Platelet-derived Wound Healing Factors Research Articles

Platelet-rich Plasma and Its Application in Clinical Trial

Platelet-rich Plasma and Its Application in Clinical Trial

Immunoglobulin G in Platelet-Derived Wound Healing Factors.

We intended to reformulate an existing platelet-derived wound healing formula to target each phase of the healing wound with the appropriate phase-specific molecules. A decreased perfusion of the skin, often associated with conditions such as thalassemia, sickle cell disease, diabetes mellitus, and chronic vascular disease, is the most common etiology of cutaneous ulcers and chronic wounds. We had previously shown that a PDWHF topically applied to a chronic nonhealing ulcer of a β-thalassemia homozygote stimulated and accelerated closure of the wound. The PDWHF was prepared from a pooled platelet concentrate of a matching blood group, consisting of a combination of platelet α-granule-derived factors. Processing of the apheresis-pooled platelets yielded various amounts of proteins (3.36 g/mL ± 0.25 (SD) (N = 10)) by the better lysis buffer method. Immunoglobulin G was found to be the most abundant α-granule-secreted protein. Equally broad quantities of the IgG (10.76 ± 12.66% (SD) (N = 10)) and IgG/albumin ratios (0.6 ± 0.4 (SD) (N = 10)) were quantified. We have developed a method using a reformulated lysis buffer followed by size exclusion chromatography and affinity chromatography to extract, identify, quantify, and purify IgG from activated platelets. IgG purification was confirmed by Western blot and flow cytometry. It was thought unlikely that the platelet IgG could be accounted for by adsorption of plasma protein, though the variable quantities could account for diversity in wound healing rates. The IgG could protect the wound even from subclinical infections and functionally advance healing. It may be useful in the management of skin ulcers in the early phase of wound healing.

One-step grafting procedure using artificial dermis and split-thickness skin in burn patients

The study aimed to achieve a one-step grafting procedure using artificial dermis and split-thickness skin. We performed simultaneous grafting of artificial dermis and skin in two severely burned patients. Artificial dermis was treated with fresh autogenous platelet-derived wound-healing factors (PDWHF), cryopreserved allogeneic cultured endothelial cells, and fibroblasts. Dermal microvascular endothelial cells and fibroblasts were obtained from a single human donor’s skin. The cultured cells were cryopreserved until use in grafting. The PDWHF was prepared from autogenous blood from each patient prior to the surgery. In two patients, the artificial dermis treated with this method was grafted to a full-thickness burn wound. Immediately after artificial dermis grafting, meshed split-thickness skin was grafted. In each case, the skin graft took well, and the skin texture was acceptable. Histological examination revealed that bovine collagen tissue remained in the dermis after surgery, indicating the success of the simultaneous grafting of the artificial dermis and the skin. The present study indicates that one-step grafting of artificial dermis and split-skin is possible when the artificial dermis is treated with PDWHF and cultured endothelial cells and fibroblasts. Level of Evidence: Level V, therapeutic study.

Tratamiento de úlceras cutáneas crónicas con plasma autólogo rico en plaquetas

BackgroundNon-healing cutaneous ulcers are increasingly frequent in dermatology consultations due to progressive population aging. Treatment involves extended care until the ulcer is healed, which, in addition to the high associated morbidity, incurs high healthcare and pharmaceutical expense and a decline in patients’ quality of life and mobility. ObjectiveTo describe in detail the treatment of cutaneous ulcers through the use of autologous platelet-derived wound healing factors. MethodsWe included five patients with 12 cutaneous ulcers with poor response to conventional treatment who were treated with this new method. ResultEarly healing of the wounds was obtained in 100% of the patients.

Regenerative Surgery: Use of Fat Grafting Combined with Platelet-Rich Plasma for Chronic Lower-Extremity Ulcers

The authors present their experience with reconstructive surgery of the lower extremity for chronic ulcers, evaluating the effects related to the use of a platelet-rich plasma combined with fat tissue. A total of 20 patients, 25 to 50 years of age (median age, 40 years), have been managed with platelet gel in the Plastic and Reconstructive Surgery Department at the "Tor Vergata," University of Rome. The patients were affected by both lower-extremity chronic ulcers and vascular disease. The authors observed that 16 of 20 chronic lower-extremity ulcers reepithelialized during an average of 9.7 weeks, with platelet releasate suspended on a collagen base (platelet-derived wound-healing factor), compared with 2 of 10 similar wounds treated with medication based on hyaluronic acid and collagen. Collectively, these data provide evidence for the clinical use of platelet technology in the healing of both soft and hard tissue wounds. Currently, plastic surgery with autogenous fat grafts can be performed for stabilization of chronic lower-extremity ulcers. The objective of this study was, through the presentation of clinical cases, to suggest a therapeutic plan formed by two sequential treatments: acquisition of platelet gel from a small volume of blood (9-18 ml) followed by the Coleman technique for reconstructing the three-dimensional projection and superficial density of tissues. The results proved the efficacy of combining these two treatments, and the satisfaction of the patients confirmed the quality of the results.

QS25. One-Step Grafting Procedure of Artificial Dermis and Split-Thickness Skin With PDWHF (Platelet Derived Wound Healing Factors) and Cultured Cells

QS25. One-Step Grafting Procedure of Artificial Dermis and Split-Thickness Skin With PDWHF (Platelet Derived Wound Healing Factors) and Cultured Cells

Outcomes of Conventional Wound Treatment in a Comprehensive Wound Center

Conventional wound care is the elementary treatment modality for treating chronic wounds. However, early treatment with topical growth factors may be needed for a subset of chronic wounds that fail to heal with good wound care alone. A prospective nonrandomized case series from a single-community outpatient wound care clinic is presented here in an effort to identify the subset of chronic wounds that may require early adjuvant intervention. There were 378 consecutive patients with 774 chronic wounds of varying etiology. All patients received 4 weeks of conventional wound care, including weekly debridement and twice-daily dressing changes. Wounds not reduced by 50 per cent volume at 4 weeks were nonrandomly treated with human skin equivalent (Apligraf), platelet-derived wound healing factor, or platelet-derived growth factor isoform BB (becaplermin gel, Regranex). A total of 601 of 774 (78%) wounds healed regardless of treatment type. The median time to heal for all wounds was 49 days (interquartile range = 26-93). More women than men healed (85% vs 71%, respectively, P < 0.0001). Diabetic wounds were as likely to heal as nondiabetic wounds (78% vs 80%, P = 0.5675). Wounds that did not heal had larger volumes and higher grade compared with wounds that healed (P < 0.0001 for both variables). The data presented here show that the majority of chronic wounds will heal with conventional wound care, regardless of etiology. Large wounds with higher grades are less responsive to conventional wound care and will benefit from topical growth factor treatment early in the treatment course.

Novel application method of artificial dermis: One-step grafting procedure of artificial dermis and skin, rat experimental study

Background Currently, to treat skin defects with artificial dermis (AD), two surgical procedures where the artificial dermis grafting and another secondary skin grafting are required. The purpose of this study was to achieve simultaneous grafting of the artificial dermis and the split-skin. To enhance the wound angiogenesis, cultured endothelial cells, fibroblasts and PDWHF (platelet derived wound healing factor) were employed. Methods The experiment consists of following two parts: (1) Investigation to obtain faster angiogenesis into the bilayer artificial dermis: full-thickness wounds created on the back of the rats were treated with the artificial dermis (Terudermis ®, with silicone sheet, TERUMO Co., Japan). Prior to the artificial dermis grafting, following four groups were established; control group (AD alone, n = 6), PDWHF group (AD treated with PDWHF, n = 6), cultured cells group (AD treated with cultured endothelial cells and fibroblasts, n = 6), combination group (AD treated with PDWHF and cultured cells, n = 6). (2) Trial of one-stage grafting of the AD and the skin: simultaneous grafting of the artificial dermis and skin was performed using the same rat model. Before making skin defects, split thickness skin were harvested. Then the skin grafting was carried out immediately after the AD grafting. To allow grafting of the skin onto the artificial dermis, the AD without silicone sheet (Terudermis ® without silicone sheet, TERUMO Co., Japan) were used. Two groups, control group (AD alone, n = 3) and treatment group (AD with PDWF and cultures, n = 3) were established. Results (1) When the artificial dermis were treated with PDWHF, cultured endothelial cells and fibroblasts, vascular invasion into the artificial dermis was observed 5 days after the surgery. (2) In the treatment group, the skin grafted immediately after the artificial dermis grafting was completely taken. Conclusions The present study revealed that treatment with PDWHF, combined with cultured endothelial cells and fibroblasts, accelerated wound angiogenesis. By this method, one-step grafting procedure of the artificial dermis and the skin is possible.

Accerelation of Wound Healing with PDWHF and Cultured Cells

Objective: Effects of platelet derived wound healing factor (PDWHF)1) with or without cultured cells on angiogenesis in treatment of defected wounds using artificial dermis (AD) were investigated in rat experimental model.Methods: Wistar strain rats were used for this study. The PDWHF was prepared from platelets. The endothelial cells and fibroblasts were prepared from thoracic aorta and back skin respectively. Two sites of 2.5 × 2.5 cm full‐thickness wounds were created on back of the animals. Artificial dermis (TERUDERMIS®, TERUMO Co, Japan) were grafted to the wounds. Prior to the AD grafts, following 4 groups were established, control group: AD alone (n = 6), PDWHF group: AD treated with PDWHF (0.1 ml/cm2)(n = 6), cultured cells group: disperse of 1 × 105 cultured endothelial cells and fibroblasts between AD and wound (n = 6) and combination group: combination of PDWHF and cultured cells (n = 6). Five days after surgery, degree of angiogenesis was evaluated by gross inspection and histological study. Evans blue perfusions test was performed to evaluate the degree of new capillary formation in the generated dermis quantitatively.Results: The combination group showed vascular invasion into AD 5 days after surgery. In this group, the absorbance of the Evans blue extracted from the grafted dermis was highest among the group.Conclusion: The result of the present study revealed that treatment with PDWHF combined with cultured endothelial cells and fibroblasts accelerate the wound angiogenesis. This method may be beneficial to shorten the period between the AD grafts and the secondary skin grafting.

P‐II‐03 Effect of Platelet Derived Wound Healing Factor (PDWHF) on the Angiogenesis in the Cases of Artificial Dermis Grafts

Aim: The effect of platelet derived wound healing factor (PDWHF) with or without cultured cells on angiogenesis in the cases of artificial dermis grafts was investigated. Methods: Wistar strain rats were used for this study. The PDWHF was prepared from platelets. The endothelial cells and fibroblasts were prepared from thoracic aorta and back skin respectively. Then 2 experimental models were designed. 1) Full‐thickness wounds were created on back of the animals. Artificial dermis (TERUDERMIS®, TERUMO Co, Japan) were grafted to the wounds. 2) Femoral artery and vein were isolated as a vascular pedicle and wrapped with a folded sheet of the AD (1 × 2 cm). Prior to AD grafts, the AD were treated with the PDWHF and/or cultured cells. Results: The combination use of the PDWHF and cultured cells showed best angiogenesis in both experimental models. Conclusions: The result of the present study revealed that treatment with PDWHF combined with cultured cells accelerate the wound angiogenesis.

P-II-03 Effect of Platelet Derived Wound Healing Factor (PDWHF) on the Angiogenesis in the Cases of Artificial Dermis Grafts

P-II-03 Effect of Platelet Derived Wound Healing Factor (PDWHF) on the Angiogenesis in the Cases of Artificial Dermis Grafts

The Effect of Thrombocytopenia on Dermal Wound Healing

The Effect of Thrombocytopenia on Dermal Wound Healing

Treatment of leg ulcers in β‐thalassaemia intermedia: use of platelet‐derived wound healing factors from the patient's own platelets

The interesting report by Josifova et al (2001), describes complete healing of an intractable ankle ulcer of nearly 20 years duration after 141 d of treatment with a combination of platelet-derived wound-healing factors (PDWHFs) in a patient with β-thalassaemia intermedia. We have used the same procedure except that the patient's own platelets were used as a source of PDWHFs in a β-thalassaemia intermedia patient with an ankle ulcer of > 4 years duration who was unresponsive to other procedures, including free skin grafting. Use of the patient's own platelets is potentially safer, and because thalassaemia intermedia patients usually have thrombocytosis it is particularly attractive. Platelets were obtained by apheresis from the patient, a 26-year-old, transfusion-independent splenectomized man with haemoglobin levels around 9 g/dl. More than 900 × 109/l platelets, were aliquoted in single-dose 2 ml vials under sterile conditions and frozen at −80°C. The preparation was thawed at room temperature before application. We applied a sterile gauze swab moistened with the thawed PDWHFs and firmly bandaged the area, every day from d 1 to d 10 and when granulation tissue developed, on alternative days (d 11 to d 32). The ulcer surface area was 13 cm2 on d 1 and complete healing was achieved at d 28 of treatment. As described by Josifova et al (2001), PDWHFs, in our case obtained from the patient's own platelets, may aid the treatment of ulcers in thalassaemia intermedia patients.

Dehiscence of Bronchial Anastomosis

Dehiscence of Bronchial Anastomosis

Mycophenolate mofetil and cyclosporin treatment for recalcitrant pyoderma gangrenosum

A 68-year-old woman with an uneventful medical history developed painful ulcers on both shins over a period of 6 months. She had large, inflamed, shallow, pretibial ulcers on both legs with a violaceous, partly bullous, and necrotic, undermined border. Diagnosis of pyoderma gangrenosum (PD) was confirmed by a biopsy specimen, which showed massive inflammatory-cell infiltration, thrombosis of dermal vessels, and leucocytoclastic vasculitis. Except for mild hyperthyroidism, no disease usually associated with PD was found. Treatment was begun with oral steroids (120 then 80 mg methylprednisolone per day) and clofazimine (200 mg/day). After 2 weeks, due to disease progression, cyclosporin was added (250 mg/day) but had to be discontinued as severe hypertension developed. Steroids were raised to 120 mg/day and cyclophosphamide therapy was started (500 mg intravenously on day 1, 50 mg by mouth on days 2–7). The lesions responded well at first, but recurrence developed after discontinuation of cyclophosphamide due to severe neutropenia. Over the next few months, the ulcers grew continuously despite treatment with high-dose intravenous immunoglobulins, dapsone, several cycles of high-dose intravenous steroids, and thalidomide 200 mg daily, each treatment having been given over several weeks. The ulcers now involved muscles and tendons of both shins. Reintroduction of cyclosporin (250 mg by mouth, this time tolerated with only mild hypertension) also had no effect. The patient was fed intravenously. The ulcers had to be debrided three times under general anaesthesia, requiring the excision of both Achilles tendons and parts of extensor muscles and tendons due to progressive necrosis. After 10 months of unsuccessful treatment, we decided to start mycophenolate mofetil (MM) 2 g/day by mouth in addition to cyclosporin and intravenous steroids. Autologous thrombocytic factors (platelet derived wound healing factors [PDWHF] Curative Technologies GmbH, Moers, Germany) were applied topically. After 14 days, the ulcers had stable borders. Healing continued until, 7 weeks after the introduction of MM, a split-thickness, meshed skin graft could be done. Healing was complete after another 4 weeks and has been now for a further 8 weeks without corticosteroids and with a reduced dose of 100 mg cyclosporin and 1 g/day MM, the latter tolerated without any side-effects. MM is effective in the prevention of renal transplant rejection. As a potent, non-competitive inhibitor of inosinmonophosphate-dehydrogenase, it influences the function of lymphocytes via the inhibition of purine synthesis. Its sideeffects are mostly restricted to mild gastrointestinal symptoms. Some evidence exists that it may also be effective in rheumatoid arthritis, psoriasis, and bullous pemphigoid. In our case, the first stable response after 10 months occurred after introduction of MM in addition to cyclosporin. Because cyclosporin monotherapy had been unsuccessful in this patient, we believe that the combination of MM and cyclosporin was the effective treatment, although application of PDWHF may have helped.

Acceleration of skin graft healing by growth factors

A special local burn wound treatment, using a healing promotion factor, attempted to achieve earlier wound closure. Clinically it is not difficult to harvest autologous human platelet-derived wound healing factor (HPDWHF) from burn patients. Another study has proved that porcine pituitary extract (PPE) could be one of the supplements for growing human keratinocytes in vitro (Wang HJ, Chen TM, Tung YM et al. Burns 1995 3). Initially, using the ELISA immunoassay, we detected titres of platelet-derived growth factor heterodimer AB (PDGF-AB) and epidermal growth factor (EGF) in HPDWHF and PPE. After elevating a 6 × 9 cm 2 flap on the backs of Sprague-Dawley (SD) rats 1 ml of heterologous HPDWHF and PPE were sprayed topically on the wound and followed by grafting six pieces of 1 cm 2 autologous full-thickness skin. The flap wound was then closed with a piece of large-pore Biobrane interposed between the skin graft wound and the flap. On postgraft day 7, the wound was reopened to measure the area of the full-thickness skin graft (FTSG) using micrographic paper; the results are reported as the percentage of graft expansion beyond the original size. In the cytokine study, we found that the concentrations of PDGF-AB dimer in HPDWHF and PPE were 5222 ± 102 and 375 ± 12 pg/ml (both values are reported after a 1:10 dilution) ( n = 3), whereas the EGF concentration was 245 ± 9 pg/ml, and undetectable in undiluted PPE ( n = 3), thereby proving that the PDGF-AB and EGF titres in the HPDWHF were significantly higher than in the PPE. Both the HPDWHF ( n = 6, with each animal grafted with six skin graft samples) and PPE ( n = 7) demonstrated significant accelerations of FTSG growth, with 14.41 ± 1.08 per cent and 13.16 ± 3.25 per cent increases in the FTSG size when compared to the sham control group 1.39 ± 3.26 per cent ( n = 5) ( P < 0.05). Comparisons between the treatment groups showed no evidence to indicate that the HPDWHF is superior to the PPE in accelerating FTSG growth ( P > 0.05). This supports the hypothesis that PDGF appears to transduce its signal through wound macrophages and may trigger the induction of positive autocrine feedback loops and synthesis of endogenous wound PDGF and other growth factors, thereby enhancing the cascade of tissue repair (Pierce GF, Mustoe TA, Altrock B et al. 1991; J Cell Biochem 1991; 45: 319–326), and the effect that PDGF-AB on wound healing is not dose dependent (Mustoe TA, Cutler NR, Allman RM et al. Arch Surg 1994; 129: 213–219). In future studies we expect to show that autologous HPDWHF and heterologous PPE will promote skin graft wound healing in burn patients since they contain a high concentration of PDGF-AB.

Diabetic wound healing and limb salvage in an outpatient wound care program.

Healing wounds and preventing amputations in diabetic patients is a challenge. Baptist Hospital in Nashville, Tennessee, developed an outpatient Wound Care Center program to treat patients with chronic wounds. This comprehensive program includes wound assessment, vascular studies, revascularization, antibiotic therapy, wound debridement, a topical growth factor solution of platelet-derived wound healing factors, protection devices, and patient education. A retrospective study of wound healing and limb salvage was conducted on 54 diabetic patients with 86 wounds. Average previous wound duration was 8 months. Amputation had previously been recommended for 15 limbs. Healing occurred in 88% of the wounds in an average of 15.8 weeks. Ninety-three percent of the limbs for which amputation had been recommended were salvaged. Healing efficacy and amputation reduction have been demonstrated in this program.

A prospective randomized trial of autologous platelet-derived wound healing factors for treatment of chronic nonhealing wounds: A preliminary report

Previous studies have suggested that topically applied platelet-derived wound healing factors (PDWHF) accelerate wound healing by stimulating angiogenesis, fibroblast proliferation, and collagen synthesis. To assess the ability of platelet factors to facilitate healing of chronic cutaneous ulcers we performed a randomized, prospective, double-blind, placebo-controlled study of topical PDWHF in 18 patients with 26 lower extremity wounds refractory to conventional therapy. Wounds were present for at least 8 weeks (mean, 5.5 ± 4.3 months). They were extensively debrided initially and were measured and photographed at weekly intervals for 12 weeks. Eight patients with nine wounds were treated with placebo solution (controls), and 10 patients with 17 wounds were treated with PDWHF (treatment group). Seventy-eight percent of patients had diabetes mellitus, 72% had occlusive peripheral vascular disease, and 28% had venous disease; distribution of these disorders was equivalent in both groups. Ankle-brachial indexes, which were often spuriously elevated, averaged 0.93 ± 0.54 in controls and 1.04 ± 0.56 in patients treated with PDWHF (p > 0.5). Mean transcutaneous oxygen tension was 37.8 ± 11.9 mm Hg in controls and 37.1 ± 9.1 mm Hg in patients treated with PDWHF. Initial wound area was larger in controls than in the patients treated with PDWHF (28.9 ± 45.2 cm2 vs 13.0 ± 14.4 cm2), but this difference was not statistically significant (p = 0.19). Three (33%) wounds (in two patients) healed in controls, and four (24%) wounds (in three patients) healed in the PDWHF group (p > 0.5). The rate of healing in controls was 1.9 ± 2.7 cm2/week. Wounds in patients treated with PDWHF increased in size during treatment, averaging −4.3 ± 12.2 cm2/week. This preliminary study suggests that treatment of chronic wounds with autologous PDWHF provides no additional benefit over traditional therapy.

A prospective randomized trial of autologous platelet-derived wound healing factors for treatment of chronic nonhealing wounds: A preliminary report

A prospective randomized trial of autologous platelet-derived wound healing factors for treatment of chronic nonhealing wounds: A preliminary report

Classification and treatment of chronic nonhealing wounds. Successful treatment with autologous platelet-derived wound healing factors (PDWHF).

Previous animal data showed that platelets contain growth factors that stimulate capillary endothelial migration (angiogenesis), fibroblast proliferation and migration, and collagen synthesis. This study utilized autologous platelet-derived wound healing factors (PDWHF) to treat 49 patients with chronic nonhealing cutaneous ulcers. Patients were classified on the basis of 20 clinical and wound status parameters to generate a wound severity index. Forty-nine patients--58% diabetic (20% with renal transplants); 16% with trauma, vasculitis, etc.; 14% with decubitus ulcers; and 6% each with venous stasis or arterial insufficiency--with a total of 95 wounds had received conventional wound care for an average of 198 weeks (range: 1-1820 weeks). After informed consent was obtained, patients received autologous PDWHF. Mean 100% healing time for all patients was 10.6 weeks. There was no abnormal tissue formation, keloid, or hypertrophic scarring. A multivariant analysis showed a direct correlation to 100% healing with initial wound size and the initiation of PDWHF therapy. This is the first clinical demonstration that locally acting growth factors promote healing of chronic cutaneous ulcers.