Bmc binder1

Regenerative medicine Bone Marrow Stem Cells Therapy

Unique Benefits • Concentrates 60 mL BMA into 6 mL BMC • >80% recovery of MNC’s from BMA • >90% Cell viability post process • In <20 minutes at point-of-care • Process up to 4 x BMA 60 mL samples simultaneously • Closed system design provides lower risk of tissue contamination

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Effective • Maintains cell viability post process Cell viability is important for successful engraftment following autologous cell transplantation. CD34+ cells were assessed by flow cytometry and no significant difference was found post vs. preprocessing.

• A full draw will consistently contain high recovery yields of >80% (MNC) mononuclear cells • Cell enrichment factors up to 9x • Contains ~ <20% RBC and small amount of plasma to reach desired harvest volume • Reproducibility +/- 5% variation between each disposable

Increased Safety • Closed, Sterile, non-pyrogenic fluid path • Minimal user-intervention • Minimizes risk of sample mix-up or contamination

Easy to Use • • • •

Automated point of care (POC) solution for bone marrow processing Target cells can be returned <20 min after sample collection Simple procedure, no specialist skills required Delivers an operator-controlled volume of autologous concentrate into a syringe for immediate use in the sterile field.

• Minimal maintenance required

• Can be used at the point-of-care in OR, or in a laboratory setting 2

Harvesting Process Step 1

Harvesting Process Step 2

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Harvesting Process Step 3

Harvesting Process Step 4

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BMC in critical limb ischemia Therapeutic angiogenesis for patients with limb ischemia by autologous transplantation of bonemarrow cells: a pilot study and a randomized controlled trial Tateishi-Yuyama, et all

Objective: To investigate safety and efficacy of autologous BMC transplantation in patients with ischemic limbs due to PAD.

Methods:     

A pilot study included 25 patients (group A) with unilateral ischemia. o Intramuscular injection of BMC was given in m. gastrocnemius of ischemic limb as well as saline injection in less ischemic limb.

Second group (group B) of patients (n = 22), with bilateral ischemia was collected. o Patients were randomly injected with BMC in one leg and with peripheral blood mononuclear cell in other leg, as control Patients monitored weekly for 4 weeks after implantation and every 4 months after that.

Their condition was assessed by ABI, transcutaneous oxygen pressure (TcO2) and pain free walking time + DSA was done 1 week before and 4 weeks after implantation Primary endpoint was safety and feasibility of treatment.

Results: 

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ABI – group A o legs injected with BMC increased from 0.34 to 0.47 at week 4 o increase of ABI was noted in 17 out of 25 patients o no changes of ABI was seen in legs injected with saline ABI – group B o ABI of the legs injected with BMC increased from 0.37 to 0.46 at week 4 (p<0.0001) o Increase in ABI was noted in 13 of 20 patients injected with BMC TcO2 – group B o TcO2 increased from 28.8 at the beginning to 46.3 at 4 weeks (p<0.0001) Group B - Legs injected with peripheral blood mononuclear cells showed much smaller increase in ABI and TcO2

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Rest pain  

In group B: in legs treated with BMC was resolved in 16 out of 20 patients, while in legs treated with peripheral blood mononuclear cells remained in 17 out of 20 pain free walking time was improved by about 3 minutes in group A and 1 minute in group B (reason for less improvement in this group is probably unilateral treatment with BMC in patients with bilateral limb ischemia)

Angiography results 

Showed striking increase in number of visible collateral vessels in 27 of 45 patients (15 in A and 12 in B group)

Conclusion 

Autologous implantation of BMC could constitute safe and effective strategy for achievement of therapeutic angiogenesis.

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BMC in diabetic, non-healing foot ulcers Treatment of non-healing wounds with autologous bone marrow cells, platelets, fibrin glue and collagen matrix Ravari et al. ď&#x201A;ˇ

reports 8 diabetic patients with resistant diabetic foot wounds that did not react on conventional therapy

Procedure: 1. Debridement of the wound was done 2. 5 ml of bone marrow cells were implanted in the wound by 1.5 cm deep injections in various sites around margin of the wound 3. 2 ml of BMC mixed with platelets and fibrin glue were applied on the wound surface to form a clot 4. 1 ml of BMC mixed with 9 ml of serum was applied on collagen matrix which was then placed on the clot 5. Paraffin gauze pads were put on the wound 6. After 3 days dressing was removed and the wound was irrigated with isotonic sodium chloride solution 7. The wound was then covered again and this procedure was repeated daily 8. Wounds were closely examined for 4 weeks

Results: The wounds of 3 patients were completely closed after 4 weeks and significant wound reduction (P<0.005) was seen in all other patients.

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BMC in Burger disease Stem cells therapy: a promising and prospective approach in the treatment of patients with severe Buerger’s disease Boda et al 

8 patients o 6 with Buerger’s disease o 1 with Arteriosclerosis obliterans o 1 with Buerger’s disease and diabetes mellitus with vascular complications

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Indications for BMC therapy: o Severe stage (Fontaine IIIb/IVa or Rutherford 5) o Rest pain, less than 50 m walking distance o In all 8 cases (11 limbs) the more painful limb was treated o No more chances remained for vascular surgery or conservative therapy

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Treatment: o BMS suspension was used as intra-muscular injection to the m. gastrocnemius of patient’s worse lower limb o Single application of 15-20 injections of 0.5-1.0 ml; 1.5-2.5 cm deep o In 2 patients therapy was repeated after reoccurrence of ulcers at the same place as before ; The effect of repeated stem cell therapy was faster, ulcers healed in a shorter time then after the first stem cell therapy

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Conclusion o Autologous BMC stem cell therapy in Buerger’s disease is effective, safe, sustained and localized

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ABI

*1 : p=0.0065; *2 : p=0.00018; *3 : p=0.00011; *4 : p=0.0031

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Clinical status before and after BMC therapy

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BONMOT-CLI: BONe Marrow Outcomes Trial in critical limb ischemia B. Amann et al.

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Investigator-initiated, multi-center, randomized, placebo-controlled, double-blind study 90 patients 1:1 randomization Screened: 805 CLI patients No revascularization possible and not healing: 111 patients Eligible: 102 patients, 8 of them declined to participate Included: 94 patients

Bone marrow stem cells were taken from pelvic bones (crista iliaca) and injected intramuscularly in 40–60 sites of the ischemic limb.

Demography/Characteristics at baseline: Stem cells (48) Diabetes 62% Smokers 59% Fontaine 4 88% Rutherford 4,9 Quality of life (EQ 5D, 0-1) 0,526 Pain (VAS 0-10) 4,5 Previous revascularization 2,1 (0-5) ABI 0,40±0,1 tcPO2 12,6±13 2 Wound area (cm ) 8,6±10

Placebo (46) 61% 54% 86% 4,9 0,576 4,8 2,0 (0-6) 0,40±0,1 14,9±11 3,9±4

p n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. p<0,05

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3 months results: Major amputation Rutherford ABI tcPO2 Wound area (cm2) Death Adverse events Quality of life (EQ 5D, 0-1) Pain (VAS)

Stem cells (48) 8,3% (4/48) -0,9 (4,0) +0,08 (0,48) +12 (25) -40% (5,2) 3 70% 0,677 4,7

Placebo (46) 15% (7/46) ±0 (4,9) ±0 (0,40) +6 (21) +32% (5,7) 5 81% 0,588 4,8

p trend p<0,05 p<0,06, trend p<0,05 p<0,05 n.s. n.s. trend n.s.

Aggregated 3 months results: Disease improved Disease same Death and/or major amputation Stem cells 24% 61% 15% Control 4% 68% 28% Disease improved: defined as patient alive, no amputation, wound closed, Rutherford/Fontaine stage improved 12 months results: Pain free walking distance Absolute walking distance Quality of life

Stem cell 169±111 (+148%) 223±257 (+116%) 77

Control 80±23 (+11%) 115±41 (+9%) 58

p p<0,001 p<0,001 p<0,05

Conclusion BMSC therapy in CLI: Is safe:  No procedural complications  No morbidity/mortality due to bone marrow harvesting and cell injection  Fast, easy to learn and affordable (bed-side procedure) Improves perfusion, wound healing, and restores walking in several patient groups:  ABI + 0.15 ( up to 0.4, tcpO2 + 20 mmHg)  “the younger the better” Probably reduces major amputations (long term results needed).

Is a good therapy for no-option patients.

Works also in lesser PAD stages (Fontaine IIb/Claudication with no simple revascularization option).

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