Treatment of leg ulcers with autologous heparinized blood 
Short therapeutic report 
TREATMENT OF LEG ULCERS WITH 
AUTOLOGOUS HEP ARINIZED BLOOD 
M. Kaštelan, I. Brajac and F. Gruber 
ABSTRACT 
Background. Human blood contains growth factors, which take part m wound healing. 
Objective. To test the hypothesis that topical application of human blood would improve ulcer healing, we have 
treated 26 leg ulcers of different etiology with repeated topical applications of human blood. 
Methods. Twenty-six ulcers were treated with autologous heparinized blood under the occlusive dressing every 
other day. Fifteen control ulcers were treated with antiseptic ointment. The duration of follow up for each 
patient was 3 weeks. The surface of the ulcers was measured by planimetry. The study was performed in 
patients at the Department of Dermatology, Rijeka. 
Results. The average surface reduction was 52.5% in ulcers treated by autologous heparinized blood. On the 
contrary, in control ulcers the average surface reduction was 18.2%. Complete closure of the ulcer was 
obtained in 3 cases (11.5%). Ulcer surface was significantly reduced in ulcers treated autologous blood 
(p=0.001), compared to controls (p=0.62). 
Conclusion. Our results suggest that autologous heparinized blood stimulates cleansing and healing of the 
ulcers. It is well tolerated and an inexpensive method in the treatment of leg ulcers. 
KEY WORDS 
lower leg, ulcers, autologous blood, heparin, topical treatment 
INTRODUCTION 
Leg ulcers are mostly caused by chronic venous or 
arterious insufficiency that leads to inadequate tissue 
oxygenation and consequently worse healing (1). 
Numbers of methods are used in the treatment of 
leg ulcers with different success. One method is 
30 
topical application of autologous heparinized blood 
(AHB) (2) It is well known that human blood 
contains growth factors, which take part in wound 
healing (3). These factors can chemotactically attract 
and activate inflammatory cells and initiate new 
expression of quiescent genes ( 4-6). 
In this paper we present 22 patients with chronic 
acta dermatovenerologica A.P.A. Vol 7, 98, No 1 
Treatment of leg ulcers with autologous heparinized blood 
Table l. Results of topical treatment of leg ulcers with autologous heparinized blood (haemotherapy) 
1 venous 9 13.73 
2 venous 7 13.95 
3 venous 10 6.64 
4 venous 9 14.17 
5 venous 9 18.82 
6 venous 9 9.08 
7 venous 9 9.74 
8 venous 8 4.43 
9 venous 7 4.87 
10 venous 3 2.88 
11 venous 8 4.65 
12 venous 9 4.21 
13 arterious 9 9.97 
14 arterious 9 3.54 
15 diabetic 9 3.76 
16 venous 8 20.82 
17 venous 8 19.49 
18 venous 9 10.41 
19 venous 8 3.10 
20 arterious 9 16.39 
21 venous 8 6.87 
22 diabetic 10 19.04 
23 arterious 9 3.76 
24 venous 8 12.40 
25 venous 9 3.99 
26 venous 9 2.44 
X 9.35 
leg ulcers of different etiology that were treated with 
repeated applications of AHB. We assumed that 
human blood applied on ulcer surface would improve 
its healing. 
PATIENTS AND METHODS 
We have treated 41 leg ulcers in 22 patients (13 
females, 9 males; mean age 73, range: 50-91) at the 
Department of Dermatology. These ulcers included 
31 venous (75.61%), 7 arterious (17.07%), and 3 
diabetic (7.32%) ulcers. Inclusion criteria were pa-
tients with ulcers caused by venous or arterial 
diseases, while exclusion criteria were ulcers associ-
ated with haematological diseases, neurologic dise-
ases, signs of acute inflammation and patients 
undergoing immunosuppressive therapy. The diagno-
sis was based on clinical examination, supported by 
Doppler's ultrasound test and in some cases also by 
plethysmography. 
32 
5.31 61.33 
0.00 100.00 
4.87 26.66 
7.75 45.31 
13 .95 25 .88 
4.87 46.37 
7.01 28.03 
0.66 85.10 
0.44 90.97 
0.00 100.00 
0.00 100.00 
4.21 0.00 
4.65 53.36 
3.54 0.00 
2.88 23.40 
3.54 83.00 
2.88 85.22 
1.32 87.32 
1.55 50.00 
14.39 12.20 
5.53 19.51 
11.95 37.24 
3.76 0.00 
7.75 37.50 
2.21 44.61 
0.66 72.95 
4.45 52.50 
Twenty-six ulcers (63.41 % ): 20 venous, 4 arterious, 
2 diabetic, were treated with AHB under the occlusive 
dressing. In 15 patients (36.59%) with two ulcers 
(11 venous, 3 arterious and 1 diabetic) one was 
treated with AHB, while the other one with antiseptic 
Figure 1. Swf ace reduction in autologous heparinized 
blood therapy treated ulcers and controls (mean 
values) 
acta dermatovenerologica A.P.A. Vol 7, 98, No 1 
Treatment of leg ulcers with autologous heparinized blood 
Table 2. Results of the therapy in control ulcers 
1 venous 9 4.43 
2 venous 9 22.15 
3 venous 9 21.48 
4 venous 9 2.44 
5 venous 9 4.65 
6 venous 9 3.10 
7 venous 9 1.77 
8 arterious 9 2.88 
9 diabetic 9 1.11 
10 venous 8 3.99 
11 venous 9 2.43 
12 arterious 9 1.99 
13 arterious 9 6.64 
14 venous 8 11.29 
15 venous 8 12.84 
X 6.88 
ointment (Betaisadona ointment - 10% Polyvinyl-
iodine complex, Mundipharma) every other day. 
Before starting the therapy all ulcers were disinfected 
with 3% hydrogen peroxide and mechanically cleaned 
with curette. Every other day the fresh AHB was 
applied to the ulcers and immediately covered with 
occlusive absorbent dressing (MetodermR). The mixture 
of 1 ml venous blood and 0.1 ml heparin (SolopakR, 
25,000 IU, Smith & Nephew, USA) was applied per 
every 4 cm2 of the ulcer's surface. 
The mean duration of ulcers was 15 months 
(range: 1-84); the average surface of haemotherapy 
treated ulcers was 9.35 cm2 (range: 2.44-20.82), 
while the average surface of the control ulcers was 
6.88 cm2 (range: 1.11-22.15). At the beginning of 
the therapy there was no significant difference in 
ulcer's surface between AHB treated group and 
controls (p=0.42). The average duration of AHB 
treatment was 16 days (8 treatments). The surface of 
the ulcers was measured by planimetry. Statistics 
were done by Student's t-test. The nature and purpose 
of the study were explained to the patients and a 
verbal informed consent was obtained from them. 
RESULTS 
Table l. presents the results of AHB treatment 
in individual patients. Complete healing and closure 
of the ulcer was obtained in 3 (11.5 % ) cases of 
acta dennatovenerologica A.P.A. Vol 7, 98, No 1 
3.99 9.93 
19.93 10.02 
20.59 4.14 
0.89 63.52 
0.33 92.90 
0.66 78.71 
1.77 0.00 
2.88 0.00 
1.11 0.00 
3.99 0.00 
0.67 72.42 
1.99 0.00 
5.54 16.56 
8.86 21.52 
11.29 12.07 
6.64 18.20 
venous ulcers. There was no reduction of ulcer's 
surface in 3 (11.5%) cases, one venous, and one 
arterious ulcer. 
The average surface reduction of AHB treated 
ulcers was 52.5%. In control ulcers the average 
surface reduction was 18.2% (Table 2). Figure l. 
shows surface reduction in AHB treated ulcers and 
controls after the therapy. As seen the ulcer's surface 
was significantly reduced after haemotherapy (p=0.001). 
At the same tirne in controls, after ointment therapy, 
the surface reduction was not statistically significant 
(p=0.62). After each treatment we noticed the 
diminution of fibronecrotic material and a rapid 
formation of new granulation tissue. At the same 
tirne in control ulcers the formation of granulation 
tissue was slower and the fibronecrotic material was 
more adherent. However, the difference between 
these two methods was not significant (p>0.05). 
In 2 patients we noticed after a few applications 
an erythema and papular exanthema surrounding the 
ulcers on the treated leg. The other patients showed 
no local or systemic side effects. 
DISCUSSION AND CONCLUSIONS 
In this study we report on topical application of 
autologous human blood in the treatment of chronic 
leg ulcers measured by surface reduction. We observed 
that AHB improved healing in the majority of ulcers 
33 
Treatment of leg ulcers with autologous heparinized blood 
cornpared to controls by stirnulating the forrnation 
of granulation tissue and increasing epithelization. 
The best results were obtained in venous ulcers. 
However, slight or no changes at all, were observed 
in ulcers caused by arterious insufficiency and diabetes, 
factors generally considered to be of bad prognostic 
significance. 
Ulcer's surface reduction after accornplished AHB 
was statistically significant. At the same tirne in 
control ulcers the surface was not significantly reduced. 
These results show that haernotherapy gives better 
results than classic treatrnent with ointrnents. The 
results between these two treatrnent rnethods were 
not significantly different (p>0.05). 
We also noticed rapid forrnation of granulation 
tissue and fast reduction of fibronecrotic material in 
cornparison to controls. This is in agreernent with 
the results of Triquet et al. (2) in which the quantity 
of new-forrned granulation tissue on the ulcer surface 
was rneasured during the topical haernotherapy 
treatrnent. According to our experience AHB is a 
successful rnethod in preparing the ulcers for skin 
grafting. 
In 3 cases in which total closure was obtained, a 
progressive epithelization, starting frorn srnall islands 
of keratinocytes in the centre of the ulcer was observed. 
Usually the epithelization starts frorn the wound edge 
what we noticed in 80% of our cases. (7) 
Our results suggest that AHB therapy stimulate 
healing and cleansing of the ulcers. This rnethod 
involves synergistic activity of growth factors, occlusive 
dressing, and heparin. These initial results suggest 
that these factors prornote and stirnulate wound 
healing. (8) Heparin, at low concentrations stimulates 
the growth of rnyocytes and fibroblasts, but is inhibitory 
at higher concentrations. (9,10) Further research will 
be necessary to explain the exact role of heparin and 
occlusive dressing during the haernotherapy. 
REFERENCES 
l. Hansson C. Studies on leg and foot ulcers. Acta 
Derm Venereol (Stockh) 1988; Suppl. 136, Thesis. 
2. Triquet E, Ruffieux P, Mainetti C, et al. Topical 
haemotherapy far leg ulcers. Dermatology 1994; 189: 
418-20. 
3. Falanga V Growth factors and wound healing. 
Dermatol Clin 1993; 4: 667-75. 
4. Deuel TF, Kawahara TS, Mustoe TA, et al. 
Growth factors and wound healing: Platelet-derived 
growth factor as a model cytokine. Ann Rev Med 
1991; 42: 567-84. 
5. Hammar H Wound healing. lnt J Dermatol liJtt3;· 
32: 6-11. 
6. McKay IA, Leigh IM. Epidermal cytokines and 
their roles in cutaneous wound healing. Er J Dermatol 
1991; 124: 513-8. 
7. Eeele H, Naeyaert JM, Goeteyn M, et al. Repeated 
cultured epidermal allografts in the treatment of 
chronic leg ulcers of various origins. Dermatologica 
1991; 183: 31-5. 
8. Steed DL. Clinical evaluation of recombinant 
human platelet-derived growth factor for the treatment 
of lower extremity diabetic ulcers. J Vase Surg 1995; 
21: 71-81. 
9. Azizkhan RG, Azizkhan JL, Zetter ER et al: MAST 
cel! Heparin stimulates the migration of capillary 
endothelial cel/s in vitro. J Exp Med 1980; 152: 931-9. 
1 O. Scott GK Proteinases and eukaryotic cel! growth. 
Comp Eiochem Physiol 1987; 87E: 1-10. 
AUTHORS' ADDRESSES 
Marija Kaštelan, MD, Departrnent of Derrnatology, Clinical Hospital Centre, Rijeka, 
Kreširnirova 42, Croatia 
Ines Brajac, MD, Departrnent of Derrnatology, Clinical Hospital Centre, Rijeka, Kreširnirova 42, Croatia 
Franjo Gruber, MD, PhD, assistant professor, Head Departrnent of Derrnatology, 
Clinical Hospital Centre, Rijeka, Kreširnirova 42, Croatia 
34 acta dermatovenerologica A.P.A. Vol 7, 98, No 1