The innovative application of digitally designed appliance for preoperative cleft lip and palate / 中华整形外科杂志

Objective@#The study presents a new method to prefabricate the nasoalveolar molding appliances for preoperative cleft lip and palate by using three-dimensional technology.@*Methods@#A long term retrospective study of 40 cases of infants with unilateral cleft lip and palate who underwent the preoperative 3D models of alveolar bone acquisition, computer aided design for the rapid prototyping process, gypsum powder printing maxillary three-dimensional entity model and install the appliance for 3-4 months (or alveolar cleft<2 mm). Simultaneously, primary rhinoplasty can be done during cleft lip repair. All patients had clinic visits three times each month.@*Results@#Deformities of infants who underwent this treatment, were significantly improved. The alveolar cleft was significantly reduced (P<0.01), while bilateral alveolar tissue volume was significantly increased (P<0.01). Alveolar bone morphology was more symmetrical than before.@*Conclusions@#The digital design and manufacture system of appliances for preoperative cleft lip and palate can be successfully applied to the preoperative orthodontic treatment. The sequence treatment of cleft lip and palate becomes better improved. It can shorten the time duration and the numbers of clinic visit on the basis of ensuring the accuracy of preoperative orthodontic effect.

Clinical analysis of mandibular reconstructions using synthetic bones based on three-dimensional printing technology in 149 cases / 上海交通大学学报（医学版）

Objective-To assess the clinical value of the computer-assisted three-dimensional reconstruction technique design and evaluate the climcal experience of manufacture artificial bone precision to repair the mandibular defect.Methods· From 2001 to 2016,163 computer-assisted reconstruction surgeries had been performed in Craniofacial Department,Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine.During six months followup,the measurement data was conducted and compared with three-dimensional CT result.Random measurement of the three key anatomical points pre-and post-operative carried out with statistical error was used to evaluate the accuracy of computer-assisted three-dimensional reconstruction in mandibular defects repairation and to investigate the clinical application value of the operation time and postoperative complication rate.Results· From July 2001 to July 2016,a total of 163 patients underwent computer-assisted three-dimensional reconstruction of artificial bone repair for mandibular defects;149 patients met the statistical criteria in which preoperative design and postoperative actual effect's average distance error (1.27±0.15) mm,operation time (2.5±1.2) h.Conclusion· Threedimensional design of artificial bone to repair the mandibular defect is a valuable technology,by relying on quantitative design and preoperative simulation to simplify the difficulty and improve the accuracy of surgery.The patients showed high satisfaction rate with low surgical complications and long-term efficacy.

The morphological study on masseter muscle following mandibular angle osteotomy / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate the morphological change of masseter after the mandibular angle osteotomy.</p><p><b>METHODS</b>Computerized tomography (CT) examination was performed on 120 patients treated by mandibular angle osteotomy before operation and at 3, 6, 12 months after operation, respectively. The pre- and postoperative masseter muscle thickness and cross-sectional area were evaluated using 3D CT images observed from 3 selected slice planes, which were paralleled with Frankfurt horizontal plane. These CT images were stored and three-dimensional reconstruction were made for calculation of masseter muscle volume through software.</p><p><b>RESULTS</b>After operation, the reduction of the masseter muscle volume and cross-sectional area was seen. The volume of the masseter at 3, 6, 12 months postoperatively decreased to 82.02%, 77.00% and 80.43% (P < 0.05). The cross-sectional area at 3, 6,12 months postoperatively decreased to 85.81%, 78.86% and 81.56% at A plane, 80.94%, 75.03% and 77.04% at B plane, and reached to 13.46%, 11.48% and 13.89% at C plane (P < 0.05, P < 0.05, P < 0.01). The masseter thickness after operation was significantly different from that before operation during the follow-up period, but not at 12 months after operation at A plane.</p><p><b>CONCLUSIONS</b>The masseter atrophy happens spontaneously after mandibular angle osteotomy, especially at the region of mandibular angle. It should be considered during surgical design.</p>

Surgical correction of craniofacial dysostosis with midface distraction osteogenesis / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of distraction osteogenesis on correction of craniofacial dysostosis.</p><p><b>METHODS</b>Le Fort III osteotomy was applied through coronal route on patients with craniofacial dysostosis such as Crouzon and Apert syndrome. The procedures included disconnecting the skeletal midface from base of cranium, setting up a RED II distraction device, and directing the device bars. The distraction was started 5 days after the surgery, with a rate of 1 mm forward per day. When midface approaching the right position, i.e. a slightly over correction of occlusion was reached, stopped distraction and kept the device for 2 - 4 months.</p><p><b>RESULTS</b>Eight cases completed all the therapy. The average blood lose was 300 ml and the average operation time was 3.5 hours. The midface had been moved averagely 9 mm forwardly and 1.5 mm downwards. The features had been improved obviously and the occlusion reached nearly normal. No serious complications occurred except for 1 case of seroma and 1 case of infection around pin on scalp. No recurrence was found in the 5 months of follow-up.</p><p><b>CONCLUSIONS</b>Midface distraction osteogenesis is propitious to teenage or severe cases of craniofacial dysostosis.</p>

Microarray analysis of dedifferentiation related gene expression of human chondrocytes cultured in vitro / 中华整形外科杂志

<p><b>OBJECTIVE</b>Dedifferentiation of chondrocytes during in vitro expansion is the major cause that limits the potential of chondrocytes for cartilage engineering. This study dissected dedifferentiation mechanism of in vitro cultured human chondrocytes by microarray analysis of gene expression changes.</p><p><b>METHODS</b>Spare human costal cartilage from ear reconstruction patients (n=3, aged 10-20) were digested with collagenase II to isolate human chondrocytes(HCC) and were expanded from P1 to P4. For microarray, RNA was isolated from the P1 and P4 cells and labeled with cy3 and cy5 fluorescence respectively as probes to hybridize a cDNA microarray( about 8300 genes). Those genes with 2-fold difference of expression were selected.</p><p><b>RESULTS</b>Microarray results showed that 56 genes were up-regulated at P4, including proteases, inflammatory factors, growth suppressor and apoptosis genes. Down-regulated 84 genes at P4 include extracellular matrices, protein anabolism, growth factors, cell cycle and cytoskeleton related genes.</p><p><b>CONCLUSIONS</b>These results provide insight into the mechanism of chondrocytes dedifferentiation. The characteristic appearance of dedifferentiation related genes expression includes increased level of stress response and inflammation. The down-regulation of anabolic metabolism related genes and up-regulation of proteases genes leaded to extra cellular matrix decrease. Decreased production of growth factors and increased apoptosis cause decreased cell proliferation of dedifferentiation chondrocytes.</p>

Experimental study of construction of tissue engineered bone ectopically by human bone marrow mesenchymal stem cells / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the possibility and mechanism of construction of tissue engineered bone with human bone marrow mesenchymal stem cells (hBMSCs) as seeding cells and partially demineralized bone matrix (pDBM) as scaffold.</p><p><b>METHODS</b>hBMSCs are cultured and mutiplified. The 4th grade hBMSCs are seeded on the pDBM, the growth and adhesion of hBMSCs on pDBM are observed under scanning electro microscope. The adhesion efficiency is assessed. The complexes are implanted in the nude mice subcutaneously, the pDBM without cells as control. The grafts are taken out on the 8th and 12th week.</p><p><b>RESULTS</b>There is new bone formation on the 8th and 12th week in complex group. There is a layer of osteoblast like cells adhered on the surface of most of the new bone, which suggest the possibility of intramembranous ossification. There is no bone formation in control group.</p><p><b>CONCLUSIONS</b>Tissue engineered bone can be constructed with hBMScs and pDBM in vivo, and the mechanism of which could be intramembranous ossification.</p>

Pilot study of using autologous bone marrow stromal cells and coral to repair canine segmental mandibular defects / 中华整形外科杂志

<p><b>OBJECTIVE</b>To repair segmental mandibular defects with autologous bone marrow stromal cells (BMSCs) engineered bone.</p><p><b>METHODS</b>Isolated BMSCs were expanded in vitro and osteogenic induced. In 12 canines, a 3 cm segmental mandibular defect at right mandible was created. 6 canine's defects were repaired with cell-scaffold constructs made from induced BMSCs and coral; others were repaired with coral as control. The engineered bone was evaluated by X-ray, CT, Dual Energy X-ray Absorptiometry (DXA), gross and histological examination, and biomechanical test post-operatively.</p><p><b>RESULTS</b>Induced BMSCs grew well on coral scaffold. At 12 weeks, X-ray showed more callus formed in experimental group, while evident scaffold duration in control group. At 32 weeks, gross observation, X-ray and CT demonstrated well bony-union in experimental group, while bony-nonunion in control group. Also DXA revealed significantly higher bone mineral density of experimental group than control group. Histologically, mature bone were commonly observed and there were bony healing in experimental group, while fibrous healing occurred in control group. Biomechanical test revealed no significant difference between experimental group and normal group.</p><p><b>CONCLUSIONS</b>Canine segmental mandibular defects can be repaired with the tissue-engineered bone generated by coral scaffold with autologous osteogenic BMSCs.</p>

Repair alveolar cleft bone defects with bone marrow stromal cells / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the feasibility of repairing alveolar cleft bone defects with bone marrow stromal cells.</p><p><b>METHODS</b>Total 7 patients of alveolar cleft were included in this study. The hBMSCs were isolated by percoll gradient centrifugation from patient's bone marrow aspirated from iliac crest. The hBMSCs were cultured in vitro and induced to become osteogenic cells in the DMEM medium containing 10% self-serum, beta-glycerophosphate (10 nmol/L) dexamethasone (10(-8) mol/L) , L-2-ascorbic acid(50 micromol/L), and 1, 25 (OH)2 VD3 (10 nmol/L). Induced hBMSCs of passage 3 were harvested and seeded onto partly demineralized allogenic bone matrix (pDBM) to form a cell-scaffold construct and in vitro co-culture for 1 week. The defects were repaired with the cell-scaffold construct. All cases were followed up for 3, 6 months post-operation as short-term evaluation and 1 to 3 years post-operation as long-term evaluation by three-dimensional computerized tomography (3D-CT) and clinical examination.</p><p><b>RESULTS</b>3D-CT demonstrated that engineered bone was formed in 3 months post-operation. Additionally, formed bone maintained stable up to 1 - 3 years without absorption.</p><p><b>CONCLUSIONS</b>Engineered bone can be used to repair clinical alveolar cleft bone defects.</p>

Induction of human mesenchymal stem cells in vitro and related identification / 中国耳鼻咽喉头颈外科

OBJECTIVE To establish the isolation and identification system of human mesenchymal stem cells(hMSCs)in vitro, and then to establish a stable cultured system in which the hMSCs can be induced to osteoblasts, chondroblasts and lipoblasts in vitro. To study the possibility if hMSCs can be used as the seed cells in bone tissue engineering.This study would lay a foundation for the clinical application of tissue engineering. METHODS 5ml bone marrow of the patient was aspirated from iliac crest. The human MSCs were isolated using Percoll gradient centrifugation. The hMSCs were cultured in DMEM medium in vitro, FACS(fluorescence axtivated cell sorter)was used to identify the phenotype of hMSCs. The hMSCs were cultured in vitro, and induced to osteoblasts, chondrocytes and lipocytes. The changes of phenotype were tested by mmunohistochemisty and molecular biology technique. RESULTS The hMSCs were isolated by Percoll gradient centrifugation from patient’s bone marrow. The expression rate of MSC markers including CD105, CD166, CD29 were(78?6)%, (43?7)%, (69? 12)% respectively. The phenotype of osteoblasts induced from hMSCs was verified by the mineralized nodes formation when cultured in vitro and Col, and OCN expression showed by immunohistochemistryand RT-PCR. The phenotype of chondrocytes induced from hMSCs was verified by type, collagen, SOX9 expression with immunohistochemical staining technique and type, collagen, aggrecan mRNA expression with RT-PCR. The lipocytes’ phenotype was verified by positive result of oil red O staining and PPAR 2mRNA expression by RT-PCR. CONCLUSION The highly pure hMSCs can be harvested by means of density gradient centrifugation, and the hMSCs have multi-potentiality of cell differentiation via induced culture in vitro. The third passage of hMSCs can be induced to express osteoplast phenotype and meet the qualitative and quantitative demands of seed cells when bone tissue engineering was used in clinical practice.

Watershed-based segmentation of histiocytic images / 生物医学工程学杂志

The task of segmenting histiocyte is a crucial step in the analysis of histiocytic images which is an important application of computer vision to histopathology. The algorithm presented in this article was composed of two steps: (1) the morph-based preprocessing; (2) the ameliorated watershed method. In the first step, the difference between histiocytes was magnified in order to increase the visibility from the view of the computer vision, and then the ameliorated terrace-flooding-simulated watershed method was used to achieve the segmentation of histiocytic images in the second step. To test the performance of the algorithm, different samples of visual quality were tested and the result figures proved successful.

Age-related changes in growth and metabolism function of human costal chondrocytes cultured in vitro / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the expressive changes of collagen I, II and aggrecan during the aging of the human costal chondrocytes cultured in vitro, in order to select the best chondrocytes for cartilage engineering.</p><p><b>METHODS</b>The human costal chondrocytes from the different passages (P1-P5) were harvested. The morphological changes and cell proliferation rate were observed, and the quantity of glycosaminoglycans (GAGs) was examined in each passage with alcian blue precipitation. The expression of collagen I, II and aggrecan was measured with immunohistochemistry and RT-PCR method.</p><p><b>RESULTS</b>From the third passage, the human costal chondrocytes were transformed into fibroblast-like cell morphology; and the GAGs content in each passage was decreased and became significantly low in the third passage. The expression of the collagen I, II was consistent in the protein level with that in the mRNA level. Before the second passage, the expression of collagen II was high while the collagen I was low. After the second passage, the expression of collagen II decreased while the collagen I increased. The expression of aggrecan was high before the third passage, and then became low in the fourth passage.</p><p><b>CONCLUSION</b>With the examinations of the cell proliferating rate and cell functions in human chondrocytes cultured in vitro, the chondrocytes of the second passage seem suitable for human cartilage engineering.</p>