The Immunologic Properties of Bone Morphogenic Protein Receptor IB Positive Subpopulation before and after Osteogenic Differentiation in Mouse Dermis.

We have previously reported that human dermal bone morphogenic protein receptor (BMPR) IB positive subpopulation had a high osteogenic differentiation potential and may be a promising cell source for allogeneic bone tissue engineering. In this study, the immunologic properties of dermal BMPR-IB+ subpopulation before and after osteogenic differentiation were reported. The results confirmed that dermal BMPR-IB+ cells possessed a similar osteogenic differentiation potential with bone marrow mesenchymal stromal cells in a mouse model. Furthermore, the expression of immune rejection-related surface antigens such as major histocompatibility class II and co-stimulatory proteins (CD40, CD80, and CD86) were absent on dermal BMPRIB+ cells. Dermal BMPRIB+ cells elicited no proliferation of allogeneic splenocytes and suppressed the proliferation of stimulated immune cells. Interestingly, osteogenic differentiation in vitro had no adverse effect on the immunological features of these cells. Most importantly, inducible NO synthase (iNOS) was involved in immunoregulatory effects by undifferentiated BMPRIB+ fibroblasts, whereas indoleamine 2,3-dioxygenase (IDO) activity was related to mediating immunomodulatory function by osteogenic differentiated BMPRIB+ fibroblasts. In conclusion, dermal BMPRIB+ cells have a low immunogenicity and possess immunosuppressive capacity before and after osteogenic differentiation in vitro, which would facilitate the allotransplantation in the future. However, mechanisms mediating immunoregulatory property between undifferentiated and osteogenic differentiated BMPRIB+ fibroblasts may be different and need further investigation.

A modified technique combining excision of the levator muscle and tarsus for blepharoptosis in Asians: a 6-year experience with 116 cases.

BACKGROUND: In patients with blepharoptosis, the function of levator muscle is insufficient or completely absent, causing blepharoptosis in various degrees. For mild or moderate blepharoptosis, levator advancement or resection is commonly performed. However, in severe cases, undercorrection results and recurrence often occur even a great length of levator muscle is resected. Because the levator muscle makes the upper eyelid move in a physiologic direction, exerting the function of residual levator muscle is still a more preferred approach for correction of blepharoptosis. This study combined tarsus resection with levator resection. The resected tarsus can offset the amount of the levator excised, making this technique applicable for severe cases. METHODS: This study included 116 patients (175 eyelids) with moderate or severe ptosis who underwent combined excision of the levator muscle and the tarsus. For cases of bilateral blepharoptosis with different levator functions between the two eyelids, surgery was performed for more severe side first and for the other side 6 months later. Postoperatively, the correction and symmetry results were evaluated and analyzed using chi-square testing by SPSS (version 10.0). RESULTS: Adequate or normal correction was achieved in 149 eyelids (85.1%). The difference in correction results did not differ significantly between moderate and severe cases. With a two-stage operation, 98 patients (84.5%) obtained good or fair asymmetry results, and no statistically significant difference existed between the bilateral and unilateral cases. CONCLUSION: The described technique appears to be effective for both moderate and severe ptosis, with better biomechanics and a satisfying aesthetic outcome.

Using human hair follicle-derived keratinocytes and melanocytes for constructing pigmented tissue-engineered skin.

BACKGROUND: Traditional tissue-engineered skin does not produce a satisfactory long-term result because it lacks natural skin pigmentation and leads to discolored cosmetically unpleasing skin that only functions to cover the body of patients. Additionally, the cell sources for tissue-engineered skin are generally derived from normal skin, which is often limited in patients with skin defects. METHODS: In this study, hair follicle melanocytes and keratinocytes were isolated from human scalp. The melanocytes were co-cultured with keratinocytes until the second passage and then purified. Purified melanocytes and keratinocytes were seeded onto the chitosan-gelatin membrane for 1 week to construct pigmented tissue-engineered skin. The pigmented skin equivalent was used to resurface the skin defect in nude mice. Four weeks after grafting, skin biopsies were harvested to take hematoxylin and eosin staining and immunohistochemistry staining of Melan-A and HLA-ABC. RESULTS: Large quantities of purified melanocytes can be obtained with co-culture method. The hematoxylin and eosin staining of repaired skin biopsy demonstrated that the tissue-engineered skin can repair skin defects successfully. Engineered skin contained pigmentation and stained positive for Melan-A and HLA-ABC, which confirmed the presence of melanocytes and its sources were of human origin. CONCLUSION: This study demonstrated the possibility of constructing pigmented tissue-engineered skin with human hair follicle-derived keratinocytes and melanocytes, which brings a promising method to make up for the deficiency of traditional tissue-engineered skin and provides an alternative treatment for depigmentation diseases.

Rapid prototyped PGA/PLA scaffolds in the reconstruction of mandibular condyle bone defects.

BACKGROUND: Craniomaxillofacial bone defects are currently reconstructed by using computer-aided design and manufacturing (CAD/CAM) processes. We have developed a novel digital medical support system that enables us to custom-make scaffolds to repair craniomaxillofacial bone defects using three-dimensional computed tomographic (CT) images and a rapid-prototyping method. METHODS: We created positive molds using CT data, CAD/CAM and a rapid prototyping method using 3D printing. Custom-made poly (glycolic acid) (PGA) and polymers poly (lactic acid) (PLA) scaffolds were prefabricated by a positive-negative mold interchange technique. A laser scanning system was used to evaluate the accuracy of the PGA/PLA scaffold. Bone marrow stem cells were incubated with the scaffold to assess biocompatibility. RESULTS: The mean error was <0.3 mm and confidence was >or=95% when the error was <1 mm. Results from in vitro cell culture demonstrated that the PGA/PLA scaffold had excellent cellular compatibility. CONCLUSIONS: This pilot study suggests that custom-made PGA/PLA scaffolds infiltrated with bone marrow stem cells may be effective for future treatment of craniomaxillofacial bone injuries.

[Reconstruction of orbital deformity after radiotherapy with transcranial orbitotomy advancement combining free flap].

OBJECTIVE: To study clinic therapeutic effect about reconstruction of severe orbital and cul-de-sac deformity after the radiotherapy with transcranial orbitotomy advancement combining cascade free flap both dorsum pedis flap and anterior tibial fascial flap. METHODS: Five cases was subjected to orbital and cut-de-sac severe deformities after both operation and radiotherapy because of retinoblastoma. The technique included transcranial orbital advancement by anterior orbital osteotomy and rigid fixed with titanic plate by coronal incision, and meanwhile incising the cul-de-sac which would be extended circumference around the central incision separation, and then designing extent of cascade flap consisted of dorsum pedis flap and anterior tibial fascial flap according to the size of cul-de-sac defect and extent of temporal depression. Then, the aforementioned two parts of cascade flap were transplanted into cul-de-sac and temple respectively. There is either the superficial temporal artery and vein or facial artery and jugular vein to chose vascular anastomosis. RESULTS: All flaps survived. After 3 to 6 months following up, the results showed satisfactory orbital contour and temporal depression improved significantly in all cases. After the conjunctival sac were fixiformed with prefabricated eye prosthesis mode about 3 months. 3 cases have good appearance with wearing eye prosthesis and the other 2 cases' appearance is poor. One of the poor appearance cases, with depressed eye socket, have orbital implant underlying conjunctival sac in secondary operation. The other one, with swallowed inferior fornix, is transplanted autogenous hard palatal mucosa into inferior fornix in secondary operation. In addition, delayed healing in donor site of dorsum pedis occurred in one of the 4 cases. CONCLUSIONS: It is a reliable procedure about reconstruction of severe orbital and cul-de-sac deformity after both the operation and radiotherapy with transcranial orbitotomy advancement combining cascade free flap both dorsum pedis flap and anterior tibial fascial flap. All deformity was corrected by one staged procedure which lessen patient's suffering and shorten patient's hospital stay and spare patient's costs.

[Microvascular nasal repair using a reversed superficial temporal artery auricular flap.].

OBJECTIVE: A reversed superficial temporal artery auricular flap was presented to explore a new method for reconstructing the defects of the distal nose by microsurgical techniques. METHODS: A reversed superficial temporal artery auricular flap had been used on fifteen patients with nasal defects, including thirteen patients with alar defects and two patients with nasal tip defects. The reversed superficial temporal vessels of the flap were anastomosed with the recipient facial vessels. The size of the flap was 2.5 cm x 2. 0 cm - 4.0 cm x 2.5 cm, the length of the vascular pedicle was 5 - 8 cm, average 6.5 cm RESULTS: The flap survived uneventfully in all fifteen patients. RESULTS: demonstrated satisfactory symmetry between the reconstructed ala and the contralateral side as well as an excellent tip projection. The donor-site defect was minimal. CONCLUSIONS: The reversed superficial temporal artery auricular flap offers an adequate length of vascular pedicle of the flap, it delivers a good solution to the problem of the vascular pedicle shortage of the proximal superficial artery auricular flap. This technique may become the top choice in the microvascular auricular transfer.

[Role of urinary basic fibroblast growth factor in differentiating hemangiomas from vascular malformation].

OBJECTIVE: To investigate a novel method to differentiate hemangioma from vascular malformation, to stage hemangiomas and to monitor the efficacy of management for hemangioma. METHODS: The urinary basic fibroblast growth factor (bFGF) concentration of 144 cases (including 69 cases of proliferating hemangiomas, 41 cases of involuting hemangiomas, 23 cases of vascular malformations and 11 negative controls) was examined using enzyme linked immunosorbent assay (ELISA). RESULTS: The differences of urinary bFGF concentration among proliferating hemangiomas, involuting hemangiomas, vascular malformations and negative control were all significant, while the differences between the latter three groups were not significant. CONCLUSIONS: Our findings suggest that examination of urinary bFGF concentration using ELISA technique is helpful in differentiating hemangioma from vascular malformation, staging hemangiomas and dynamically monitoring the efficacy of treatment for hemangiomas. Our results probably shed new light on the potential pathogenesis of hemangiomas and vascular malformation.

Reverse anterior tibial artery flap for reconstruction of foot donor site.

The foot offers numerous useful options for hand reconstruction. Hallux transfer, dorsalis pedis flap, second toe transfers, and toe joint transfers offer good functional results in reconstructed hands. However, when the donor site is repaired with skin grafts, delayed wound healing, scarring, and contractures often result. Poor cosmesis of the donor site and altered gait are the main drawbacks of the procedures. The authors propose a new concept of primary reconstruction of the donor foot using a reverse-flow anterior tibial flap from the same leg. Two flaps are raised from the same anterior tibial vessel system in continuity as a distal free flap for hand reconstruction and as a proximal reverse-flow pedicled flap to resurface the donor defect. This technique allows good flap reconstruction of the foot donor site, reducing morbidity and limiting the operation to the same limb. The authors report their experience of 33 cases. There were no failures. Primary wound healing was achieved in the foot donor site, with acceptable cosmesis and satisfactory function.

[The expression of activin A and transforming growth factor-beta 1 during rabbit mandibual distraction osteogenesis].

OBJECTIVE: To examine the expression of activin A (ACT A) and transforming growth factor-beta 1 (TGF-beta 1) during mandibular lengthening and elucidate the difference between the role of ACT A and TGF-beta 1 during mandibular distraction osteogenesis. METHOD: Skeletally mature white new zealand rabbits were established right mandibular distraction osteogenesis model. The regenerating tissue of animals' lengthened mandibes were harvested at different time points to have immunohistochemistric research of ACT A, TGF-beta 1 protein and analysis ACT A, TGF-beta 1 mRNA by using RT-PCR semiquantitative mean. RESULTS: AT the end of latency period day, positive stain of ACT A were found in the osteoblasts while positive stain of TGF-beta 1 was found in mesenchymal cells. At the end of distraction phase, fibrosis tissue had no stain of ACT A, but had strong stain of TGF-beta 1. At the period of fixation days of 20 days, both cytoplasm of osteoblasts and extracellular matrix in primary mineralization front were strongly stained of ACT A. The osteoblasts, osteoid and osteocytes in peripheral new bone zone were moderately stained of ACT A. TGF-beta 1 had strongly positive stained in fibrosis zone and weekly positive stained in primary mineralization front and peripheral new bone zone. There were also broad activin A stains in cytoplasm of osteoblasts, osteoid and cytoplasm of ACT A, TGF-beta 1 in osteocytes after distraction for 30 days. Activin A mRNA began to express at the end of latency period. Expression for activin A mRNA increased gradually along with the beginning of distraction and at the peak in distraction of 10 days and 20 days, while TGF beta 1 mRNA increased at the peak at the end of latency period. CONCLUSION: ACT A and TGF beta 1 have different role during rabbit mandibular distraction osteogenesis.

[Indentification of differentially expressed genes in proliferating and involuting hemangiomas by cDAN microarray analysis of gene-expression profiles].

OBJECTIVE: To investigate the differentially expressed genes of proliferating and involuting hemangiomas by cDNA microarray analysis of gene-expression profiles in an effort to identify the key disease-related genes. METHODS: Samples were processed from total RNA and purified to mRNA, which was reverse-transcripted and hybridized onto Biodoor Genechip expression microarrays. Analyses were performed to determine the consensus pattern of gene expression in the proliferating and involuting stages of the same hemangioma and the changes in the expression level. RESULTS: In proliferating hemangioma, 79 genes were overexpressed, and 115 genes were underexpressed in comparison with the involuting hemangioma. Some cytokines and growth factors such as neurotensin, Nov, CYR6, keratinocyte growth factor, interleukin-10 were overexpressed in proliferative hemangioma. In involuting hemangioma, apoptotic factors such as bcl-2 binding component, cytochrome C were overexpressed. The overexpression of Nov, CYR6, c-myc implied that angiogenesis and oncogenes might participate in the pathogenesis of hemangiomas. Mitochondria activated apoptotic passage (cytokines, bcl-2, cytochrome C) and Wnt/beta-catenin passage(Frizzled, beta-catenin, c-myc) were involved. CONCLUSION: The development of hemangiomas may be the results of imbalance of cell proliferation and apoptosis.

[Staging and management of extracranial arteriovenous malformations].

OBJECTIVE: Congenital arteriovenous malformations(AVM) are considered to be the most difficult and challenging problems in the treatment of hemangiomas and vascular lesions. This study focused on the natural history, the clinical classification, the choice and effectiveness of various treatments. METHODS: This retrospective review included 83 patients with extracranial arteriovenous malformations, who were referred to our department over the past 6 years. The anatomic patterns, clinical staging, respective treatments, influential factors of endovascular treatment, causes of recurrence were analyzed. RESULTS: According to clinical manifestations, arteriovenous malformations were categorized as three clinical stages: the quiescence, the expansion and the decompensation stages. Most AVMS in the quiescence stage only require endovascular treatment while those in the decompensation stage require surgical resection. Angiography was performed not only for diagnosis of AVM but also as an initial therapeutic step in the form of embolization, which might be the only means to some AVM without surgical possibility or necessity. CONCLUSION: The new concept of staging and management is expected to be helpful for diagnosis and treatments of AVM.