Objective To study the adhesion characteristic in vitrobetween porous biphasic calcium phosphate(BCP) nanocomposite and bone marrow mesenchymal stem cells (MSCs) that have been induced and proliferated. Methods MSCs obtained from SD ratbone marrow were in vitro induced and proliferated. After their osteoblastic phenotype were demonstrated, MSCs were seeded onto prepared porous BCP nanocomposite(experiment group)and common porous hydroxyapatite (control group). Their adhesion situation was analyzed by scanning electron microscope. The initial optimal cell seeding density was investigated between new pattern porous BCP nanocomposite and MSCs by MTT automated colormetric microassay method. Results The differentiation of MSCs to osteoblastic phenotype were demonstrated by the positive staining of mineralized node, alkaline phosphatase (ALP) and collagen typeⅠ, the most appropriate seeding density between them was 2×106／ml. The maximal number which MSCs could adhere to porous BCP nanocomposite was 1.28×107/cm3. Conclusion MSCs can differentiate to osteoblastic phenotype.The MSCs were well adhered to porous BCP nanocomposite.
Objective To establish a scaffold model from heterogeneoussmall blood vessels. Methods Caudal arteries from 34 Wistar rats( average length 12.08±1.69 cm) were made into acellular blood vessel scaffolds. Some scaffoldswere observed by electron microscope, and others were transplanted to the cut ends of ear central arteries of male Japanese big ear white rabbits. Results Average external diameter was 0.74±0.08 mm in proximal, and 0.55±0.08 mm in distal end of rat caudal arteries. The small blood vessel scaffolds had shin wall whichwas white and soft, composed of fibrous tissues without cells. On the intima surface the fibrous tissues were arrayed densely in a grid-like pattern. After transplantation, the blood flow was reserved, and kept flowing freely in 24 hours. The pulsation of the transplanted artery was accessible and no blood leakage wasfound.Conclusion The natural scaffolds are composed of fibrous tissues, and can sustain the artery pulse pressure for 24 hours. It is better to suture the blood vessels by sleeve anastomosis.
【摘要】 目的 探討經鼻胃鏡在食管狹窄擴張和支架置入術中的作用。 方法 2007年10月-2009年3月對28例癌性食管狹窄以及并發食管-氣管瘺的患者行PENTAX（EG-1580K）超細經鼻電子胃鏡檢查，計算插入深度，并進行擴張和支架治療，在胃鏡直視下調節輸送器內支架上端的位置，觀察擴張效果、支架放置成功率、定位的準確性以及并發癥情況。 結果 全部患者均行擴張治療，效果良好，支架一次性放置全部成功，定位準確，自膨滿意，最狹窄處的內徑由（4.8±1.2）mm擴至（12.5±1.5）mm，食管氣管瘺被覆蓋治療效果好，患者的吞咽困難評級由3.25±0.58降至0.94±0.59。 結論 在食管狹窄擴張和支架置入術中應用經鼻超細胃鏡患者依從性好，方法簡便，安全有效。【Abstract】 Objective To explore the role of nasal endoscopy in the esophagostenosis expansion and esophageal stent. Methods PENTAX (EG-1580K) ultrafine nasal endoscopy was used in 28 patients (October 2007-March 2009) with esophageal cancer complicated with esophagostenosis and fistula to check the e-calculated insertion depth, stent expansion and the average diameter expansion. The endoscopy was carried under the direct vision with the location of the top bracket in order to ensure the accurate stent placement, all without X-ray assist. The success rates of stent placement, positioning accuracy, as well as complications were evaluated. Results The expansion treatments were successful in all patients with one-off operation. The narrowest part increased from (4.9±1.6) mm to (12.7±1.5) mm, and the esophageal fistula was covered. The rating of dysphagia decreased from 3.15±0.68 to 0.91±0.49. Conclusion Nasal endoscopy is simple, safe and effective in the treatment of ultrafine expansion for esophagostenosis and stent implantation with good compliance.
OBJECTIVE: To sum up the clinical results of bio-derived bone transplantation in orthopedics with tissue engineering technique. METHODS: From January 2000 to May 2002, 52 cases with various types of bone defect were treated with tissue engineered bone, which was constructed in vitro by allogeneous osteoblasts from periosteum (1 x 10(6)/ml) with bio-derived bone scaffold following 3 to 7 days co-culture. Among them, there were 7 cases of bone cyst, 22 cases of non-union or malunion of old fracture, 15 cases of fresh comminuted fracture of bone defect, 4 cases of spinal fracture and posterior route spinal fusion, 3 cases of bone implant of alveolar bone, 1 case of fusion of tarsotarsal joint. The total weight of tissue engineered bone was 349 g in all the cases, averaged 6.7 g in each case. RESULTS: All the cases were followed up after operation, averaged in 18.5 months. The wound in all the case healed by first intention, but 1 case with second intention. Bone union was completed within 3 to 4.5 months in 50 cases, but 2 cases of delayed union. Six cases were performed analysis of CD3, CD4, CD8, ICAM-1 and VCAM-1 before and after operation, and no obvious abnormities were observed. CONCLUSION: Bio-derived tissue engineered bone has good osteogenesis. No obvious rejection and other complications are observed in the clinical application.
Objective To locate sinoatrial node (SAN) in suckl ing pigs, to develop a rel iable method for isolation, purification and cultivation of SAN cells and to observe the compatibil ity of SAN cells and Col I fiber scaffold. Methods Five newborn purebred ChangBaiShan suckl ing pigs (male and female), aged less than 1-day-old and weighing 0.45-0.55 kg, wereused. Multi-channels electrophysiological recorder was appl ied to detect the original site of atrial waves. Primary SAN cells harvested from that area were cultured by the conventional culture method and the purification culture method including differential velocity adherent technique and 5-BrdU treatment, respectively. Atrial myocytes isolated from the left atrium underwent purified culture. Cell morphology, time of cell attachment, time of unicellular pulsation, and pulsation frequency were observed using inverted microscope. The purified cultured SAN cells (5 × 105 cells/mL) were co-cultured with prewetted Col I fiber scaffold for 5 days, and then the cells were observed by HE staining and scanning electron microscope (SEM). Results The atrial waves occurred firstly at the area of SAN. The purified cultured SAN cells were spindle, triangular, and irregular in morphology, and the spindle cells comprised the greatest proportion. Atrial myocytes were not spindle-shaped, but primarily triangular and irregular. The proportion of spindle cells in the conventional cultured SAN cells was decreased from 73.0% ± 2.9% in the purified cultured SAN cells, to 44.7% ± 2.3% (P lt; 0.01), and the proportion of irregular cells increased from 7.0% ± 1.7% in the purified cultrued SAN cells to 36.1% ± 2.6% (P lt; 0.01) . The proportion of the triangular cells in the purified and the conventional cultured SAN cells was 20.0% ± 2.1% and 19.2% ± 2.5%, respectively (P gt; 0.05). At 5 days after co-culture, HE staining displayed lots of SAN cells in Col I fiber scaffold, and SEM demonstrated conglobate adherence of the cells to the surface and lateral pore wall of scaffold, mutual connections of the cell processes, or attachment of cells to lateral pore wall of scaffold through pseudopodia. Conclusion With accurate SAN location, the purification culture method containing differential velocity adherent technique and 5-BrdU treatment can increase the proportion of spindle cells and is a rel iable method for the purification and cultivation of SAN cells. The SAN cells and Col I fiber scaffold have a good cellular compatibil ity.
目的：探討國產自膨式鎳鈦記憶合金食管支架在食管、賁門良惡性狹窄及食管瘺中的臨床應用并觀察療效。方法:收集我院2005年11月～2008年8月采用胃鏡輔助置入鎳鈦記憶合金食管支架的患者108例，其中男90例，女18例，年齡37~88歲；包括食管癌性狹窄41例，賁門癌性狹窄5例，放療后狹窄1例，肺癌壓迫致食管狹窄1例，食管、賁門癌術后吻合口頑固性狹窄21例，食管癌術后復發致狹窄6例，食管瘺33例。結果：108例患者共置入食管支架116枚，一次性置入成功率100%，置入后患者吞咽梗阻的癥狀有不同程度的改善，嗆咳的癥狀基本消失。結論：食管支架置入操作簡單、安全，成功率高，是治療食管良惡性狹窄及封堵瘺口的有效方法。
Objective To construct a new type of self-assembling peptide nanofiber scaffolds—RGDmx, and to study the cell compatibility of the new scaffolds and the proliferation and chondrogenic differentiation of precartilaginous stem cells(PSCs) in scaffolds. Methods PSCs were separated and purified from newborn Sprague Dawley rats by magnetic activated cell sorting and indentified by immunohistochemistry and immunofluorescent staining. The RGDmx were constructed by mixing KLD-12 and KLD-12-PRG at volume ratio of 1 ∶ 1. PSCs at passage 3 were seeded into the KLD-12 scaffold (control group) and RGDmx scaffold (experimental group). The proliferation of PSCs in 2 groups were observed with the method of cell counting kit (CCK) -8 after 1, 3, 7, and 14 days after culture. The RGDmx were constructed by mixing KLD-12-PRG and KLD-12 at different volume ratios of 0, 20%, 40%, 60%, 80%, and 100% and the prol iferation of PSCs was also observed. The complete chondrogenic medium (CCM) was used to induce chondrogenic differentiation of PSCs in different scaffolds. The differentiation of PSCs was observed by toluidine blue staining and RT-PCR assay. Results PSCs were separated and purified successfully, which were identified by immunohistochemistry and immunofluorescent staining methods. The results of CCK-8 showed that the absorbance (A) value in the experimental group increased gradually and reached the highest at 7 days; the A value in the experimental group was significantly higher than that in the control group at 7 days and 14 days (P lt; 0.05). Meanwhile, the A value in the RGDmx scaffold with a volume ratio of 40% was significantly higher than those in others (P lt; 0.05). After 14 days of induction culture with CCM, the toluidine blue staining results were positive in 2 groups; the results of RT-PCR showedthat the expression levels of collagen type II and the aggrecan in the experimental group were significantly higher than those in the control group (P lt; 0.05). Conclusion The self-assembling peptide nanofiber scaffold—RGDmx is an ideal scaffold for tissue engineer because it has good cell compatibility and more effective properties of promoting the differentiation of PSCs to chondrocytes.
Numerical simulation of stent deployment is very important to the surgical planning and risk assess of the interventional treatment for the cardio-cerebrovascular diseases. Our group developed a framework to deploy the braided stent and the stent graft virtually by finite element simulation. By using the framework, the whole process of the deployment of the flow diverter to treat a cerebral aneurysm was simulated, and the deformation of the parent artery and the distributions of the stress in the parent artery wall were investigated. The results provided some information to improve the intervention of cerebral aneurysm and optimize the design of the flow diverter. Furthermore, the whole process of the deployment of the stent graft to treat an aortic dissection was simulated, and the distributions of the stress in the aortic wall were investigated when the different oversize ratio of the stent graft was selected. The simulation results proved that the maximum stress located at the position where the bare metal ring touched the artery wall. The results also can be applied to improve the intervention of the aortic dissection and the design of the stent graft.
Objective To establish a model for studying on mechanical responses of osteoblasts seeded in 3 dimensional(3D) scaffold. Methods Fifty pieces of bioderived cancellous bones, whose holes were 500 to 800 μm and density was 0.36 to 0.45g/cm3, were obtained as the scaffolds. They were cultured with the third passage suspension of Wistar rat. Twenty-four of the 50 scaffolds were constructed under apparent strain sine waveform with amplitude of 1 000 με, frequency of 3 Hz, and duration of 3 min/d, as experimental group. The other scaffolds were control group. After 3day coculture, osteoblasts were observed with scanning electron microscope. The proliferation of the osteoblasts was checked by MTT on scheduled date. Results Scanning electron microscopic observation showed that osteoblasts ttached and spread on the trabeculae, which presented the validity of the model under proper mechanical condition. Experiment showed that mechanical environment promoted theproliferation of osteoblasts. The observation of proliferation of osteoblasts showed that the quantity of osteoblasts in the experimental group was higher than that in the control group 1,4,8,12,16,20,24, and 28 days after culturing. Therewas significant difference between the two groups 12,16,20,24,and 28 days afterculturing(P＜0.05). Conclusion The establishment of the model can facilitate the study of mechanical responses of osteoblasts under different conditions.
OBJECTIVE: To prepare chitosan-gelatin/hydroxyapatite (CS-Gel/HA) composite scaffolds, and to investigate the influence of components and preparing conditions to their micromorphology. METHODS: The CS-Gel/HA composite scaffolds were prepared by phase-separation method. Micromorphology and porosity were detected by using scanning electron microscope and liquid displacement method respectively. RESULTS: Porous CS-Gel/HA composite scaffolds could be prepared by phase-separation method, and their density and porosity could be controlled by adjusting components and quenching temperature. CONCLUSION: The study suggests the feasibility of using CS-Gel/HA composite scaffolds for the transplantation of autogenous osteoblasts to regenerate bone tissue.