Application Of Autologous Stem Cells Research Articles

Human amniotic mesenchymal stromal cells promote bone regeneration via activating endogenous regeneration.

Rationale: The effectiveness of stem cell based-therapy for bone regeneration has been demonstrated; yet, clinical application of autologous stem cells is still limited by invasive acquisition, long culture processes, and high cost. Besides, it remains controversial whether autologous stem cells could directly participate in tissue repair after differentiation. Thus, increasing allogeneic stem cells have been developed into drugs to indirectly activate endogenous regeneration and induce tissue regeneration. Human amniotic mesenchymal stromal cells (HAMSCs) have been extensively studied, showing multiple regulatory functions, but mechanisms of HAMSCs in promoting bone regeneration are remain unclear.Methods: Proteome profile of HAMSCs and their functions on vascularized bone regeneration were investigated in vitro, while rabbit cranial defect model was used to further detect the effects of bone formation in vivo.Results: HAMSCs secrete many osteogenic, angiogenic, and immunomodulatory cytokines. In vitro, HAMSCs can promote human bone-marrow mesenchymal stromal cells (HBMSCs) migration and osteogenic differentiation; promote the capillary-tube formation of human umbilical vascular endothelial cells (HUVECs), induce HUVECs migration and pro-angiogenic genes expression, and promote M2 macrophage polarization. Further, in vivo studies suggested that transplanted HAMSCs could survive and induce M2 macrophages to secrete bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) in rabbits' skull defects at an early stage, and, in turn, promote more new bone formation.Conclusion: HAMSCs have good biocompatibility and paracrine function to promote bone repair by stimulating endogenous regeneration.

Autologous Hematopoietic Stem Cell Infusion for Myelosuppression after Bcma-CART Therapy in Relapsed Myeloma Patients

Abstract Background: The adoption of B cell mature antigen (BCMA) targeted chimeric antigen receptor (CAR) T cells is demonstrating promise prospects in refractory relapsed multiple myeloma (RRMM), however the persistent severe myelosuppression remains a serious complication. Autologous hematopoietic stem cell infusion facilitates the recovery of hematopoietic function and has contributed to efficacy and safety in clinical trials with BCMA-specific CAR T cells. In this phase I clinical study of BCMA-specific CAR T cell therapy for RRMM, we applied autologous hematopoietic stem cell(auto-HSC) infusion to promote the hematopoietic recovery which may improve the efficacy of CAR T therapy. Methods: Eligible relapsed or treatment refractory MM patients had received lymphodepleting chemotherapy of FC regimen((Fludarabine 25 mg/m2 daily for 3 days, Cyclophosphamide 500 mg/m2 daily for 2 days)) before infusion of BCMA CAR T cells at a dose of 1.0×107 cells. We performed autologous hematopoietic stem cell infusion (2.1-3.6×108/kg for MNC cells and 1.8-2.6×106/kg for CD34+ cells) in patients with Ⅳ degree of myelosuppression sustained more than 30 days, and observe hematopoietic recovery of the patient. Results: A total of 23 patients developed grade Ⅳ myelosuppression. Seven of patients suffered grade Ⅳ myelosuppression for more than 30 days received autologous hematopoietic stem cell infusion, and all 7 patients recovered hematopoietic within 14 days after stem cell reinfusion. Patient 1 had myelosuppression after CART therapy, which was characterized by severe thrombocytopenia and manifested as bleeding. Auto-HSCs were infused on the 37th day after CART cell therapy. Hematopoiesis gradually recovered 4 days after infusion, platelets increased and returned to normal 13 days after auto-HSCs infusion. Patient 2 and 5 developed granulocytopenia after CART therapy. They were infused auto-HSCs on the 32nd and 33rd day after CART cell therapy respectively. Leukocytes of patient 2 and patients 5 gradually increased 2 days and 3 days after infusion, returned to normal 7 days and 9 days after auto-HSCs infusion, respectively. Patient 3 and 4 suffered IV degree of myelosuppression, referred as leukopenia, anemia and thrombocytopenia. Stem cell was infused on day 31st and 33rd after CART therapy, erythrocyte and platelet increased 3 days after reinfusion for patient 3, as for patient 4, erythrocyte and platelets increased 3 days and 5 days after infusion, respectively. Patient 6 developed severe pancytopenia, auto-HSCs were infused on 32 days after CART therapy, leukocytes gradually increased 3 days after infusion. Leukocyte and platelet increased to normal 6 and 11 days after stem cell infusion. Patient 7 is an elderly relapse who intend to receive auto-HSCT, however, owing to his advanced age, CART therapy was preferred. Grade IV myelosuppression occurred after CART cell infusion, patient had undergone neutropenia and severe. Auto-HSCs was infused on the 32nd day after CART cell therapy, granulocyte and erythrocyte gradually increased 5days after infusion, recovered 9 daysand14days after stem cell infusion. Conclusions: Our results demonstrate application of autologous stem cells in the CAR-T therapy, whose effect was similar to the role of hematopoietic stem cell infusion in transplantation after high-dose chemotherapy, which could promote the reconstruction of bone marrow hematopoietic, avoid serious infection and hemorrhage. It suggests that autologous stem cell infusion is also a worthwhile option when patients suffer from severe and sustained myelosuppression after treatment with CAR-T cells. Disclosures No relevant conflicts of interest to declare.

Diabetes mellitus type 2 reduces the viability, proliferation, and angiogenic marker of adipose-derived stem cells cultured in low-glucose anti-oxidant-serum supplemented medium

Introduction: Hyperglycemia in diabetic patients induces elevated pro-inflammatory cytokine production, resulting in cellular damage, which may affect the regenerative function of mesenchymal stem cells (MSCs), such as adipose-derived stem cells (ADSCs). Identifying the effect of diabetes on ADSCs and optimization of culture conditions is therefore an important starting point for the application of autologous stem cells to improve clinicial outcomes. The aim of this study was to investigate the effect of diabetes on ADSCs that cultured in low-glucose anti-oxidant-serum supplemented medium.
 Methods: In this study, freshly isolated stromal vascular fraction (SVF) and expanded ADSCs were compared between diabetic and non-diabetic donors. SVF were isolated from the abdominal fat, and total viable cells and viability were estimated. Fresh SVF were cultured in low-glucose (100 mg/dL) culture medium supplemented with an anti-oxidant and fetal bovine serum (complete culture medium) at a low density for 14 days for the colony formation unit-fibroblast (CFU-F) assay. The remaining SVF were expanded to obtain ADSCs in the complete culture medium, which were evaluated based on MSCs surface marker expression and three lineage differentiation potential. Diabetic and non-diabetic ADSCs were compared with respect to population doubling time and viability after serial passage.
 Results: Total viable counts (0.97 +/- 0.39 x 109 cells/10 mL of adipose tissue, 0.56 +/- 0.39 x 109 cells/10 mL of adipose tissue, p=0.02, independent t-test), but not viability (98.63 +/- 1.12%, 98.20 +/- 1.21%, p= 0.38, independent t-test), were significantly higher for SVF cells from adipose tissues of non-diabetic donors than diabetic donors. Fewer CFU-F were obtained from cultured diabetic SVF than from non-diabetic SVF. Diabetic and non-diabetic ADSCs had similar differentiation potency and CD73 (99.44 +/- 0.34%, 97.15 +/- 5.37%, p= 0.21, Mann-Whitney U test) and CD90 (97.30 +/- 2.86%, 95.06 +/- 6.32%, p= 0.90, Mann- Whitney U test) expression, but significantly fewer diabetic ADSCs expressed CD105 or endoglin, a marker for angiogenesis (89.91 +/- 7.14%, 57.90 +/- 21.36% for non-diabetic and diabetic groups, p< 0.001, Mann-Whitney U test). Diabetic ADSCs tended to exhibit slower proliferation (4.43 +/- 2.70 days, 3.04 +/- 0.55 days, p= 0.27 in passage 2 (P2); 3.95 +/- 1.55 days, 2.96 +/- 0.91 days, p= 0.21 in P3, independent t-test) and lower viability than those of non-diabetic ADSCs (77.65 +/- 10.61%, 87.13 +/- 10.06%, p= 0.25 in P2; 82.70 +/- 8.07%, 91.15 +/- 3.77%, p= 0.04 in P3, independent t-test). Culture in low-glucose anti-oxidant-serum supplemented medium did not improve CD105 expression (65.14 +/- 5.86%, 71.06 +/- 10.27%, 64.05 +/- 10.04%, p= 0.70, for P1, P2, and P3, respectively, repeated measure ANOVA) and cell proliferation (p= 0.50 for P2 vs. P3, paired t-test) of diabetic ADSCs.
 Conclusions: Overall, diabetes reduced CD105 expression and ADSCs proliferation, suggesting that the angiogenic potency of diabetic ADSCs is reduced. The diabetic ADSCs in this study were also more prone to cell death caused by handling technique compared to non-diabetic ADSCs. Therefore, more advanced culture techniques should be applied to expand ADSCs from diabetic patients to achieve expected clinical outcomes.

ЕКСПЕРИМЕНТАЛЬНА МОДЕЛЬ ЛІКУВАННЯ ХРОНІЧНОЇ КРИТИЧНОЇ ІШЕМІЇ ТКАНИН НИЖНЬОЇ КІНЦІВКИ З ВИКОРИСТАННЯМ КУЛЬТУРИ СТОВБУРОВИХ КЛІТИН, ОДЕРЖАНИХ З ЖИРОВОЇ ТКАНИНИ

Вступ. Захворювання периферійних артерій (ЗПА) є важливою медико-соціальною проблемою, що потребує розробки нових видів терапії, зокрема, використання аутологічних стовбурових клітин.
 Мета. Вивчити морфофункціональний стан нижніх кінцівок кролів, у яких моделювали хронічну критичну ішемію тканин нижніх кінцівок (ХКІНК) та застосовували культуру аутологічних стовбурових клітин, одержаних з їх жирової тканини (ADSCs).
 Матеріали і методи. У дослідження включені 18 кролів-самиць, розподілених на 3 групи: псевдооперованих (І), оперованих, яким вводили ізотонічний розчин NaCl (ІІ), оперованих, яким вводили ADSCs (ІІІ). Морфофункціональний стан тазових кінцвок оцінювали за даними дуплексної флоуметрії, функціональних тестів та морфологічного дослідження.
 Результати. У тварин ІІІ групи спостерігали часткове відновлення функції тазової кінцівки та покращення стану м’язової тканини, чого не було у тварин ІІ групи. В ІІІ групі відзначене припинення прогресування некротичних процесів та часткова епітелізація трофічних виразок.
 Висновки. Використання ADSCs при моделюванні ХКІНК у кролів сприяло відновленню функції тазової кінцівки, що проявлялося якісними й кількісними змінами морфофункціональних показників через 5 тиж після введення стовбурових клітин.
 Ключові слова: хронічна критична ішемія тканин нижніх кінцівок; захворювання периферійних артерій; ADSCs; експеримент.

ASCOT: Autologous Bone Marrow Stem Cell Use for Osteoarthritis of the Thumb-First Carpometacarpal Joint.

Background:The first carpometacarpal joint (CMCJ) in the hand is a commonly affected joint by osteoarthritis. It causes significant thumb base pain, limiting functional capacity. Microfracturing and application of autologous stem cells has been performed on large joints such as the knee but has never been evaluated for use in the smaller joints in the hand. Our aim was to determine the potential benefit of microfracturing and autologous bone marrow stem cells for treatment of osteoarthritis of the first CMCJ in the hand.Methods:All inclusion criteria were satisfied. Preoperative assessment by the surgeon, physiotherapist, and occupational therapist was performed. The first CMCJ was microfractured and the Bone Marrow Stem Cells were applied directly. Postoperatively, the patients were followed up for 1 year.Results:Fifteen patients met inclusion criteria; however, 2 patients were excluded due to postoperative cellulitis and diagnosis of De Quervain's tenosynovitis. The mean scores of the 13-patient preoperative and 1 year follow-up assessments are visual analog score at rest of 3.23–1.69 (P = 0.0292), visual analog score on activity of 7.92–4.23 (P = 0.0019), range of motion 45.77o–55.15o (P = 0.0195), thumb opposition score 7.62–9.23 (P = 0.0154), Disability of the Arm, Shoulder and Hand score of 51.67–23.08 (P = 0.0065). Strength improved insignificantly from 4.7 kg preoperatively to 5.53 kg at 12 months (P = 0.1257). All patients had a positive Grind test preoperatively and a negative test after 12 months.Conclusions:This innovative pilot study is a new approach to osteoarthritis of the thumb.

Abstract 65: ASCOT- Autologous use of bone marrow derived Stem Cells for Osteoarthritis of the Thumb-1st CMCJ

PURPOSE: The 1st Carpometacarpal Joint (CMCJ) in the hand is a commonly affected joint by Osteoarthritis (OA). It causes significant thumb base pain, limiting functional capacity. Surgical techniques are fraught with complications. Conservative management slows disease progression but does not reverse pre-existing damage. Microfracturing and application of Autologous Stem Cells has been performed on large joints such as the knee but has never been evaluated for use in the smaller joints in the hand. Our aim is to determine the potential benefit of Autologous Bone Marrow Stem Cells (BMSC) for treatment of osteoarthritis of the 1stCMCJ in the hand. METHODS: All inclusion criteria were satisfied and patients were consented. Pre-operative assessment by the surgeon, Physiotherapist (PT) and Occupational Therapist (OT) was performed. Intra-operatively, BMSC were isolated from the hip via BMAC. The 1st CMCJ was opened and the joint surface microfractured. The BMSC were applied directly to the microfractured articular surface. Post-operatively the patients were followed up at regular intervals by the same surgeon, PT and OT. The assessments performed included: the visual analogue score (VAS) at rest and with activity, range of motion (ROM) in palmar abduction, functional DASH, strength test by lateral pinch, kapandji opposition score and the Grind Test. Statistical analysis was done using Prism Software, comparing preoperative scores and scores 6 months later using a paired T-test. RESULTS: All patients had a positive Grind test preoperatively and a negative test after 6 months. Statistically significant improvements were seen (Means scores with P values) VAS at rest 3.25 to 1.425 (P=0.0138), VAS on activity 7.7 to 3.9 (P=0.0021), ROM 46o to 53o (P= 0.0095) and DASH 47.23 to 18.7 (P=0.0046). Strength and Kapandji score improved insignificantly at 6 months. There were no significant postoperative complications. Patients were satisfied with the procedure. CONCLUSIONS: This innovative pilot study is a new approach to OA of the thumb. It is a safe procedure with no post op complications. ASCOT 2 will be designed to determine the exact role of BMSC in this procedure with a control group receiving microfracture alone.

Regenerative capacity of autologous stem cell transplantation in elderly: a report of biomedical outcomes.

The occurrence of chronic diseases such as neurological, metabolic and cardiovascular degenerative disorders increases with age. Cell therapy is an emerging approach to the treatment of these conditions. Of particular interest is the application of autologous stem cells because it eliminates post-transplantation immune rejection and there are less ethical concerns associated with their use. The regenerative capacity of stem cells harvested from elderly people is however controversial. In this review, we analyze if self-renewal potential, differentiation capability and expression of stemness genes in stem cells collected from elderly patients validate their application in clinical trials and examine the results.

Autologous Stem Cell Application in Periodontal Regeneration Technique (SAI-PRT) Using PDLSCs Directly From an Extracted Tooth···An Insight.

Periodontal regeneration represents the ultimate goal of periodontal therapy. The current regenerative techniques have limited success rates especially in advanced periodontal defects. Currently the research is focused on novel cell-based approaches for periodontal regeneration to overcome the limitations of existing treatment. The human clinical trial on stem cells based periodontal regeneration is promising. The plethora of animal studies provide sound evidence to support the belief that periodontal ligament stem cells (PDLSCs) can be used for periodontal regeneration. The direct application of autologous periodontal stem cells in treatment of intrabony defects is attempted for the first time in periodontal literature. Stem cell Application in Periodontal Regeneration Technique (SAI-PRT) using direct PDLSCs has overcome the limitations and concerns of ex- vivo stem cell culture methods like high cost, technique sensitivity, loss of stemness during cell passage, genetic manipulation and tumorigenic potential. Clinical feasibility, success and cost effectiveness over currently available techniques are encouraging. The clinical utility of this novel idea is recommended.

Co-Culture Systems of Human Sweat Gland Derived Stem Cells and Peripheral Nerve Cells: An in Vitro Approach for Peripheral Nerve Regeneration

Background/Aims: The treatment of peripheral nerve lesions still represents a clinical challenge. Several approaches such as novel biomaterials for nerve guides, addition of growth factors or cellular supplements moved in the focus of research. Especially the application of autologous stem cells is highly promising for future applications. Human sweat gland derived stem cells (hSGSCs) represent an easy accessible source of autologous adult stem cells and did already show a beneficial effect in dermal wound healing. Methods: In this study, the effect of hSGSCs on neurite outgrowth of primary adult or prenatal Dorsal root ganglia (DRG) neurons was analysed in an indirect co-culture model. Additionally, direct co-cultures with hSGSCs as a feeder layer were performed. Results: Adult and prenatal DRG neurons showed increased neurite outgrowth after 24 h co-culture with hSGSCs. The outgrowth increased significantly by the factors 5.6 and 2.6 respectively. Direct co-cultures revealed neurite alignment along the hSGSCs orientation. Conclusion: The paracrine influence of hSGSCs on neurite outgrowth, but also their ability to operate as a feeder layer with guidance properties shows great potential for future applications in peripheral nerve regeneration.

Portal vein embolization and application of autologous stem cells in patients with primary unresectable liver tumours

Only 15-20 % of patients with liver tumours can undergo radical surgery. Insufficient future liver remnant volume (FLRV) is one of the main causes of tumours unresectability. Portal vein embolization (PVE) together with administration of haematopoietic stem cells (HSC) may expand the operability of primary unresectable liver tumours. In this pilot study, the authors reported on five patients (1 hepatocellular carcinoma, 4 colorectal cancer metastases) with FLRV <30 %, who underwent PVE on the side of the tumour with a subsequent application of HSC to the non-embolized branch of portal vein. PVE with HSC application was without any complications. In three patients, a sufficient increase of FLRV occurred within 2-4 weeks followed by a liver resection. All patients were between 5-12 months after the surgery in good condition; one of them was diagnosed with pulmonary metastasis after nine months that was successfully treated with laser metastasectomy. In one patient with hepatocellular carcinoma, an increase of FLRV and progression of the tumour in the liver occurred following the PVE with administration of HSC and the patient was treated only symptomatically. Despite an adequate increase of FLRV, severe intraabdominal adhesions hampered liver resection in one patient. Combination of PVE with HSC administration appeared to be a promising method that stimulated growth of FLRV with a subsequent possibility of an early radical liver resection. The issue is a danger of tumour progression in the liver parenchyma following the PVE with HSC. The current randomized study should answer these questions (Tab. 1, Fig. 4, Ref. 38).

70th Birthday symposium of Prof. Dr. Riederer: autologous adult stem cells in ischemic and traumatic CNS disorders

Ischemic and traumatic insults of the central nervous system both result in definite chronic disability, only to some extent responsive to rehabilitation. Recently, the application of autologous stem cells (fresh bone marrow-derived mononuclear cells including mesenchymal and hematopoietic stem cells) was suggested to provide a strategy to further improve neurological recovery in these disorders. During the acute phase, stem cells act mainly by neuroprotection with prevention of apoptosis, whereas during the chronic situation they provide neurorestoration by transdifferentiation and/or the secretion of neurotrophic factors. To reach these goals, in the acute phase, stem cells (10 million mononuclear cells per kg body weight) might be best applied intravenously, as during the first 7 days after the lesion, the blood-brain barrier permits passage of cells from the blood into the brain or the spinal cord. In the more chronic situation, though, those cells might be applied best intrathecally by lumbar puncture. Based on the reported results so far, it seems justified to develop well-designed clinical double-blind trials in chronic spinal cord injury and ischemic stroke patients, as efficacy and safety concerns might not be answered by preclinical studies.

Stem-cell-capturing collagen scaffold promotes cardiac tissue regeneration

Stem cell based therapy is coming of age. Besides stem cell transplantation, it has been a goal to use native autologous stem cells for tissue regeneration. However, the recruitment of native autologous stem cells at the targeting site has not been sufficient which limits the clinical application of autologous stem cells. Biomaterials have been increasingly used in tissue repair. They not only serve as scaffolds for cell proliferation, differentiation, and also provide guidance for 3-D reestablishment. In this study, we have attempted to enrich autologous stem cells at the wound site through a stem-cell-capturing collagen scaffold by conjugating with a stem cell specific antibody. Sca-1 is a common surface marker of hematopoietic, cardiac and skeletal muscle stem cells. Due to the interaction of antibody and antigen, Sca-1 positive cells could be enriched to the functional collagen scaffold both in vitro and in vivo. When the functional collagen scaffold is transplanted into C57/BL6 mouse as a patch to repair a surgical heart defect, the regeneration of cardiomyocytes has been observed. Thus, the collagen scaffolds covalently conjugated with stem cell specific antibody could be an effective approach to promote tissue regeneration.

Selected Abstracts from the “Best of Europe” Session of the 2nd Congress of the European Association of Societies of Aesthetic Plastic Surgery (EASAPS)

Selected Abstracts from the ‘‘Best of Europe’’ Session of the 2nd Congress of the European Association of Societies of Aesthetic Plastic Surgery (EASAPS) Best of Europe in Aesthetic Plastic Surgery 2009 (Editorial Coordinator: Cristino Suarez Lopez de Vergara). Strahil Efremov • Chavdar Botev • Ivan Tonev • Milcho Mincheff • James Henderson • Marketa Duskova • Dirk F. Richter • Andrea Grisotti • Emeri Jean-Francois • Toma Mugea • Jose Carlos Parreira • Ercan Cihandide • E. E. Aydin • A. Uzunismail • Juan Monreal • Alain Bzowski • Hassan Tazzi • Cristino Suarez Lopez de Vergara Springer Science+Business Media, LLC and International Society of Aesthetic Plastic Surgery 2010 7. Use of Autologous Stem Cells for Stimulation of Neoangiogenesis in Plastic Surgery Strahil Efremov, Chavdar Botev, Ivan Tonev, Milcho Mincheff (Medical Center ‘‘Liliya’’, Sofia, Bulgaria) Recent years have witnessed an increase in the therapeutic use of stem cells in both animal models and humans. Embryonic stem cells have shown great promise in regenerative medicine due to their capability to differentiate into mature stem cells of all lineages. Their use, however, has been hampered by the possibility of them forming tumors and for religious reasons. Adult stem cells are easier to obtain and several different ‘‘stem’’ cells such as tissue-specific, endothelial precursor, or ‘‘embryonic’’ -like cells have been described. These can be obtained from different sources such as bone marrow, peripheral blood, fat tissue, and cord blood. We have found that the mononuclear fraction from both adult bone marrow and fat tissue contains stem cells (endothelial precursors) whose application leads to neoangiogenesis and better nutrition of the affected tissue. In this presentation we will discuss two cases. The first is a young female who suffered a trauma after an explosion that totally destroyed her right ear and the basin of the right a. temporalis. The lack of adequate blood supply resulted in two unsuccessful otoplasties using implants. The application of autologous stem cells at the surgery site a month prior to and during otoplasty, however, led to neoangiogenesis at the affected area and a successful third otoplasty. The second case involves a female with a breast defect that was resolved successfully by the application of a mixture of autologous fat tissue and bone marrow-derived stem cells. 9. The Cosmetic Surgery Postcode Lottery James Henderson (Addenbrooke’s Hospital, Cambridge, UK) Introduction: There is wide variation of availability for many treatments, leading to a ‘‘postcode lottery’’ for health-care provision in England. The aim of this work was to evaluate the extent of the inequality of funding for the plastic surgery procedures which are viewed by PCTs as cosmetic. Materials and methods: All PCTs in the UK were asked for their funding policies for cosmetic procedures, including breast reduction and augmentation, removal of implants, mastopexy, abdominoplasty, face-lift, blepharoplasty, rhinoplasty, pinnaplasty, body lifting, surgery for gynecomastia, and tattoo removal. Results: Details of policies were received from 122 PCTs (82% response rate). Guidelines varied widely; some refuse all procedures while others allow a range. Different and sometimes contradictory rules governing symptoms, body S. Efremov C. Botev I. Tonev M. Mincheff J. Henderson M. Duskova D. F. Richter A. Grisotti E. Jean-Francois T. Mugea J. C. Parreira E. Cihandide E. E. Aydin A. Uzunismail J. Monreal A. Bzowski H. Tazzi C. S. Lopez de Vergara (&) EASAPS—Cardiff 2009, Wales, UK e-mail: cristinosuarez@gmail.com 123 Aesth Plast Surg (2010) 34:551–554 DOI 10.1007/s00266-010-9560-4

Application of autologous stem cells in treatment of lower extremity disorders

Application of autologous stem cells in treatment of lower extremity disorders