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Reverse transcription-polymerase chain reaction detection of collagen transcripts in healing human wounds.

The aim of this study was to analyse the expression of COl1A1, COl1A2 and COl3A1 in 6 mm diameter punch biopsies obtained from human wounds. Total RNA was isolated from biopsies taken from Sacrococcygeal pilonidal sinus excision cavities at weekly intervals between surgery and clinical closure. cDNAs were generated from the RNA using reverse transcriptase and polymerase chain reaction (PCR) amplifications performed with oligonucleotide primer pairs specific for regions of the COl1A1, COl1A2 and COl3A1 genes. The expression of these three genes was demonstrated throughout the course of healing on 36 biopsies taken from nine patients between surgery and clinical closure. Amplification bands demonstrated on cDNAs generated from 6 mm diameter biopsies were comparable in intensity and specificity with those generated from 50 mg excised scar tissue and cultured fibroblasts. The RT-PCR technique described here allows the rapid 'routine' detection of specific gene expression in 6 mm biopsies obtained from healing wounds.

Biopsy↗

[Tumescent local anesthesia in proctologic surgery].

INTRODUCTION: Tumescent local anesthesia (TLA) was introduced to facilitate liposuction and other procedures mainly in plastic surgery. The aim of our study was to evaluate if this new technique can also be safely applied in proctologic surgery. METHODS: The following 50 proctologic procedures were performed using TLA as day surgery cases: perianal vein thrombectomy (n=15), subanodermal fistula resection (n=7), anal polyp resection (n=7), fissurectomy ( n=6), perianal abscess revision (n=6), excision of anal skin tags (n=3), hemorrhoidectomy for thrombosed piles (n=3), intersphincteric fistula extirpation (n=1), pilonidal sinus (n=1), and perianal tumor excision (n=1). The subcutaneously infused solution contained 200 ml of Ringer's solution, 50 ml of 2% Mepivacain, and 2.5 ml of epinephrine diluted 1:10,000. RESULTS: During an average application time of 14.3+/-4.9 min, a mean of 93.1+/-40.7 ml tumescent solution was infused. This dosage resulted in complete pain relief after 18.3+/-5.0 min. Local anesthesia was maintained up to 14 h postoperatively, with a mean duration of 6.7+/-1.9 h. Two patients developed moderate hematomas which required no further treatment. CONCLUSIONS: Our pilot study demonstrates that slow infusion tumescent local anesthesia can be widely and safely applied in proctologic surgery and enables the performance of these procedures on an outpatient basis.

Abscess↗

Is routine cavity drainage necessary in Karydakis flap operation? A prospective, randomized trial.

PURPOSE: Different surgical techniques for pilonidal disease have been described in the literature. In this study our aim was to evaluate the influence of routine cavity drainage in the Karydakis flap technique. METHODS: Fifty patients with pilonidal sinus who underwent the Karydakis flap operation were evaluated prospectively. The patients were assigned randomly into two groups-with and without suction drainage of the cavity-and the effects of drains were studied in terms of wound complications, hospital stay, and recurrence rate. RESULTS: There was no significant difference between groups in term of length of hospital stay. Complication rate was 20 percent and the complications were caused exclusively by fluid collections. Wound infection, dehiscence, or failure was not observed in any of the patients. There has been no recurrence in any of the patients during the follow-up period. There was a significant increase in the number of fluid collections in patents without a suction drain. CONCLUSION: The present study indicates that routine cavity drainage reduces the incidence of fluid collection after the Karydakis flap operation.

Adolescent↗

[Excision and immediate suture technic in the treatment of pilonidal fistula. Our experience].

The authors report their personal experience of the "excision and primary suture" operating technique in the surgical treatment of pilonidal sinus. A rapid recovery by postoperative day 10 was achieved in 58 out of 60 cases treated (96.7%). Dehiscence of the surgical wound was observed in 2 cases (3.3%) following ischemic lesion caused by decubitus of the cutaneous margins and healing occurred by second intenti. No cases of short- or long-term recidivation were observed. The advantages of this method are the early return to working activities, minor patient discomfort and the reduced risk that the surgical would might become infected. Three basic stages for the successful outcome of this type of surgery have been identified as follows: 1) Although and accurate tricotomy of the sacro-coccigeal region. Antibiotic therapy is started about 2 hours before surgery. 2) The precise execution of the surgical technique. 3) The continuation of antibiotic therapy until postoperative day 7. Compressive medication is removed on postoperative day 4. If these three basic stages are respected, no cases of recidivation will occur.

Adolescent↗

[Sacro-coccygeal cyst: surgical techniques and results].

69 patients with a chronic inflamed pilonidal sinus were reviewed. They were operated either with a deep excision to sacrum (37 patients) or with the Lord Millar technique (32 patients). 66 patients (87%) were controlled at a mean follow-up of 3.7 years after the operation. The socio-economical advantage of the Lord Millar operation is demonstrated by the shorter hospital stay (4.9 vs 7.6 days) and the shorter period of disability (14 vs 26 days). 5 recurrent episodes were observed among the 31 patients followed-up after an excision (16%) and 7 among the 29 followed-up after a Lord Millar operation (24%). In the later group one recurrence was seen in the sub-group of the 16 patients regularly epilated after the operation (6%) and 6 in the sub-group of the 13 who were not controlled (46%). We conclude that the Lord Millar operation is an alternative to the deep excision operation provided the patients is motivated enough to come regularly to the control for an epilation.

Follow-Up Studies↗

The role of lymphocytes in human dermal wound healing.

BACKGROUND: Animal experimental data indicate a requirement for functionally active T lymphocytes to allow optimal healing of dermal wounds. Little evidence exists to confirm that this is the case in humans. Lymphocyte involvement in regulation of healing is probably mediated by release of secreted cytokines/growth factors, and we hypothesize that the cytokine profile requirement will be modulated as healing progresses. OBJECTIVES: As this is likely to be reflected in lymphocyte subset changes over the course of normal healing, we investigated the immunophenotype of lymphocyte subpopulations during wound healing. METHODS: Sequential biopsies were taken over 42 days from the margin of 12 wounds healing by secondary intention after pilonidal sinus excision. Serial biopsy sections were analysed by immunohistochemistry using lymphocyte-specific monoclonal antibodies, and lymphocytes were counted microscopically. RESULTS: Within 42 days, the mean decrease in wound volume was 87.5%. This was accompanied by significant changes in the wound margin lymphocyte population. Total numbers (mean +/- SEM) of T lymphocytes decreased from 36.8 +/- 9.8 cells per field at inclusion in the study to 25.9 +/- 3.0 immediately prior to wound closure, with a concomitant increase in B lymphocytes from 1 +/- 0.4 to 9.5 +/- 3.6 cells per field. The CD4/CD8 T-lymphocyte ratio fell from an initial level of 3.6 +/- 0.3 to 2.1 +/- 0.3 (mean +/- SEM) prior to closure. CONCLUSIONS: These data indicate that human wound-associated lymphocyte populations are modulated during healing; the increase in numbers of CD8+ T-suppressor lymphocytes is in accordance with previous animal data, indicating a role for these cells in downregulating healing as the wound closes. This study also documents an associated increase in B lymphocytes and healing of human wounds, with an as yet undefined role.

Antibodies, Monoclonal↗

Reconstruction of the natal cleft with a perforator-based flap.

Recurrent chronic conditions of the natal cleft, such as pilonidal sinuses, are difficult problems to treat. The deep natal cleft and the rolling effect of one buttock surface over the other contribute to the high recurrence rate. Wide excision of the affected area is the treatment of choice; to cover the defect many techniques have been described that flatten the natal cleft and shift the suture line away from the midline to try to reduce recurrence. These techniques include Z-plasties, rhomboid flaps, V-Y flaps, gluteus maximus myocutaneous flaps and others. Five patients with recurrent natal-cleft problems were operated on over a 1 year period. After wide excision of the natal cleft, the defect was reconstructed with a parasacral perforator-based flap. Preoperatively, a perforator situated superolaterally to the defect is identified with a Doppler probe; the flap is then designed horizontally around the perforator and, after elevation, is rotated 908 over the defect. The mean hospital stay was 5 days. Follow-up ranged from 3 months to 15 months, with no signs of recurrence. We believe this to be a simple and reliable technique, with the advantages of placing the scars away from the midline and flattening the natal cleft - factors that help to prevent recurrence.

Adult↗

Free flap failure in a patient with a long standing, infected, squamous cell carcinoma.

A 40-year-old patient presented with a long history of a pilonidal sinus, which had been operated on several times during the last 20 years. On clinical examination the patient had a large tumour in the sacral and perineal region, with involvement of the rectal wall. General surgeons first attempted to excise the tumour with wide healthy margins, and close the wound by local flaps. After partial flap necrosis and wound dehiscence, the patient underwent a reconstruction with a free latissimus dorsi myocutaneous flap. During the anastomosis it was noted that the recipient vessel walls were brittle, mainly at the arterial site, so the arterial anastomosis had to be done three times. Despite this the artery thrombosed again 12 hours later. Biopsy specimens were taken from the anastomotic sites and studied under light microscopy. There were signs of acute intramural inflammation, with many polymorphonuclear leukocytes present in microabscesses, and spots of necrosis in the elastic layer at the site of the recipient artery. In conclusion, the long lasting infection was considered to be the main factor that caused the anastomosis to fail, leading to thrombosis, through alteration of the vessel walls. The anomalies in the vessel walls were found at some distance from the clinically diseased area, further than is usually found in acute infection. The use of primary arteriovenous vein graft anastomosis can be made on undamaged vessels, and possibly a less traumatic anastomosis such as the "sleeve" type, should be considered for similar cases.

Abscess↗

Human wound contraction: collagen organization, fibroblasts, and myofibroblasts.

The closure of ungrafted sacrococcygeal pilonidal sinus excisional wounds was studied in 15 patients. Wound punch biopsies were taken on a regular basis, and histologic sections were made. To document changes, computer-assisted morphometric image analysis was employed. Initial average wound depth was 37.8 +/- 4.6 mm, and complete closure (0 wound depth) was reached by 68 days. Wound contraction contributed 88 percent to wound closure, whereas the deposition of scar only contributed 12 percent. Maximum cells density within granulation tissue was reached by day 18. Myofibroblasts, identified by alpha-smooth muscle actin immunostaining, first appeared on day 11. Unlike those observed in laboratory animals, myofibroblasts were a minor cell population of granulation tissue, never exceeding 10 percent of the cells. The pattern of collagen fiber organization was documented by polarized light microscopy of Sirius red-stained sections. Early granulation tissue collagen fibers demonstrated a fine greenish birefringence, whereas more mature granulation tissue collagen fibers were thicker, displaying orange-yellowish birefringence. Myofibroblasts were associated exclusively with thicker collagen fibers, whereas fibroblasts were associated with both fine and thick collagen fibers. It is proposed that human wound contraction involves a volume change whereby normal dermal and adipose tissues are pulled into the defect by forces generated within fibroblasts.

Actins↗

Image-guided surgery for anal fistula in a 0.5T interventional MRI unit.

PURPOSE: To determine whether MR-guided anorectal surgery is feasible, and to develop techniques for MR-guided anal fistula surgery. MATERIALS AND METHODS: Six patients with pilonidal sinus (PNS), and 21 with suspected anal fistulae were operated on in the GE Signa SPIO 0.5T interventional MRI unit. Procedures were performed with magnet-safe Lockhart-Mummery fistula probes. Preprocedural and intra-operative MRI (IOMRI) techniques were used to identify the extent of the fistula tracts and septic foci, and to ensure the adequacy of the surgical procedure. RESULTS: IOMRI demonstrated the PNS lesions and the adequacy of excision. Imaging failed to demonstrate a fistula in two patients, as confirmed by surgical examination. No images were obtained in one patient due to his size (weight in excess of 100 kg). In 18 patients a fistula tract or abscess was demonstrated and IOMRI was used to assess the adequacy of the surgical procedure. In three patients this demonstrated incomplete drainage of septic foci, which was not obvious on inspection of the surgical field. We believe that in these patients IOMRI prevented an incomplete procedure and the potential requirement for a second operation. Further surgery was performed to rectify this situation. The fistula tract was laid open in 13 patients, and a Seton drain was inserted in five. CONCLUSION: MRI-guided surgery for anal fistula is feasible. IOMRI demonstrates the exact anatomy of the tracts and abscesses, and confirms that all have been adequately treated. We believe it may become particularly useful in surgery for recurrent and complex anal fistulae, and may lead to fewer recurrences.

Adult↗