K.-J. Walgenbach

Plasmid Gene Delivery to Human Keratinocytes Through a Fibrin-Mediated Transfection System

We have developed a matrix-mediated transfection system to deliver plasmids to human keratinocytes. The matrix is a soluble, self-hardening fibrin matrix (Tissucol), Baxter) that has been used clinically. Recently it has been shown that full thickness burn wounds can be successfully treated with a keratinocyte fibrin glue suspension. Further, it has been demonstrated that hEGF transfected cells accelerate wound healing. In this study, we inoculated the matrix with the hEGF expression plasmid and resuspended the matrix with either cultured or noncultured human keratinocytes. We obtained successful transfection rates of these cells (up to a 100-fold increase compared to controls containing no EGF expression plasmid) in vitro. After transplantation to full thickness wounds on athymic mice we were able to show a 180-fold increase in EGF concentration compared to controls, which persisted over the entire 7-day monitored period, decreasing from 180 to 20 pg/mL at day seven. This unique approach indicates the possible utility to combine a matrix for cell transplantation with a transfection system to release therapeutic proteins in vitro and in vivo.

Hemangiosarcoma of the left hand in a patient with the rare combination of Maffucci's and Stewart Treves syndrome.

We describe a patient with the previously unseen combination of Maffuccis and Stewart Treves syndrome who presented with an angiosarcoma of the hand. Maffuccis syndrome is characterized by the presence of multiple enchondroma and soft tissue hemangioma. The syndrome is a rare nonhereditary condition with a usual onset in childhood. Malignant transformations are a common feature of this syndrome. In 1948, Stewart and Treves first described six cases of lymphangiosarcoma after radical mastectomy. This syndrome is an unusual form of angiosarcoma occuring as a complication of lymphedema. Chronic lymphedema and lymphangiectasia preceding lymphangiosarcoma may not only be induced by radical mastectomy with axillary lymph node dissection and postoperative radiation therapy. Posttraumatic, congenital or spontaneous chronic lymphedema may also be associated with lymphangiosarcoma. A time interval of many years seems to be required before malignant transformation develops. Generally the syndrome has a very poor prognosis. Both syndromes described above are of a rare frequency. We report this case because of prior unknown coincidence of both syndromes.We describe a patient with the previously unseen combination of Maffucci’s and Stewart Treves syndrome who presented with an angiosarcoma of the hand. Maffucci’s syndrome is characterized by the presence of multiple enchondroma and soft tissue hemangioma. The syndrome is a rare nonhereditary condition with a usual onset in childhood. Malignant transformations are a common feature of this syndrome. In 1948, Stewart and Treves first described six cases of lymphangiosarcoma after radical mastectomy. This syndrome is an unusual form of angiosarcoma occuring as a complication of lymphedema. Chronic lymphedema and lymphangiectasia preceding lymphangiosarcoma may not only be induced by radical mastectomy with axillary lymph node dissection and postoperative radiation therapy. Posttraumatic, congenital or spontaneous chronic lymphedema may also be associated with lymphangiosarcoma. A time interval of many years seems to be required before malignant transformation develops. Generally the syndrome has a very poor p...

Human Recombinant EGF Protein Delivered by a Biodegradable Cell Transplantation System

We have previously shown a new approach to expand cultured human keratinocytes and reconstitute the epidermis in full-thickness wounds using a new microsperical transport system. This was a new approach to increase the cell yield for seeding without altering the anchoring proteins by enzymatic steps. That time we used Cytodex 3 which failed to be degraded and induced an inflammatory reaction in a t-cell-deficient organism. Therefore, we have investigated another microcarrier consisting of PLGA, which is a well-known carrier material for cell culture and transplantation. After coating the PLGA carrier with gelatine the seeding time of viable cells reached 4 h and the cell gain after 7 days of spinner culture was 16-fold. At 14 days after transplantation, we could detect a new stratified epithelium in our full-thickness wound healing model. Because cytokines play a major role in wound healing, we loaded this carrier material with different concentrations of rhEGF, showing a dose dependent release of the protein in vitro and in vivo. This result might lead to a different approach in the treatment of wounds.

Management of hypovascularized wounds not responding to conventional therapy by means of free muscle transplantation.

Background: Chronic ulceration as a complication of arteriosclerotic disease, venous congestion or diabetes mellitus is still a serious clinical problem, resulting in immobilization, extended hospitalization and cost-intensive treatment. Other than standard conservative treatment protocols or early amputation, microsurgical free transfer of well vascularized muscle tissue onto chronic wounds can induce angiogenesis and improve wound healing even in the hypovascularized wound. Patients: From 1993–1999 we treated 12 patients (mean age: 46 years) with vascular ulcers of the lower extremity with free muscle or fasciocutaneous tissue transfer. Results: The average hospitalization was 51.4 days. The perioperative mortality was zero. In one patient with factor V deficiency a partial flap necrosis occurred. Two revisions of the micro anastomoses had to be performed. Two seromas occurred at the donor site. No secondary flap loss was observed. Extremity or stump length preservation was achieved in all cases. Conclu...BACKGROUND Chronic ulceration as a complication of arteriosclerotic disease, venous congestion or diabetes mellitus is still a serious clinical problem, resulting in immobilization, extended hospitalization and cost-intensive treatment. Other than standard conservative treatment protocols or early amputation, microsurgical free transfer of well vascularized muscle tissue onto chronic wounds can induce angiogenesis and improve wound healing even in the hypovascularized wound. PATIENTS From 1993-1999 we treated 12 patients (mean age: 46 years) with vascular ulcers of the lower extremity with free muscle or fasciocutaneous tissue transfer. RESULTS The average hospitalization was 51.4 days. The perioperative mortality was zero. In one patient with factor V deficiency a partial flap necrosis occurred. Two revisions of the micro anastomoses had to be performed. Two seromas occurred at the donor site. No secondary flap loss was observed. Extremity or stump length preservation was achieved in all cases. CONCLUSIONS Optimal postoperative treatment with physiotherapy and orthopaedic shoe support is important. If all these factors are present and if the patient is highly motivated a reintegration into normal life can be achieved.

Infiltration of tumescent anesthetic solution by a modified wound irrigation system.

Abstract. An easily set-up, efficient infiltration system for subcutaneous infiltration of tumescent anesthetic solution is presented. It can be contructed from parts of a wound irrigation system usually available in surgical units. Thereby it is an economical means of precise and convenient infiltration procedures.

Management therapierefraktärer hypovaskularisierter Wunden durch freie Muskelverpflanzung

Background: Chronic ulceration as a complication of arteriosclerotic disease, venous congestion or diabetes mellitus is still a serious clinical problem, resulting in immobilization, extended hospitalization and cost-intensive treatment. Other than standard conservative treatment protocols or early amputation, microsurgical free transfer of well vascularized muscle tissue onto chronic wounds can induce angiogenesis and improve wound healing even in the hypovascularized wound. Patients: From 1993–1999 we treated 12 patients (mean age: 46 years) with vascular ulcers of the lower extremity with free muscle or fasciocutaneous tissue transfer. Results: The average hospitalization was 51.4 days. The perioperative mortality was zero. In one patient with factor V deficiency a partial flap necrosis occurred. Two revisions of the micro anastomoses had to be performed. Two seromas occurred at the donor site. No secondary flap loss was observed. Extremity or stump length preservation was achieved in all cases. Conclu...BACKGROUND Chronic ulceration as a complication of arteriosclerotic disease, venous congestion or diabetes mellitus is still a serious clinical problem, resulting in immobilization, extended hospitalization and cost-intensive treatment. Other than standard conservative treatment protocols or early amputation, microsurgical free transfer of well vascularized muscle tissue onto chronic wounds can induce angiogenesis and improve wound healing even in the hypovascularized wound. PATIENTS From 1993-1999 we treated 12 patients (mean age: 46 years) with vascular ulcers of the lower extremity with free muscle or fasciocutaneous tissue transfer. RESULTS The average hospitalization was 51.4 days. The perioperative mortality was zero. In one patient with factor V deficiency a partial flap necrosis occurred. Two revisions of the micro anastomoses had to be performed. Two seromas occurred at the donor site. No secondary flap loss was observed. Extremity or stump length preservation was achieved in all cases. CONCLUSIONS Optimal postoperative treatment with physiotherapy and orthopaedic shoe support is important. If all these factors are present and if the patient is highly motivated a reintegration into normal life can be achieved.