Effectiveness and Safety of Adipose-Derived Regenerative Cells for the Treatment of Critical Lower Limb Ischemia

Study Overview
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Brief Summary
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Adipose-derived regenerative cells (ADRC) will be extracted from lipoaspirate by enzymatic digestion. 10 mL of autologous ADRC suspension injected intramuscularly, close to the site of muscle injury. All patients will receive cell therapy. This is a single arm study with no control.

Detailed Description
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Fat tissue obtainment: Subjects will undergo liposuction under local anesthesia. In this procedure, Ringer's solution with the anesthetic lidocaine and vasoconstrictor adrenaline infused into the adipose compartment to minimize blood loss and contamination of the tissue by peripheral blood cells. 15 minutes later a hollow blunt-tipped 3 mm cannula introduced into the subcutaneous space through small (0.5 cm) incision. The cannula attached to syringe and under gentle suction moved through the adipose compartment, mechanically disrupting the fat tissue. Aspirate volume - approximately 150-200 cc. Procedure time - 30 minutes. ADRC isolation: Harvested adipose tissue will be processed according to patent pending technology based on enzymatic digestion, washing and concentration of cell pellet in 10.5 ml of normal saline. Obtained ADRC divided into 2 portions. First portion (0.5 mL) used for counting, viability and sterility assessment. The second portion (10 ml) placed into sterile syringe for injection. Autologous ADRC administration 10 mL of autologous ADRC suspension will be injected intramuscularly, close to the site of muscle injury. 10 to 20 injections (0.5 to 1.0 mL each) will be performed so as to infiltrate the injured muscle.

Official Title
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Effectiveness and Safety of Adipose-Derived Regenerative Cells for the Treatment of Critical Lower Limb Ischemia

Conditions
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Critical Limb Ischemia, Arteriosclerosis Obliterans, Peripheral Arterial Disease, Thromboangiitis Obliterans, Diabetic Angiopathies

Intervention / Treatment
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* Procedure: ADRC injection
* Other: ADRC isolation
* Procedure: Liposuction


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Patients with critical lower limb ischemia, with resting pain wich can't be adequately managed by opioid analgetics and/or foot ulcers or necrosis, with II-4, III-5 и IV-6 stages of chronic arterial failure according to Rutherford's classification Ankle-brachial index less than 0.4 and/or ТСрО2 less than 30 mm Hg Patients with lower limbs' arteries lesions revealed by angiography uneffectiveness or impossibility of limb's revascularisation or patient's refusal of the procedure Patient is familiar with Participant information sheet Patient signed informed consent form Non-inclusion Criteria: Contraindications for local anesthesia or history of allergy for local anesthetics Systemic glucocorticoid and/or immunosuppressant therapy Any condition that might limit compliance (dementia, mental disorders, narcotic drug or alcohol abuse etc.) Participation in other clinical trials (or administration of investigational drugs) during 3 months prior inclusion Patients with malignant tumors including postoperative period, patients receiving chemotherapy and/or radiotherapy Clinically significant abnormalities in results of laboratory tests Patient received anticoagulants at least 12 hours prior the liposuction Medical history of heterotopic ossifications Patients prescribed for glycoprotein inhibitors treatment Exclusion Criteria: Patient's refusal from the further participation in trial Chronic kidney disease IV- V stages (creatinine clearance < 30 mL/min estimated by Cockcroft-Gault formula) Confirmed syphilis, HIV, hepatitis B or C infections Dropout Criteria: Indications for the amputation of the limb

Ages Eligible for Study
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Minimum Age: 50 Years
Maximum Age: 85 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: ADRC injection<br>Subjects will undergo liposuction under local anesthesia. Lipoaspirate will be processed to isolate and concentrate Adipose-derived regenerative cells (ADRC). After isolation 10 mL of autologous ADRC suspension will be injected intramuscularly, close to the site of muscle injury. 10 to 20 injections will be performed so as to infiltrate the injured muscle | Procedure: ADRC injection<br>* 10 mL of autologous ADRC suspension will be injected intramuscularly<br>Other: ADRC isolation<br>* ADRC will be isolated according to patent pending technology based on enzymatic digestion, washing and concentration of cell pellet in 10 ml of normal saline.<br>Procedure: Liposuction<br> <br> |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Serious adverse events | Frequency, type and severity of serious adverse events (SAE) | 4 weeks after injection of ADRC suspension | 
| Serious adverse reactions | Frequency, type and severity of serious adverse reactions (SAR) | 4 weeks after injection of ADRC suspension | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes of ankle-brachial index | Influence of intervention on ankle-brachial index. | Follow up to completion (24 weeks after intervention) | 
| Changes of hemodynamics in lower extremity - 1 | Influence of intervention on blood flow velocity assessed by arterial duplex scanning. | Follow up to completion (24 weeks after intervention) | 
| Changes of hemodynamics in lower extremity - 2 | Influence of intervention on pulsatility index assessed by arterial duplex scanning. | Follow up to completion (24 weeks after intervention) | 
| Changes of hemodynamics in lower extremity - 3 | Influence of intervention on resistance index assessed by arterial duplex scanning. | Follow up to completion (24 weeks after intervention) | 
| Metabolic state of targeted tissues measurements | Changes of transcutaneous oxygen tension (ТсРО2) in injured limb assessed by transcutaneous oximetry. | Follow up to completion (24 weeks after intervention) | 
| Quality of life monitoring - 1 | Quality of life estimated by validated questionnaire: the Short Form (SF-36). | Follow up to completion (24 weeks after intervention) | 
| Quality of life monitoring - 2 | Quality of life estimated by validated questionnaire:Peripheral Artery Questionnaire. | Follow up to completion (24 weeks after intervention) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Ischemic ulcers, Critical lower limb ischemia, Atherosclerosis obliterans, Diabetic angiopathies, Thromboangiitis obliterans, Peripheral arterial disease, Indirect revascularisation, Intramuscular injection, Ankle-brachial index, Transcutaneous oxygen tension, Stromal vascular fraction (SVF), Adipose-derived regenerative cells (ADRC), Adipose tissue, Stem cells