Autologous Bone Marrow Cells Research Articles

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HomeCirculation ResearchVol. 120, No. 2In This Issue Free AccessResearch ArticlePDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessResearch ArticlePDF/EPUBIn This Issue Ruth Williams Ruth WilliamsRuth Williams Search for more papers by this author Originally published20 Jan 2017https://doi.org/10.1161/RES.0000000000000134Circulation Research. 2017;120:245SMC Reprogramming to Resident Progenitor Cells (p 296)Download figureDownload PowerPointMajesky et al discover an unexpected origin of vascular progenitor cells.The adventitia, which forms the outer layer of blood vessels, is a collagen-rich tissue that has been traditionally thought to provide only structural support to the vessel. But recent research has shown that the adventitia contains a variety of dynamic cell types, such as the vascular progenitor cells. The origin of these progenitors was unclear, but Majesky and colleagues have now discovered, quite by accident, the source of at a least a subpopulation of these cells in mice. The team was studying how, after injury, the smooth muscle cells (SMCs) within the medial layer of blood vessels dedifferentiate and migrate to the innermost intimal layer. Unexpectedly, the team observed that SMCs also migrate to the adventitia, where they lost their SMC characteristics and gained stem cell markers. Further fate-mapping experiments confirmed that SMCs do indeed give rise to a subpopulation of adventitia-residing progenitors. These SMC-derived progenitors were multipotent, giving rise to several cell types in vitro and in matrigel implants in mice. The team also found that the transition from SMC to progenitor was regulated by the transcription factor Klf4. Further studies are required to assess how these SMC-derived progenitors contribute to vascular homeostasis and pathology and how these cells could be targeted to treat vascular diseases, say the authors.CD34+ Cells for Post STEMI LV Dysfunction (p 324)Download figureDownload PowerPointQuyyumi et al report the results of a phase II stem cell trial for treating myocardial infarction.Nearly 20% of patients aged 45 and over who suffer a heart attack die within a year. Treatments to improve outcomes are desperately needed, and one possible therapeutic approach that has received much recent attention is coronary infusion of bone marrow-derived cells. Indeed, over 2600 patients have received autologous bone marrow cells in a variety of trials, but the outcomes have been inconsistent. It has been suggested that such unpredictable results may be due to variable bone marrow cell constituents, and that enriching for cells that express the stem cell marker CD34 may provide cell populations with greater clinical benefit. Indeed, a phase I trial of treatment with autologous CD34-positive bone marrow cells improved outcomes in heart attack victims. Now, Quyyumi and colleagues report that some patients enrolled in a randomized, double-blind phase II trial of CD34 cells also fared better than control subjects—with improved ejection fractions and reduced infarct sizes after 6 months. Improvement was seen in patients who were given a high-dose of cells (over 20 million), but low-dose recipients were indistinguishable from controls. The results confirm the safety of autologous CD34+ cell infusions and suggest that high doses of these cells may have some clinical benefit. Based on these results, the authors suggest that further randomized controlled trials should be performed to examine the efficacy of bone marrow-derived CD34+ cells.Mesenchymal Stem Cells for Heart Failure (p 332)Download figureDownload PowerPointButler et al present the results of an intravenous stem cell therapy trial for nonischemic cardiomyopathy.Nonischemic cardiomyopathy is a contractile dysfunction not caused by coronary artery disease (CAD) or myocardial infarction (MI). Nevertheless, as with CAD- or MI-induced heart failure, patients with nonischemic cardiomyopathy could potentially benefit from stem cell therapy. To date, cell therapies for any form of heart failure have been administered directly to the heart—because cell engraftment was considered necessary for clinical effect. However, some effects may derive from secreted factors, such as the anti-inflammatory factors produced by mesenchymal stem cells (MSCs). Butler and colleagues have, therefore, performed the first trial of intravenously administered stem cells for heart failure, giving 22 patients with nonischemic cardiomyopathy infusions of allogenic MSCs. While the team noted no difference in the occurrence of death or hospitalization between test and control subjects, patients receiving the MSCs had reduced systemic inflammation and could walk greater distances after 3 months. An inverse association between inflammation and left ventricle ejection fraction was also noted. These promising early results now pave the way for further follow up trials, say the authors. Previous Back to top Next FiguresReferencesRelatedDetails January 20, 2017Vol 120, Issue 2 Advertisement Article InformationMetrics © 2017 American Heart Association, Inc.https://doi.org/10.1161/RES.0000000000000134PMID: 30520342 Originally publishedJanuary 20, 2017 PDF download Advertisement

Safety and Effectiveness of Bone Marrow Cell Concentrate in the Treatment of Chronic Critical Limb Ischemia Utilizing a Rapid Point-of-Care System.

Critical limb ischemia (CLI) is the end stage of lower extremity peripheral vascular disease (PVD) in which severe obstruction of blood flow results in ischemic rest pain, ulcers and/or gangrene, and a significant risk of limb loss. This open-label, single-arm feasibility study evaluated the safety and therapeutic effectiveness of autologous bone marrow cell (aBMC) concentrate in revascularization of CLI patients utilizing a rapid point-of-care device. Seventeen (17) no-option CLI patients with ischemic rest pain were enrolled in the study. Single dose of aBMC, prepared utilizing an intraoperative point-of-care device, the Res-Q™ 60 BMC system, was injected intramuscularly into the afflicted limb and patients were followed up at regular intervals for 12 months. A statistically significant improvement in Ankle Brachial Index (ABI), Transcutaneous Oxygen Pressure (TcPO2), mean rest pain and intermittent claudication pain scores, wound/ ulcer healing, and 6-minute walking distance was observed following aBMC treatment. Major amputation-free survival (mAFS) rate and amputation-free rates (AFR) at 12 months were 70.6% and 82.3%, respectively. In conclusion, aBMC injections were well tolerated with improved tissue perfusion, confirming the safety, feasibility, and preliminary effectiveness of aBMC treatment in CLI patients.

91 - Efficacy of Combined Treatment with Bone Marrow Cell Transplantation and S-Allyl-Glutathione Administration for CCl4-Induced Hepatic Fibrosis

Object It is an important challenge to prevent and/or inhibit the progression of hepatic fibrosis. Autologous bone marrow cell infusion therapy is used clinically for the treatment of decompensated liver cirrhosis as advanced medical treatment B in Japan. However, it is necessary to collect a large amount of bone marrow fluid containing mesenchymal stem cells and macrophages at least twice under general anesthesia. Macrophages, especially M2 polarized macrophages have an important role in the progression of hepatic fibrosis. We found that s-allyl-glutathione (SAG) controls the polarization of macrophages and inhibits hepatic fibrosis. The aim of this study was to evaluate the enhancing anti-fibrotic effect of combined treatment with bone marrow cell (BMC) transplantation and administration of SAG on CCl4-induced hepatic fibrosis in rats. Methods Male Wistar rats (5-weeks-old) were intraperitoneally injected with 0.5 µg/g body weight of CCl4 twice a week for 8 weeks. BMCs (3.0×10 6) were infused once through the tail vein (BMC group) 4 weeks after starting CCl4 treatment. Some rats were administered 200 mg/kg/day of SAG (SAG group) every day after the first two weeks of CCl4 administration. Others received combined treatment with BMC and SAG (BMC+SAG group). After 9 weeks of CCl4 injections, rats were sacrificed and the blood and liver were analyzed. Results Four weeks of CCl4 injections established liver fibrosis. The BMC, SAG, and BMC+SAG groups showed significantly improved indices of fibrosis (hepatic hydroxyproline content, mRNA of collagen 1A1, and histological analysis). In particular, collagen 1A1 mRNA was markedly decreased by 80% in the BMC+SAG group compared with 40% in the SAG or BMC alone groups. Conclusion Combination therapy of BMC and SAG improved CCl4-induced liver fibrosis. These findings suggest that the combination therapy might be effective and convenient for the treatment of established fibrosis.

Status of and candidates for cell therapy in liver cirrhosis: overcoming the "point of no return" in advanced liver cirrhosis.

The treatment of liver cirrhosis is currently being standardized and developed specifically to reduce activation of hepatic stellate cells (HSCs), inhibit fibrosis, increase degradation of matrix components, and reduce activated myofibroblasts. Cell therapy can be applied in the treatment of liver cirrhosis; however, the characteristic features of this therapy differ from those of other treatments because of the involvement of a living body origin and production of multiple cytokines, chemokines, matrix metalloproteinases (MMPs), and growth factors. Thus, cell therapies can potentially have multiple effects on the damaged liver, including alleviating liver cirrhosis and stimulating liver regeneration with affecting the host cells. Cell therapies initially involved autologous bone marrow cell infusion, and have recently developed to include the use of specific cells such as mesenchymal stem cells and macrophages. The associated molecular mechanisms, routes of administration, possibility of allogeneic cell therapy, and host conditions appropriate for cell therapies are now being extensively analyzed. In this review, we summarize the status and future prospects of cell therapy for liver cirrhosis.

Characteristics of responders to autologous bone marrow cell therapy for no-option critical limb ischemia

BackgroundThe present study investigated factors associated with therapeutic benefits after autologous bone marrow cell (BMC) therapy in patients with “no-option” critical limb ischemia (CLI).Methods and resultsSixty-two patients with advanced CLI (Rutherford category 5 or 6) not eligible for revascularization were randomized to treatment with 40 ml of autologous BMCs (SmartPreP2) by local intramuscular (n = 32) or intra-arterial (n = 30) application. The primary endpoint was limb salvage and wound healing at 12 months. Seven patients (11 %) died during the follow-up from reasons unrelated to stem cell therapy. The BMC product of patients with limb salvage and wound healing (33/55) was characterized by a higher CD34+ cell count (p = 0.001), as well as a higher number of total bone marrow mononuclear cells (BM-MNCs) (p = 0.032), than that of nonresponders (22/55). Patients with limb salvage and wound healing were younger (p = 0.028), had lower C-reactive protein levels (p = 0.038), and had higher transcutaneous oxygen pressure (tcpO2) (p = 0.003) before cell application than nonresponders. All patients with major tissue loss at baseline (Rutherford 6 stage of CLI, n = 5) showed progression of limb ischemia and required major limb amputation. In the multiple binary logistic regression model, the number of applied CD34+ cells (p = 0.046) and baseline tcpO2 (p = 0.031) were independent predictors of limb salvage and wound healing. The number of administrated BM-MNCs strongly correlated with decreased peripheral leukocyte count after 6 months in surviving patients with limb salvage (p = 0.0008).ConclusionPatients who benefited from autologous BMC therapy for “no-option” CLI were treated with high doses of CD34+ cells. The absolute number of applied BM-MNCs correlated with the improvement of inflammation. We hypothesize that the therapeutic benefit of cell therapy for peripheral artery disease is the result of synergistic effects mediated by a mixture of active cells with regenerative potential. Patients at the most advanced stage of CLI do not appear to be suitable candidates for cell therapy.Trial registrationThe study was approved and registered by the ISRCTN registry. Trial registration: ISRCTN16096154. Registered: 26 July 2016.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0379-z) contains supplementary material, which is available to authorized users.

Sustained quality of life improvement after intracoronary injection of autologous bone marrow cells in the setting of acute myocardial infarction: results from the BONAMI trial.

Cardiac cell therapy is a promising treatment for acute myocardial infarction (AMI), leading to cardiac function improvement. However, whether it translates into quality of life (QoL) improvement is unclear. We hypothesized that administration of bone marrow cells (BMC) to patients with AMI improves QoL. In the multicenter BONAMI trial (NCT00200707), patients with reperfused AMI and decreased myocardial viability were randomized to intracoronary autologous BMC infusion (n=52) or state-of-the-art therapy (n=49). QoL data, derived from the Minnesota Living with Heart Failure questionnaire (MLHFQ), were obtained 1, 3, and 12months after AMI and analyzed using a Rasch-family model. Using this model, QoL improved over time in the BMC group (p=0.025) but not in the control group. Furthermore, the BMC-group patients displayed a better QoL than the control-group patients at 3 and 12months post-AMI (p=0.034 and p=0.003, respectively). These findings were not detected when analyzing MLHFQ data using a standard method. Cardiac function, myocardial viability, mortality, and number of major adverse cardiac events did not differ between treatment groups. Our results suggest that BMC therapy can improve QoL, stressing the need for confirmation trials and for systematic QoL assessment in cardiac cell therapy trials .

LONG-TERM RESULTS OF TRANSMYOCARDIAL LASER REVASCULARIZATION COMBINED WITH IMPLANTATION OF AUTOLOGOUS BONE MARROW MONONUCLEAR FRACTION IN THE TREATMENT OF CHRONIC ISCHEMIC HEART DISEAS

Aim. Clinical and instrumental assessment of long-term results of autologous bone marrow cells (BMC) implantation in laser channels in surgery of ischemic heart disease with diffuse and distal coronary disease.Materials and Methods. In the period of 2007–2008 35 CHD patients with diffuse and distal coronary disease underwent BMC implantation in laser channels during coronary artery bypass grafting (CABG). The control group consisted of 29 patients. All patients in this group underwent only direct myocardial revascularization (DMR). In the long-term period we examined only 30 patients of the first group. Clinical and instrumental assessment of the method efficacy was carried out in 2 weeks, 6 months and 6 years after surgery.Results. FC (NYHA) mean value in indirect revascularization group significantly decreased from 2.57 ± 0.61 preoperatively to 1.77 ±0.66 in6 months after surgery (p = 0.043). After 6 years FC (NYHA) was not significantly changed – 1.84 ± 0.42 (p = 0.053). Perfusion scintigraphy revealed a slight decrease of stable perfusion defect (SPD) in the immediate postoperative period, a more pronounced reduction of SPD in 6 months after surgery. The average value of SPD before surgery was 20.46 ± 10.75%, in 2 weeks after the operation – 19.07 ± 9.69%, in 6 months after surgery – 15.22 ± 9.49%. In the long-term period (6 years) SPD was 14.8 ± 8.43% (p = 0.047). A similar pattern was observed in the analysis of transient perfusion defect: baseline – 30 ± 2.2%, in 6 months – 15 ± 1.3%, in the long term period – 20 ± 6.1% (p = 0.047). The average value of left ventricular ejection fraction (LVEF) before surgery was 55 ± 10.4%, in 2 weeks after the operation – 55.7 ± 9.3%, in 6 months – 56.7 ± 10%, after 6 years – 54 ± 12%. The dynamics is unauthentic (p = 0.068).Conclusion. BMC implantation in laser channels in addition to CABG is a safe and effective method of surgical treatment in case of CABG inability. The effect of indirect revascularization is formed in the first 6 months after surgery and remains at the same level for 6 years.

Autologous bone-marrow mesenchymal cell induced chondrogenesis (MCIC).

Degenerative and traumatic articular cartilage defects are common, difficult to treat, and progressive lesions that cause significant morbidity in the general population. There have been multiple approaches to treat such lesions, including arthroscopic debridement, microfracture, multiple drilling, osteochondral transplantation and autologous chondrocyte implantation (ACI) that are currently being used in clinical practice. Autologous bone-marrow mesenchymal cell induced chondrogenesis (MCIC) is a single-staged arthroscopic procedure. This method combines a modified microfracture technique with the application of a bone marrow aspirate concentrate (BMAC), hyaluronic acid and fibrin gel to treat articular cartilage defects. We reviewed the current literatures and surgical techniques for mesenchymal cell induced chondrogenesis.

High-dose chemotherapy and autologous bone marrow or stem cell transplantation versus conventional chemotherapy for women with early poor prognosis breast cancer.

Overall survival rates are disappointing for women with early poor prognosis breast cancer. Autologous transplantation of bone marrow or peripheral stem cells (in which the woman is both donor and recipient) has been considered a promising technique because it permits use of much higher doses of chemotherapy. To compare the effectiveness and safety of high-dose chemotherapy and autograft (either autologous bone marrow or stem cell transplantation) with conventional chemotherapy for women with early poor prognosis breast cancer. We searched the Cochrane Breast Cancer Group Specialised Register, MEDLINE (1966 to October 2015), EMBASE (1980 to October 2015), the World Health Organization's International Clinical Trials Registry Search Platform, and ClinicalTrials.gov on the 21 October 2015. Randomised controlled trials (RCTs) comparing high-dose chemotherapy and autograft (bone marrow transplant or stem cell rescue) versus chemotherapy without autograft for women with early poor prognosis breast cancer. Two review authors selected RCTs, independently extracted data and assessed risks of bias. We combined data using a Mantel-Haenszel fixed-effect model to calculate pooled risk ratios (RRs) and 95% confidence intervals (CIs). We assessed the quality of the evidence using GRADE methods. Outcomes were survival rates, toxicity and quality of life. We included 14 RCTs of 5600 women randomised to receive high-dose chemotherapy and autograft (bone marrow transplant or stem cell rescue) versus chemotherapy without autograft for women with early poor prognosis breast cancer. The studies were at low risk of bias in most areas.There is high-quality evidence that high-dose chemotherapy does not increase the likelihood of overall survival at any stage of follow-up (at three years: RR 1.02, 95% CI 0.95 to 1.10, 3 RCTs, 795 women, I² = 56%; at five years: RR 1.00, 95% CI 0.96 to 1.04, 9 RCTs, 3948 women, I² = 0%; at six years: RR 0.94, 95% CI 0.81 to 1.08, 1 RCT, 511 women; at eight years: RR1.17, 95% CI 0.95 to 1.43, 1 RCT, 344 women; at 12 years: RR 1.18, 95% CI 0.99 to 1.42, 1 RCT, 382 women).There is high-quality evidence that high-dose chemotherapy improves the likelihood of event-free survival at three years (RR 1.19, 95% CI 1.06 to 1.34, 3 RCTs, 795 women, I² = 56%) but this effect was no longer apparent at longer duration of follow-up (at five years: RR 1.04, 95% CI 0.99 to 1.09, 9 RCTs, 3948 women, I² = 14%; at six years RR 1.04, 95% CI 0.87 to 1.24, 1 RCT, 511 women; at eight years: RR 1.27, 95% CI 0.99 to 1.64, 1 RCT, 344 women; at 12 years: RR 1.18, 95% CI 0.95 to 1.45, 1 RCT, 382 women).Treatment-related deaths were much more frequent in the high-dose arm (RR 7.97, 95% CI 3.99 to 15.92, 14 RCTs, 5600 women, I² = 12%, high-quality evidence) and non-fatal morbidity was also more common and more severe in the high-dose group. There was little or no difference between the groups in the incidence of second cancers at four to nine years' median follow-up (RR 1.25, 95% CI 0.90 to 1.73, 7 RCTs, 3423 women, I² = 0%, high-quality evidence). Women in the high-dose group reported significantly worse quality-of-life scores immediately after treatment, but there were few statistically significant differences between the groups by one year.The primary studies were at low risk of bias in most areas, and the evidence was assessed using GRADE methods and rated as high quality for all comparisons. There is high-quality evidence of increased treatment-related mortality and little or no increase in survival by using high-dose chemotherapy with autograft for women with early poor prognosis breast cancer.

Joint-preserving regenerative therapy for patients with early-stage osteonecrosis of the femoral head.

Osteonecrosis of the femoral head is an intractable disease often occurring in patients aged 30–40 years that can cause femoral head collapse, pain, and gait disturbance. Background factors, including corticosteroid use, alcohol intake, and idiopathic causes, have been indicated. It is estimated that 70–80 % of osteonecrosis patients experience femoral head collapse, for which total hip arthroplasty is considered the most effective treatment, even in young patients. Thus, there is a crucial need for developing a minimally invasive regenerative therapy as a preventive surgery for femoral head collapse: this has been an important area of research in the past decades. Core decompression, the most popular minimally invasive surgery for osteonecrosis of the femoral head, has been used for a long time; however, it has been insufficient to prevent femoral head collapse. For further improvement in therapeutic efficacy, cell transplantation and the use of artificial bone and growth factors have been proposed in addition to core decompression. Since 2000, newer therapies such as autologous bone marrow cell transplantation and the embedding of metal implant rods have been developed in Europe and the USA; however, these approaches have yet to become a global standard. This practical review summarizes applied state-of-the-art regenerative therapy-based core decompression. We introduce the clinical application of recombinant human fibroblast growth factor (rhFGF)-2-impregnated gelatin hydrogel for patients with precollapse osteonecrosis of the femoral head. Radiography and computed tomography have confirmed bone regeneration inside the femoral heads around the region of rhFGF-2 gelatin hydrogel administration. With further development, the minimally invasive method, which can be expected to promote bone regeneration in necrotic areas, could become a useful early-stage treatment for osteonecrosis of the femoral head. Patients can resume their daily routine soon after surgery, and the procedure is inexpensive. As such, it is a promising regenerative therapy that can be actively employed in osteonecrosis of the femoral head before femoral head collapse.

Disc Regeneration Using MSC Transplanted via the Endplate Route

Introduction Stem cell based intervertebral disc (IVD) regeneration is quickly moving toward clinical applications.1 However, many aspects need to be investigated to routinely translate this therapy to clinical applications, in particular, the most efficient way to deliver cell to the IVD. Cells are commonly delivered to the IVD through the annulus fibrosus (AF) injection. However, recent studies have shown serious drawbacks of this approach2–4 suggesting that intradiscal injection through the AF route itself is not completely innocuous and may disable the treatments to therapeutic agents delivered. As an alternative we have described and tested a new surgical approach to the IVD via the endplate-pedicles (transpedicular approach). The Purpose of the study was to test MSCs/hydrogel transplantation for IVD regeneration in a grade IV preclinical model of IDD on large size animals via the transpeducular approach5 with cell dose escalation. Material and Methods Adult sheep ( n = 18) underwent bone marrow aspiration for autologous MSC isolation and expansion. MSC were suspended in autologous PRP and conjugated with Hyaluronic Acid and Batroxobin at the time of transplant (MSCs/hydrogel). Nucleotomy was performed via the transpedicular approach under fluoroscopy6 in four lumbar IVDs and that were injected with 1) hydrogel, 2) Low doses of MSC/hydrogel [5x106 cell/ml] 3) High doses of MSC/hydrogel [1x107 cell/ml], 4) no injection (CTRL). The endplate tunnel was sealed using a polyurethane scaffold.7 X-ray and MRI were performed at baseline and 1,3,6,12 months. Disc height and MRI indexes were calculated at each time point. Disc macro- and micro-morphology were analyzed at each time point. Results The MRI index showed a significant decrease in the untreated group, the disc injected with hydrogel and those injected with low MSC dose compared with healthy discs in all time points. The discs treated with high dose of MSC showed maintenance of the MRI index compared with the healthy disc. Morphologically, the grade of degeneration evaluated using the Thompson grade system8 was in agreement with the grades observed at the MRI. The High MSC dose treated discs demonstrated abundant cartilage formation at 3 months, and to a lesser extent at 6 and 12 months. For the carrier and low MSC dose treated groups, however, there was less proteoglycan matrix. Conclusion An effective dose of autologous MSC [1x107 cell/ml] delivered via the alternative transpedicular approach regenerates the NP in a preclinical model of grade IV IDD. These data highlight as the disc regeneration can be achieved maintaining the AF intact via the end-plate route. This preclinical study has high translational value as large animal model with the long fallow up were used, MSCs were expanded in GMP facility simulating the clinical scenario, and the hydrogel were composed of clinically available drags and materials. This study bring a significant contribution toward the translation of regenerative therapies for the biological restoration of degenerative changes in the IVD, which is crucial to improve present clinical treatment and life quality of several patients. Acknowledgment The support of the Italian Ministry of Instruction, University and Research Grant (PRIN-200938NT8Z), the Young Investigator Research Grant of the Italian Ministry of Health (GR-2010–2318448) and the BIOSPINA Award of the Italian Society of Spine Surgery (SICV&GIS) are gratefully acknowledged. References Orozco L, Soler R, Morera C, Alberca M, Sánchez A, García-Sancho J. Intervertebral disc repair by autologous mesenchymal bone marrow cells: a pilot study. Transplantation 2011;92(7):822–828 Carragee EJ, Don AS, Hurwitz EL, Cuellar JM, Carrino JA, Herzog R. 2009 ISSLS Prize Winner: Does discography cause accelerated progression of degeneration changes in the lumbar disc: a ten-year matched cohort study. Spine 2009;34(21):2338–2345 Korecki CL, Costi JJ, Iatridis JC. Needle puncture injury affects intervertebral disc mechanics and biology in an organ culture model. Spine 2008;33(3):235–241 Vadalà G, Sowa G, Hubert M, Gilbertson LG, Denaro V, Kang JD. Mesenchymal stem cells injection in degenerated intervertebral disc: cell leakage may induce osteophyte formation. J Tissue Eng Regen Med 2012;6(5):348–355 Vadalà G, Russo F, Pattappa G, et al. The transpedicular approach as an alternative route for intervertebral disc regeneration. Spine 2013;38(6):E319–E324 Vadalà G, De Strobel F, Bernardini M, Denaro L, D'Avella D, Denaro V. The transpedicular approach for the study of intervertebral disc regeneration strategies: in vivo characterization. Eur Spine J 2013;22(Suppl 6):S972–S978 Vadala G, et al. A nucleotomy model with intact annulus fibrosus to test intervertebral disc regeneration strategies. Tissue Eng Part C Methods 2015;21(11):1117–1124 Thompson JP, Pearce RH, Schechter MT, Adams ME, Tsang IK, Bishop PB. Preliminary evaluation of a scheme for grading the gross morphology of the human intervertebral disc. Spine 1990;15(5):411–415

Translation of cell therapy into clinical practice: validation of an application procedure for bone marrow progenitor cells and platelet rich plasma.

Tissue regeneration can be improved by local application of autologous bone marrow derived progenitor cells (BMSC) and platelet rich plasma (PRP). However, there is a lack of standardized application procedures for clinical use. Therefore, a technique in accordance with the guidelines for advanced therapies medical products of the European Medicine Agency was developed and established. In detail, a process for the isolation and formulation of autologous bone marrow cells (BMC) and PRP in a clinical setting was validated. To investigate the influence of storage time and temperature on gel formation and gel stability, different concentrations of BMC were stored with and without additional platelets, thrombin and fibrinogen and analyzed over a period of 28 days. In addition, cell vitality using a live-dead staining and migration ability of human mesenchymal stem cells (hMSC) in the gel clot was investigated. For an optimized stable gel clot, human BMC and PRP should be combined with 10% to 20% fibrinogen (9 mg/mL to 18 mg/mL) and 5% to 20% thrombin (25 I.E. to 100 I.E.). Both freshly prepared and stored cells for 1 to 7 days had a stable consistence over 28 days at 37°C. Different platelet concentrations did not influence gel clot formation. The ratio of living cells did not decrease significantly over the observation period of 5 days in the live-dead staining. The study identified an optimal gel texture for local application of BMC and PRP. Seeded hMSC could migrate therein and were able to survive to initiate a healing cascade.

MP65-04 A NOVEL REGENERATIVE TREATMENT FOR FEMALE STRESS URINARY INCONTINENCE: LONG-TERM OUTCOME OF INITIAL THREE CASES UNDERGOING PERIURETHRAL INJECTION OF AUTOLOGOUS ADIPOSE-DERIVED REGENERATIVE CELLS

METHODS: ISD was created in female NHPs by cutting and then cauterizing the pudendal innervation to the urinary sphincter complex. Chronic fibrotic, muscle poor ISD developed within 6 months post-surgery. Partial bone marrow transplantation with autologous Lenti-GFP bone marrow cells (BMCs) was performed and NHPs (n1⁄46/ experimental condition) were then given sphincter injections of LentiMcherry-labeled 5 million autologous skMPCs, or SDF-1a. Tissues were removed 6 months later and analyzed for sphincter content of muscle, collagen, BMCs, skMPCs and SDF-1a. RESULTS: When compared to previously studied NHPs with acute ISD (Badra et al, J Urol. 2013;190(5):1938), there was less expression of BMCs andSDF-1a in the urinary sphincter complexes ofNHPswith chronic ISD (p<0.05). This correlated with reduced effectiveness of skMPCs to restore sphincter muscle, innervation and vascularization in chronic vs. almost complete restoration in acute ISD (p<0.05). That injection of SDF-1a restores sphincter structure and function in chronic ISD further supports a role of the SDF-1a pathway in sphincter regeneration. CONCLUSIONS: The underlying reason(s) by which SDF-1a can, but skMPC cannot, stimulate sphincter regeneration in chronic ISD remains unclear, but could be related to the reduced capacity of skMPCs to stimulate SDF-1a mediated cell homing to the sphincter complex in chronic ISD. Providing SDF-1a to a fibrotic urinary sphincter may represent a novel approach to treat chronic ISD.

Various Cell Populations Within the Mononuclear Fraction of Bone Marrow Contribute to the Beneficial Effects of Autologous Bone Marrow Cell Therapy in a Rodent Stroke Model

Cell-based therapies including bone-marrow derived mononuclear cells (MNCs) are now widely being studied because of their pleotropic effects and promising results to improve recovery after stroke in animal models. Unlike other types of cell therapies, MNCs is a mixture of lymphoid, myeloid, erythroid, and stem cell populations. Which cell population(s) accounts for the beneficial effects of MNCs in stroke recovery is unclear. In this paper, we employed a mouse stroke model with middle cerebral artery occlusion (MCAo), and used positively and negatively sorted autologous MNCs by MACs to determine which fractions of the MNCs contribute to their beneficial effects. We evaluated the benefits of neurofunctional recovery produced by individual cell lineages within MNCs in a long-term observation study up to 28days after stroke. Mortality and modulation of inflammation were also compared among different sub-populations. We further studied the impact of neurotoxicity posed by activated microglia in the presence of different cell lineages within MNCs. We concluded that myeloid cell lineage and stem cell/progenitors appeared to be important components within MNCs that contribute to improved outcomes after stroke.

Efficiency of Cell Therapy in Liver Cirrhosis.

We studied safety and clinical efficacy of transplantation of autologous bone marrow cell in complex therapy of 158 patients with chronic hepatitis and cirrhosis of the liver. The efficiency of cell therapy was assessed in 12 months after single injection of the cells. The positive response (alleviation of liver cirrhosis or stabilization of the pathological process) was observed in 70% cases. The efficacy of therapy correlated with the severity and etiology of the disease and was maximum in patients with Child-Pugh class A (in 82.5% cases) and class B liver cirrhosis (in 79% cases); in patients with class C liver cirrhosis, the positive response was achieved in 42.5% cases. In 39 patients, ultrasonic examination performed in 3 years after transplantation revealed no focal lesions or ectopic ossification foci.

Abstract TMP32: Intra-arterial ALD401 Cell Therapy is Associated With Reduction in Stroke Volume at 90 Days in a Subset of the RECOVER-stroke Trial

Background: Stroke is a leading cause of major long-term disability worldwide. Despite availability of IV tPA and mechanical thrombectomy, over half of treated patients have significant disability. Cell based therapy is a promising in pre-clinical studies in improving stroke recovery. Brain infarct volume is a major predictor of stroke outcome and may decrease with this treatment. We hypothesized that patients treated with intra-arterial stem cell therapy will have a significant reduction in stroke volume compared to non-treated patients. Methods: The RECOVER-Stroke trial is a randomized, sham-controlled study to determine the safety and efficacy of processed autologous bone marrow cells (ALD-401) injection via intracarotid infusion in ischemic stroke patients. Ischemic stroke patients were double-blind randomized into 2 groups, receiving ALD-401 injection or sham procedure between 9-19 days post anterior circulation ischemic stroke. All enrolled patients at 1 of 8 sites in the trial were included in this analysis. Two blinded physicians used OSIRIX software to analyze stroke volume on DWI/ 24h CT brain and MRI at 90, 180, 365 days post intervention. Volumes were compared between the 2 groups at these timepoints. Results: A total of 11 patients at our center (out of 48 multicenter patients) were included in this study, 6 patients receiving sham procedure and 5 patients receiving ALD-401. The two groups did not show any significant difference in their baseline variables including age, Mean baseline stroke volumes were not significantly different between ALD-401 (103.9±96.2ml) and sham groups (70.2±46ml), p=0.53. The stroke volume reduction in the ALD-401 was 22.9±15.5ml vs. 4.04±12.5ml in the sham group (p=0.046). On day 180 and 365, both groups did not show any further significant stroke volume reduction from day 90. Conclusion: In this exploratory analysis from one center in the multi-center RECOVER-Stroke trial, intra-arterial stem cell injection with ALD-401 was associated with significant reduction in stroke volume at 90 days post therapy compared to sham procedure in subacute ischemic stroke patients.

Novel Technique of Achilles Tendon Reconstruction Using Semitendinosus and Gracilis Tendon. Preliminary Report.

Achilles tendon ruptures are a common clinical problem. While there are many techniques of Achilles tendon reconstruction, there is little evidence that any of them is clearly superior to the others. The most common techniques rely on augmentation of the plantar tendon, peroneus brevis tendon and flexor hallucis longus tendon. The article describes a novel procedure for Achilles tendon reconstruction using the hamstring grafts, Endobutton stabilization, collagen membrane and the administration of autologous bone marrow cells to the region of the reconstructed tendon. This technique was used by the author (with the good early results) in so-called "difficult cases" as a "salvage procedure". The use of a hamstring autograft is safe and does not further affect the injured foot or induce an autoimmune reaction.

Umbilical Cord Mesenchymal Stromal Cell With Autologous Bone Marrow Cell Transplantation in Established Type 1 Diabetes: A Pilot Randomized Controlled Open-Label Clinical Study to Assess Safety and Impact on Insulin Secretion.

To determine the safety and effects on insulin secretion of umbilical cord (UC) mesenchymal stromal cells (MSCs) plus autologous bone marrow mononuclear cell (aBM-MNC) stem cell transplantation (SCT) without immunotherapy in established type 1 diabetes (T1D). Between January 2009 and December 2010, 42 patients with T1D were randomized (n = 21/group) to either SCT (1.1 × 10(6)/kg UC-MSC, 106.8 × 10(6)/kg aBM-MNC through supraselective pancreatic artery cannulation) or standard care (control). Patients were followed for 1 year at 3-month intervals. The primary end point was C-peptide area under the curve (AUC(C-Pep)) during an oral glucose tolerance test at 1 year. Additional end points were safety and tolerability of the procedure, metabolic control, and quality of life. The treatment was well tolerated. At 1 year, metabolic measures improved in treated patients: AUCC-Pep increased 105.7% (6.6 ± 6.1 to 13.6 ± 8.1 pmol/mL/180 min, P = 0.00012) in 20 of 21 responders, whereas it decreased 7.7% in control subjects (8.4 ± 6.8 to 7.7 ± 4.5 pmol/mL/180 min, P = 0.013 vs. SCT); insulin area under the curve increased 49.3% (1,477.8 ± 1,012.8 to 2,205.5 ± 1,194.0 mmol/mL/180 min, P = 0.01), whereas it decreased 5.7% in control subjects (1,517.7 ± 630.2 to 1,431.7 ± 441.6 mmol/mL/180 min, P = 0.027 vs. SCT). HbA1c decreased 12.6% (8.6 ± 0.81% [70.0 ± 7.1 mmol/mol] to 7.5 ± 1.0% [58.0 ± 8.6 mmol/mol], P < 0.01) in the treated group, whereas it increased 1.2% in the control group (8.7 ± 0.9% [72.0 ± 7.5 mmol/mol] to 8.8 ± 0.9% [73 ± 7.5 mmol/mol], P < 0.01 vs. SCT). Fasting glycemia decreased 24.4% (200.0 ± 51.1 to 151.2 ± 22.1 mg/dL, P < 0.002) and 4.3% in control subjects (192.4 ± 35.3 to 184.2 ± 34.3 mg/dL, P < 0.042). Daily insulin requirements decreased 29.2% in only the treated group (0.9 ± 0.2 to 0.6 ± 0.2 IU/day/kg, P = 0.001), with no change found in control subjects (0.9 ± 0.2 to 0.9 ± 0.2 IU/day/kg, P < 0.01 vs. SCT). Transplantation of UC-MSC and aBM-MNC was safe and associated with moderate improvement of metabolic measures in patients with established T1D.

Preliminary Results of Multiple Epiphyseal Drilling and Autologous Bone Marrow Implantation for Osteonecrosis of the Femoral Head Secondary to Sickle Cell Disease in Children.

Sickle cell disease (SCD) is the most common cause of osteonecrosis of the femoral head (ONFH) during childhood. The natural history of ONFH in SCD is poor with progression to femoral head collapse and deformity that ultimately may lead to hip osteoarthritis. Multiple epiphyseal drilling with autologous bone marrow implantation may enhance the mechanism of osteogenic repair. The purpose of this study was to describe early clinical and radiographic outcomes in pediatric patients with ONFH secondary to SCD after multiple epiphyseal drilling and AMBI. Nine boys and 2 girls (14 hips) with a mean age of 12.7 years (range, 9.7-18 y) at the time of surgery were evaluated. All patients were followed for at least 1 year after surgery and the mean duration of follow-up was 25 months (range, 12-47 mo). Clinical outcome was assessed by the pain domain from the Children's Hospital Oakland Hip Evaluation Scale and by hip range of motion. The extent of femoral head involvement was assessed by measuring the arc of necrosis and the amount of collapse on anteroposterior and frog lateral radiographs. The University of Pennsylvania system was used to assess the osteonecrosis stage. Wilcoxon signed-rank tests were used to compare radiographic and clinical variables. Compared with preoperative measures, multiple epiphyseal drilling with AMBI resulted in significant improvement in pain (P=0.0010), internal hip rotation (P=0.0005), external hip rotation (P=0.0010), and flexion (P=0.0015). The amount of lateral collapse (P=0.4609), anterior collapse (P=0.4258), and the arc of necrosis (P=0.2251) were not significant after surgery. The majority of the hips had either improvement in the Steinberg stage of disease (4/14 hips) or no further progression (7/14 hips) at the latest follow-up. There were no surgical complications. Multiple epiphyseal drilling with autologous bone marrow cell implantation allows for short-term clinical improvement and limits radiographic progression of ONFH associated with SCD in children with minimal morbidity.

Injection of Human Cord Blood Cells With Hyaluronan Improves Postinfarction Cardiac Repair in Pigs.

Recent clinical trials using autologous bone marrow or peripheral blood cells to treat myocardial infarction (MI) show controversial results, although the treatment has a good safety profile. These discrepancies are likely caused by factors such as aging, systemic inflammation, and cell processing procedures, all of which might impair the regenerative capability of the cells used. Here, we tested whether injection of human cord blood mononuclear cells (CB-MNCs) combined with hyaluronan (HA) hydrogel improves cell therapy efficacy in a pig MI model. A total of 34 minipigs were divided into 5 groups: sham operation (Sham), surgically induced-MI plus injection with normal saline (MI+NS), HA only (MI+HA), CB-MNC only (MI+CB-MNC), or CB-MNC combined with HA (MI+CB-MNC/HA). Two months after the surgery, injection of MI+CB-MNC/HA showed the highest left ventricle ejection fraction (51.32%±0.81%) compared with MI+NS (42.87%±0.97%, p<.001), MI+HA (44.2%±0.63%, p<.001), and MI+CB-MNC (46.17%±0.39%, p<.001) groups. The hemodynamics data showed that MI+CB-MNC/HA improved the systolic function (+dp/dt) and diastolic function (-dp/dt) as opposed to the other experimental groups, of which the CB-MNC alone group only modestly improved the systolic function (+dp/dt). In addition, CB-MNC alone or combined with HA injection significantly decreased the scar area and promoted angiogenesis in the infarcted region. Together, these results indicate that combined CB-MNC and HA treatment improves heart performance and may be a promising treatment for ischemic heart diseases. This study using healthy human cord blood mononuclear cells (CB-MNCs) to treat myocardial infarction provides preclinical evidence that combined injection of hyaluronan and human CB-MNCs after myocardial infarction significantly increases cell retention in the peri-infarct area, improves cardiac performance, and prevents cardiac remodeling. Moreover, using healthy cells to replace dysfunctional autologous cells may constitute a better strategy to achieve heart repair and regeneration.