Objective To compare the effect of the composite skin graft consisting of spl it-thickness skin grafts (STSGs) and porcine acellular dermal matrix (PADM) with STSGs only, and to histologically observe the turnover of the PADM in rats. Methods Twenty female Sprague-Dawley rats, weighing 200-225 g, were included. The size of 4.0 cm × 2.5 cm PADM was implanted into hypoderm of the left side of Sprague-Dawley rats’ back. After 10-14 days, the size of 4.0 cm × 2.5 cm full-thickness skin defects were made on the left to expose the PADM under the skin and the same size of full-thickness skin defects were made on the right of the rats’ back. The excised full-thickness skin was made to STSGs about 0.2 mm by drum dermatome. The defects were grafted with composite skin (STSGs on the PADM, experimental group) and STSGs only (control group). The survival rate, the constraction degree of grafts, and the histological change in grafts area were observed at 2, 4, 8, and 20 weeks after operation. Results At 2 weeks after STSGs (0.2 mm) placed on vascularized PADM, STSGs and PADM adhered together and the composite skin had a good survival. The control group also had a good survival. Histological observations showed that STSGs and PADM grew together, neutrophil ic granulocytes and lymphocytes infiltrated in the PADM and some macrophages around the PADM. Fibrous connective tissues were filled under the STSGs in control group. At 4-8 weeks after transplantation, the composite skin had a good survival and the composite skin was thick, soft, and elastic. STSGs survived almost totally in control group, but the grafts were thin. Histological observations showed that inflammatory reactions of PADM faded gradually in experimental group; scar tissues formed under the STSGs in control group. At 20 weeks after transplantation, composite skin was flat, thick, and elastic in experimental group, but the STSGs were thinner and less elastic in control group. Histological observations showed that histological structures of the PADM were similar to the dermal matrix of rats, and the results showed that the collagen matrix of PADM was gradually replaced by the rats’ collagen matrix. Scar tissues were filled under the STSGs in control group. Wound heal ing rates of experimental group were lower than those of control group at 4 and 8 weeks (P﹤0.05); wound contraction rates of experimental group had lower tendency than those of control group, but showing no significant differences (P gt; 0.05). Conclusion Coverage wound with composite skin which composed of STSGs and PADM could improve wound heal ing qual ity; the composite skin is thicker and better elastic than STSGs only. The collagen matrix of PADM is gradually replaced by rats’ collagen matrix.
Objective Human acellular amniotic membrane (HAAM) contains collagens, glucoproteins, proteinpolysaccharide,integrin, and lamellar, which can supply rich nutrition to cell prol iferation and differentiation. To explore the possibil ity of HAAM with adi pose-derived stem cells (ADSCs) as a good engineered skin substitute for repairing skin defect. Methods Primary ADSCs were obtained from inguinal fat of 30 healthy 4-month-old SD rats, male or female, weighing 250-300 g, and cultured in vitro and purified. The 3rd passage ADSCs were used to detect CD44, CD49d and CD34 by immunocytochemistry staining. After physical and trypsin preparation, the HAAM was observed by HE staining and scanning electron microscope(SEM) respectively. ADSCs were seeded on epithel ial side of HAAM at the density of 2 × 105/cm2, cocultured, and observed by SEM at different time. MTT test was used to detect viabil ity of cells that seeded on HAAM, the group without HAAM was used as control. Thirty SD rats were made models of full-thickness skin wound and randomly divided into three groups (A, B, and C). Wound was repaired with HAAM/ADSCs composites in group A, with HAAM in group B, and with gauze as control in group C. The rats underwent postoperative assessment of wound heal ing rate and histological observation at the 1st, 2nd, and 4th weeks. Results HE staining showed that the 3rd passage ADSCs was spindle-shaped with an ovoid nucleus which located in the middle of cell; the immunocytochemistry staining showed positive result for CD44 and CD49d and negative result for CD34. There were no residues of cells in the HAAM by HE staining. SEM showed that there were different structures at the two sides of HAAM；one side had compact reticular structure and the other side had fibrous structure. After 3 days of co-culture, ADSCs showed good growth on HAAM; the cells were closely packed onto the HAAM, attached firmly and prol iferated to confluence on the stromal surface of HAAM. MTT test showed that the cells on the HAAM grew well and had b prol iferation vital ity. There was no significant difference between ADSCs cultured in the HAAM and control group (P gt; 0.05). One, 2, 4 weeks after graft, there were significant differences in wound heal ing rate between group A and groups B, C (P lt; 0.05), between group B and group C (P lt; 0.05). HE staining showed that wound healed faster in group A than in groups B, C. Cytokeratin 19 (CK19) immunohistochemical statining showed that there were more CK19 positive cells in group A than in groups B, C. Conclusion The graft of HAAM with ADSCs plays an effective role in promoting the repair of full-thickness skin wound
Objective To investigate an operative method of repairing large skin defect of the forearm and the hand. Methods From July 2003 to September 2008, 11 patients with large skin defect of the forearm and the hand were repaired using bilateral groin flaps in complex with abdominal flaps, including 7 males and 4 females aged 17-55 years old (average33.5 years old). Among the 11 cases, 5 were caused by carding machine and 4 by traffic accident, and the interval between injury and operation was 90 minutes to 6 hours (average 3.5 hours); 2 cases suffered from severe cicatricial contracture deformity in the late stage of burn injury, and the interval between injury and operation was 7 months and 19 months, respectively. The size of skin defect ranged from 42 cm × 12 cm to 60 cm × 16 cm. The flaps harvested during operation was 45.0 cm × 10.5 cm - 62.0 cm × 18.0 cm in size. Pedicle division of the combined flaps was performed 4 weeks after operation. The donor site wound was repaired by direct suturing in 7 cases and by free skin grafting in 4 cases. Results All flaps survived. All incisions healed by first intention. The donor site wound all healed by first intention. Skin graft all survived. All patients were followed up for 2 months to 3 years. The flaps were soft in texture, full in contour, and normal in color. Sensory recovery of the flaps was evaluated according to the Criteria of UK Medical Research Council (1954), 4 cases were in grade S1, 6 in grade S2, and 1 in grade S3. Hand function was assessed by the Criteria of Chinese Hand Surgery Society, 7 cases were graded as excellent, 2 as good, 2 as poor, and the excellent and good rate was 81.8%. Conclusion Combined use of bilateral groin flaps and abdominal flap is an effective approach to repair large skin defect of the forearm and the hand due to its simple operative procedure and satisfying effect.
To introduce the experience and comprehend of appl ication of many kinds of skin flaps in repair of heel skin and soft tissue defect. Methods From August 1993 to April 2007, 242 cases of skin and soft tissue defect on heel were treated. There were 157 males and 85 females aged 27-76 years. There were 35 cases of unstable scar, 46 cases ofchronic ulcer, 57 cases of squamous cell carcinoma, 72 cases of mal ignant melanoma and 32 cases of injury. The defect area ranged from 2 cm × 2 cm to 8 cm × 14 cm, wound was with low-grade infection in 51 cases. The course of disease was 1 hour to 5 years. The lateral calcaneal flaps (34 cases), the dorsum pedis flaps (15 cases), the medial plantar artery island flaps (108 cases), the sural neurovascular flaps (36 cases), the saphenous neurocutaneous vascular flaps (26 cases) and free (myocutaneous) skin flaps (23 cases) were used to repair heel wounded surface according to specific location of heel wounded surface, range of skin and soft tissue defect. The size of the (myocutaneous) skin flap was 3.0 cm × 2.5 cm to 15.0 cm × 9.0 cm. The donor area was directly sutured or covered with skin grafts. Results In 242 cases, 235 cases achieved heal ing by first intention, 5 cases had partial necrosis of flaps, 2 cases had mild infection. The donor area healed by first intention. A total of 217 patients were followed up for 1 month to 60 months. The color of flaps were normal and the texture of flaps were softer than that of normal heel tissue; the flaps were wearable, the shape of flaps were satisfactory. The patients can walk after 6 months of operation, andthe gait was normal. In 118 cancer patients, no local tumor recurrence occurred, and distant metastatic lesions were observe in 22 mal ignant melanoma patients. In 32 cases of mal ignant melanoma followed up 60 months, no distant metastatic lesions were found. Conclusion In base of following the primary disease treatment, heel function reconstruction and contour structural feature remodel ing, we adopted some kind of island or free (myocutaneous) skin flap can be used to repair heel wound. The ideal effect in heel function reconstruction and contour structural feature remodel ing were obtained.
Objective The amniotic carrier complex membrane, which contains bFGF and vitamin C (VitC) and is loaded with BMSCs, is planted into the deeply-partial wounds of rabbits. To explore its influence on the epidermis renascence and regenerating speed in the process of the dermis restore. Methods BMSCs were isolated from the marrows of 24 healthy3-month-old New Zealand rabbits, male or female, weighing 1.0-1.5 kg. The BMSCs were cultured in vitro and purified, and then amniotic carrier complex membrane was prepared, whose size was 4.52 cm2. Three deep-partial wounds, with the area of about 3.14 cm2, were produced on the back of each rabbit. All the wounds were randomly divided into 3 groups: group A, group B and group C. Group A was the experimental group in which the amniotic carrier complex membrane was planted, including 1 ml BMSCs, 10 mL bFGF (0.2 mg/L) and 10 mL VitC (0.02 g/L). In group B, the amniotic carrier complex membrane was planted, including only 1 mL BMSCs. In group C, the amniotic carrier complex membrane alone was planted. After the operation, general observation was conducted. At postoperative 7, 14 and 21 days, respectively, the observation by HE, Masson, Van Giesonr staining and immunohistochemical staining of collagen type I was performed. The ink perfusion method was performed to evaluate the velocity and the qual ity of the wound heal ing after the transplantation. Results All the wounds obtained good heal ing. At 14 days after the operation, the ratio of wound heal ing was 60%, 41% and 23% in groups A, B and C, respectively. At 21 days after the operation, the the ratio of wound heal ing was 99%, 90% and 81% in groups A, B and C, respectively. There were significant differences between any two groups (P lt; 0.05). The depth of the newborn dermis, the number of the active collagen type I mascul ine cells and the number of the blood vessels in group A were better and more than in group B. And those in group B were better and more than in group C. At the exterior area of the newborn dermis, there was lots of regenerated epidermis from the peripheral normal skin, which in group A was better than in group B, and in group B was better than in group C. onclusion The amniotic carrier complex membrane transplanted to deep-partial wounds, which is appended withBMSCs, bFGF and VitC, can accelerate repair and reconstruction of the dermis. There has an optimal time of the renascence and regeneration of the epidermis in the process of dermis repair.
Objective To investigate the feasibility of using the porcine small intestinal submucosa (SIS) as a kind of the new tissue engineered materials to repair the rat full skin defect. Methods Twenty-eight 6-week-old SD rats weighing 300-350 g were selected in this experimental study. Two 2-cm-diameter round full skin defects were made on the rat back. The upper round defect was used as the blank group, which had no coverings, and the lower round defect was used as the SIS group. SIS that had been produced earlier was transplanted in the defected area. At 3 days, 1, 2, 3, 4, 6 and 8 weeks after the transplantation, the observation was made on the repaired skin conditions, the HE stain, and the repaired skin proportion. Results There was no infection in the two groups. The repairing speed in the SIS group was faster than that in the blank group at 2, 3, 4 and 6 weeks after the transplantation. The skin repaired by SIS was soft and elastic in texture, which had the same high level as the normal skin. The scar tissues in the SIS group were thinner than those in the blank group. The repaired skin proportions at 1, 2, 3, 4, 6 and 8 weeks after the transplantation were 15.72%±3.64%, 43.81%±4.87%, 65.35%±5.63%, 87.95%±4.78%，96.90%±6.89% and 100%, respectively in the SIS group, and 13.42%±5.63%，58.74%±4.48%，76.50%±5.23%，92.30%±5.75% and 100%, respectively in the blank group. Therewas a statistically significant difference between the two groups at 1, 2, 3 and 4 weeks after the transplantation(P＜0.05). Under the microscope, the SIS-repaired skin was observed to have more keratinocytes and collagen tissues, whichwas familiar to the normal skin.Conclusion Porcine SIS can be used as a new kind of the tissue engineered materials to repair the full skin defect.
Objective To observe the clinical outcome of primary repair of the tissue defects of the Achilles tendon and skin by thigh anterolateral free flap and free iliotibial tract. Methods From January 2000 to January 2005, the thigh anterolateral free flap and the iliotibial tract were used to primarily repair the defects of the Achilles tendon and skin in 11 patients (7 males and 4 females, aged 6-45 years). The defects of the skin and Achilles tendon were found in 6 patients, and the defects of the Achilles tendonand skin accompanied by the fracture of the calcaneus were found in 5 patients.The defect of the Achilles skin was 6 cm×5cm-14 cm×8 cm in area. The defect of the Achilles tendon was 511 cm in lenth. The skin flap was 11 cm×6 cm-17 cm×11 cm in area.The iliotibial tract was 7-13 cm in length and 3-5 cm in width. The medial and lateral borders were sutured to from double layers for Achilles tendon reconstruction. The woundon the donor site could be sutured directly in 5 patients, and the others could be repaired with skin grafting. Results After operation, all the flaps survived and the wound healed by first intention. The followup of the 11 patients for 6 mouths-4 years (average, 30 months) revealed that according to Yin Qingshui’s scale, the result was excellent in 6 patients, goodin 4, and fair in 1. The excellent and good rate was 99%. The results showed a significant improvement in the “heel test” and the Thompson sign, and both were negative. No complications of ulceration on the heel and re-rupture of the Achiles tendon occurred. Conclusion The primary repair of the tissue defects of the Achilles tendon and skin by free grafting of the anterolateral femoral skin flap and the iliotibial tract is an effective surgical method.
Objective To report the clinical result of the improvedisland skin flap with distallybased sural nerve nutrient vessels in repairing skin defect in the heel, ankle or foot. Methods From August2004 to April 2005, 15 patients with skin defect in the heel, ankle or foot at distal part were treated by the improved island skin flap with distally-based of sural nerve nutrient vessels. Of 15 flaps, 12 were simplex flaps and 3 were complex flaps. These flap area ranged from 7 cm×6 cm to 11×8 cm. The donor sites were sutured directly and covered with free flap. Results All flaps survived without flap swelling and disturbance of blood circulation. The wounds of donor and recipient sites healed by first intention. The followup period ranged from 3 to 6 months. The texture of flap was soft and the color of flap was similar to that of normal skin. The foot function was excellent. Conclusion The improved island skin flap with distally-based sural nerve nutrient vessels is an ideal skin flap for repairing skin defect in the heel, ankle or foot distal part in clinical. The operation is simple and need not to anastomose blood vessel.
Objective To introduce the application of the pedicled anterolateral thigh flap transferring for coverage of the oversized skin defect of the hand. Methods The pedicled anterolateral thigh flap was transferred to cover the large skin defects of the hands or the skin defects of theabdomen after the abdominal flap transferred to the hand in 5 male patients aged 16-44 years from April 2002 to August 2005. The injured sites were as follows:4 right hands and 1 left hand, including 2 hands injured by a machine and 3 hands injured by burning.The mechanically injured patients underwent an operation within 6 hours after the injury. The burned patients were reconstructed by the flap transferring 4-7 days after the burn when the decayed tissues could be clearly indentified.The areas of the hand defects were 12.19 cm×18.22 cm.The areas of the pedicled anterolateral thigh flaps were 7.12 cm×16.24 cm. The areas of the abdominal flaps were 13.20 cm×19.23 cm.The pedicles were separated 3 weeks after the repairing operation. Results All the flaps survived well and there was no vascular crisis, with the wound healing of the first intention. The skin defects of the hand were covered completely. Five patients were followed up for 6-12 months. The texture of the flaps was soft and the flaps had a good blood circulation. Of the patients, 3 underwent the finger exclusion and degreasing operation 47 months after operation. All the flaps of the hands had protective sensation, which could meet the requirement of the daily life. Conclusion The pedicled anterolateral thigh flap can provide the large coverage for the skin defects of the hands. The risk of the operation can be greatly decreased by obviation of the vessel anastomosis. It can be an optimal choice for themanagement of the oversized skin defects of the hands.
Objective To study the allograft effect of two kinds of tissue engineered oral mucosa lamina proprias on skin fullthickness wounds. Methods The cultured Wistar rat oral mucosa fibroblasts (OMF) were incorporated into collag en or chitosancollagen to construct the tissue engineered oral mucosa laminaproprias, and then the OMFs were labeled with BrdU. The fullthickness round skin defects were made with a round knife (diameter, 0.8 cm) on the backs of 36 Wistar rats (2125 weeks old), which were divided into 2 experimental groups: the fibroblastpopulated collagen lattices (FPCL) group (grafted by FPCLs) and the fibroblastpopulated chitosan collagen lattices (FPCCL) group (grafted by FPCCLs), and the control group (only covered with gauges). All the wounds were observed by the naked eyes or the light microscope, and were measured 4, 7, 14, and 21 days postoperatively. Results There were no infection during the wound healing period. At 7 days after the grafting, the wounds in the 3 groups were covered by scab and/or gauze; at 14 days, the gauze and scab on the wounds in the three groups were all replaced by the new epidermis naturally except one scab each in the FPCCL group and the control groups,which was replaced at 17 days.All the centers of the new epidermis were measurable as the pink red points. At 21 days, all the new skins were smooth without hairs, and their color was similar to the normal one. At 4, 7, and 14 days,there was an indication that the wound diameters became significantly smaller in the three groups; but after the 14th day, there was no significant indication of this kind. At 7 days, the wound diameter in the FPCL group was significantly smaller than that in the FPCCL group and the control group (Plt;0.01). Under the lightmicroscope, at 4 days postoperatively, the decayed tissue on the surfaces of the recipient wounds in the FPCL group and the FPCCL group was separated from the lower granular tissue in which there were many inflammatory cells, fibroblasts, and new vessels. There was a similar-phenomenon in the control group. Each skin wound in the three groups was only partly keratinocyted at 7 days postoperativel y. The recipient wounds were wholly keratinocyted with when rete ridges observed at 14 and 21 days, but in the control group the wounds were keratinocyted with no rete ridges. Fibers in the new dermis were thin. The OMFs with Brdu appeared in the granular tissue and new dermis at 4, 7, 14, and 21 days postoperatively, which could be illustr ated by the immunohistochemical staining. The positive OMFs and the granular tissue joined in the repair of the skin defe cts without any allergic reaction during the period of the wound healing. Conclusion The oral mucosa fibroblasts as the new seed cells can join i n the repair of the skin defects effectively and feasibly. The fibroblastpopul ated collagen lattices and the fibroblastpopulated chitosan collagen lat tices can repair skin defects effectively and feasibly, too. And the quality of the new skins was better in the two experimental groups than in the control group.