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1.
Exp Ther Med ; 18(6): 4575-4582, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31807147

RESUMO

Objective measurement is important for diagnosing congenital or acquired auricular abnormalities and the evaluation of therapeutic efficacy. However, methods applied in the past were mostly inaccurate and unreliable. The present study aimed to introduce five standardized indices for auricle measurement and present a highly precise and reliable methodology combining three-dimensional (3D) scanning techniques and the Materialise Mimics software for the evaluation of auricle sizes. A total of 20 normal ears were measured independently by four surgeons using the standardized digital method with 3D scanning technique and the traditional manual method. Parameters of the auricle, including the length and width, arc length, cranioauricular height and angle were measured using the Mimics software. Paired t-test, Wilcoxon signed rank test and intra-class correlation coefficients (ICC) were performed on the data to assess the precision, uniformity and observer independence of the method. Pearson's product moment correlation was calculated to assess the correlation between auricle length and width in addition to the correlation between cranioauricular height and angle. No significant differences were indicated between measurements of five auricular parameters made by two surgeons using the digital method. However, significant differences were found using the manual method (P<0.01). ICC values derived from digital measurements ranged from 0.901 to 0.987, whereas those derived from manual measurements ranged from 0.526 to 0.807. These results suggested that the standardized digital method was replicable and reliable compared with the traditional manual method. Pearson's coefficient analysis showed that there was a significant correlation between cranioauricular height and angle (P<0.05), but no correlations were found between the height and width of the auricle (P>0.05). Taken together, data from the present study suggested that measurements of the length and width, arch length, and cranioauricular height and angle of auricles using the standardized digital method combining 3D scanning with the Mimics software were comprehensive, precise, convenient, repeatable and reliable.

2.
EBioMedicine ; 28: 287-302, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29396297

RESUMO

Microtia is a congenital external ear malformation that can seriously influence the psychological and physiological well-being of affected children. The successful regeneration of human ear-shaped cartilage using a tissue engineering approach in a nude mouse represents a promising approach for auricular reconstruction. However, owing to technical issues in cell source, shape control, mechanical strength, biosafety, and long-term stability of the regenerated cartilage, human tissue engineered ear-shaped cartilage is yet to be applied clinically. Using expanded microtia chondrocytes, compound biodegradable scaffold, and in vitro culture technique, we engineered patient-specific ear-shaped cartilage in vitro. Moreover, the cartilage was used for auricle reconstruction of five microtia patients and achieved satisfactory aesthetical outcome with mature cartilage formation during 2.5years follow-up in the first conducted case. Different surgical procedures were also employed to find the optimal approach for handling tissue engineered grafts. In conclusion, the results represent a significant breakthrough in clinical translation of tissue engineered human ear-shaped cartilage given the established in vitro engineering technique and suitable surgical procedure. This study was registered in Chinese Clinical Trial Registry (ChiCTR-ICN-14005469).


Assuntos
Cartilagem da Orelha/cirurgia , Procedimentos Cirúrgicos Reconstrutivos , Regeneração , Animais , Biópsia , Criança , Microtia Congênita/patologia , Microtia Congênita/terapia , Cartilagem da Orelha/transplante , Feminino , Seguimentos , Humanos , Masculino , Camundongos , Engenharia Tecidual , Tecidos Suporte
3.
J Plast Reconstr Aesthet Surg ; 69(10): 1436-44, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27496290

RESUMO

BACKGROUND: During microtia reconstruction, the intraoperative design of the cartilage framework is important for the appearance and symmetry of the bilateral auricles. Templates (traditionally, the X-ray film template) are usually utilized to complete the task, which can provide cues regarding size, cranioauricular angle and positioning to the surgeons. With a combination of three-dimensional (3D) scanning and additive manufacturing (AM) techniques, we utilized two different ear-shaped templates (sheet moulding and 3D templates) during the fabrication of 3D-customized autologous cartilage frameworks for auricle reconstruction. METHODS: Forty unilateral microtia patients were included in the study. All the patients underwent auricle reconstruction using the tissue-expanding technique assisted by the new AM templates. Images were processed using computer-aided design software and exported to print two different AM ear-shaped templates: sheet moulding and 3D. Both templates were assisted by the 3D framework fabrication. The 3D images of each patient's head were captured preoperatively using a 3D scanner. X-ray film templates were also made for the patients. The lengths and widths of the contralateral auricles, X-ray film and sheet moulding templates were measured in triplicate. The error of the template and the contralateral auricle were used to compare the accuracy between the two templates. RESULTS: Between January and May 2014, 40 unilateral microtia patients aged 6-29 years were included in this study. All patients underwent auricle reconstruction using autogenous costal cartilage. The sterilized AM templates were used to assist in the framework fabrication. The operative time was decreased by an average of 15 min compared with the method assisted by the X-ray film template. Postoperative appearance evaluation (based on five indexes: symmetry, length, width, cranioauricular angle and the substructure of the reconstructed ear) was performed by both the doctors and the patients (or their parents). Follow-up (ranging from 9 to 18 months) showed that all of the patients obtained satisfactory results with life-like 3D configuration and symmetric cranioauricular angle. The follow-up showed that no surgery-related complications occurred. Comparing the accuracy of the X-ray film and sheet moulding templates, the average errors of length were 1.8 mm ± 1.44 mm and 0.39 mm ± 0.35 mm, respectively, and the average width errors were 1.32 mm ± 0.88 mm and 0.3 mm ± 0.47 mm, respectively. The new sheet moulding template was more accurate than the X-ray template. CONCLUSIONS: The new sheet-moulding template is much more accurate than the traditional X-ray film template. Framework fabrication assisted by accurate 3D and informative AM templates contributed to individualized cartilage framework fabrication and satisfactory results.


Assuntos
Cartilagem/transplante , Microtia Congênita/cirurgia , Pavilhão Auricular/cirurgia , Procedimentos Cirúrgicos Reconstrutivos , Retalhos Cirúrgicos , Expansão de Tecido/métodos , Adolescente , Adulto , Criança , China , Projeto Auxiliado por Computador , Feminino , Humanos , Imagem Tridimensional/métodos , Masculino , Satisfação do Paciente , Procedimentos Cirúrgicos Reconstrutivos/instrumentação , Procedimentos Cirúrgicos Reconstrutivos/métodos , Cirurgia Plástica/métodos , Resultado do Tratamento
4.
J Craniofac Surg ; 27(2): e178-81, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26854778

RESUMO

An anterolateral thigh myo-adipofascial flap was used in the definitive management of a patient presented with chronic infective process associated with a large fronto-nasal defect. Unfortunately, the risk of free flap transfer failure emerged when intraoperative dissection showed absence of a reliable ipsilateral superficial temporal artery as the recipient artery. This rare incident happened at the stage whereby the anterolateral thigh flap was nearly completely raised with a distal perforator in situ. In this article, the authors presented an innovative strategy to salvage the flap by transforming the flap into a modified composite flap based on the retrograde blood flow principle. To the best of our knowledge, this is the first report of using such a technique in reconstructive microsurgery. This successful salvage strategy has clinical application and could potentially minimize free flap transfer failure.


Assuntos
Retalhos de Tecido Biológico/transplante , Retalho Miocutâneo/transplante , Retalho Perfurante/transplante , Procedimentos Cirúrgicos Reconstrutivos/métodos , Tecido Adiposo/transplante , Adulto , Fístula Cutânea/cirurgia , Fáscia/transplante , Seguimentos , Retalhos de Tecido Biológico/irrigação sanguínea , Osso Frontal/lesões , Humanos , Complicações Intraoperatórias , Masculino , Microcirurgia/métodos , Retalho Miocutâneo/irrigação sanguínea , Fraturas Orbitárias/cirurgia , Retalho Perfurante/irrigação sanguínea , Complicações Pós-Operatórias/cirurgia , Fluxo Sanguíneo Regional/fisiologia , Terapia de Salvação , Transplante de Pele/métodos , Fraturas Cranianas/cirurgia , Coxa da Perna/cirurgia , Sítio Doador de Transplante/cirurgia , Resultado do Tratamento
5.
Front Med ; 5(1): 61-9, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21681676

RESUMO

Tissue engineering aims to produce a functional tissue replacement to repair defects. Tissue reconstruction is an essential step toward the clinical application of engineered tissues. Significant progress has recently been achieved in this field. In our laboratory, we focus on construction of cartilage, tendon and bone. The purpose of this review was to summarize the advances in the engineering of these three tissues, particularly focusing on tissue regeneration and defect repair in our laboratory. In cartilage engineering, articular cartilage was reconstructed and defects were repaired in animal models. More sophisticated tissues, such as cartilage in the ear and trachea, were reconstructed both in vitro and in vivo with specific shapes and sizes. Engineered tendon was generated in vitro and in vivo in many animal models with tenocytes or dermal fibroblasts in combination with appropriate mechanical loading. Cranial and limb bone defects were also successfully regenerated and repaired in large animals. Based on sophisticated animal studies, several clinical trials of engineered bone have been launched with promising preliminary results, displaying the high potential for clinical application.


Assuntos
Osso e Ossos , Cartilagem , Tendões , Engenharia Tecidual/tendências , Animais , Materiais Biocompatíveis , Modelos Animais , Regeneração , Engenharia Tecidual/métodos
6.
Neurosurgery ; 68(3): 691-704, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21311297

RESUMO

BACKGROUND: Intracerebral hemorrhage (ICH) represents at least 15% of all strokes in the Western population and a considerably higher proportion at 50% to 60% in the Oriental population. OBJECTIVE: To investigate whether administration of bone marrow stem cells (BMSCs) overexpressing glial cell line-derived neurotrophic factor (GDNF) provides more efficient neuroprotection for rats with ICH and neurons exposed to hypoxia/reoxygenation. METHODS: Primary rat BMSCs were transfected with rat GDNF gene using virus vector (GDNF/BMSCs) and blank virus plasmid (BVP/BMSCs). Primary rat cortical neurons of rats were exposed to hypoxia and then reoxygenated with GDNF/BMSCs (GDNF/BMSCs group) or BVP/BMSCs (BMSCs group) treatment for 12 hours and 1, 2, 3, and 5 days. Hoechst 33258 staining was used to evaluate apoptosis. GDNF/BMSCs, BVP/BMSCs, and saline (GDNF/BMSCs, BMSCs, and control groups) were injected into the right striatum 3 days after rat ICH induced by injecting collagenase. Modified neurological severity scores and hematoxylin and eosin staining were performed to evaluate neurological function and lesion volume at 1 and 2 weeks after transplantation. Immunostaining was used to observe differentiation of grafted cells (neurofilament-200 for neurons, glial fibrillary acidic protein for astrocytes). The GDNF level and apoptosis were evaluated by Western blotting and terminal deoxynucleotidyl transferase dUTP nick-end labeling, respectively. RESULTS: The GDNF/BMSCs group had significantly lowered apoptosis compared with the BMSCs group at the given time. The GDNF/BMSCs group had significantly improved functional deficits and reduced lesion volume compared with the BMSCs group. Stable GDNF expression in the GDNF/BMSCs group was detected at the given time in the host brain. The neurofilament-positive grafted cells in the GDNF/BMSCs group were more numerous than in the BMSCs group. The GDNF/BMSCs group had significantly decreased apoptotic cells compared with the BMSCs group. CONCLUSION: These results suggest that GDNF/BMSCs provide better neuroprotection for rats with ICH and neurons exposed to hypoxia/reoxygenation.


Assuntos
Hemorragia Cerebral/metabolismo , Hemorragia Cerebral/terapia , Terapia Genética/métodos , Fator Neurotrófico Derivado de Linhagem de Célula Glial/uso terapêutico , Transplante de Células-Tronco Mesenquimais/métodos , Neurônios/metabolismo , Oxigênio/metabolismo , Animais , Hipóxia Celular , Células Cultivadas , Terapia Combinada , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Células-Tronco Mesenquimais/metabolismo , Neurônios/patologia , Ratos , Ratos Wistar , Resultado do Tratamento
7.
Biomaterials ; 31(36): 9406-14, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21056466

RESUMO

In vivo niche plays an important role in determining the fate of exogenously implanted stem cells. Due to the lack of a proper chondrogenic niche, stable ectopic chondrogenesis of mesenchymal stem cells (MSCs) in subcutaneous environments remains a great challenge. The clinical application of MSC-regenerated cartilage in repairing defects in subcutaneous cartilage such as nasal or auricular cartilage is thus severely limited. The creation of a chondrogenic niche in subcutaneous environments is the key to solving this problem. The current study demonstrates that bone marrow stromal cells (BMSCs) could form cartilage-like tissue in a subcutaneous environment when co-transplanted with articular chondrocytes, indicating that chondrocytes could create a chondrogenic niche to direct chondrogenesis of BMSCs. Then, a series of in vitro co-culture models revealed that it was the secretion of soluble factors by chondrocytes but not cell-cell contact that provided the chondrogenic signals. The subsequent studies further demonstrated that multiple factors currently used for chondroinduction (including TGF-ß1, IGF-1 and BMP-2) were present in the supernatant of chondrocyte-engineered constructs. Furthermore, all of these factors were required for initiating chondrogenic differentiation and fulfilled their roles in a coordinated way. These results suggest that paracrine signaling of soluble chondrogenic factors provided by chondrocytes was an important mechanism in directing the in vivo ectopic chondrogenesis of BMSCs. The multiple co-culture systems established in this study provide new methods for directing committed differentiation of stem cells as well as new in vitro models for studying differentiation mechanism of stem cells determined by a tissue-specific niche.


Assuntos
Células da Medula Óssea/citologia , Diferenciação Celular , Condrócitos/citologia , Condrogênese , Coristoma , Células Estromais/citologia , Animais , Proteínas Morfogenéticas Ósseas/metabolismo , Comunicação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Condrócitos/transplante , Condrogênese/efeitos dos fármacos , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Fator de Crescimento Insulin-Like I/metabolismo , Camundongos , Camundongos Nus , Solubilidade/efeitos dos fármacos , Células Estromais/metabolismo , Células Estromais/transplante , Tela Subcutânea/efeitos dos fármacos , Tela Subcutânea/metabolismo , Sus scrofa , Tecidos Suporte , Fator de Crescimento Transformador beta1/metabolismo
8.
Biomaterials ; 31(13): 3564-71, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20153525

RESUMO

Adipose-derived stem cells (ASCs) are considered as a promising cell source for cartilage regeneration. However, the heterogeneity of this cell source may affect their ability in cartilage formation. It is therefore necessary to establish an efficient method for isolating the cells that have chondrogenic potential. To date, no specific markers have been reported to be able to isolate such a cell population from human adipose tissue. In recent studies, endoglin (CD105) has been known as a relatively specific marker for identifying mesenchymal stem cells, but no studies show it is related to chondrogenic potential of human ASCs. In this study, human cells from adipose tissue were isolated, cultured, and sorted according to CD105 expression. The sorted cells were then subjected to adipogenic, osteogenic, and chondrogenic induction to confirm their multi-potentiality. In adipogenic conditions, CD105- cells showed stronger Oil Red staining and higher expression of adipose-specific genes compared to CD105+ cells. By contrast, CD105+ cells exhibited better osteogenic potential with stronger Alizarin Red staining and higher expression of osteogenic specific genes than CD105- cells. Noticeably, CD105+ cells also exhibited a much stronger chondrogenic potential than CD105- cells, with stronger collagen II staining and higher gene expression of collagen II and aggrecan. Most importantly, CD105+ cells could form a homogeneous cartilage-like tissue when seeded into a biodegradable scaffold and cultured in chondrogenic media for 8 weeks. These results indicate that sorting of ASC subpopulation with CD105 as a marker may allow better in vitro chondrogenesis and thus provide an important implications for cartilage regeneration and reconstruction using autologous cells from adipose tissue.


Assuntos
Tecido Adiposo/citologia , Antígenos CD/imunologia , Condrogênese , Receptores de Superfície Celular/imunologia , Células-Tronco/citologia , Tecido Adiposo/imunologia , Sequência de Bases , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Primers do DNA , Endoglina , Citometria de Fluxo , Humanos , Reação em Cadeia da Polimerase , Células-Tronco/imunologia
9.
Asian J Androl ; 12(2): 196-202, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20023691

RESUMO

Smoothened (SMO) is an important member of the Hedgehog signaling pathway. We constructed a specific recombinant lentiviral vector for RNA interference, targeting the SMO gene (NM_005631) to observe its effect on SMO expression, cell proliferation and the cell cycle in the human androgen-sensitive prostate cancer cell line, LNCaP, and in the androgen-independent prostate cancer cell line, PC3. Four siRNA sequences were designed and inserted into a lentiviral vector pGCSIL-GFP to construct four recombinant vectors. The vector with the highest interfering efficiency was co-transfected with packaging vectors (pHelper1.0 and pHelper2.0) in 293T cells to assemble lentivirus particles by liposome for infecting LNCaP and PC3 cell lines, respectively. The expression level of SMO mRNA, tumor cell proliferation and cell cycle were measured by quantitative realtime polymerase chain reaction (qRT-PCR), 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay and flow cytometry, respectively. Sequence results showed that recombinant lentiviral vectors were constructed successfully. pGCSIL-GFP-723 had the highest interfering efficiency, named Lv-SIL-SMO723 after co-transfection, with which LNCaP and PC3 cell lines were infected. Compared with the control groups, results showed significantly decreased (P < 0.05) SMO mRNA expressions of LNCaP and PC3, lower mean percentage of S-phase cells and higher mean percentage of G(2)/M phase cells, as well as obviously slow proliferation (P < 0.01) of LNCaP in the infected group. Yet, the proliferation of PC3 was not altered (P > 0.05). In conclusion, the recombinant lentivirus particles were able to suppress SMO expression, regulate the cell cycle in the LNCaP and PC3 cell lines and markedly inhibit proliferation of LNCaP cells but not PC3 cells.


Assuntos
Proliferação de Células , Vetores Genéticos , Lentivirus/genética , Neoplasias da Próstata/patologia , Interferência de RNA , Receptores Acoplados a Proteínas-G/genética , Sequência de Bases , Ciclo Celular , Linhagem Celular Tumoral , Primers do DNA , Citometria de Fluxo , Humanos , Masculino , Receptor Smoothened
10.
Biotechnol Lett ; 31(5): 639-46, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19169885

RESUMO

TGF-beta1 plays a necessary and important role in the induction of chondrogenic differentiation of bone marrow stromal cells (BMSCs). In this study, porcine BMSCs were infected with a replication-deficient adenovirus expression vector carrying the hTGF-beta1 gene. The transduced BMSCs were cultured as pelleted micromasses in vitro for 21 days, seeded onto disk-shaped PGA scaffolds for 3 days and subsequently implanted into the subcutaneous tissue of mice. BMSCs transduced with AdhTGF-beta1 expressed and secreted more hTGF-beta1 protein in vitro than those of the control group. Histological and immunohistological examination of the pellets revealed robust chondrogenic differentiation. Tissues made from cells transduced with AdhTGF-beta1 exhibited neocartilage formation after 3 weeks in vivo. The neocartilage occupied 42 +/- 5% of the total tissue volume which was significantly greater than that of the control group. Furthermore, there was extensive staining for sulfated proteoglycans and type II collagen in the AdhTGF-beta1 group compared to controls, and quantification of GAG content showed significantly greater amounts of GAG in experimental groups. The results demonstrate that transfer of hTGF-beta1 into BMSCs via adenoviral transduction can induce chondrogenic differentiation in vitro and enhance chondrogenesis in vivo.


Assuntos
Adenoviridae/genética , Medula Óssea , Condrogênese , Células Estromais/fisiologia , Transdução Genética , Fator de Crescimento Transformador beta1/biossíntese , Animais , Células Cultivadas , Implantes Experimentais , Camundongos , Suínos , Fator de Crescimento Transformador beta1/genética
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