thickness, tissue remodeling, uneventful gestation, improved pregnancy and live birth rate. Need for Surrogacy in Non-Responsive Thinendometrium May Be Re-Evaluated. Role of hematopoetic stem cell research and knowledge generated is representing reliable strategy in IVF that may be routinely used in clinicalpractice. Trial registration number Not applicable

Intra-articular injection of bone marrow aspirate concentrate (BMAC) or adipose-derived stem cells (ADSCs) for knee osteoarthritis: a prospective comparative clinical trial

BackgroundWe determined whether autologous mesenchymal stem cells (MSCs) injections provide clinical and functional improvements in knee osteoarthritis (KOA) patients, and whether the results differ between autologous bone marrow cells (BMAC) and adipose-derived stromal cells (ADSCs).MethodsBetween January 2021 and April 2022, 51 patients undergoing intra-articular injection of BMAC and 51 patients undergoing intra-articular injection of ADSCs were prospectively recruited. The Kellgren and Lawrence (K–L) classification was used to grade the severity of osteoarthritis. Knee Injury and Osteoarthritis Outcome Score (KOOS), Oxford Knee Score (OKS), and visual analog scale (VAS) were collected for all 102 patients in the previous week before the procedures, and at the one and 6 months from injection.ResultsKnee KOOS scores, knee OKS scores, and VAS pain scores changed in similar ways in the two treatment groups. Both treatment groups demonstrated significant improvement pre-procedure to post-procedure in knee KOOS scores (p < 0.0001), knee OKS scores (p < 0.0001), and VAS pain scores (p < 0.0001). Patients with K–L grade 2 showed better functional and clinical outcomes than patients with K–L grades 3 and 4 (p < 0.0001).ConclusionBoth intra-articular BMAC and ADSC injections significantly improved pain and functional outcomes at 6-month follow-up in patients with KOA. The difference between BMAC and ADCSs groups as tissue sources of MSCs was not statistically significant in terms of clinical and functional outcomes.

Efficacy of Activated Autologous Hematopoietic Stem Cell Transplantation in the Treatment of Acute Myeloid Leukemia

To compare the efficacy of activated autologous bone marrow and peripheral blood hematopoietic stem cell transplantation (Auto-HSCT) and matched sibling donor allogeneic hematopoietic stem cell transplantation (MSD-HSCT) for the first complete remission of adult acute myeloid leukemia (AML-CR1). For 86 adult patients with first complete remission of AML who underwent auto-HSCT (41 cases) and MSD-HSCT (45 cases) in our hospital from June 2012 to June 2020, the patients were treated with modified MAC ［Malflane 160 mg/(m2·d), -3 days, Ara-C 2 g/(m2·2), -3 days 21∶00, -2 days 9∶00, CTX 60 mg/(kg·d),-3 d, -2 d］, the stem cells were activated by IL-2 (1 000 U/ mL), IFN-α (100 U/ mL) and IFN-γ (100 U/ml). The overall survival (OS), leukemia free survival (LFS), cumulative incidence of recurrence (CIR) and non-recurrence mortality (NRM) of patients with different types of transplantation were compared. The 3-year OS rates of Auto-HSCT group and MSD-HSCT group were 75% and 69.5%, and the 3-year LFS rates were 70.6% and 82.4%, respectively. There was no statisticaly significant difference in the 3-year OS rates of low risk, medium risk and high risk patients in the Auto-HSCT and MSD-HSCT group (90.2% vs 87.5%, 68.4% vs 68.8%, 28.6% vs 53.3%), the LFS rates of low risk, medium risk and high risk patients in the auto-HSCT and MSD-HSCT group were 90.2% and 87.5%(P=0.838), 71.8% and 91.7%(P=0.184), 0 and 67.5%(P=0.027), respectively. The NRM of Auto-HSCT and MSD-HSCT group were 4.9% and 20% (P=0.036), and CIR were 24.4% and 13.3% (P=0.188). Univariate analysis showed that the survival time of patients was significantly correlated with the number of CR courses and disease risk stratification (P=0.005, P=0.000). Cox multivariate analysis showed that disease risk stratification was an independent risk factor affecting OS (P=0.001). For adult patients with primary AML-CR1, Auto-HSCT is safe and effective. In the absence of sibling donor, Auto-HSCT can be regarded as an effective post-remission treatment for patients with intermediate risk AML-CR1.

Role of Vascular Endothelial Growth Factor-C during Stem Cell Therapy Using Autologous Bone Marrow Mononuclear Cells in Patients with Lower Limb Lymphedema

Introduction: Vascular endothelial growth factor-C (VEGF-C) is the primary lymphangiogenic factor that stimulates lymphangiogenesis by signaling via specific receptor, vascular endothelial growth factor receptor 3 (VEGFR3). This study was conducted to evaluate the change in the level of VEGF-C before and after autologous bone marrow mononuclear cell transplantation for treatment of Lower limb lymphedema. Patient and methods: Forty patients with lower limb lymphedema were divided into two groups. Group I included 20 patients with chronic lower limb lymphedema who underwent autologous bone marrow mononuclear cell transplantation. Group II included 20 patients with chronic lower limb lymphedema who were exposed only to compression therapy as a control group. VEGF-C level in the diseased limbs was measured in both groups at the beginning of the study then 3 and 6 months respectively. Results: Group I included 20 patients, 8 patients were male (40%) and 12 patients were females (60%) with mean age 29.5 ± 12.15 while group II included 20, 10 patients were male (50%) and 10 patients were females (50%) with mean age 39.5 ± 11.5. In group I, the specimens were taken at 3 and 6 months after transplantation showed a marked decrease in the VEGF-C level with statistically significant p value, 0.02 and 0.001 respectively. In group II the level of VEGF-C after compression therapy alone at 3 and 6 months interval showed fluctuation with statistically non-significant p value, 0.64 and 0.55 respectively. Conclusion: VEGF-C is essential for regulation of lymphangiogenesis. The level of VEGF-C was found elevated in patients with lymphedema and decrease after autologous mononuclear bone marrow cells, however these results were statically non-significant.

Single-Surgery Co-Implantation of Autologous Primary Articular Chondrocytes with Bone Marrow Cells in the Treatment of Articular Cartilage Lesions: Observations from the Operating Room on Tissue Input and Cell Output.

Single-surgery variants of autologous chondrocyte implantation to repair cartilage are emerging, but practical data on such procedures are scarce. We set out to describe a 1-stage autologous chondrocyte co-implantation procedure and include quantitative characteristics of the biopsy tissues collected and of the cells obtained from those tissues in the operating room. Data concerning patient age, articular cartilage lesion size and location, as well as measurements of cartilage biopsy mass, bone marrow aspirate volume, and the cell yields harvested from those biopsies were intraoperatively collected for 141 patients. The mean patient age was 35.7 ± 9.8 years, and the mean total lesion size was 4.0 ± 3.1 cm2. Cartilage biopsy mass ranged from 0.19 to 2.0 gram and provided a mean yield of 1.23 × 106 chondrocytes/gram. Bone marrow aspirate volume ranged from 7.2 to 62 milliliters and provided a mean yield of 7.18 × 106 mononuclear cells/mL. The cell yields did not correlate with patient age, which ranged from 12 to 57 years. The mean primary chondrocyte supply was 272 thousand per cm2 of lesion, ranging from 10.4 thousand to 1.07 million per cm2. The total cell supply was kept at 9 million cells per cm2 of lesion by adding mononuclear cells to the chondrocytes. The mean tissue processing time was 100 ± 19 minutes, which was frequently used to perform concurrent interventions. Single-surgery co-implantation of clinically relevant numbers of autologous primary articular chondrocytes and bone marrow cells is feasible for a wide range of ages and lesion sizes.

Our experience in cell therapy in surgical problems

Aim: To study the effect and efficacy of autologous bone marrow cell therapy in various surgical pathologies of varied etiology which were refractory to conventional methods of management. Material and method: Ethical committee clearance and informed consent of patients were taken. Two groups of each surgical pathologies were selected randomly. The patients in study group were treated by conventional methods followed by autologous bone marrow cell application locally, whereas patients in control group were treated by conventional methods only. The patients were then followed post-operatively and analyzed by statistical methods. Result: Post-operatively the patients in study group experienced faster recovery by appearing good quality granulation tissue and rapid epithelisation and there was also significant reduction in local pain. Biopsy showed improved fibroblast, macrophages, neovascularisation and less necrosis per HPF in study group as comparison to control group. Conclusion: Autologous bone marrow cell therapy is a novel method in the management of various surgical pathologies.

Therapeutic effect of autologous bone marrow cells injected into the liver under the guidance of B‑ultrasound in the treatment of HBV‑related decompensated liver cirrhosis.

The 5-year mortality rates associated with decompensated liver cirrhosis (DLC) can reach 50%, which suggests that this condition poses a serious health risk. In previous studies conducted by our group, autologous bone marrow nucleated cells (ABMNCs) were used to treat HIV-positive patients with DLC through the right omental vein; however, trauma and poor compliance were encountered. In the present study, the percutaneous liver approach to inject ABMNCs under the guidance of B-ultrasound was employed for the treatment of DLC. A total of 108 patients with DLC were retrospectively divided into the routine drug treatment group (control group; 30 cases), the right omental vein infusion of ABMNCs group (observation group 1; 38 cases) and the B-ultrasound-guided liver injection of ABMNCs group (observation group 2; 40 cases). After treatment, the liver synthesis (prothrombin time, albumin and ascites) and secretion functions (total bilirubin) in observation groups 1 and 2 were significantly improved compared with those of the control group (P<0.01) and the bone marrow function was also significantly improved compared with that of the control group (P<0.01). While, the bone marrow function (white blood cell, platelet, and hemoglobin) in observation group 1 was significantly improved compared with that of observation group 2 at the end of treatment (P<0.01). After a 1-year follow-up, the case fatality rate was 2.5% (1/40) in observation group 2, which was significantly lower than the 20% fatality rate (6/30) recorded in the control group (P<0.05). The injection of ABMNCs into the liver under the guidance of B-ultrasound was significantly better than conventional drug therapy in treating DLC. This approach has obvious advantages such as no hospitalization, minimal trauma, rapid recovery and good compliance, all of which make it worthy of application in primary hospitals.

P389: BISPECIFIC ANTIBODIES ACTIVATE AUTOLOGOUS LEUKEMIA-REACTIVE T CELLS IN ACUTE MYELOID LEUKEMIA

Background: Acute myeloid leukemia (AML) is a highly immunosuppressive disease with a high risk of relapse. This makes the finding of effective immunotherapy-based treatments important, but also difficult. T cell-engaging bispecific antibodies (TCEs) are designed to bind to both T cells and target cells, activate the T cells and thus induce an immune response against the leukemia cells without employing the MHC-T cell receptor (TCR) interaction. Several therapies of this kind are under development for treatment of AML. Surprisingly, both in vitro and in vivo, a subset of these TCE immunocoached T cells (ICTs) can subsequently target autologous AML cells and mediate efficient TCR-MHC-dependent AML cell killing in the absence of the TCE. This points to that a) a portion of the T cells present in an AML are leukemia-reactive but have been immunosuppressed - a suppression that can be reversed by the addition of a TCE, b) that this anti-AML T cell reactivation may play an important part of the mechanism of action of AML-targeted TCEs and c) that by understanding this reactivation mechanism, we will be able to design improved AML immunotherapies in the future. Aims: To understand the mechanisms by which a TCE induces reactivation of leukemia-reactive T cells (LRTs) in AML. Methods: T cells in biobanked AML patient bone marrow samples were activated with either an anti-CD3 x anti-CD123 TCE or a conventional anti-CD3 antibody for 5 days before isolating the ICTs. ICT assay: ICTs were washed to remove the TCE, and isolated ICTs were co-cultured with additional autologous bone marrow cells at different E:T ratios. Killing efficiency and T cell expansion were determined over several days using flow cytometry. Trogocytosis assay: ICTs were co-cultured with stained additional autologous AML cells for 1 h and T cell populations of interest were isolated using FACS. T cell repertoires were analyzed by targeted RNAseq of the TCR. Results: ICTs generated with a TCE kill autologous AML cells efficiently without further addition of the TCE. Adding additional TCE at this stage also does not increase the AML killing efficiency of the ICTs. ICTs generated with an activating anti-CD3 antibody show a similar AML killing efficiency and a similar trogocytotic behavior as TCE ICTs. Most of the TCR clones present in the AML bone marrow pre-stimulation were also represented among the ICTs and trogocytotic ICTs (ICTs that are directly targeting the AML cells after either TCE or anti-CD3 stimulation; Figure 1), suggesting that the bulk of T cells in an AML bone marrow represent heterogeneous immunosuppressed LRTs which can be equally well reactivated by a TCE or an anti-CD3 antibody. Trogocytotic ICTs generated from both types of T cell activation have a similar TCR distribution. ICTs derived from the bone marrow target autologous AML cells more efficiently than ICTs from peripheral blood. Image:Summary/Conclusion: Similarities between TCE ICTs and CD3 ICTs in regards to TCR distributions and killing efficiencies suggest that the TCE reactivates leukemia-reactive T cells via a CD3-dependent mechanism rather than a more complex proximity mechanism. Our data further suggests that T cells in the AML bone marrow are highly heterogeneous and leukemia targeted to a very high degree. In summary, AML bone marrow ICTs are targeting and killing leukemic cells efficiently in a polyclonal fashion and our results highlight that boosting this mechanism can leverage the clinical impact of AML TCEs and be utilized in other novel AML immunotherapy approaches.

Vertebral Body Adherent Allogeneic Mesenchymal Stromal Cells Increase in Vivo Perfusion and Improve Muscle Function in Diabetic Mouse Models of Critical Limb-threatening Ischemia

Critical limb-threatening ischemia (CLTI) is a health care priority; since 2010, there has been a 50% increase of non-traumatic major amputations. We have previously shown that autologous bone marrow cell therapy is not effective in preventing amputations on patients with poor options for revascularization. Allogeneic mesenchymal stromal cells (MSCs) from young healthy donors may have greater therapeutic potential, particularly in diabetic populations. Here, we describe the effects of intramuscular injection of vertebral body adherent (vBA)-MSCs in restoring limb perfusion, muscle regeneration and limb function in two diabetic CLTI mouse models.

Repeat infusion of autologous bone marrow cells in progressive multiple sclerosis – A phase I extension study (SIAMMS II)

BackgroundDuring the safety and feasibility ‘Study of Intravenous Autologous Marrow in Multiple Sclerosis (SIAMMS)’, intravenous infusion of autologous marrow was well tolerated. The efficacy of the approach is being explored in a placebo-controlled randomised controlled trial (ACTiMuS, NCT01815632) but it is not known whether repeated infusions will be required to optimise benefit. The objective of the current study was to explore the safety and feasibility of repeat treatment with intravenous autologous bone marrow for patients with progressive multiple sclerosis (MS). Methods‘SIAMMS II’ was a prospective, single centre phase I extension study in which participants in the SIAMMS study were offered repeat bone marrow harvest and infusion of autologous, unfractionated bone marrow as a day-case procedure. The primary outcome measure was number of adverse events and secondary outcome measures included change in clinical rating scales of disability, global evoked potential and cranial magnetic resonance imaging (MRI). ResultsIn total, 4 of the 6 participants in the SIAMMS study had repeat bone marrow harvest and infusion of filtered autologous marrow as a day case procedure which was well tolerated. There were no serious adverse effects. Additional outcome measures including clinical scales, global evoked potentials and cranial MRI were stable. ConclusionSIAMMS II demonstrates the safety and feasibility of repeated, non-myeloablative autologous bone marrow-derived cell therapy in progressive MS.

Transcoronary stem cell transfer and evolution of infarct-related artery atherosclerosis: evaluation with conventional and novel imaging techniques including Quantitative Virtual Histology (qVH).

It has been suggested that infarct-related artery (IRA) atherosclerosis progression after stem cell transcoronary administration might represent a stem-cell mediated adverse effect. To evaluate, using conventional (quantitative coronary angiography, QCA, intravascular ultrasound - IVUS) and novel (quantitative virtual histology - qVH) tools, evolution of IRA atherosclerosis following transcoronary stem cell transfer. QCA, IVUS, VH-IVUS and qVH were performed in 22 consecutive patients (4 women) aged 59 years (data provided as median) undergoing a distal-to-stent infusion of 2.21 × 106 CD34+CXCR4+ autologous bone marrow cells via a cell delivery-dedicated perfusion catheter at anterior AMI day 7. Imaging was repeated at 12 months. This was a substudy of Myocardial Regeneration by Intracoronary Infusion of Selected Population of Stem Cells in Acute Myocardial Infarction (REGENT) Trial (NCT00316381). 18.2% subjects showed absence of distal-to-stent angiographic/IVUS atherosclerotic lesion(s) at baseline and no new lesion(s) at 12-months. In the remaining cohort, there were 28 lesions by QCA (32 by IVUS) at baseline and no new lesion(s) at follow-up. Three fibroatheromas evolved (2 to calcified fibroatheroma and 1 to a fibrocalcific lesion); other plaques maintained their stable (low-risk) phenotypes. Diameter stenosis of QCA-identified lesions was 29.5 vs. 26.5% (p = 0.012, baseline vs. 12-months). Gray-scale IVUS showed reduction in area stenosis (33.8 vs. 31.0%, p = 0.004) and plaque burden (66.27 vs. 64.56%, p = 0.009) at 12-months. Peak fibrotic plaque content increased from 70.41% to 75.0% (p = 0.004). qVH peak confluent necrotic core area and minimal fibrous cap thickness remained stable (0.64 vs. 0.59 mm2, p = 0.290, and 0.15 vs. 0.16 mm, p = 0.646). This study, using a range of classic and novel imaging techniques, indicates lack of any stimulatory effect of transcoronary stem cell transfer on coronary atherosclerosis. Whether, and to what extent, a moderate reduction in plaque burden and stenosis severity at 12-months results from optimized pharmacotherapy and/or stem cell transfer requires further elucidation.

Polarized Autologous Macrophages (PAM) Can Be a Tumor Vaccine

Immunotherapy is currently recognized as one of the most promising anticancer strategies. In the tumor microenvironment, tumor-associated macrophages are mainly M2-type macrophages with tumor-promoting effects. Therefore, the reprogramming of tumor-associated macrophages from M2 to M1 type is a potential strategy for cancer therapy. We have previously shown the anticancer effects of implantable allogeneic M1 macrophages in mice. Here, we further engineered autologous mouse bone marrow cells into M1 macrophages and then embedded them into a sodium alginate gel to prepare an implantable immunotherapeutic agent (M1@Gel). We demonstrate that M1@Gel repolarizes M2 macrophages to M1 type and activates the immune responses in mice. As a result, M1@Gel can potently inhibit the tumor recurrence in mice after the surgical tumor removal. These results suggest that the implantation of autologous M1 macrophages might be a promising strategy for preventing postoperative tumor recurrence. We envisage that the employment of polarized autologous macrophages as a tumor vaccine might be translated into clinic.

Autologous Bone Marrow Cell Infusion for the Treatment of Decompensated Liver Cirrhosis Patients With Type 2 Diabetes Mellitus.

This study aimed to indicate whether autologous bone marrow cell infusion (ABMI) via the right omental vein (ROV) could have a regulatory effect on decompensated liver cirrhosis (DLC) patients with type 2 diabetes mellitus (T2DM). For this purpose, 24 DLC patients with T2DM were divided into observation group (n=14) and control group (n=10). Patients in the observation group were given ABMI through the ROV and right omental artery (ROA), and cases in the control group received ABMI through the ROV. At 1, 3, 6, and 12months after ABMI, it was revealed that the prothrombin time, the total bilirubin levels, and the amount of ascites were significantly lower, while the serum albumin levels in the two groups were markedly higher compared with those before ABMI (p<0.01), and there was no significant difference between the two groups at each time point (p>0.05). The fasting blood glucose and glycosylated hemoglobin levels at 6 and 12months after ABMI in the two groups significantly decreased compared with those before ABMI (p<0.05 or p<0.01), while the decreased levels in the observation group were more obvious than those in the control group at each time point (p<0.01). The amount of insulin in the observation group at 3, 6, and 12months after ABMI was significantly less than that before ABMI in the control group (p<0.01). In summary, ABMI showed a significant therapeutic efficacy for DLC patients with T2DM through ROV and ROA.

Contemporary Outcomes of Distal Lower Extremity Bypass for Chronic Limb-Threatening Ischemia and A Model-Based Comparison With Autologous Bone Marrow-Derived Cell Therapy for Limb Preservation

The standard of therapy for chronic limb-threatening ischemia (CLTI) is revascularization. However, for patients for whom below-the-knee bypass is indicated, an autologous great saphenous vein (GSV), the optimal conduit, might not be available. For these patients, an alternative conduit, such as a synthetic or cryopreserved biologic graft, will be necessary. Contemporary outcomes of distal bypass with these suboptimal conduits have not been well described, and recent advances in novel nonoperative management such as autologous bone marrow cell (BMC) therapy warrant a comparative analysis as a potential alternative for limb preservation.