The effects of a single dose of 5-fluorouracil on keloid scars: a clinical trial of timed wound irrigation after extralesional excision.

The possibility of altering the pathophysiology of keloid scars was investigated in 11 patients, using a single application of 5-fluorouracil solution for 5 minutes after extralesional excision was performed. Similar excisional wounds treated with phosphate-buffered saline for 5 minutes served as synchronous controls. An objective scoring system and subjective assessment were made to assay the change in the quality of the wound-healing and scar tissue produced by this treatment. A keloid scar score was used at regular time intervals after treatment to assess the quality of scar produced, thereby enabling the treated and control scars to be clinically compared. Biopsies were taken of the control and treated scars 1 month after treatment; the biopsy specimens were then subjected to immunohistochemical analysis as well as a functional assessment of cultured keloid fibroblasts. The immunohistochemical antigens assayed were Ki-67 (also called MIB-1; a marker of cell proliferation); vascular cell adhesion molecule-1 (a marker of inflammation); transforming growth factor beta-1 (a factor involved in scarring) and CD-68 (a macrophage-specific marker). Fibroblast-populated collagen lattices provided a functional assessment of fibroblast contraction. All treated and control wounds healed without any dehiscence or infection. The keloid scar score revealed that there was a perceived improvement in condition for those treated with 5-fluorouracil, compared with the control specimens, during the 6-month follow-up period in the five patients who attended all their clinic appointments; data on later recurrence are not complete as yet. The wounds treated with 5-fluorouracil produced scars that had a significant (p < 0.01) reduction in all the markers assayed, apart from CD-68. Functionally, the keloid fibroblasts from three of five of the treated patients showed reduced contractile capacity. This pilot study demonstrates that a "single-touch" technique with 5-fluorouracil can produce a change in the characteristics of the healing keloid wound after extralesional excision. Long-term studies are required to elucidate the correct dosage and time of exposure to improve the efficacy of this potential treatment.

Role of phenytoin in wound healing--a wound pharmacology perspective.

Topical agents used for the enhancement of wound healing are designed to act locally and, therefore, do not undergo classic systemic metabolic modification. This commentary reviews the potential role of a vulnerary agent, phenytoin, (PHT), from a wound pharmacology perspective. This agent may have the potential to alter the dynamics of wound healing, suggesting a therapeutic use for the stimulation of chronic wounds. Oral PHT therapy is used widely for the treatment of convulsive disorders, and about half the patients treated develop gingival overgrowth as a side-effect. This apparent stimulatory effect has prompted its assessment in wound healing. Investigations into the mechanisms of gingival overgrowth also provide clues to its action in wound healing, and important similarities and differences are discussed. It appears also that both gingiva and skin are important extrahepatic sites for xenobiotic metabolism, and analysis of the biochemical mechanisms should lead to the design of safer analogues for wound healing. On the other hand, differences between the pharmacokinetics of topical PHT in these tissue situations indicate that different formulations are required for gingival and cutaneous wound healing and during the changing course of wound healing itself.

Phenytoin reduces the contraction of recessive dystrophic epidermolysis bullosa fibroblast populated collagen gels.

Recessive dystrophic epidermolysis bullosa (RDEB) is a group of genetic disorders in which blistering occurs below the basement membrane, in many cases resulting in extensive scar formation, contractures and mitten deformities. Our aim was to compare quantitatively the contraction forces generated by normal and RDEB fibroblats and to investigate the effect of Phenytoin (5,5-diphenyl-2,4-imidazolidinedione, sodium salt; PHT). PHT is an anticonvulsant agent, that causes fibrosis as a side effect. This study utilised conventional untethered fibroblast populated collagen lattice contraction and a quantitative force measurement instrument, the culture force monitor (CFM). The RDEB cell lines were hypercontractile, generating 2.5 times the force of normal fibroblasts, though they appeared morphologically normal. In untethered collagen gels PHT (20 micrograms/ml) significantly reduced contraction of both normal and RDEB fibroblasts over 7 days. Pre-treatment of RDEB cells for 5 days also produced a 40% reduction in contraction as measured in the CFM. One suggested mechanism of PHT action is through inhibition of matrix metalloproteinase activity, but the similar effects of PHT and Colchicine (an inhibitor of microtubule polymerisation) in the CFM, indicate that it may act on contraction through disruption of microfilaments and changes to cell shape. These findings show that isolated RDEB fibroblasts retain the hypercontractile features of many of the patient's lesion sites and imply that local application of PHT may have a therapeutic effect in controlling contraction.

Balanced mechanical forces and microtubule contribution to fibroblast contraction.

Fibroblast locomotion is thought to generate tractional forces which lead to contraction and reorganisation of collagen in tissue development and repair. A culture force monitor device (CFM) was used to measure changes in force in fibroblast populated collagen lattices, which resulted from cytoskeletal reorganisation by cytochalasin B, colchicine, vinblastine, and taxol. Microfilament disruption abolished contraction forces, microtubule disruption elicited a new peak of contraction, while taxol stabilisation of microtubules produced a gradual fall in measured force across the collagen gel. Based on these measurements, it is suggested that the cell can be viewed as an engineering structure in which residual intracellular forces, from contractile microfilaments, exert compressive loading on microtubular elements. This microtubular structure appears to act as a "balanced space frame" (analogous to an aeroplane chassis), maintaining cell shape and consequently storing a residual internal tension (RIT). In dermal fibroblasts this hidden RIT was up to 33% of the measurable force exerted on the collagen gel. Phenotypic differences between space frame organisation and RIT levels could explain site and pathological variations in fibroblast contraction.