EUROPEAN TISSUE REPAIR SOCIETY
JOINT MEETING IN BORDEAUX |
EUROPEAN TISSUE REPAIR SOCIETY JOINT MEETING IN BORDEAUX |
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POSTER WINNERS ABSTRACTS from the JOINT MEETING of
the ETRS and WHS Remittance Spectroscopy in Monitoring of Wound Healing in vivo Liebold K1,2, Faßler D2,
Schmidt W-D2, Kühn T2 and Wollina
U1. Purpose: The wound healing process of chronic venous and mixed arterio-venous ulcers is still a challenging problem in modern medicine. We report on the development and the adaptation of an objective method, the remittance spectroscopy of human skin, to determine important factors affecting wound healing, especially the cutaneous per-fusion in skin adjacent to ulcers. Methods: The spectroscopic measurements were performed with fibre optic diode-array spectrometers (Carl ZEISS, Germany) working in the visible and near infrared region. Wavelength ranged from 300 to 1700 nm. Patients with venous and mixed arterio-venous ulcers were enrolled in the study. The readings were taken in the skin surrounding ulcers and above granulation tissue. The ulcer healing was determined by clinical findings like the rate of change in wound area and the state of granulation and epithelia-lisation. Results: The remittance of skin showed absorption peaks of haemoglobin between 425 and 575 nm. Rapid wound healing lead to an increased perfusion with local higher blood volume and concentration of oxyhaemoglobin in the skin adjacent to ulcers. These facts caused changes in haemoglobin absorption especially in oxyhaemoglobin absorption peaks. Conclusion: Remittance spectroscopy monitoring of chronic ulcers seems to be a helpful method to determine the cutaneous perfusion as a critical factor of wound healing. Chemotherapy for Breast Cancer down regulates Fibronectin Production in Human Acute Wounds without under-expression of a5ß1 Integrin Banerjee D, Ameen H, Barrett-Lee P*,
Harding K, Moore K and Jones D Aim: To evaluate the potential effects of adjuvant chemotherapy for breast cancer on acute wound healing at the cellular level. Methods: In this prospective study, we have studied healing in an in vivo wound model. Patients with invasive, node-positive carcinoma of the breast, receiving Cyclophosphamide, Methotrexate and 5-FU (CMF) were biopsied on day eight and day thirteen of the first cycle. The first (3mm) biopsy created a wound in healthy skin at a cosmetically acceptable site away from the original operative area and the subsequent 6mm biopsy included the healing wound. Biopsy tissue was snap frozen in liquid nitrogen and serial 6m cryostat sections were mounted on microscope slides pre-coated in poly-L-Lysine and stored at ‑20°C prior to immunohistochemical staining using monoclonal antibodies specific for a5b1, Integrin and Fibronectin. The degree of positive staining was scored 1+ to 5+. The results were compared against biopsies from six age-matched healthy volunteers. Results: A total of twelve patients were studied (age range 34–56). All wounds were considered clinically healed by the end of three weeks. Wound re-epithelialisation was delayed and the provisional wound matrix was more friable. Keratinocyte a5b1 Integrin expression was unaffected in the patients receiving CMF. However, the level of Fibronectin within the provisional wound matrix was found to be less than the controls. Conclusions: (1) Chemotherapy has no demonstrable effect on the keratinocyte expression of a5ß1 Integrin. (2) The down regulation in the level of Fibronectin may account for the delay in wound re-epithelialisation and friability of wound matrix. Immunohistochemical localisation of types I, III, IV and V collagens in a bioengineered living skin construct grafted onto nude mice Guerret S, Govignon EJ*, Hartmann DJ,
and Ronford V*. Type I collagen is a biomaterial largely used in tissue engineering and is clinically approved. It acts as a regenerative template in which the implanted collagen is progressively degraded and replaced by new cell-synthetised tissue. Apligraf™, a bioengineered living skin tissue, is composed of a bovine collagen lattice containing living human fibro-blasts overlaid with a fully differentiated epithelium made of human keratinocytes. The epithelial part of the tissue construct provides an immediate biological coverage and the collagen lattice containing fibroblasts provides the dermal template. In this report, we investigated the rate of Apligraf™ remodelling when grafted onto nude mice. Apligraf™ was applied to cover full thickness wound on the back of athymic mice. Wounds were biopsied and examined at timed intervals up to six months after grafting. The distribution of type I, III, IV and V collagens was studied by immunohistochemistry using species cross-reacting or species-specific antibodies for mouse, bovine or human proteins. Persistence of the cells present in the tissue construct was revealed with antibodies directed against human specific markers such as human involucrin for keratinocytes and human vimentin for fibroblasts. Experiments done on the nude mice demonstrated that human cells were still present up to six months after grafting. Bovine type I collagen persisted for several months but was progressively invaded by a murine neovascular-isation, as shown by mouse type IV collagen immunostain-ing. Some amount of type I and III human collagen was secreted by human fibroblasts. We observed the presence of type IV and type V human collagen close to the epithelium layer. These results have shown: (1) living cells present in the tissue construct were synthetically active at least six months after grafting, (2) the use of species-specific antibodies has enabled the evaluation of the progressive replacement of bovine collagen by the host tissue. |
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