Document Type : Original Article
Authors
1 Department of plastic surgery and burn, Faculty of medicine, Al-Azhar University, Cairo, Egypt
2 Faculty of Science, Al-Azhar University, Cairo, Egypt
Abstract
Keywords
INTRODUCTION
Chronic wounds constitute a real challenge for the clinicians as they do not respond to traditional treatment and they are not rare.1 During wound healing, mesenchymal stem cells (MSCs) participate in all stages of healing and are settled to persist at the wound site even after the completion of this process.2
Management of chronic wound varies from traditional wound dressings to skin substitutes, hyperbaric oxygen, negative pressure wound therapy, novel platelet-rich plasma (PRP), nanofat injection and Stem cell therapy.3
Stem cell therapies have emerged as a progressive field of research because of their potential for the treatment of such wounds. Bone marrow-derived stem cells (BMSCs) represent a heterogeneous population from the non-blood-forming fraction of bone marrow it represents a novel approach in management of chronic skin injuries.4
Platelet-rich plasma (PRP) is known to be a rich source of cytokines and growth factors that are important for healing and re-epithelialization of chronic wounds.5
The work aimed to compare the results of clinical application of bone marrow derived stem cells and Platelet-rich plasma for treatment of chronic wounds.
PATIENTS AND METHODS
Study Design:
A prospective randomized comparative study included 30 patients complaining of chronic wounds with average surface area less than 10 cm2 divided into 2 groups; group (A) were managed by injection with BMSCs and group (B) were managed by injection with PRP. The study was done at Al-Azhar University Hospitals, Center for Genetic Engineering at faculty of science, Al-Azhar University and Elsalam specialized hospital with approval of the ethical committee.
Inclusion criteria: Patients with chronic wound "duration of the wound > 3 months", average surface area of the wound is less than 10 cm2 and age between 15 and 60 years.
Exclusion criteria: Patient in critical condition (e.g., shock, debilitating disease, and serious infections), significant co-morbidities, history of any hematological disease, post radiation wounds, psychiatric disorders or pregnant female.
Preoperative:
Clinical assessment: Careful history taking, general condition, local examination of the chronic wound and measurement of the wound surface area.
Investigation: CBC, PT, PTT, INR and HBA1C "for diabetic patients"
Stem Cell Preparation:
The posterior iliac spine was sterilized, and local anesthesia was injected to the skin and periosteum of that area. Bone marrow was aspirated under complete aseptic conditions on preservative-free heparin.
Stem cells were then isolated from bone marrow aspirates. Mononuclear cells were resuspended in complete culture medium. After 24 hours, non-adherent cells were discarded, and of the adherent cells, the spindle-shaped cells were morphologically evaluated. The cell population was characterized by typical immunophenotyping, fibroblast-like morphology and ability to differentiate.
Preparation of PRP:
30 mL of the whole blood was taken from the patient under existence of ACD-A solution. The mixing rate was 9:1 in volume. Eight 5-mL syringes were prepared via cutting their finger- holders using scissors.
4 ml of the blood was put into each syringe and then centrifuged. The rotation speed was 3000 rpm for 3 minutes to separate red blood cells (RBCs) from plasma. The syringes were taken out from the centrifuge then arranged on a holder.
After centrifugation a 3-way cock and an extension tube were connected together, and the syringe was attached to the other end of the extension tube. Four syringes were prepared for the second centrifugation by the same previous method. 1 microgram of PGE1 diluted in 0.05 mL of saline was added to each syringe. After that the syringe was connected to the 3-way cock then the plasma was aspirated. The second centrifugation was done for 15 minutes at 4000 rpm. The supernatant was discarded leaving 0.65 mL which was mixed with the sediment using a vortex mixer. At the end, we got 0.65 mL of PRP solution from 16 mL of the blood.
Injection: At outpatient clinic: Sterilization, injection of local anesthesia, debridement of the wound and injection was done from wound edges to the center at circumferential manner. Debridement and Injection was done once for all the patients of both groups.
Follow up:
We followed up the patients for two months.
Digital photographs of the wounds were taken pre-operatively, 3 days postoperatively then weekly for two months. Wound healing rate was calculated as follows:
(The original wound surface area – residual wound surface area)/the original wound surface area × 100%.
RESULTS
Group A (BMSCs) |
Group B (PRP) |
Chi square test |
|||||
No |
% |
No |
% |
x2 |
p value |
||
Sex |
Female |
4 |
26.7% |
7 |
46.7% |
1.292 |
0.256 |
Male |
11 |
73.3% |
8 |
53.3% |
|||
Age |
Mean ± SD |
39.20 |
10.28 |
40.20 |
13.31 |
0.230 |
0.820 |
Table 1: Shows demographic data in both groups.
Group A |
Group B |
Independent t test |
||||
Mean |
SD |
Mean |
SD |
t |
p value |
|
Duration of the wound in months |
11.33 |
16.91 |
5.47 |
5.30 |
-1.282 |
0.210 |
Surface area of the wound in cm2 |
4.93 |
1.00 |
5.60 |
1.92 |
1.169 |
0.253 |
Table 2:Shows wound duration in months and surface area in cm2.
Group A |
Group B |
Chi square test |
|||||
No |
% |
No |
% |
x2 |
p value |
||
Cause of the wound |
Diabetic foot |
4 |
26.7% |
4 |
26.7% |
1.234 |
0.745 |
Post burn ulcer |
3 |
20.0% |
4 |
26.7% |
|||
Traumatic wound |
5 |
33.3% |
6 |
40.0% |
|||
Venous ulcer |
3 |
20.0% |
1 |
6.7% |
|||
Site of wound |
Rt arm |
0 |
0.0% |
2 |
13.4% |
20.800 |
0.235 |
Dorsum of lt foot |
2 |
13.4% |
2 |
13.4% |
|||
Sole of lt foot |
0 |
0.0% |
1 |
6.7% |
|||
Sole of rt foot |
0 |
0.0% |
1 |
6.7% |
|||
Dorsum of rt foot |
2 |
13.4% |
5 |
33.4% |
|||
Dorsum of lt hand |
0 |
0.0% |
1 |
6.7% |
|||
Dorsum of rt hand |
1 |
6.7% |
0 |
0.0% |
|||
Dorsum of rt big toe |
0 |
0.0% |
1 |
6.7% |
|||
Planter surface of lt big toe |
1 |
6.7% |
0 |
0.0% |
|||
Lt knee |
1 |
6.7% |
0 |
0.0% |
|||
Lt leg |
5 |
33.3% |
0 |
0.0% |
|||
Lt thigh |
1 |
6.7% |
0 |
0.0% |
|||
Rt heel |
1 |
6.7% |
0 |
0.0% |
|||
Rt knee |
1 |
6.7% |
0 |
0.0% |
|||
Rt leg |
0 |
0.0% |
2 |
13.3% |
Table 3:Shows the cause and site of the wound in both groups.
% of improvement
Diabetic foot
Post burn ulcer
Traumatic wound
Venous ulcer
One way ANOVA
Mean
SD
Mean
SD
Mean
SD
Mean
SD
f
P value
after 3 days
11.75
3.40
12.67
2.08
11.00
1.22
7.00
2.00
3.673
0.047
after 1 wk
27.00
6.98
35.67
4.51
28.40
3.05
15.00
4.36
9.333
0.002
after 2 wk
50.25
12.61
64.33
5.03
52.80
5.81
28.67
6.35
9.964
0.002