An interventional study of mesenchymal stem cells in Diabetic Peripheral Neuropathy, sponsored by Cairo University. Completed. Open to participants aged 18 Years to 45 Years. Per ClinicalTrials.gov, last updated 2018-07-03.
Sponsored by Cairo University · Not applicable, Interventional, and Treatment
A debilitating consequence of diabetes mellitus (DM) is neuropathy which globally affects between 20 -30% of diabetic patients and up to 50% in other studies. The incidence of diabetic neuropathy (DN) is estimated to be up to 45% for type 2 diabetic patients and 59% for type 1diabetic patients in USA.(DN) is the most common complication of DM.The pathophysiology of DN is promoted by several risk factors: micro vascular disease, neural hypoxia, and hyperglycemia-induced effects.At the molecular level, the primary cause of diabetic complications is known to be hyperglycemia, which disrupts cellular metabolism by the formation of reactive oxygen species (ROS).In the aspect of nerve functions, ROS formation increases neuron's susceptibility to damage. In addition, hyperglycemia impedes production of angiogenic and neurotrophic growth factors, which are necessary for normal function of neurons and glial cells and maintenance of vascular structure.No definitive disease-modifying treatments have been to reverse DN. The current treatment focuses on tight glycemic control which can reduce potential risk factors for further nerve damage and DN-associated pain management.In many studies, deficiency of neurotrophic factors and lack of vascular support have been regarded as key factors in the development DN.Mesenchymal stem cells (MSCs) are particularly attractive therapeutic agents because of their ability to self-renew, differentiate into multi lineage cell types, and locally secrete angiogenic cytokines, including basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) .These factors were reported to prompt neovascularization and have support for neural regeneration.It was plausible that MSCs may also be an effective therapeutic agent for the DN treatment through the paracrine effects of bFGF (Shibata et al., 2008) and VEGF and their potential to differentiate into neural cells such as astrocytes, oligodendrocytes , and Schwann cells.The adherent nature of MSCs makes them easy to expand in culture and an attractive candidate to use in cell therapy.Therefore, cell therapy has recently emerged as an attractive therapeutic strategy to meet the needs of both neurotrophic and vascular deficiencies of DN.Proper diagnosis of DN depends on the pattern of sensory loss, reflex test, electrodiagnostic studies, and imaging
Objectives This study will be conducted to investigate the effects of Mesenchymal stem cells (MSC) transfusion on diabetic peripheral neuropathy in diabetic patient. (MSCs) have been reported to secrete various cytokines that exhibit angiogenic and neuro supportive effects.
Study Design Experimental interventional study. phase II clinical trial
Ethical committee approval (was it ethically approved by the department) Internal medicine department Yes
Study Methods
Population of study \& disease condition (e.g women with hepatitis, .........) Diabetic patients(type I, type II) with documented peripheral neuropathy as determined by impaired nerve conduction
Inclusion criteria:
(Type I, type II) diabetic patients age range (18-45) years, with diabetic peripheral neuropathy proved by clinical assessment and nerve conduction who did not receive treatment for diabetic peripheral neuropathy.
Exclusion criteria:
Decompensated cardiac, renal or liver disease. Associated autoimmune diseases Associated endocrinal diseases Pregnancy, usage of contraceptive pills or steroids.
Methodology in details The study will be conducted on patients with diabetic peripheral neuropathy collected from internal medicine department(inpatient and out patients, males and females)
All subjects of this study will be submitted to the following :( preparatory visit before (MSCs) transfusion visit.)
Plasma biochemical blood measurements will be determined by standard laboratory procedures in the central lab at clinical pathology department, Kasr Alaini hospital)
Kidney functions in the form of serum creatinine.
To avoid infection: During bone marrow aspiration, procedure will be done under complete aseptic precautions, placed in sterile tubes containing pre-servative-free heparin (Sigma-Aldrich, St. Louis, USA) Separation and processing of the sample will be done under good manufacture procedure (GMP): Bone Marrow Aspirate (BMA) will be withdrawn under good sterilization of the skin in an isolated area. Processing of the sample will be done in the laminar air flow; all supplies are disposable and sterile.
Technique:
Separation of mononuclear cells:
The bone marrow aspirate will be diluted at a ratio of 6:1 with phosphate buffer saline (PBS) with 2 mM EDTA (30 ml BM aspirate+ 5 ml PBS/EDTA buffer). The MNCs will be separated under aseptic conditions using a Ficoll. Hypaque desity gradient (density 1.077, GibcoBRL, Grand Islan, NY, USA) by centrifugation at 1800 rpm for 20 min then the MNCs will be plated in 40 ml alpha-modi-field Eagle's medium (αMEM), serum free media; mesencult(Mesenchymal stem cell culture),penicillin (100 U/ml),streptomycin(10 mg/ml),0.5 ml amphotericin B(all from Gibco BRL) and 10 ng/ml basic fibroblast growth factor (b-FGF) (R\&D system, Minneapolis, MN) and will be incubated at 370 c in a humidified atmosphere containing 5% CO2 (Digirolamo et al.1999).after one day ,non adherent cells will be cultured in the presence of Mesenchymal media for 3 weeks changed every 1 week (cambrex Bioscience ,Nottingham, uk). After reaching 80% confluence the MSCs will be placed in 10 ml saline and will be infused intravenously
Flow cytometry Surface expression of MSCs using anti- (CD271, CD34, CD73, CD90, CD105, CD29) monoclonal antibodies (mAbs) will be analyzed using flow cytometry. MSCs (2X105 cells) will be suspended in PBS containing 1% BSA and will be stained with flurochrome -conjugated mAbs for 20 minutes on ice (anti-mouse mAanti-CD 271, CD34 CD73, CD90, CD105,CD29; BD Bioscience, MN, USA).flow cytometric analysis will be performed using a FACSCaliber (BD bioscience)equipped with cell Quest software.10000 cells will be passed in front of the laser for each sample. Each sample will be analyzed in duplicate. A cut off value at 20% will be set to categorize samples as positive.
Mesenchymal stem cells will be identified by morphology and immunophenotyping in the central lab at clinical pathology department, Kasr Alaini hospital( stem cell lab).
Mesenchymal stem cells transfusion slowly intravenous will be applied after these procedures for the patients after taking their approval and informed consent.
Follow up 3 months after Mesenchymal stem cells transfusion by fasting blood glucose level, 2 hours postprandial, C-peptide, Hb A1C, (bFGF), (VEGF) and nerve conduction at kasr Alaini hospital departments as mentioned before.
Possible Risk (mention if there is any risk or not) Anaphylaxis Infection
Primary outcomes (Most important outcomes to be assessed)
1- Effect of mesenchymal stem cells transfusion on diabetic peripheral neuropathy regarding improvement of clinical symptoms like pain, sensory loss and improvement of nerve conduction.
Secondary outcome parameters (other outcomes to be assessed)
Sample size (number of participants included) 10 diabetic patients with diabetic peripheral neuropathy .
Source of funding (is there any source of funds or not) Faculty Of Medicine, Cairo University.
Time plan (when to start/ when expected to finish/ when to publish) At least 20 months
1,003 studies on the registry are indexed under Peripheral Nervous System Diseases; 177 are open to participants now.
This study's enrollment of 10 is below the median of 60 across 768 interventional studies indexed under Peripheral Nervous System Diseases.
Browse Peripheral Nervous System Diseases studies →Cairo University is the lead sponsor of 4,780 studies on the registry; 1,427 are open to participants now.
Of its 36 completed or terminated interventional studies of FDA-regulated products, 5 (14%) have results posted.
Counted across the registry records on this site, refreshed daily.
Exclusion Criteria:
The BM aspirate will be diluted at 6:1 ratio with phosphate buffer saline with 2 ml EDTA (30 ml BM aspirate+ 5 ml PBS/EDTA buffer).MNCs will be separated under aseptic conditions using a Ficoll. Hypaque desity gradient by centrifugation at 1800 rpm for 20 min then the MNCs will be plated in 40 ml(αMEM), serum free media; mesencult(MSCs culture),penicillin (100 U/ml),streptomycin(10 mg/ml),0.5 ml amphotericin B(all from Gibco BRL) and 10 ng/ml basic fibroblast growth factor (b-FGF)(R\&D system, Minneapolis, MN) and will be incubated at 370 c in a humidified atmosphere containing 5% CO2 .after one day ,nonadherent cells will be cultured in the presence of Mesenchymal media for 3 weeks changed every week. After reaching 80% confluence the MSCs will be placed in 10 ml saline and infused IV.
Genetic: mesenchymal stem cells
collection of stem cells by bone marrow biopsy from iliac crest, then culture for 1 month , then IV transfusion on 2 sessions to the same patient
Measurement of b-FGF, v-EGF MEASURED BY ELISA
measurement of b-FGF and v-EGF MEASURED BY ELISA before (at zero), and after at (7 days, 90) days after stem cell transfusion to measure the effect of stem cell and its role in nerve regeneration
Time frame: zero ( before) , 7 DAYS, 90 days
Change of Nerve Conduction Velocities of Nerves Affected Measured by Nerve Conduction Study.
Measuring nerve conduction velocities(NCV) in m/sec upper and lower limbs nerves(sensory and motor) lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory upper limb nerves: ulnar nerve as motor and sensory and compare at base line(zero day) and 90 days after stem cells transfusion
Time frame: base line(zero dya), 90 days after stem cells transfusion.
Change of Nerve Conduction Latency of Nerves Affected Measured by Nerve Conduction Study
Measuring nerve conduction latency in msec of upper and lower limbs nerves(sensory and motor) lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory upper limb nerves: ulnar nerve as motor and sensory and compare at base line and 90 days after stem cells transfusion
Time frame: base line(zero dya), 90 days after stem cells transfusion .
Change of Nerve Conduction Amplitude of Nerves Affected Measured by Nerve Conduction Study.
Measuring nerve conduction amplitudes in uv of upper and lower limbs nerves(sensory and motor). lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory . upper limb nerves: ulnar nerve as motor and sensory. and compare at base line and 90 days after stem cells transfusion
Time frame: base line(zero dya), 90 days after stem cells transfusion
Change of Levels of Fasting Blood Sugar and 2 Hours Post Prandial at Base Line ( Zero Day ) and After (90 Days) After Stem Cells Transfusion
fasting, 2 hours postprandial blood sugar measurement before at base line (zero day) and after (90 days) stem cells transfusion as a follow up and comparing the values.
Time frame: base line (zero day) and 90 days after stem cells transfusion
Change of Levels of Glycated Haemoglobin( HA1C) After Stem Cells Transfusion Measured in Percent %
Blood tests before and after stem cells(90 days) transfusion and comparing the values in percent % which is reflecting the patient blood sugar control in the previous 3 months
Time frame: at base line (zero day) and 90 days after stem cells transfusion
| Milestone | Mesenchymal Stem Cells Transfusion in Diabetic Neuropathy |
|---|---|
| Started | 10 |
| Completed | 10 |
| Not completed | 0 |
measurement of b-FGF and v-EGF MEASURED BY ELISA before (at zero), and after at (7 days, 90) days after stem cell transfusion to measure the effect of stem cell and its role in nerve regeneration
| pg/ml | Measurement of b-FGF and v- EGF MEASURED BY ELISA |
|---|---|
| b-FGF ZERO | 30.2 ± 16.7 |
| b-FGF 7 DAYS | 55.4 ± 12.3 |
| b-FGF 90 days | 30.3 ± 14.8 |
| v- EGF ZERO | 428.7 ± 125 |
| v- EGF 7 DAYS | 601.8 ± 141.8 |
| v-EGF 90 DAYS | 371.5 ± 121.9 |
Measuring nerve conduction velocities(NCV) in m/sec upper and lower limbs nerves(sensory and motor) lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory upper limb nerves: ulnar nerve as motor and sensory and compare at base line(zero day) and 90 days after stem cells transfusion
| m/sec | Nerve Conduction Velocities of Nerves |
|---|---|
| Tibial Motor Nerve conduction velocity zero | 43.6 ± 8.5 |
| Tibial motor nerve conduction velocity 90 days | 44.4 ± 6.6 |
| common peroneal motor nerve velocity at zero day | 43.8 ± 10.3 |
| common peroneal motor nerve velocity at 90 day | 45.2 ± 12.3 |
| Ulnar Motor Nerve conduction velocity at zero day | 56.0 ± 8.7 |
| Ulnar Motor Nerve conduction velocity at 90 days | 54.2 ± 10.6 |
| Ulnar sensory Nerve conduction velocity zero day | 52.0 ± 16.3 |
| Ulnar sensory Nerve conduction velocity at 90 days | 51.2 ± 12.4 |
| Sural Nerve conduction Velocity at zero day | 21.1 ± 22.7 |
| Sural Nerve conduction Velocity at 90 days | 25.5 ± 27.0 |
Measuring nerve conduction latency in msec of upper and lower limbs nerves(sensory and motor) lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory upper limb nerves: ulnar nerve as motor and sensory and compare at base line and 90 days after stem cells transfusion
| msec | Nerve Conduction Latency of Nerves |
|---|---|
| Tibial Motor nerve conduction latency zero day | 4.4 ± 1.9 |
| Tibial Motor nerve conduction latency 90 days | 4.1 ± 1.3 |
| common peroneal motor nerve latency at zero day | 4.4 ± 1.8 |
| common peroneal motor nerve latency at 90 day | 4.2 ± 1.1 |
| Ulnar Motor Nerve conduction latency at zero day | 2.9 ± 0.8 |
| Ulnar Motor Nerve conduction latency at 90 day | 2.9 ± 0.7 |
| Ulnar sensory Nerve conduction latency zero day | 3.0 ± 1.0 |
| Ulnar sensory Nerve conduction latency 90 days | 2.7 ± 0.8 |
| Sural Nerve conduction latency at zero day | 2.2 ± 2.4 |
| Sural Nerve conduction latency at 90 days | 2.5 ± 2.8 |
Measuring nerve conduction amplitudes in uv of upper and lower limbs nerves(sensory and motor). lower limb nerves : tibial , common peroneal(CP) as motor and sural nerve as sensory . upper limb nerves: ulnar nerve as motor and sensory. and compare at base line and 90 days after stem cells transfusion
| uv | Nerve Conduction Amplitudes of Nerves |
|---|---|
| Tibial Motor nerve Amplitude zero day | 3.8 ± 4.0 |
| Tibial Motor nerve Amplitude at 90 days | 3.4 ± 3.0 |
| common peroneal motor nerve amplitude at zero day | 2.2 ± 1.9 |
| common peroneal motor nerve amplitude at 90 days | 2.2 ± 1.2 |
| Ulnar Motor Nerve conduction amplitude zero day | 5.4 ± 1.9 |
| Ulnar Motor Nerve conduction amplitude at 90 days | 6.3 ± 1.8 |
| Ulnar sensory Nerve conduction amplitude zero day | 23.1 ± 16.5 |
| Ulnar sensory Nerve conduction amplitude at 90 day | 21.3 ± 11.6 |
| Sural Nerve conduction amplitude at zero day | 5.2 ± 5.8 |
| Sural Nerve conduction amplitude at 90 day | 7.3 ± 8.6 |
fasting, 2 hours postprandial blood sugar measurement before at base line (zero day) and after (90 days) stem cells transfusion as a follow up and comparing the values.
| mg/dl | Blood Tests Measured Before and After Stem Cell Transfusion |
|---|---|
| fasting blood sugar zero | 211.30 ± 61.92 |
| fasting blood sugar 90 days | 145.70 ± 37.56 |
| 2 hours post postprandial blood sugar Zero | 291.50 ± 106.56 |
| 2 hours post postprandial blood sugar 90 days | 190.30 ± 56.42 |
Blood tests before and after stem cells(90 days) transfusion and comparing the values in percent % which is reflecting the patient blood sugar control in the previous 3 months
| percent % | HA1C Measurment Before and Afterr Stem Cell |
|---|---|
| HA1C Level Zero | 9.12 ± 3.42 |
| HA1C Level 90 days | 7.96 ± 1.89 |
Collected over 3 months. Non-serious events are listed at a 0% frequency threshold.
| Group | Deaths | Serious | Other |
|---|---|---|---|
| Mesenchymal Stem Cells Transfusion in Diabetic Neuropathy | — | 0/10 (0%) | 0/10 (0%) |
blood pressure all patients were normal blood pressure measurement routine lab investigations were normal to exclude other co-morbidites
| Age, Categorical(Participants) | Mesenchymal Stem Cells Transfusion in DPN Patients |
|---|---|
| <=18 years | 0 |
| Between 18 and 65 years | 10 |
| >=65 years | 0 |
| Age, Continuous(years) | Mesenchymal Stem Cells Transfusion in DPN Patients |
|---|---|
| Mean | 41.20 ± 4.59 |
| Sex: Female, Male(Participants) | Mesenchymal Stem Cells Transfusion in DPN Patients |
|---|---|
| Female | 4 |
| Male | 6 |
| Region of Enrollment(participants) | Mesenchymal Stem Cells Transfusion in DPN Patients |
|---|---|
| Egypt | 10 |
No study locations are listed for this record.
Plan to share: Undecided
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Cairo University