A new technique to resurface wounds with composite biocompatible epidermal graft and artificial skin.

BACKGROUND: The incorporation of cultured epidermal autograft on the neodermis of artificial skin (Integra, Integra LifeSciences, Plainsboro, NJ) has been met with some difficulties. A new engraftment technique to resurface the wounds with Integra and composite biocompatible epidermal graft (CBEG) has been successfully applied on three patients for elective reconstructive procedures. METHODS: A small skin biopsy was taken from the normal edge of the lesion for keratinocytes and dermal fibroblast cultures 2 weeks before surgery. When sufficient cells were grown, the patient was admitted for the excision of the lesions or scars. The wounds of the patients, ranging from 125 to 250 cm2, were covered with Integra. When the neodermis of the Integra was fully vascularized, the silicone membrane of the Integra was removed and replaced with the CBEG, which consisted of autologous keratinocytes cultivated on a hyaluronate-derived membrane (Laserskin; Fidia Advanced Biopolymers, Abano Terme, Italy)) using human dermal fibroblasts as a feeder layer. RESULTS: Clinically, there was good initial "take" of the CBEGs in these three patients, ranging from 50% to 100%. Biopsy specimens of the grafted wounds were taken 1 to 3 weeks after the application of the CBEGs. Epithelialization was noted in all patients. CONCLUSION: This engraftment technique has several advantages. The CBEG is much easier to handle than the conventional cultured epidermal autograft. It eliminates the invasive second procedure for skin harvesting, with resulting pain and scarring. The application of the CBEG can be easily performed at the bedside.

Dermal fibroblasts do not enhance the graft take rate of autologous, cultured keratinocyte suspension on full-thickness wounds in rats.

Dermal fibroblasts are known to play an important role in wound healing. In this study, cultured autologous keratinocyte suspension was applied with fibrin glue to the full-thickness wounds in rats (N = 20). Histological analysis on day 14 showed regenerated epithelium in 10 wounds (50%). Keratinocytes were also premixed with allogeneic dermal fibroblasts in a ratio of 3:1 and 5:1 before application to other full-thickness wounds (N = 20) with fibrin glue. Regeneration of epithelium was observed in 10 (50%) and 9 (45%) wounds respectively. Acute inflammatory reaction and mild to moderate proliferation of fibroblasts in the subepithelial layer of the allogeneic fibroblasts were noted. The addition of dermal fibroblasts to keratinocytes/fibrin glue does not enhance the take rate of the cultured keratinocyte suspension.

A new system for the cultivation of keratinocytes on acellular human dermis with the use of fibrin glue and 3T3 feeder cells.

The growth of human keratinocytes on human acellular dermis in 4 different culture systems was compared. Epidermis was completely separated and removed from dermis after skin samples had been soaked in 0.1% trypsin at 4 degrees C for 1 week. Forty pieces of saline-washed dermis, 1 cm2 each, were randomized into 4 groups: in group A, human keratinocytes that had undergone 2 to 3 cell passages were seeded (30 x 10(4) cell/cm2) onto the dermis and sprayed with a thin layer of fibrin glue and proliferative 3T3 feeder cells that had been growing separately on the culture dish; in group B, the dermis was only sprayed with fibrin glue; in group C, the dermis was treated with 3T3 cells only; and in group D, the dermis was not sprayed with anything. The dermis samples in all groups were raised on a grid to provide an air-liquid culture system. Histology results of the composite grafts at 2 weeks were assessed as having either scanty colonies of keratinocytes (SCK), continuous stratified epithelium (CSE), or no observable keratinocyte growth. Eight out of the ten dermis samples (80%) in group A demonstrated CSE, and 30% of the samples in group B showed SCK. There were 10% SCK and 20% CSE in group C, and in group D, 30% SCK and 10% CSE were found. The good results in group A indicated that the fibrin glue facilitated the seeding efficiency of the keratinocytes on the dermis and that the vital factors released from the 3T3 feeder cells enhanced the growth and differentiation of the keratinocytes. This model provides an optimal system for the cultivation of keratinocytes on acellular dermis.

Diagnostic and prognostic accuracy of the exercise electrocardiogram in patients with preexisting right bundle branch block.

The value of exercise testing in patients with right bundle branch block (RBBB) is uncertain. A retrospective review of 3609 patients who underwent exercise testing identified 163 (4.5%) with preexisting RBBB. After excluding those with coronary artery bypass graft(s), 133 patients remained and 48 (36%) had a prior myocardial infarction. Angiograms were available for 30 (23%) patients. After a mean follow-up of 33 +/- 23 months, seven patients had a fatal or nonfatal myocardial infarction. Twenty five (19%) patients had > or = 1 mm of non-upsloping ST depression in leads V5 or V6. With angiographic disease or previous myocardial infarction used as endpoints of coronary artery disease, the exercise test had a sensitivity of 27% (15/56), a specificity of 87% (67/77), and a predictive accuracy of 62% (82/133), (chi 2 = 4.04, p = 0.04). There were 24 deaths, a 10% annual mortality rate. Univariate analysis of clinical, exercise, and angiographic data revealed that nonsurvivors had a lower peak systolic blood pressure, a lower exercise capacity in METS, and a higher prevalence of coronary artery disease (p = 0.0001, p = 0.02, p = 0.03, respectively). Left ventricular ejection fraction and the amount of additional ST depression during exercise did not differ significantly (p = NS). Receiver operating characteristic curve analysis revealed that systolic blood pressure (area = 0.741, z = 5.22, p < 0.001) and exercise capacity (area = 0.66, z = 3.12, p = 0.009) were predictive of mortality, whereas additional ST depression during exercise (area = 0.588, z = 0.70, p = 0.24) was not.(ABSTRACT TRUNCATED AT 250 WORDS)