A Modified LeClerc Procedure with Oblique Osteotomy and V-shaped Notch for Habitual Dislocation of the Temporomandibular Joint.

INTRODUCTION: In patients with habitual dislocation of the temporomandibular joint, anterior dislocation readily occurs with mouth opening in the normal range. METHODS: This short paper describes our novel procedure, modified zygomatic arch osteotomy reported by LeClerc et al., with oblique osteotomy to create a V-shaped notch at the tip for more accurate locking. CONCLUSION: The technique obtained favorable results without the recurrence of TMJ dislocation.

Utilization of Platelet-Rich Plasma for a Fistula With Subcutaneous Cavity Following Septic Bursitis: A Case Report.

In platelet-rich plasma (PRP) therapy, various growth factors and cytokines released the α-granules contained in platelets after activation can potentially enhance wound healing by delivering. We report a patient in whom treatment with PRP, prepared using a syringe-centrifugation-system PRP kit (KYOCERA Medical PRP Kit), for a fistula following bursitis of the lateral malleolus, which could not be healed with conventional wound therapy, led to successful healing. A 58-year-old man was on dialysis for type II diabetes and chronic renal failure. In the left lateral malleolus, septic bursitis developed, leading to a refractory fistula with a subcutaneous cavity measuring 4 × 3 cm, which persisted for more than 2 months. Platelet-rich plasma was prepared using the KYOCERA Medical PRP Kit (KYOCERA Medical Corporation, Osaka, Japan) and infused into the cavity twice to close it. After this procedure, the cavity size reduced, but the orifice and subcutaneous cavity were not closed. Therefore, additional PRP therapy was conducted after 10 weeks of the first PRP session. Complete closure was achieved 13 weeks after the first PRP therapy. In the present case, PRP was prepared using the KYOCERA Medical PRP Kit, and wound healing of a fistula with subcutaneous cavity following bursitis of the lateral malleolus was successfully cured. The KYOCERA Medical PRP Kit was useful, because PRP could be prepared simply and inexpensively using the syringe-centrifugation system.

Platelet and growth factor concentrations in activated platelet-rich plasma: a comparison of seven commercial separation systems.

Platelet-rich plasma (PRP) is blood plasma that has been enriched with platelets. It holds promise for clinical use in areas such as wound healing and regenerative medicine, including bone regeneration. This study characterized the composition of PRP produced by seven commercially available separation systems (JP200, GLO PRP, Magellan Autologous Platelet Separator System, KYOCERA Medical PRP Kit, SELPHYL, MyCells, and Dr. Shin's System THROMBO KIT) to evaluate the platelet, white blood cell, red blood cell, and growth factor concentrations, as well as platelet-derived growth factor-AB (PDGF-AB), transforming growth factor beta-1 (TGF-ß1), and vascular endothelial growth factor (VEGF) concentrations. PRP prepared using the Magellan Autologous Platelet Separator System and the KYOCERA Medical PRP Kit contained the highest platelet concentrations. The mean PDGF-AB concentration of activated PRP was the highest from JP200, followed by the KYOCERA Medical PRP Kit, Magellan Autologous Platelet Separator System, MyCells, and GLO PRP. TGF-ß1 and VEGF concentrations varied greatly among individual samples, and there was almost no significant difference among the different systems, unlike for PDGF. The SELPHYL system produced PRP with low concentrations of both platelets and growth factors. Commercial PRP separation systems vary widely, and familiarity with their individual advantages is important to extend their clinical application to a wide variety of conditions.

Platelet-rich plasma releasate promotes angiogenesis in vitro and in vivo.

Platelet-rich plasma (PRP) is a plasma fraction in which several growth factors are concentrated at high levels. In recent years, the biological effects on various cells of the active soluble releasate that is isolated following platelet activation of PRP [PRP-releasate (PRPr)] have been reported. The purpose of this study was to determine the angiogenic effects of human PRPr in vitro and in vivo. PRPr was prepared from human whole blood using the double spin method and was activated with CaCl2 and autologous thrombin. PRPr stimulated proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs) and in vivo angiogenesis-inducing ability in nude mice. PRPr led to the phosphorylation of Erk1/2 and Akt in HUVECs, and the induction of proliferation and migration by PRPr was suppressed by PRPr inhibitors PD98059 and LY294002. PRPr induces angiogenesis in vitro and in vivo, and the present findings suggest that the mechanism for this is activation of the ERK and phosphatidylinositol-3-kinase-Akt pathways. Our results demonstrate that PRPr is a promising autologous source for therapeutic angiogenesis in treating cardiovascular disease.

Effects of glycolic acid chemical peeling on facial pigment deposition: evaluation using novel computer analysis of digital-camera-captured images.

BACKGROUND: Chemical peeling is becoming increasingly popular for skin rejuvenation in dermatological cosmetic medicine. However, the improvements seen with chemical peeling are often very minor, and it is difficult to conduct a quantitative assessment of pre- and post-treatment appearance. AIMS: We report the pre- and postpeeling effects for facial pigment deposition using a novel computer analysis method for digital-camera-captured images. PATIENTS/METHODS: Glycolic acid chemical peeling was performed a total of 5 times at 2-week intervals in 23 healthy women. We conducted a computer image analysis by utilizing Robo Skin Analyzer CS 50 and Clinical Suite 2.1 and then reviewed each parameter for the area of facial pigment deposition pre- and post-treatment. Parameters were pigmentation size and four pigmentation categories: little pigmentation and three levels of marked pigmentation (Lv1, 2, and 3) based on detection threshold. Each parameter was measured, and the total area of facial pigmentation was calculated. RESULTS: The total area of little pigmentation and marked pigmentation (Lv1) was significantly reduced. On the other hand, a significant difference was not observed for the total area of marked pigmentation Lv2 and Lv3. CONCLUSION: This suggests that glycolic acid chemical peeling has an effect on small facial pigment disposition or has an effect on light pigment deposition. As the Robo Skin Analyzer is useful for objectively quantifying and analyzing minor changes in facial skin, it is considered to be an effective tool for accumulating treatment evidence in the cosmetic and esthetic skin field.

Adipose-derived regenerative cell (ADRC)-enriched fat grafting: optimal cell concentration and effects on grafted fat characteristics.

BACKGROUND: To overcome the absorption of traditional fat grafting, techniques for adipose-derived regenerative cell (ADRC)-enriched fat grafting are currently being adapted for practical application. The Celution800/CRS (Cytori Therapeutics, San Diego, CA) has enabled rapid grafting of the patient's own freshly harvested ADRCs without requiring a culturing step. However, the optimal cell concentration and the effects of ADRCs on the characteristics of grafted fat after free fat grafting remain unclear. METHODS: ADRCs were isolated and purified from human fat tissue using the Celution800/CRS. Animals that received fat grafting without the addition of ADRCs were designated the control group (group A). The number of ADRCs per grafted fat volume (mL) was adjusted to 3 × 105, 1.5 × 106, and 3 × 106 cells/mL (groups B, C, and D, respectively), mixed with free fat, and transplanted as ADRC-enriched fat grafting. These mixtures were transplanted subcutaneously into BALB/C Jcl-nu/nu mice. The volume of grafted fat was determined 5 months after transplantation, and histological assessments were performed. RESULTS: ADRC-enriched fat grafting resulted in decreased fat absorption and the formation of greater numbers of new blood vessels in the grafted fat. The optimal ADRC concentration in this study was found to be 3 × 105 cells/mL (group B), with higher concentrations resulting in increased cyst and fibril formation in the grafted fat. CONCLUSIONS: This study used the Celution800/CRS for free fat grafting and demonstrated that the concentration of transplanted ADRCs affected the engraftment and quality of the grafted fat.

Effects of platelet-rich plasma on proliferation and myofibroblastic differentiation in human dermal fibroblasts.

Platelet-rich plasma (PRP) is plasma that is produced from autologous blood, and its usefulness in plastic surgery and dermal wound healing has garnered attention in recent years. The aim of this study was to investigate the effects of PRP and platelet-poor plasma on the proliferation and differentiation of skin fibroblasts into myofibroblasts and on wound contraction using Western blotting, immunofluorescence staining, and collagen gels containing an embedded fibroblast model. PRP promotes proliferation of human dermal fibroblasts. PRP addition enhanced the expression of alpha-smooth muscle actin protein, a myofibroblast marker, as shown by immunofluorescence staining and Western blotting. PRP-treated groups demonstrated more marked contraction in the collagen gel model than the platelet-poor plasma and vehicle groups. PRP promotes proliferation, causes the differentiation of human dermal fibroblasts into myofibroblasts and promotes wound contraction, thus providing a potential therapeutic agent for skin wound healing.

Effects of transforming growth factor-beta1 on cell motility, collagen gel contraction, myofibroblastic differentiation, and extracellular matrix expression of human adipose-derived stem cell.

Human adipose-derived stem cells (ASCs) are adult pluripotent stem cells, and their usefulness in plastic surgery has garnered attention in recent years. Although, there have been expectations that ASCs might function in wound repair and regeneration, no studies to date have examined the role of ASCs in the mechanism that promotes wound-healing. Transforming growth factor-beta1 (TGF-ß1) is a strong candidate cytokine for the triggering of mesenchymal stem cell migration, construction of extracellular matrices, and differentiation of ASCs into myofibroblasts. Cell proliferation, motility, and differentiation, as well as extracellular matrix production, play an important role in wound-healing. We have evaluated the capacity of ASCs to proliferate and their potential to differentiate into phenotypic myofibroblasts, as well as their cell motility and collagen gel contraction ability, when cultured with TGF-ß1. Cell motility was analyzed using a wound-healing assay. ASCs that differentiated into myofibroblasts expressed the gene for alpha-smooth muscle actin, and its protein expression was detected immunohistochemically. The extracellular matrix expression in ASCs was evaluated using real-time RT-PCR. Based on the results, we conclude that human ASCs have the potential for cell motility, extracellular matrix gene expression, gel contraction, and differentiation into myofibroblasts and, therefore, may play an important role in the wound-healing process.

Bone regeneration with BMP-2 and hydroxyapatite in critical-size calvarial defects in rats.

BACKGROUND: This study describes the effect of bone formation by BMP-2 (bone morphogenetic protein-2), a bone formation inducer, with or without hydroxyapatite (HAP) application to critical-size defects in rat calvarial bone. MATERIAL AND METHODS: Twenty male Wistar rats were divided into four groups of 5 animals each: control, HAP, BMP, and mixed BMP/HAP. A Critical-size defect of 4mm was made using a trephine in the calvarial bone and, after that, BMP and/or HAP was applied to the defect according to the grouping. Defects were evaluated radiographically and histologically using ImageJ color analyzer software at 4 weeks postoperatively. RESULTS: The histological data were more precise than the radiologic data due to the white color of the porous-type HAP material. The highest radiopacity was noted in the mixed BMP/HAP group (162.07±9.06), followed by the HAP group (133.15±21.8), then the BMP group (100.79±8.27), and, lastly, the control group (54.45±8.39). After subtracting the white background and using ImageJ for histological analysis, the highest rate of osteochondrogenesis was in the mixed BMP/HAP group (85.29%±8.21), and then the BMP group (77.34%±7.39), followed by the HAP group (59.82%±11.23), and, lastly, the control group (40.27%±7.44). Differences in the values between groups were then analyzed using confidence intervals (CI) of 95 and 99%. CONCLUSION: Within 4 weeks, the mixed BMP/HAP group showed the highest level of bone induction, especially compared to the BMP group, but this was non-significant; even with a 95% CI, the result was negative. This reveals that BMP alone can be applied, with a final result the same as that seen in the mixed BMP/HAP group. BMP and HAP, both being osteoinducting agents, even though they differ from a material classification point of view, have a positive effect on osteogenesis.

Platelet-rich plasma promotes epithelialization and angiogenesis in a splitthickness skin graft donor site.

Autologous platelet-rich plasma contains multiple growth factors. We performed a side-by-side (half-side) test between the platelet-rich plasma (PRP)-treated and control (untreated) sides of a split-thickness skin graft donor site, and compared the number of days until epithelialization and pain during gauze change. On day 13 after surgery, we performed punch biopsy on the two sides and for adjacent normal skin tissue and compared the epidermal thickness and numbers of collagen fibers and newly formed vessels in the dermis by H&E staining, elastica van Gieson staining, and α-smooth muscle actin (α-SMA) immunostaining. Epithelialization progressed more rapidly, pain during gauze change was milder, and the epidermal thickness and number of newly formed vessels in the dermis were significantly greater on the PRP-treated side. This study revealed that PRP promotes epithelialization and angiogenesis of split-thickness skin graft donor sites.

Effect of C3 transferase on human adipose-derived stem cells.

Human adipose-derived stem cells (ASCs) are adult pluripotent stem cells, which have the ability to differentiate into fat, cartilage, bone, or nerves that can be applied in tissue engineering. On the other hand, the exoenzyme C3 transferase (C3) is a Rho inhibitor. Once in the cytosol, the cell-penetrating moiety is released, thereby allowing C3 transferase to freely diffuse intracellularly and inactivate RhoA, RhoB, and RhoC, but not related GTPases such as Cdc42 or Rac1. In this study, we investigated ASC cytoskeletal changes induced by the addition of C3 employing immunofluorescence staining, changes in alpha-smooth muscle actin (a-SMA) gene expression employing real-time RT-PCR, and the Rho-inhibitory effect employing the pull-down assay. C3 significantly reduced stress fiber disruption and a-SMA expression 24 h after its addition at a concentration of 1 µg/ml, and it also reduced the Rho activity level. While the correlation of the occurrence can be assumed, it requires further examination to verify it. C3 may be an effective inhibitor of intracellular signal transmission in ASC cytoskeletal control involving Rho.

A novel method to measure conspicuous facial pores using computer analysis of digital-camera-captured images: the effect of glycolic acid chemical peeling.

BACKGROUND/PURPOSE: Chemical peeling is becoming increasingly popular for skin rejuvenation in dermatological esthetic surgery. Conspicuous facial pores are one of the most frequently encountered skin problems in women of all ages. This study was performed to analyze the effectiveness of reducing conspicuous facial pores using glycolic acid chemical peeling (GACP) based on a novel computer analysis of digital-camera-captured images. METHODS: GACP was performed a total of five times at 2-week intervals in 22 healthy women. Computerized image analysis of conspicuous, open, and darkened facial pores was performed using the Robo Skin Analyzer CS 50. RESULTS: The number of conspicuous facial pores decreased significantly in 19 (86%) of the 22 subjects, with a mean improvement rate of 34.6%. The number of open pores decreased significantly in 16 (72%) of the subjects, with a mean improvement rate of 11.0%. The number of darkened pores decreased significantly in 18 (81%) of the subjects, with a mean improvement rate of 34.3%. CONCLUSION: GACP significantly reduces the number of conspicuous facial pores. The Robo Skin Analyzer CS 50 is useful for the quantification and analysis of 'pore enlargement', a subtle finding in dermatological esthetic surgery.

Clinical outcome of surgical treatment for periorbital basal cell carcinoma.

Basal cell carcinoma (BCC) has a predilection for the periorbital region, which is a special, prominent, cosmetic, functional area to protect the eyeball. For squamous cell carcinoma and melanoma, extensive resection with reconstruction is performed. In contrast, for BCC, resection is often confined to a small to medium-sized area, necessitating higher-quality reconstructive surgery. We analyze the surgical outcomes of treatment for periorbital BCC, and evaluate reconstruction method after resection. Forty-nine patients with periorbital BCC had surgery in our hospital over 20 years. Age, gender of the patients, and size, localization, and histology of the tumor, and surgical procedures, and their early and late complications were analyzed retrospectively. BCC was most frequently occurred in the lower lid (55%), followed by inner canthus (19%), upper lid (17%), and outer canthus (9%). The histologic classifications were solid (80%), morphea (7%), mix (7%), superficial (2%), keratotic (2%), and adenoid (2%). Recurrence of the tumor was observed in 2 advanced cases in patients treated with resection of the tumor including surrounding tissue 5 mm from the margin. A rotation advancement cheek flap procedure was most frequently applied. Horizontal shift of the skin was most effective to prevent postoperative lagophthalmos. BCC occurred most frequently in the lower lid within the periorbital area. Rotation advancement of cheek flap with horizontal shift of the skin is most effective procedure in both appearance and function of the eyelid.

Proliferation-promoting effect of platelet-rich plasma on human adipose-derived stem cells and human dermal fibroblasts.

BACKGROUND: This study evaluated changes in platelet-derived growth factor (PDGF)-AB and transforming growth factor (TGF)-beta1 release from platelets by platelet-rich plasma activation, and the proliferation potential of activated platelet-rich plasma and platelet-poor plasma on human adipose-derived stem cells and human dermal fibroblasts. METHODS: Platelet-rich plasma was prepared using a double-spin method, with the number of platelets counted in each preparation stage. Platelet-rich and platelet-poor plasma were activated with autologous thrombin and calcium chloride, and levels of platelet-released PDGF-AB and TGF-beta1 were determined by enzyme-linked immunosorbent assay. Cells were cultured for 1, 4, or 7 days in serum-free Dulbecco's Modified Eagle Medium supplemented with 5% whole blood plasma, nonactivated platelet-rich plasma, nonactivated platelet-poor plasma, activated platelet-rich plasma, or activated platelet-poor plasma. In parallel, these cells were cultured for 1, 4, or 7 days in serum-free Dulbecco's Modified Eagle Medium supplemented with 1%, 5%, 10%, or 20% activated platelet-rich plasma. The cultured human adipose-derived stem cells and human dermal fibroblasts were assayed for proliferation. RESULTS: Platelet-rich plasma contained approximately 7.9 times as many platelets as whole blood, and its activation was associated with the release of large amounts of PDGF-AB and TGF-beta1. Adding activated platelet-rich or platelet-poor plasma significantly promoted the proliferation of human adipose-derived stem cells and human dermal fibroblasts. Adding 5% activated platelet-rich plasma to the medium maximally promoted cell proliferation, but activated platelet-rich plasma at 20% did not promote it. CONCLUSIONS: Platelet-rich plasma can enhance the proliferation of human adipose-derived stem cells and human dermal fibroblasts. These results support clinical platelet-rich plasma application for cell-based, soft-tissue engineering and wound healing.

Bone tissue engineering using human adipose-derived stem cells and honeycomb collagen scaffold.

Human adipose-derived stem cells (ASCs) have the capacity to regenerate and the potential to differentiate into multiple lineages of mesenchymal cells. The aim of this study was to investigate the possibility of using honeycomb collagen scaffold to culture ASCs in bone tissue engineering. The osteogenic capacity of ASCs in vitro, was confirmed by histology and measuring the expression of cbfa-1. After that, ASCs were cultured for up to 14 days in the honeycomb scaffold to allow a high density, three-dimensional culture. Scanning electron microscopy data showed that the scaffold was filled with the grown ASCs, and calcification, stained black with von Kossa, was confirmed. Furthermore, The ASC-loaded honeycomb collagen scaffolds cultured for 14 days were subcutaneously transplanted into nude mice, and excised after 8 weeks. Bone formation in vivo was examined using HE stain, von Kossa stain, and osteocalcin immunostain. Those histological views showed significant positive stains in the samples of osteogenic medium in the three types of stain. These results suggest that this carrier is a suitable scaffold for ASCs and will be useful as a three-dimensional bone tissue engineering scaffold in vitro and in vivo.

Analysis of osteochondro-induction using growth and differentiation factor-5 in rat muscle.

Growth and differentiation factor-5 (GDF-5) belongs to the TGF-beta super family, and reportedly plays an important role in cartilage development and differentiation. In this study, we implanted GDF-5 in rat leg muscle, and evaluated its in vivo osteochondro-inducing activity by histological and X-ray examinations. GDF-5 (0, 100, 300, and 500 microg) and the carrier type I collagen were mixed, and the mixture was implanted into rat leg muscle. Three weeks later, the site of implantation was examined by soft X-ray, and examined histologically. The GDF-5 0 and 100 microg groups showed no osteochondro-induction. The GDF-5 300 microg group showed aggregates of cartilage and some bone tissue in the carrier. The GDF-5 500 microg group revealed bone and no cartilage. This is the first report of the dose-dependent effect of GDF-5 inducing osteochondrogenesis or osteogenesis.