Use of Adipose-Derived Mesenchymal Stem Cells to Increase Viability of Composite Grafts.

OBJECTIVE: Chondrocutaneous composite grafts figure among the reconstruction alternatives for alar rim defects resulting from tumor resection and trauma. The major problem with composite grafts is the limited graft survival area. In the present study, the authors aimed to increase the survival area of composite grafts by utilizing the ability of stem cells to promote neovascularization which is crucial in composite graft viability. METHODS: The study included 36 adult Wistar Albino rats, which were allocated to 6 groups. Groups 1, 2, and 3 were the groups in which the grafts were implanted immediately after the defect was formed, and Groups 4, 5, and 6 were those in which grafts were adapted 4 days after the defect was formed. Composite grafts of 1 × 1 cm containing both the cartilage and the skin were prepared from 1 ear, and after forming punctures and incisions on the cartilage, the grafts were adapted to the 1 × 1 cm defects on the back. The backs of the rats in groups 1 and 4 were injected with adipose-derived stem cell (ADSC), those in groups 2 and 5 with medium solution, while the rats in Groups 3 and 6 did not receive any injection. The procedures were followed by histopathological and scintigraphic evaluations. RESULTS: An evaluation of the statistical results showed that composite graft survival areas of the group treated with stem cells increased significantly, in comparison with control and medium groups. When scintigraphic evaluations were considered, it was seen that the group treated with stem cells had significantly higher radioactive substance retention than the control group. Histopathological examination demonstrated that microscopic survival rates in the stem cell group were higher than those in the control group. Green fluorescent protein (GFP) was used in the experiment to tag adipose tissue-derived stem cells. Immunofluorescence staining studies showed less apoptosis and fewer GFP (+) stem cells in the composite grafts of the stem cell group. However, apoptosis was more severe in the control and medium groups which also had decreased vascularity in the graft. DISCUSSION: As the authors have shown in the present study, ADSCs have favorable effects on the viability of composite grafts. They have increased the survival rate of the grafts to a considerable extent. As a clinical implication of this experimental study, the authors think that in the patient of auricular and nasal defects involving the cartilage and the skin, injection of the ADSC and the adaptation of composite grafts 4 days after the preparation of the receiving bed may increase the composite graft viability rates. Thus, it has been found that if the composite grafts are implanted 4 days after stem cell injection, the injection of adipose tissue-derived mesenchymal stem cells is useful in enhancing the survival of composite grafts.

Use of Adipose-Derived Mesenchymal Stem Cells to Accelerate Neovascularization in Interpolation Flaps.

OBJECTIVE: Interpolation flaps are commonly used in plastic surgery to cover wide and deep defects. The need to, wait for 2 to 3 weeks until the division of the pedicle still, however, poses a serious challenge, not only extending treatment and hospital stay, but also increasing hospital expenses. To solve this problem, we have aimed to use the angiogenic potential of stem cells to selectively accelerate neovascularization with a view to increasing the viability of interpolation flaps and achieving early pedicle removal. MATERIALS AND METHODS: A total of 32 rats were allocated to 2 groups as control (N = 16) and experiment (N = 16). The cranial flaps 6 × 5 cm in size located on the back of the rats were raised. Then, a total suspension containing 3 × 10(6) adipose-derived mesenchymal stem cells (ADSC) tagged with a green fluorescent protein (GFP) was injected diffusely into the distal part of the flap, receiving bed, and wound edges. In the control group, only a medium solution was injected into the same sites. After covering the 3 × 5 cm region in the proximal part of the area where the flap was removed, the distal part of the flap was adapted to the uncovered distal area. The pedicles of 4 rats in each group were divided on postoperative days 5, 8, 11, and 14. The areas were photographed 7 days after the pedicles were released. The photographs were processed using Adobe Acrobat 9 Pro software (San Jose, CA) to measure the flap survival area in millimeters and to compare groups. Seven days after the flap pedicle was divided, the rats were injected with 250 mCi Tc-99 mm (methoxy-isobutyl-isonitrie) from the penile vein, and scintigraphic images were obtained. The images obtained from each group were subjected to a numerical evaluation, which was then used in the comparison between groups. The flaps were then examined by histology to numerically compare the number of newly formed vessels. Neovascularization was also assessed by microangiography. In addition, radiographic images were obtained by mammography and evaluated quantitatively. RESULTS: An evaluation of statistical results revealed a significant increase in the flap survival area of the group on stem cell treatment in comparison to the control group. In scintigraphic examinations, the rate of radioactive substance retention was significantly higher in the stem cell group, relative to the control group. Histopathologic examination showed that the capillary density in the stem cell group was higher than that in the control group. Green fluorescent protein had been used to label ADSC in the experiment and it was found by immunofluorescence staining that endothelial samples of control animals did not have GFP (+) cells, whereas all the animals in the experiment group had GFP (+) cells. The comparison of microangiographic images of the experiment and control groups demonstrated significantly elevated vascularity in the former, relative to the latter. DISCUSSION: It has been established in the current study that ADSC injection worked well in speeding up the neovascularization of interpolated flaps and reducing the time of pedicle division. It seems possible to minimize the morbidity of interpolated skin flaps with mesenchymal stem cell therapy at an appropriate dose and for an appropriate length of time.

Adipose-Derived Stem Cells Improve Survival of Random Pattern Cutaneous Flaps in Radiation Damaged Skin.

BACKGROUND: Tissue ischemia and necrosis following surgery after radiotherapy on the skin and subcutaneous tissue are well known to all reconstructive surgeons. Nevertheless, there has been no report so far on local effects of adipose-derived stem cells (ADSCs) on random flap survival elevated in an irradiated rat dorsum. In this experimental study, we aimed to identify the effect of adipose tissue-derived stem cell injection on random flap survival in irradiated tissues. METHODS: Adipose-derived stem cells were isolated from the groin region of Sprague-Dawley rats and expanded ex vivo for 3 passages. Animals were divided into 2: irradiated and nonirradiated and then again into ADSC injected and noninjected groups altogether 4 groups. After elevation of caudally based dorsal random skin flaps (10  cm long and 3  cm wide), Green fluorescent protein labeled ADSCs were then injected to the base of the pedicle. Radiotherapy was 20  Gy single dose applied during 8 weeks before surgery. At postoperative day 7, flap viability measurement and tissue harvest for histologic and immunocytochemical assessment were performed in all groups. RESULTS: We have observed increased flap viability in ADSCs injected irradiated group compared with control radiation group with small but not statistically significantly increase in vessel count per field. Mean survival rate of the flaps in groups A, B, C, and D were 40.46%, 60.07%, 40.90%, and 56.13%, respectively. There was a statistically significant vessel count difference between group B and group A and also with group D (P < 0.001). CONCLUSIONS: These findings suggest that ADSCs have a potential for enhancing the blood supply of random pattern skin flaps after radiation injury. This mechanism might be both neovascularization and vasodilation along with endothelial repair. Further studies are needed.

Utility of the omentum in sacral reconstruction following total sacrectomy due to recurrent and irradiated giant cell tumour of the spine.

Reconstruction of the lumbosacral region after surgical excision of irradiated and recurrent spinal giant cell tumours remains a challenging problem. In this case report, we describe the use of the pedicled omentum flap in reconstruction of an irradiated and infected wide sacral defect of a 19-year-old male patient. The patient had radiotherapy and subsequent wide surgical resection after recurrence of the tumour. A myocutaneous flap from the gluteal area had failed previously. Local flap options could not be used because of the recent radiotherapy to the gluteal area. Since the patient had a laparotomy for tumour resection and a colostomy, abdominal muscles were not considered reliable for reconstructive procedures. A pedicled omentum flap was chosen as a reconstructive option because of its rich blood supply, large surface area, and angiogenic capacity. This report aims to describe the use of the pedicled omentum flap for reconstruction of the lumbosacral area following surgical resection of a spinal tumour, when gluteal and abdominal flap options for reconstruction are jeopardised.