Faculty Bibliography

Subcutaneous fat has emerged as an alternative tissue source for stromal/stem cells in regenerative medicine. Over the past decade, international research efforts have established a wealth of basic science and preclinical evidence regarding the differentiation potential and regenerative properties of both freshly processed, heterogeneous stromal vascular fraction cells and culture expanded, relatively homogeneous adipose-derived stromal/stem cells. The stage has been set for clinicians to translate adipose-derived cells from the bench to the bedside; however, this process will involve "development" steps that fall outside of traditional "hypothesis-driven, mechanism-based" paradigm. This concise review examines the next stages of the development process for therapeutic applications of adipose-derived cells and highlights the current state of the art regarding clinical trials. It is recommended that the experiments addressing these issues be reported comprehensively in the peer-review literature. This transparency will accelerate the standardization and reproducibility of adipose-derived cell therapies with respect to their efficacy and safety.

Mesenchymal and hematopoietic tissues are important reservoirs of adult stem cells. The potential of tissue resident mesenchymal stem cells (MSCs) to differentiate into cells of mesodermal and ectodermal lineages has been reported previously. We examined the hypothesis that adherent adipose tissue resident mesenchymal stem cells (ASCs) are capable of generating cells with hematopoietic characteristics. When cultured in differentiation media, clonally isolated ASCs develop into cells with hematopoietic attributes. The hematopoietic differentiated cells (HD) express early hematopoietic (c-kit, PROM1, CD4) as well as monocyte/macrophage markers (CCR5, CD68, MRC1, CD11b, CSF1R). Additionally, HD cells display functional characteristics of monocyte/macrophages such as phagocytosis and enzymatic activity of α-Naphthyl Acetate Esterase. HD cells are also responsive to stimulation by IL-4 and LPS as shown by increased CD14 and HLA-DRB1 expressions and release of IL-2, IL10, and TNF. Taken together, this study characterizes the potential of ASCs to generate functional macrophages in vitro, and therefore paves way for their possible use in cell therapy applications.

Reconstruction following oncologic resection in the head and neck is complex due to large surgical defects left after removal of skin, subcutaneous, and skeletal structures. It is essential to adequately fill the defect as well as provide an acceptable tissue match in terms of tone, texture, thickness and contour. A 55-year-old male presented with an advanced melanoma in the right pre-tragal area. Surgical resection was performed including a total auriculectomy. A tunnelled right supraclavicular artery island (SAI) flap was used to repair the surgical defect. A Doppler probe ensured adequate circulation within the flap, especially in the distal tip. Reconstruction using the SAI flap after oncologic ear resection reduced operating room time, required less technical expertise, and provided excellent tissue match compared to more traditional methods of surgical defect reconstruction including free flaps, local flaps, and pedicled myocutaneous flaps. Successful use of the SAI flap in this case further expands the flaps versatility. We recommend that the reconstructive surgeon consider the SAI flap when presented with challenging infratemporal fossa and lateral skull base cases.

OBJECTIVES/HYPOTHESIS: AlloDerm (LifeCell Corp., Branchburg, NJ) is commonly employed for reconstruction of ablative soft tissue and mucosal defects following surgical resections. Although devoid of growth factors, AlloDerm may serve as an adhesive matrix for binding of growth factors, increasing local angiogenesis, and wound healing. We hypothesized that AlloDerm would enhance angiogenesis and might be altered with autologous blood products to enhance initiation of the angiogenic response. METHODS: We used a human placental vein in a fibrin-thrombin clot-based angiogenesis model. Four groups, human placental vein (HPVM), HPVM with AlloDerm, HPVM with AlloDerm plus platelet-poor plasma, and HPVM with AlloDerm plus platelet-rich plasma were evaluated. Endothelial cell growth was evaluated visually (40x). Hematoxylin and eosin staining and immunofluorescent staining for growth within the AlloDerm matrix were also performed. To assess human umbilical vein endothelial cell (HUVEC) sites of attachment to AlloDerm, we incubated HUVEC cells with AlloDerm for a period of 2 weeks and evaluated attachment with anti-factor VIII immunofluorescence. RESULTS: Angiogenic initiation decreased in the combined placental vein with AlloDerm group (P < .0001 at day 7, 14, 21). Additionally, initiation in the AlloDerm plus platelet-poor plasma group was significantly better than the AlloDerm alone group when placentas 2 and 3 were compared (P < .0001). On hematoxylin and eosin staining and immunofluorescent factor VIII staining, no endothelial growth into the AlloDerm was noted in the samples analyzed. CONCLUSIONS: AlloDerm may be enriched with platelet-poor plasma to stimulate greater initiation and wound healing; however, AlloDerm inhibits angiogenic initiation in this model.

BACKGROUND: The supraclavicular artery island flap is a useful regional option in head and neck reconstruction. Previous studies have recorded pedicle length, caliber, and ink injection studies of the supraclavicular artery. This study presents a three- and four-dimensional appraisal of the vascular anatomy and perfusion of the supraclavicular artery island flap using a novel computed tomographic technique. METHODS: Ten supraclavicular artery island flaps were harvested from fresh cadavers. Each flap was injected with contrast media and subjected to dynamic computed tomographic scanning using a GE Lightspeed 16-slice scanner. Static computed tomographic scanning was also undertaken using a barium-gelatin mixture. Images were viewed using both General Electric and TeraRecon systems, allowing the appreciation of vascular territory (three-dimensional) and analysis of perfusion flow (four-dimensional). RESULTS: The entire skin paddle was perfused in the majority (nine of 10) of flaps. One of the flaps was perfused only 50 percent. In this case, the pedicle artery was found to be much smaller than the other flap pedicles. Direct linking vessels and recurrent flow by means of the subdermal plexus were found to convey the flow of contrast between adjacent perforators. This explains how perfusion extends to adjacent perforators by means of interperforator flow, and how perfusion is maintained all the way to the distal periphery of the flap. CONCLUSIONS: Using this imaging technique, the authors elucidated the vascular anatomy of the supraclavicular artery island flap. This study confirms previous clinical findings that the supraclavicular artery island flap is a reliable option and gives surgeons new information for future flap refinement.

Ear mesenchymal stem cells (EMSCs) represent a readily accessible population of stem-like cells that are adherent, clonogenic, and have the ability to self-renew. Previously, we have demonstrated that they can be induced to differentiate into adipocyte, osteocyte, chondrocyte, and myocyte lineages. The purpose of the current study was to characterize the growth kinetics of the cells and to determine their ability to form colonies of fibroblasts, adipocytes, osteocytes, and chondrocytes. In addition, the immunophenotypes of freshly isolated and culture-expanded cells were evaluated. From 1 g of tissue, we were able to isolate an average of 7.8 x 10(6) cells exhibiting a cell cycle length of approximately 2-3 days. Colony-forming unit (CFU) assays indicated high proliferation potential, and confirmed previously observed multipotentiality of the cells. Fluorescence-activated cell sorting (FACS) showed that EMSCs were negative for hematopoietic markers (CD4, CD45), proving that they did not derive from circulating hematopoietic cells. The FACS analyses also showed high expression of stem cell antigen-1 (Sca-1) with only a minor population of cells expressing CD117, thus identifying Sca-1 as the more robust stem cell biomarker. Additionally, flow cytometry data revealed that the expression patterns of hematopoietic, stromal, and stem cell markers were maintained in the passaged EMSCs, consistent with the persistence of an undifferentiated state. This study indicates that EMSCs provide an alternative model for in vitro analyses of adult mesenchymal stem cells (MSCs). Further studies will be necessary to determine their utility for tissue engineering and regenerative medical applications.

BACKGROUND: The supraclavicular island flap has been used successfully for difficult facial reconstruction cases, providing acceptable results without using microsurgical techniques. The authors use this regional flap in reconstructing various head and neck oncologic defects that normally require traditional regional or free flaps to repair surgical wounds. METHODS: A pedicled supraclavicular artery flap was used to reconstruct head/neck oncologic defects. Complications and functional outcomes were assessed. RESULTS: Head and neck oncologic patients underwent tumor resection followed by immediate reconstruction using a supraclavicular artery island flap. Ablative defects included neck, tracheal-stomal, mandible, parotid, and pharyngeal walls. All flaps (n = 18) were harvested in less than 1 hour. All ablative wounds and donor sites were closed primarily and did not require additional surgery. Major complications included a complete flap loss when the vascular pedicle was inadvertently divided and pharyngeal leaks. The leaks resolved without surgical intervention, and both patients regained the ability to swallow using their neo-esophagus. Minor complications included donor-site wound dehiscence and cellulitis. None of the patients reported functional donor-site morbidity. CONCLUSIONS: This thin flap is easy and quick to harvest, has a reliable pedicle, and has minimal donor-site morbidity. It is now the authors' flap of choice for many common head and neck reconstructive problems. Early experience using the supraclavicular artery island flap suggests that it is an excellent flap option for head and neck oncologic disease patients.