Immune Tolerance of Human Dental Pulp-Derived Mesenchymal Stem Cells Mediated by CD4⁺CD25⁺FoxP3⁺ Regulatory T-Cells and Induced by TGF-β1 and IL-10

PURPOSE: Most studies on immune tolerance of mesenchymal stem cells (MSCs) have been performed using MSCs derived from bone marrow, cord blood, or adipose tissue. MSCs also exist in the craniofacial area, specifically in teeth. The purpose of this study was to evaluate the mechanisms of immune tolerance of dental pulp-derived MSC (DP-MSC) in vitro and in vivo. MATERIALS AND METHODS: We isolated DP-MSCs from human dental pulp and co-cultured them with CD4⁺ T-cells. To evaluate the role of cytokines, we blocked TGF-β and IL-10, separately and together, in co-cultured DP-MSCs and CD4⁺ T-cells. We analyzed CD25 and FoxP3 to identify regulatory T-cells (Tregs) by fluorescence-activated cell sorting (FACS) and real-time PCR. We performed alloskin grafts with and without DP-MSC injection in mice. We performed mixed lymphocyte reactions (MLRs) to check immune tolerance. RESULTS: Co-culture of CD4⁺ T-cells with DP-MSCs increased the number of CD4⁺CD25⁺FoxP3⁺ Tregs (p<0.01). TGF-β or/and IL-10 blocking suppressed Treg induction in co-cultured cells (p<0.05). TGF-β1 mRNA levels were higher in co-cultured DP-MSCs and in co-cultured CD4⁺ T-cells than in the respective monocultured cells. However, IL-10 mRNA levels were not different. There was no difference in alloskin graft survival rate and area between the DP-MSC injection group and the non-injection group. Nonetheless, MLR was reduced in the DP-MSC injected group (p<0.05). CONCLUSION: DP-MSCs can modulate immune tolerance by increasing CD4⁺CD25⁺FoxP3⁺ Tregs. TGF-β1 and IL-10 are factors in the immune-tolerance mechanism. Pure DP-MSC therapy may not be an effective treatment for rejection, although it may module immune tolerance in vivo.

Mega Fat Graft for Breast Augmentation

After numerous experiments with free-fat transfer since 1893, many promising methods and results of large-volume fat graft have been published recently. A common disadvantage was the time of the procedure and a lack of proof of efficiency. This method was evaluated in a prospective clinical study with 100 patients, the overall number of transplantations amounting to 342 treated breasts. Indications were general lack of breast volume, either genuine or acquired in the course of surgical procedures. The fat was harvested with the water jet assisted method, which consists in general of the harvest of very small fat particles by means of water-assisted liposuction (body-jet, human med AG, Germany) and reinjection of the fat after separation from superfluous water by means of the Lipo-Collector. All procedures were performed in a standardized pattern, measurements were taken preoperatively, at day 1 postop, after 1 week, 4 weeks, 3months, 6months. An 3D imaging of the breasts was taken preoperatively and 6 months postoperatively, the longest follow-up is 24 months. Operation time was 2 hours on average. In every case a definite increase of the volume was observed. There was no macroscopic oily cyst. The volume control of 75 aesthetic patients by 3D image could verify a permanent take rate of 62+/-11% of the grafted fat. In aesthetic patients generally 2 (80%) fat-grafted procedures with an average gain in volume of 1/2 bra cup size or 100~150 ml) per procedure were required. After implant removal, satisfaction was usually reached after only a single procedure.

A Modified MACS Lift Combined with Composite Flap Elevation

Composite face lift has been accepted as the best technique for getting satisfactory outcome in surgical rejuvenation. As operations that ensure fast recovery and minimal complication were favored, MACS lift has been regarded as one of the innovative solutions. We experienced that nasolabial fold and neck wrinkle was hardly improved with conventional MACS lift. Therefore, we devised a new modification of composite face lift and MACS lift to achieve both outstanding improvement and fast recovery. From May, 2004 to April, 2008, we operated 49 cases with our technique. They were between 35 and 67 years old. Follow up ranged from 3 to 26 months. Fifteen of 49 patients had longer than 1 year follow up were evaluated with photos and patients interview. Eleven patients were satisfied with the results 12 months after operation. However, two patients complained of unimproved marionette line and remaining two patients had irregularity on the skin. We experienced one hematoma immediately after operation, however most of the patients went to their usual daily lives in one or two weeks after operation. Our operation had several outstanding advantages and long lasting outcome. We, therefore, concluded that this technique is an effective surgery to improve aged face.

Clinical Application of Maggots in Chronic Difficult Wounds

PURPOSE: Since ancient times, larval therapy has been applied to help wound healing. Its use has recently been rediscovered, and the interest in this therapy has been increased in clinical practice, as well as in research. The objective of this study is to assess the efficacy of maggot therapy for the treatment of intractable, chronic wounds. METHODS: Twenty patients, suffering mostly from chronic wounds were treated using maggots of greenfly (Phaenicia sericata). Sterile maggots were administered to the wound twice a week. The causes of the development of wounds were diabetic foot(14), trauma(3), plate exposure(1), and unknown origin(2). RESULTS: Complete debridement was achieved in 10 wounds; in 8 wounds, the debridement was partially achieved; 2 patients escaped. Four patients were healed without any additional surgical procedures but fourteen patients were treated with skin graft. Pain, reported by six patients, was relatively well controlled with oral analgesics. CONCLUSION: We found larval therapy to be effective for chronic wounds. Maggot therapy is a relatively rapid and effective treatment, particularly in necrotic wounds or the wounds resistant to conventional treatment.

Treatments of Phyllodes Tumor in Breast and Immediate Breast Reconstruction

PURPOSE: There have been limited reports on breast reconstruction after excision of phyllodes tumor. This paper reports four patients who had immediate reconstruction of the breast following excision of phyllodes tumor. METHODS: We retrospectively reviewed the medical records of 14 patients from March 2000 to March 2008. Clinical data were analyzed including age, presenting symptoms and signs, type of surgery and metastasis. RESULTS: The mean age was 38.6 years. The mean follow-up period was 40.6 months. Reconstruction was performed with latissimus dorsi musculocutaneous flap in 3 patients and transverse rectus abdominis musculocutaneous flap in 1 patient. Other cases were covered with skin graft or primary repair. 2 local recurrent cases were noted. CONCLUSION: The breast affected by phyllodes tumor must undergo complete excision. Followed by mastectomy, immediate reconstruction of breast improved cosmetic results, and allowed a wider surgical excision margin of tumor.