Andrej Ring

Extracorporeal shock waves improve angiogenesis after full thickness burn

OBJECTIVE Extensive wounds of burn patients remain a challenge due to wound infection and subsequent septicemia. We wondered whether extracorporeal shock wave application (ESWA) accelerates the healing process. The aim of the study was to analyze microcirculation, angiogenesis and leukocyte endothelium interaction after burns by using ESWA with two types of low intensity. METHODS Full-thickness burns were inflicted to the ears of hairless mice (n=51; area: 1.3 mm(2)). The mice were randomized into five groups: (A) low-energy shock waves after burn injury (0.04 mJ/mm(2)); (B) very low-energy shock waves after burn injury (0.015 mJ/mm(2)); (C) mice received burns but no ESWA (control group); (D) mice without burn were exposed to low-energy shock waves; (E) mice without burns and with no shock wave application. Intravital fluorescent microscopy was used to assess microcirculatory parameters, angiogenesis and leukocyte behavior. ESWA was performed on day 1, 3 and 7 (500 shoots, 1 Hz). Values were obtained straight after and on days 1, 3, 7 and 12 post burn. RESULTS Group A showed accelerated angiogenesis (non-perfused area at day 12: 5.3% vs. 9.1% (group B) and 12.6% (group C), p=0.005). Both shock wave groups showed improved blood flow after burn compared to group C. Shock waves significantly increased the number of rolling leukocytes compared to the non-ESWA-treated animals (group D: 210.8% vs. group E: 83.3%, p=0.017 on day 7 and 172.3 vs. 90.9%, p=0.01 on day 12). CONCLUSION Shock waves have a positive effect on several parameters of wound healing after burns, especially with regard to angiogenesis and leukocyte behaviour. In both ESWA groups, angiogenesis and blood flow outmatched the control group. Within the ESWA groups the higher intensity (0.04 mJ/mm(2)) showed better results than the lower intensity group. Moreover, shock waves increased the number of rolling and sticking leukocytes as a part of an improved metabolism.

Leiomyosarcoma of intravascular origin - a rare tumor entity: clinical pathological study of twelve cases

BackgroundLeiomysarcoma of intravascular origin is an exceedingly rare entity of malignant soft tissue tumors. They are most frequently encountered in the retroperitoneum arising from the inferior vena cava and are scarcely found to arise from vessels of the extremities. These tumors were analysed with particular reference to treatment outcome and prognosis. The aim of this article is to broaden the knowledge of the clinical course of this rare malignancy.MethodDuring 2000 and 2009 twelve patients were identified with an intravascular origin of a leiomyosarcoma. Details regarding the clinical course, follow-up and outcome were assessed with focus on patient survival, tumor relapse and metastases and treatment outcome. 3 year survival probability was calculated using Kaplan-Meier method.ResultsVascular leiomyosarcomas accounted for 0.7% of all malignant soft tissue tumors treated at our soft tissue sarcoma reference center. The mean follow up period was 38 months. Tumor relapse was encountered in six patients. 6 patients developed metastatic disease. The three year survival was 57%.ConclusionVascular leiomysarcoma is a rare but aggressive tumor entity with a high rate of local recurrence and metastasis.

Influence of anti-inflammatory and vasoactive drugs on microcirculation and angiogenesis after burn in mice

OBJECTIVE The treatment of burns remains a challenge. Besides the administration of physiological saline, local disinfection and symptomatic medications, no causal therapy is known to accelerate angiogenesis and wound healing. The aim of this study was to investigate the influences of dilatative and anti-inflammatory acting drugs on microcirculation, angiogenesis and leukocyte behavior, which had shown positive effects in former burn studies. METHODS The ears of male hairless mice (n=47) were inflicted with full thickness burns using a hot air jet. Then the affects of five intraperitoneal injections of either acetylsalicylic acid (ASA), isosorbide dinitrate, prostaglandin E1 (PGE1) or sodium chloride (each administered to one of four corresponding study groups), on microcirculation, leukocyte-endothelial interaction and angiogenesis were investigated over a 12 day period using intravital fluorescent microscopy. RESULTS Angiogenesis was slightly improved by PGE1 (0.3 vs. 1.3% non-perfused area in other groups on day 12, p=0.029). Additionally, blood flow increased and rolling leukocytes decreased compared to other groups. The ASA-group showed best functional vessel density and lowest leukocyte-adhesion. The often described posttraumatic expansion of tissue damage could not be observed in either group. CONCLUSION Prostaglandin E1 improved angiogenesis, increased the blood flow and reduced the number of rolling leukocytes. ASA had positive influences on functional vessel density, edema formation and the number of sticking leukocytes. However, it reduced the blood flow. Overall, out of all the drugs tested, prostaglandin seems to have the greatest positive impact on microcirculation and angiogenesis in burns.

Intravital Pathophysiologic Comparison of Frostbite and Burn Injury in a Murine Model

BACKGROUND The breakdown of skin microcirculation and the leukocyte-endothelium interaction are assumed to play key roles in the pathophysiology of burn and frostbite injuries. Available data on frostbite and burn injuries were collected using different experimental models and setups, which limits direct comparisons of these thermal traumata significantly. To determine pathophysiologic similarities and differences, two comparable in vivo frostbite and burn models were used to assess microcirculatory and angiogenetic changes in burn and frostbite injuries. MATERIALS AND METHODS Either deep partial thickness no-touch burns or frostbite injuries were inflicted to the ears of hairless mice (n = 40) by a hot or cold gas jet (117.0 ± 2.1°C for 1 s and -195.8 ± 2.7°C for 1.5 s, respectively) resulting in a necrotic, nonperfused area of about 1.56 ± 0.28 mm2. Intravital fluorescent microscopy was used in combination with fluorescent dyes in order to assess the microcirculation, angiogenesis, and leukocyte-activity over a 12-d period. RESULTS The angiogenesis occurred significantly faster after frostbite than after burn (16.4% ± 4.5% versus 30.6% ± 2.8% nonperfused area, compared with the baseline value on d 7 (P = 0.009)). The loss of functional vessel density was significantly more pronounced after frostbite (57.6% ± 2.2% versus 89.2% ± 4.9% (P < 0.001)). However, the area recovered faster. The edema formation, as a parameter for endothelial integrity, was significantly more pronounced and lasted longer after frostbite, compared with the burn injury, and reached its maximum level on d 7 after trauma (162.4% ± 4.2% versus 142.% ± 5.9%; P = 0.007). In contrast to the rolling leukocytes, which showed the same increase on d 1 and then a subsequent decrease in both groups, the number of adherent leukocytes after the burn was markedly higher on d 1 (480% versus 167%; P = 0.001) but decreased much faster. The number of adherent leukocytes after frostbite remained significantly higher than those of the burn group during the entire observation. CONCLUSION The comparison of analogous intravital burn and frostbite models indicates that despite the similarities, decisive microcirculatory differences in extension and recovery from these two types of thermal trauma exist.

Orthogonal polarization spectral imaging: a tool for assessing burn depths?

The estimation of burn depths is still a challenge, and even experienced surgeons often fail. In search of an objective method for differentiation between deep- and partial-thickness burns, we investigated the use of orthogonal polarization spectral (OPS) imaging to visualize the microcirculation in burn wounds.Twenty-seven burned patients were included in the study, 81 burn areas were investigated at day 1 and 4 post burn. The final therapy was compared with the therapy that would have been carried out as a consequence of the OPS imaging result. The patients were treated conservatively with dressings or surgically with necrectomy and split skin grafts.A comparison of the estimation of the burn depths of OPS imaging with the final therapy showed a correlation of 76.5%, about 5% less than the clinical assessment. It has proved a useful tool in particular for the prognosis as to whether the burn will heal spontaneously within 14 days or not.The presented OPS-Imaging device provides additional qualitative and quantitative information about the perfusion of the skin and therefore facilitates decisions about the follow on therapy. It is not an alternative to an experienced burn surgeon but provides important additional information.

Determination of Microcirculatory Changes and Angiogenesis in a Model of Frostbite Injury In Vivo

BACKGROUND The breakdown of skin microcirculation and the leukocyte-endothelium interaction are assumed to play a key role in the pathophysiology of frostbite injuries. However, little is known as yet. The aim was to develop an in vivo frostbite model to monitor microcirculatory changes and angiogenesis after frostbite injury. MATERIALS AND METHODS Deep partial thickness frostbite injuries were inflicted with a no-touch-technique to the ears of hairless mice (n=9). To this end, a gas jet of nitrogen vapor (T=-195,8±2.7°C) was delivered onto an area of 1.9 mm(2) for 1,5 s. Intravital fluorescent microscopy in combination with FITC-dextran and Rhodamin 6G as fluorescent dyes was used to assess microcirculatory changes, leukocyte behavior, and angiogenesis during the 14 d of wound healing. RESULTS The area of no perfusion decreased significantly over the observed period, and perfusion was almost completely restored due to angiogenesis by d 14 (day 1: 1.89 [mm(2)]±0.44SEM, d 14: 0.02±0.01). No post-traumatic extension of the trauma could be observed. Edema formation increased significantly up to d 7. The number of adherent leukocytes showed a significant increase during the first 7 d. Functional vessel density showed a significant post-frostbite decrease to 60% of the baseline value. CONCLUSIONS This novel frostbite model provides a simple and nonetheless highly effective technique of creating locally limited reproducible frostbite injuries using a no touch technique. Tissue damage can be fully attributed to the thermal trauma, and the model allows repetitive intravital fluorescent microscopy of the microcirculation, leukocyte-endothelium interaction, and angiogenesis.

Improved Neovascularization of PEGT/PBT Copolymer Matrices in Response to Surface Modification by Biomimetic Coating

PEGT/PBT (polyethylene glycol terephthalate/polybutylene terephthalate) copolymer matrices with three different surface coatings [calcium-phosphate (Ca-P), collagen, and gas plasma] were placed into dorsal skinfold chambers of 24 balb/c mice. Untreated PEGT/PBT matrices served as the controls. The basal surfaces of the implants directly contacted the striated skin muscle. Neovascularization of the implants was analyzed by intravital fluorescence microscopy. Microcirculatory observations were performed in the surrounding skin muscle, at the border zone of the implant, and in the center of the implant. The functional vessel density (FVD; mm/mm2), as the length of perfused microvessels per observation area, was measured by computer-assisted analysis. The FVD served as the parameter of neovascularization. At the end of the protocol, histological observation of hematoxylin/eosin-standard-stained sections was performed by light microscopy. The FVD in the center of the implant on day 8 was only observed in gas-plasma-coated (8.8 ± 10.2 mm/mm2) and Ca-P-coated implants (0.8 ± 2.0 mm/mm2). None of the other groups showed perfused microvessels in the center of the implant on day 8 (p < 0.05). The FVD values in the center of the gas-plasma-coated and the Ca-P-coated implants were 20.7 ± 8.2 and 19.2 ± 15.5 mm/mm2 as compared with 7.1 ± 17.4 and 7.7 ± 5.9 mm/mm2 for collagen-coated and untreated implants on day 16. The histological examination confirmed the profound microvascular ingrowth into the matrix pores of the gas-plasma-treated and the Ca-P-coated copolymer matrices in the center of the implants. The study showed that the ingrowth of microvessels into PEGT/PBT matrices can be accelerated by Ca-P coating and gas plasma treatment in the dorsal skinfold chamber in mice.

Repetitive extracorporeal shock wave applications are superior in inducing angiogenesis after full thickness burn compared to single application

OBJECTIVE Burn wounds remain a challenge due to subsequent wound infection and septicemia, which can be prevented by acceleration of wound healing. The aim of the study was to analyze microcirculation and leukocyte endothelium interaction with particular focus on angiogenesis after full-thickness burn using three different repetitions of low energy shock waves. METHODS Full-thickness burns were inflicted to the ears of hairless mice (n=44; area: 1.6±0.05 mm2 (mean±SEM)). Mice were randomized into four groups: the control group received a burn injury but no shock waves; group A received ESWA (0.03 mJ/mm2) on day one after burn injury; group B received shock waves on day one and day three after burn injury; group C ESWA on day one, three and seven after burn injury. Intravital fluorescent microscopy was used to assess microcirculatory parameters, angiogenesis and leukocyte interaction. Values were obtained before burn (baseline value) immediately after and on days 1, 3, 7 and 12 after burn. RESULTS Shock-wave treated groups showed significantly accelerated angiogenesis compared to the control group. The non-perfused area (NPA) is regarded as a parameter for angiogenesis and showed the following data on day 12 2.7±0.4% (group A, p=0.001), 1.4±0.5% (group B, p<0.001), 1.0±0.3% (group C, p<0.001), 6.1±0.9% (control group). Edema formation is positively correlated with the number of shock wave applications: day 12: group A: 173.2±9.8%, group B: 184.2±6.6%, group C: 201.1±6.9%, p=0.009 vs. control: 162.3±8.7% (all data: mean±SEM). CONCLUSION According to our data shock waves positively impact the wound healing process following burn injury. Angiogenesis showed significantly improved activity after shock wave application. In all three treatment groups angiogenesis was higher compared to the control group. Within the ESWA groups, double applications showed better results than single application and three applications showed better results than single or double applications.

Surface modification by glow discharge gasplasma treatment improves vascularization of allogenic bone implants.

Sufficient induction of blood vessel ingrowth decisively influence transplant functionality. In this study, microvascular response to transplants of surface modified bone substitutes were assessed in vivo. The surface modification of allogenic bone substitutes (dehydrated human femoral head) was achieved in a double‐conductive low‐pressure gasplasma reactor (Ar2/O2, 13.65 MHz, 1,000 W, 5 Pa). The modified bone substitutes (n = 10) as well as untreated bone substitutes serving as controls (n = 10) were placed into the dorsal skinfold chamber of female balb/c mice (n = 10). Dynamic assessment of microcirculatory parameters was performed using intravital fluorescence microscopy during an implantation period of 10 days. The angiogenic response was found markedly accelerated in gasplasma‐treated bone. Compared to untreated implants, the gasplasma‐activated bone substitutes showed significantly higher microvascular density on days 5 and 10. The quantification of the microvascular diameters, red blood cell velocity, and microvascular permeability displayed stable perfusion and vascular integrity of the newly developed blood vessels throughout the 10‐day observation period. The surface activation via cold low‐pressure glow discharge gasplasma supports the vascular integration of allogenic bone by earlier induction of the angiogenesis.

Thoracic Wall Reconstruction after Tumor Resection

Introduction Surgical treatment of malignant thoracic wall tumors represents a formidable challenge. In particular, locally advanced tumors that have already infiltrated critical anatomic structures are associated with a high surgical morbidity and can result in full-thickness defects of the thoracic wall. Plastic surgery can reduce this surgical morbidity by reconstructing the thoracic wall through various tissue transfer techniques. Sufficient soft-tissue reconstruction of the thoracic wall improves quality of life and mitigates functional impairment after extensive resection. The aim of this article is to illustrate the various plastic surgery treatment options in the multimodal therapy of patients with malignant thoracic wall tumors. Materials and methods This article is based on a review of the current literature and the evaluation of a patient database. Results Several plastic surgical treatment options can be implemented in the curative and palliative therapy of patients with malignant solid tumors of the chest wall. Large soft-tissue defects after tumor resection can be covered by local, pedicled, or free flaps. In cases of large full-thickness defects, flaps can be combined with polypropylene mesh to improve chest wall stability and to maintain pulmonary function. The success of modern medicine has resulted in an increasing number of patients with prolonged survival suffering from locally advanced tumors that can be painful, malodorous, or prone to bleeding. Resection of these tumors followed by thoracic wall reconstruction with viable tissue can substantially enhance the quality of life of these patients. Discussion In curative treatment regimens, chest wall reconstruction enables complete resection of locally advanced tumors and subsequent adjuvant radiotherapy. In palliative disease treatment, plastic surgical techniques of thoracic wall reconstruction provide palliation of tumor-associated morbidity and can therefore improve patients’ quality of life.