Extracellular matrix proteins of human epidermal keratinocytes and feeder 3T3 cells.

Cultures of human epidermal keratinocytes obtained from adult epidermis were initiated using irradiated BALB/3T3 cells as feeder layers. At different stages of confluence of the epidermal islands, feeder cells were removed and the extracellular matrix proteins of both pure component cells and cocultures were analyzed biochemically and by immunochemical methods and compared to those of skin fibroblasts of the same donors. The keratinocytes synthesized and secreted fibronectin and small amounts of laminin and type IV collagen. In addition, a nondisulfide-linked collagenous polypeptide (Mr = 120,000) was synthesized by the keratinocytes and was confined to the cell layers. Collagenous polypeptides with Mr = 120,000 were also synthesized by organ cultures of epidermal tissue and were detected in its acid or detergent extracts but again no secretion to culture medium was found. The Mr = 120,000 collagen had biochemical and immunological properties distinct from those of types I-V collagens. In immunofluorescence of keratinocyte cultures, fibronectin staining was prominent in the lining marginal cells of the expanding periphery of the epidermal cell islands but was not detected in the terminally differentiating cells in the upper layers of stratified colonies. Very little type IV collagen was found deposited in pericellular matrix form by the keratinocytes. In contrast, the mouse 3T3 feeder cells were found to produce both type IV collagen and laminin in addition to the previously identified connective tissue glycoproteins of fibroblasts, interstitial procollagens, and fibronectin. Basement membrane collagen of the 3T3 cells was found deposited as apparently unprocessed procollagen alpha 1(IV) and alpha 2(IV) chains. The production in culture conditions of basal lamina glycoproteins by the fibroblastic feeder cells may promote the attachment and growth of the cocultured keratinocytes.

Late results of skin-sparing mastectomy followed by immediate breast reconstruction.

BACKGROUND: Skin-sparing mastectomy (SSM) facilitates optimal immediate breast reconstruction (IBR) by preserving the inframammary fold and most of the breast skin. Concerns persist that SSM might increase the rate of local recurrence as the surgical approach is less extensive. Patients who had SSM and IBR over 10 years at a single institution were reviewed. METHODS: A total of 207 consecutive women who underwent SSM and IBR from 1992 to 2001 were included in the study. The patient records were analysed retrospectively and follow-up data were included. RESULTS: Postoperative complications included native skin flap necrosis (10.1 per cent), haematoma (10.6 per cent), infection (3.4 per cent), anastomotic thrombosis (5.3 per cent) and hernia (2.6 per cent). During a mean follow-up of 70 months, 5.8 per cent of patients with stage 0-2 disease developed a locoregional recurrence, although none of these later had a systemic recurrence. The rate of locoregional recurrence was 31 per cent (five of 16) in women with stage 3 breast cancer. CONCLUSION: SSM followed by IBR was both surgically and oncologically safe, especially for early-stage breast cancer.

The importance of a multidisciplinary group in the treatment of soft tissue sarcomas.

In 1987, a multidisciplinary soft tissue sarcoma (STS) group was established and a treatment protocol was set up. By 1993, there were 193 patients with a diagnosis of STS of the superficial trunk or extremities. 134 patients were referred with primary (stage M0) tumours and treated with curative or palliative intention. Nine amputations were performed. 94 (70%) patients were treated with wide or compartment surgery (n = 62) or marginal surgery combined with postoperative radiotherapy (n = 32). According to the protocol, these patients had received adequate treatment. 18 patients have recurred locally (13%) (median follow-up: 36 months). 12 were salvaged. 33 had metastases. The estimated 3-year survival, local control and disease-free survival rates are 79, 87 and 69%, respectively. These results compare favourably with previously published results from this hospital and from a nationwide study. The improved results emphasise the importance of a multidisciplinary STS group.

Bone marrow induced osteogenesis in hydroxyapatite and calcium carbonate implants.

In this experimental study, blocks of natural coral (calcium carbonate) and its structurally similar derivate in the form of hydroxyapatite (calcium phosphate) were implanted in rat latissimus dorsi muscle with autogenous bone marrow to compare their bone-forming capability. A block without marrow placed in the opposite latissimus muscle served as a control. The animals were killed at 3, 6 and 12 weeks and, in the hydroxyapatite group, also at 24 weeks. The sections were analysed histologically and histomorphometrically. Bone was found only in implants containing bone marrow. Bone formation was significantly (p < 0.05) higher in coral than in hydroxyapatite implants at 3 weeks (10.8% versus 4.8%) and at 12 weeks (13.7% versus 6.3%, bone/total original block area). At 12 weeks all the coral implants had lost their original structure, and the cross-sectional area of the block had diminished to 40% of the original area.

Compressive strength of calcium carbonate and hydroxyapatite implants after bone-marrow-induced osteogenesis.

Natural coral and structurally similar porous hydroxyapatite (HA) have been used as bone substitutes. They are not osteoinductive but bone formation can be induced by marrow cells, even in extraosseal sites. In our previous study we induced bone formation in porous coral and HA after having implanted the materials in intramuscular pockets in rat. New bone formed only in HA or coral implants soaked with marrow cells; fibrous tissue ingrowth alone was observed in the controls (without marrow). In the present study we examined the effect of tissue ingrowth on the mechanical properties of coral and HA implants obtained in a similar process to that used before. At 12 weeks the compressive strength of HA was higher in the marrow group than in the controls; it was also higher than that of the wet unimplanted material. The HA blocks did not show resorption. Coral resorbed quickly and lost its compressive strength, which was originally higher than in HA. At three weeks the marrow group was stronger than the control specimens. After six weeks only the marrow group, but not the controls, could be tested. Bone ingrowth seemed to maintain the strength of the coral implant even if it was dissolving. The mechanical strength of both materials was comparable to that of cancellous bone.

Overrepresentation of 1q21-23 and 12q13-21 in lipoma-like liposarcomas but not in benign lipomas: a comparative genomic hybridization study.

Twenty lipomatous tumors, including eight lipoma-like liposarcomas and 12 benign lipomas, were analyzed using comparative genomic hybridization (CGH). DNA sequence copy number changes detected in five lipoma-like liposarcomas (mean, 1.1 aberrations/tumor; range, 0-2) consisted of gains of 12q13-21 (five tumors) and 1q21-23 (four tumors). Two of the tumors showed high-level amplification at 12q14-21 and one tumor at 1q21-22. No copy number changes were found in lipomas. Overrepresentation of 1q and 12q sequences was a recurrent finding in lipoma-like liposarcomas but not in lipomas. Thus, CGH may help in the differential diagnosis of low-grade or borderline adipose neoplasms.

A cytogenetic study of malignant fibrous histiocytoma.

We report the results of cytogenetic analysis of malignant fibrous histiocytoma of soft tissue (MFH). Seven of 12 successfully cultured MFHs had complex clonal aberrations, including translocations, deletions, and unidentifiable marker chromosomes. Telomeric associations were observed in five and the double minute phenomenon in four of seven MFHs with abnormal karyotypes. In one case (a storiform-pleomorphic MFH, grade IV) with a complex polyploid karyotype, two clonal ring chromosomes were present, one interpreted as r(19)(p13q13), one unidentified. In two tumors, clonal structural rearrangements of chromosome 1 were seen: del(1)(q21) in a storiform-pleomorphic MFH, grade IV, and add (1)(q21 or q32), t(1;10)(p22;q22) in a myxoid MFH, grade I. The remaining five MFHs had normal karyotypes, but in two of them nonclonal, structural aberrations were found. The modal chromosome number in the studied MFHs varied widely, but the majority of tumors with abnormal karyotypes had polyploid chromosome complements (five of seven cases). Our results confirm many of the previous findings and indicate that double minutes (dmins) may be more frequent in MFH than previously reported.

Cytogenetic study of extraskeletal mesenchymal chondrosarcoma. A case report.

Extraskeletal mesenchymal chondrosarcoma (EMC) is a rare and highly malignant type of chondrosarcoma of soft tissue origin. We performed a cytogenetic study on a patient with EMC. Cytogenetic analysis revealed the tumor karyotype: 48-49,XX, t(4;9)(q23;q22), add(10)(q?26), +16, ?del(19)(p13), +1-2mar[cp12] / 48-50,idem, t(1;20)(q21;q13), +mar[cp6] / 46,XX [7].

Prognostic value of tumour vascularity in primary melanoma.

To investigate the prognostic value of tumour vascularity we studied 84 patients with primary melanomas ranging in tumour thickness (Breslow) from 0.37 to 7 mm and in depth of tumour infiltration (Clark) from II to V. Vascularization was assessed by immunohistochemistry with a CD-31 antibody recognizing endothelial cells. The CD-31-positive vessels were counted and the degree of vascularization was correlated with the survival of the patients. In addition, the relationship between blood vessel density and some histopathological data is discussed. In our study, the multivariate Cox model showed that the only independent variable in disease-free survival was tumour thickness (Breslow classification) and the only one in overall survival was depth of tumour infiltration (Clark classification). In disease-free survival, tumour thickness (Breslow classification) was a clear prognostic factor (P = 0.004) after 4 years' follow-up, as were depth of tumour infiltration (Clark classification) (P = 0.04) and ulceration (P = 0.04). In overall survival, tumour vascularity was the strongest prognostic factor at 4 years, high vascularity being associated with a good prognosis (P = 0.06). Clark classification was also a prognostic factor (P = 0.02) in overall survival. We conclude that high vascularization is associated with a better prognosis but is not an independent prognostic indicator.

Haemostatic disturbances in burned patients during early excision and skin grafting.

Bleeding is a major problem during early excision of burned skin. Therefore, 13 severely burned adult patients operated on during the first week after the trauma were studied. Blood loss was replaced with crystalloids, colloids and packed red cell concentrates (PRC). After ten infused PRCs, four fresh frozen plasma (FFP) units were given and thereafter one FFP unit with one PRC unit. Arterial blood samples were drawn before anaesthesia (SO), during operation after every four units of PRC transfusion (S1-4), 4 h postoperatively (S5) and on the first postoperative morning (S6). Prothrombin time (%) and activated partial thromboplastin time (s) were abnormal before operation (median values 67%, range 22-99% and 44 s, range 30-86 s, respectively). Prothrombin time decreased during operation and reached the critical level for normal haemostasis at S2. Thrombelastography showed decreased clot formation rate and impaired fibrin platelet interaction peri- and postoperatively. Fibrinogen and factor VIII activity were high preoperatively (median 6.1 g/l and 253%) and the critical values for normal haemostasis were not reached. Burned patients have a consumption coagulopathy which, in combination with haemodilution during operation, results in a clinically significant deficiency of coagulation factors II, VII and X, in spite of reactive elevation of coagulation factor VIII and fibrinogen.

Panoramic zonography in evaluation of recurrent basal cell carcinoma of the face.

Nine patients with recurrent, facial basal cell carcinoma were followed up both clinically and radiographically using a panoramic technique. A cylindrical image track was used for panoramic radiography. Five patients had bony destruction due to tumour re-occurrence, four patients had bony defects caused by surgery. The confirmation of the results by surgery and/or follow-up shows that the spread of basal cell carcinoma into bone can be diagnosed using a panoramic technique.

Anti-melanoma antibodies bind preferentially to diploid metastases in immunoscintigraphy.

Immunoscintigraphy with 99Tcm-labelled anti-melanoma monoclonal antibody F(ab')2- fragments was performed in 23 patients with histologically verified metastatic melanoma. Immunoscintigraphy was positive in 14 patients and all known metastases were detected in eight patients, five of whom had only one lesion. Lesion localization and detectability were as follows: 12/13 (92%) cutaneous and subcutaneous, 11/14 (79%) lymph node, 5/7 (71%) bone, 3/6 (50%) lung and 1/5 (20%) abdominal metastases were visualized. Despite its high specificity--no false positive immunoscintigrams--the low sensitivity of this method in detecting deep metastases hampers its usability. The false negative results were not due to lack of antigen expression as positive immunostaining results were observed also in specimens from patients with negative immunoscintigrams. Flow cytometric analysis of the metastases revealed that in 7/8 (88%) patients with diploid tumours had positive immunoscintigrams but only 7/15 (47%) patients with aneuploid tumours. These results show that the diagnostic accuracy of melanoma immunoscintigraphy can be improved by selecting patients not only by testing for the antigen but also on the basis of DNA analysis of an accessible lesion.

Delayed breast reconstruction.

Immediate reconstruction is becoming increasingly popular because it gives a better aesthetic result for breast reconstruction, and is more economical and convenient for patients than delayed reconstruction. Delayed breast reconstructions are still needed in 80% of mastectomized patients under 40 years and in half the women under 54 years--at least in Finland. This article discusses the technique of and contraindications for the free transverse rectus abdominis myocutaneous (TRAM) flap technique based on the author's experience with over 300 free TRAM operations. The pedicled TRAM and the deep inferior epigastric artery (DIEA) perforator flap are also discussed.

Return to employment after burn.

The increase in the survival rate of burned patients has stressed the need to study their rehabilitation. The purpose of our study was to characterize the factors influencing such patients' return to work. We conducted a mail survey among 316 patients aged 15-65 years, treated at Töölö Hospital Burns Unit between 11 November 1988 and 31 December 1994. Of 175 participants, 130 (74 per cent) were men and 45 (26 per cent) women. The mean TBSA was 14.0 per cent, mean FT 6.4 per cent and the mean time of hospital treatment (TOT) 17.5 days. Statistical significance was calculated by Mann-Whitney U test, Kruskall-Wallis one-way analysis of variance and Chi-square test as appropriate, with a probability level of 0.05. In the study, 54 per cent of patients whose burn area was 1-10 per cent, returned to work within 2 months. No difference was found between patients who had hand burns and those who had burn injuries in other parts of the body. Patients who did not return to work were significantly older (mean age 45 years) than those who did (mean ages varying from 33 to 36 years; P < 0.05). Total body surface area burned (TBSA), FT, TOT, age and employment status at the time of injury were the factors predicting the resumption of working ability after burn injury.

Suitability of the scapular flap for reconstructions of the foot.

Eighteen patients with mainly a traumatic soft-tissue defect of the foot underwent reconstruction with a microvascular free scapular flap. Of the 17 successful transfers, 13 were to the weight-bearing parts of the foot. The stability and contour of the flaps were assessed after an average follow-up time of 3 years (range 1 to 5 years). The thicknesses of the scapular donor site and flap and the recipient site were measured by an ultrasound technique. The resistance of the flap to shear was measured with a dynamometer. The ultrasound measurements aided in refining our operative technique. In early cases, the flap thickness after transfer could be more than double what it was in the donor area. With proper tightening, the thickness could be reduced, with improvement in contour but no increase in soft-tissue stability or shear resistance of the flap. Without proper tightening, the scapular flap tended to be redundant when transferred to the foot. For good results, the patient should be lean, since the optimal thickness of the scapular donor site was less than 6 mm and the maximum thickness should not exceed 8 to 10 mm. The differences in shear resistance between the flaps were not associated with the soft-tissue stability of the reconstruction. The relative laxity of the flap on the plantar surface was found by several patients to be subjectively unpleasant. Although good contour could be achieved when covering the plantar heel, the tendency of the flap to develop abrasions and superficial breakdowns made it unsatisfactory for covering this area.(ABSTRACT TRUNCATED AT 250 WORDS)

A prospective study of changes in muscle dimensions following free-muscle transfer measured by ultrasound and CT scanning.

A retrospective study demonstrated that noninnervated free-muscle flaps do not lose bulk when evaluated at a mean of 41 months. The purpose of the present study was to evaluate changes in muscle bulk in noninnervated free-muscle transfers prospectively. This study included 22 flaps (17 latissimus dorsi, 4 rectus abdominis, and 1 gracilis). The thickness of the muscle was measured by ultrasonography preoperatively and 2 and 6 weeks and 3, 6, 9, 12, 15, 18, and 23 months postoperatively. The volume of the muscle was measured by computed tomographic (CT) scan preoperatively and 2 and 6 weeks and 3, 6, and 9 months postoperatively. Postoperative data were normalized to the preoperation measurements. The results demonstrated that the thickness of the muscle increased by a mean of 2.4 times (range 0.9 to 3.9) compared with the initial thickness in a 2-week period (p < 0.05), 2.0 times (range 0.9 to 4.2) in 6 weeks (p < 0.05), 1.7 times (range 0.8 to 4.2) in 3 months (p < 0.05), 1.5 times (range 0.6 to 3) in 6 months (p < 0.05), and 1.2 times (range 0.4 to 2.8) in 9 months (not significant). Thereafter, the mean thickness was the same as the initial thickness. CT scan measurements of the muscles confirmed the ultrasound findings. Our prospective study of free-muscle flaps found significant swelling that peaks at 2 weeks and extends until 6 months after the operation. This study also demonstrated that ultrasound evaluation of thickness gives the same conclusion as volumetric measurement by CT scanning.

Immediate breast reconstruction.

Immediate breast reconstruction (IBR) in conjunction with mastectomy for cancer or high risk of breast cancer is safe from an oncological point of view. The cosmetic outcome can be excellent, especially when performing mastectomy by sparing the skin of the breast and reconstructing the breast mound with autogenous tissue. The majority of women at their working age are willing to have their breast reconstructed. Patients with newly diagnosed cancer undergoing major surgery need extra support compared with those undergoing late reconstructions. Immediate reconstructions with one operation, one hospital stay and one sick leave are economically favourable by diminishing the demand of delayed reconstructions. Preference in patient selection for IBR should be in patients with good prognoses like those with diffuse non-invasive cancer and those with a considerable risk to develop breast cancer. Women with axillary-node negative invasive cancer and women with late local recurrences in a breast earlier conservatively treated are also suitable for IBR. If needed, oncological treatments can be given after IBR, although radiotherapy after pure implant reconstructions is not recommended. In order to give all eligible patients an equal opportunity to have IBR, treatment of breast patients should be centralised to hospitals with a team comprising breast cancer surgeons, pathologists, radiologists, and plastic surgeons.

Prognosis of primary melanoma.

MATERIALS AND METHODS: To investigate the prognosis of primary melanoma, we studied a Finnish population of 298 primary melanoma patients, the majority with stage I or II tumours. The median clinical follow-up (4.8 years) was acquired from the patients' records, and the overall survival thereafter was collected from patient registries. The median follow-up for overall survival was 9.5 years. RESULTS: The overall survival rate was 66.8%. 24.5% developed metastasis, 17.8% died of melanoma, and 15.4% died of some other cause. Surgical margins had no effect on survival. In univariate analysis the most significant prognostic factors for disease-free and overall survival were stage of tumour (p < 0.0001), thickness of tumour (p < 0.0001), depth of tumour invasion (p < 0.0001) and tumour ulceration (p = 0.0005, p < 0.0002). Ulceration was an unfavorable prognostic marker. Younger patients had better survival outcomes than older ones (p = 0.04). Accordingly, in the multivariate Cox model the independent prognostic factors for both disease-free and overall survival were stage of tumour and thickness of tumour. Tumour location on trunk was an independent adverse prognosticator for overall survival. CONCLUSION: We conclude that the prognosis of primary melanoma has improved in Finland in the last decades being in line with a global tendency.

Hereditary breast cancer and handling of patients at risk.

The identification of BRCA1 and BRCA2, the two known genes causing a dominantly inherited susceptibility for breast and ovarian cancer has allowed genetic testing and identification of high risk individuals in a proportion of breast cancer families. In the future, when both the surveillance methods and prophylactic measures will be further developed this will have even more important clinical value in the management of breast cancer families. To date, as prophylactic mastectomy and/or oophorectomy have been shown to offer a significant risk reduction, these should be considered at least for known mutation carriers. Before considering this, patients should be referred for genetic counseling including risk assessment and genetic testing. Identification of a mutation in the family facilitates carrier detection by allowing predictive testing of healthy individuals. In mutation positive families, a negative test result for an individual has great value as it releases from coping with high risk of cancer and from intensive surveillance. When prophylactic surgery is considered, young age is an important determinant. A skin-sparing mastectomy with implant or autologous tissue transfer is the reconstruction method of choice. Other options like surveillance or chemoprevention can be accepted, but their uncertainty should be pointed out.

Granulocyte kinetics in burns.

Granulocytes, predominantly polymorphonuclear neutrophils, are the major components of the body's defense against acute infections. Starting from pioneering work done 20 years ago by Alexander and coworkers, neutrophil dysfunction after major thermal injury has been well documented. Defects in neutrophil function have been recognized as being serum-borne. Additionally, a review of granulocyte function in burns was presented in 1984, at the Conference on the Immune Consequences of Thermal and Traumatic Injuries. Because of methodologic difficulties, however, our knowledge of granulocyte kinetics has increased very slowly. Although the bone marrow produces 1.5 billion granulocytes per kg body weight per day and the marrow granulocyte reserve is enormous, the hematopoietic tissues, along with the release, distribution, and possible role of granulocytes in the pathophysiology of burns, have not been studied thoroughly. This article presents a current review of our knowledge of granulocyte kinetics in burns and the production and possible fate of granulocytes in tissues.