Objective To investigate whether the implanted myoblasts with the soluble carriers can improve the repairing efficiency for theseverelycryodamaged tibialis anterior muscles. Methods The skeletal myoblasts were isolated from the newborn SD rats by the use of the enzyme digestion. They were purified and serially subcultivated; the subcultivated myoblasts of the 3rd generation were marked with BrdU. The severelycryodamaged tibialis anterior muscle models were established from 84 SD rats aged 5 months. They were randomly divided into 4 groups, including Group A1 (the implanted myoblasts with the carriersF12 containing 0.1% sodium hyaluronate), Group A2 (the implanted myoblasts, with the carriersF12 that did not contain 0.1% sodiumhyaluronate), Group B1 (the implanted carrier solution containing 0.1% sodium hyaluronate, but with no myoblasts), and Group B2 (with no carrier solution or myoblasts). Six rats were killed at the following time points: at 2, 5 and 9 days,and 2, 4, 8 and 12 weeks after operation; the immunohistochemical and the Mallory staining studies were performed for an evaluation on the repairing efficiencyfor the severelycryodamaged tibialis anterior muscles. By the imaging analysis, the number of the survived cells in each group was compared at 2 days, and the area ratio of the collagen fiber in each group was also compared at 8 weeks. Results The BrdU immunohistochemical staining showed that the number of the remaining implanted cells was significantly greater in Groups A1 than in Group A2, the migrating area of the myoblasts was greater, the distribution of the cells was more uniform, the cell differentiating potential was undestroyed, the repairing efficiency for the severelycryodamaged tibialis anterior muscles was significantly improved. There was no bluestained nucleus at each time point in Group B. The Mallory staining showed that the fibrous degeneration inthe tissue repairing process was significantly inhibited in Groups A1, A2 and B1; the inhibition was most obvious in Group A1, and next in Group A2. The imaging analysis indicated that at 2 days after operation, the number of the survived cells was significantly-greater in Group A1 than in Group A2 (Plt;0.05). At 8 weeks after operation, the collagen fiber was the least in Group A1, less in Group A2, more in Group B1,and the most in Group B2 (Plt;0.05). Conclusion The implanted myoblasts can significantly improve the repairing efficiency for the severelycryodamaged muscle tissues, and the implanted carrier solution containing 0.1% sodium hyaluronate can improve the implanting efficiency for the myoblasts.
ObjectivesTo systematically review the patients’ satisfaction of implant supported mandibular dentures and conventional dentures.MethodsPubMed, EMbase, The Cochrane Library, CNKI, WanFang and VIP databases were searched to collect randomized controlled trials (RCTs) on patients’ satisfaction of implant supported mandibular dentures (IODs) and conventional dentures (CDs) from inception to November 31st, 2016. Two reviewers independently screened literature, extracted data and evaluated the risk of bias of the included studies. Then, meta-analysis was conducted using RevMan 5.3 software.ResultsSeven randomized-controlled trials were identified. The results of meta-analysis showed that compared with CDs group, IODs group had a relatively higher level of patients’ satisfaction (SMD=1.11, 95%CI 0.79 to 1.43, P<0.001) and oral health quality of life (MD=–12.41, 95%CI –18.96 to –5.86,P<0.001).ConclusionsThe current evidence suggests that IODs may be a better choice for mandibular edentulous patients than CDs. Due to the limitations of the quality and quantity of the included studies, the above conclusions still require larger sample and high quality research to verify.
Objective To investigate the influence of infarcted myocardial construction by umbilical cord blood mesenehymal stem cells（MSC） with induced to myo-derived stem cells and implanted into infarcted myocardium. Methods Thirty-six adult mongrel canines were randomly divided into MSC transplant group and control group （18 each group）. Transplant group： the umbilical cord blood MSC differented to myo-derived stem cells induced by 5-azacytidine（5-aza） were implanted into the acute myocardial infarct site via the ligated left anterior descending （LAD） artery. Control group： administer the same volume of IMDM culture medium containing 0. 02% 4,6- diamidino-2-phenylindone （DAPI）. At 2, 4, 8 weeks after implantation, the change of basic myocardial construction and the distribution of desmin were observed by using Nagar-Olsen staining and immunohistology respectively. Results With less fusing, the arrangement of gelatine fibers and elastic fibers were in order in transplant group,and they were partly fused in control group by Nagar-Olsen staining. The expression of desmin of infarcted myocardial cell in transplant group was much higher than those in control group （P〈0. 01）. No significant difference was detected in the expression of desmin in normal site of both groups （P〉0. 05）. Conclusion There is an protective effect on the basic myocardial construction in infarcted myocardium after the umbilical cord blood MSC was differented to myo-derived stem cell by induced with 5-aza in vitro and implanted into the acute myocardial infarction.
Objective To explore the effective autologous bone marrow stem cell dosage for treatment of severe lower limb ischemia. Methods From December 2003 to December 2004, 22 cases of bilateral lower limb ischemia were treated with autologous bone morrow cell transplantation. All the patients were randomly divided into two groups according to ischemia degree. In group A(severe ischemia side), the amount of transplanted autologous bone marrow cells was more than 1×108, and ingroup B(mild ischemia side), the amount was less than 1×105. A series of subjective indexes, such as improvement of pain, cold sensation and numbness, and objective indexes, such as increase of ankle/brachial index (ABI) and transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing were used to evaluate the effect of autologous bone marrow stem cells implantation. Results The rates of pain relief were 90.0% in group A and 16.7% in group B (Plt;0.01); the rates of cold sensation relief were 90.5% in group A and 5.3% in group B（Plt;0.01）;the improvement of numbness was 62.5% in group A and 9.1% in group B（Plt;0.01）. Increase of ABI was 31.8% and 0 in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Increase of TcPO2was 94.4% and 11.1% in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Twelve cases of angiography showed rich new collateral vessels in 100% of the limbs in group A while no remarkable new collateral vessel in group B. The amputation rates were 4.5% in group A and 27.3% in group B（Plt;0.05） at 4 weeks after implantation. The rate of improvement of foot wound healing was 75% in group A and there was no changein wound healing in group B after 4 weeks of implantation. Conclusion The effectiveness of autologous bone marrow stem cell implantation depends on the number of implanted stem cells. Effectiveness is expected in most patients if the implanted stem cell is more than 1×108, whereas there would be little effect if the cell number is less than 1×105.
Objective To investigate the possibility of repairing defected tendon with a tissue engineering tendon, combined culture of allogenous tenocyte and derived tendon. Methods Macaca tenocytes labelled by BrdU were seeded on the derived tendon. The flexor digitorum profundus of five fingers of left hand in 15 Macaca mulatta were resected and made 2.5cm defects as experimental model. They were divided into three groups according to repair methods (Group A: Combined culture of derived tendon materials and alloggenous tendon cells; Group B; Derived tendon materials; Group C; Autograft). In different stages, the labeled BrdU of tendon cells were observed. Results In Groupo A, after iin vivo implantation, the tenocytes could proliferate and synthesize collagen; the new tissue was white and glossy and the collagen fibers fused to form dense tendon structure as several weeks passed. Twelve weeks after implantation, the tenocytes still survived and synthesized collagen, the results of labelled cells were positive by immunothistochemical methods. By scanning electron microscopic observation, the tenocytes arraged regularly and evely among the derived tendon; the collagen fibers formed a network and its main direction was accord with that of the derived tendon. Normal nucleus, nucleolus, and cell organelles were seen under transmission electron microscope. Conclusion Combined culture of tenocytes with derived tendon is able to make tendon like tissue. The structure of tissue engineering tendon in similar to that of normal tendon.
The aim of this experiment was to study the osteogenesis in vivo of allogenic osteoblast combined culture with calcium phosphate composites. The osteoblasts were obtained by enzymatic digestion of periosteum from fibula subcultured to 13 generations, the cells were combined culture with hydroxyapatite and biphasic calcium phosphate. Subseguently, the composite was implanted into rabbits subcutaneously or intramuscularly. The blank material was implanted in the contralateral side as control. Four weeks later, all animals were sacrificed. All the implants were examined by gross observation, histological examination and EDXA. The results showed: 1. obvious ingrowth of connective tissue with very little inflammatory reaction; 2. new bone formation in the composites with deposit of Ca and P on the surface of osteoblast, but none in the blank materials; 3. no significant difference of new bone formation between the different sites of implantation or different materials, but those implanted intramuscularly had lamellae form of new bone while those implanted subcutaneously had only mineralization of extracellular matrix. The conclusion were: 1. the composites are biocompatible with prior osteogenesis property; 2. periosteal-derived allogenic osteoblasts obatined by enzymatic digestion could survive following implantation with bioactivity; 3. rich blood supply might be advantageous to new bone formation and its maturation.
Objective To evaluate the clinical result of using osseointegratedimplants to retain removable orbital prostheses in repairing orbital defects. Methods Two patients with orbital defects caused by orbitaltumor were treated. Each of them got 4 implants. After average 6 months, we performed the secondary operation. After 7 weeks, we took impressions to make the implant-retained prostheses. The magnetic attachment was adopted. The prostheses were made of polysiloxane material(Factor II,Lakeside,Ariz). Results Both of the patients got the successful facial prostheses and were followed up 2 and 8 years respectively. All the implants were integrated well. There were no apparent inflammatory reactions in the soft tissue around percutaneous implants. The patients were satisfied with the facial appearance. Conclusion Implant-retained orbital prostheses are safe and effective in repairing orbital defects.
ObjectiveTo explore the antitumor effect of tumor vaccine fused from dendritic cells (DC) and Walker-256 cancer cells on implanted liver cancer in rats and the related mechanism of inhibition for tumor angiogenesis. MethodsWalker-256 cancer cells and mature DC were fused by 50% polyethylene glycol method for preparation of DC-Walker-256 fusion vaccines. Implanted liver cancer models were established through operations on healthy male SD rats at the age of 6－8 weeks. All the rats were divided into four groups, and rats in each group were injected subcutanely with fusion vaccine (group), mixed cultured cells (group), simple DC (group), and PBS (blank control group), respectively. On 28 d after making model, the rats were put to death, the tumor was observed and pathological essays were prepared. All rats’ spleens were collected and prepared into lymphocyte to detect antigenic specificity cytotoxic T lymphocyte (CTL) by enzymelinked immunosorbent spot (ELISPOT) method. The expressions of VEGF, ANG-1, ANG-2, and MVD were detected by immunohistochemistry. ResultsThe numbers of rats survived in the fusion vaccine group, mixed culture cells group, simple DC group, and blank control group was 8, 5, 6, and 3, respectively. The rats in the other three groups except for fusion vaccine group were manifested as inaction, anorexia, and gloomy fur in some degree as well as ascites. The tumorigenesis was found in all survival rats except for two in the fusion vaccine group. The weight of liver tumors of rats in the fusion vaccine group 〔(32.4±9.2) g〕 was significantly lighter than that in the mixed culture cells group 〔(67.3±5.1) g, P=0.031〕, simple DC group 〔(75.0±8.3) g, P=0.019〕, and blank control group 〔(86.6±10.5) g, P=0.008〕, respectively. The number of tumorspecific CTL of rats in the fusion vaccine group was also significantly higher than that in the other three groups (P=0.019, P=0.025, and P=0.001, respectively). The MVD of tumor tissue in the fusion vaccine group was (24.12±2.32) vessels/HP, which was significantly lower than that in the mixed culture cells group 〔(40.34±1.29) vessels/HP, P=0.025〕, simple DC group 〔(42.36±3.16) vessels/HP, P=0.035〕, and blank control group 〔(56.48±5.16) vessels/HP, P=0.006〕, respectively. The MVD of tumor tissue in the mixed cultured cells group and simple DC group was similar (P=0.165), however, which was significantly lower than that in the blank control group (P=0.040 and P=0.043). The positive rate of VEGFA protein expression was 23.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (42.5%, P=0.031), simple DC group (61.3%, P=0.019), and blank control group (89.6%, P=0.003), respectively. The positive rate of VEGF-A protein expression in the mixed cultured cells and simple DC groups was similar (P=0.089), however, which was significantly lower than that in the blank control group (P=0.027 and P=0.038). The positive rate of ANG-1 protein expression in the fusion vaccine group (43.2%) was not different from that in the mixed culture cells group (46.3%, P=0.292), simple DC group (51.3%, P=0.183), or blank control group (49.6%, P=0.179), respectively, and the difference of pairwise comparison in latter three groups was not significant (P=0.242, P=0.347, and P=0.182). The positive rate of ANG2 protein expression was 19.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (62.3%, P=0.007), simple DC group (67.3%, P=0.005), and blank control group (71.6%, P=0.004), respectively, however, the difference of pairwise comparison in latter three groups was not significant (P=0.634, P=0.483, and P=0.379). ConclusionFused vaccine can induce CD8+ CTL aiming at tumor cells and establish the effective antitumor immunity in vivo and also downregulate the level of VEGF and ANG-2 to suppress tumor angiogenesis and thereby achieve the purpose of curing tumor.
Objective To investigate the effect of iodine-125 on inhibiting breast cancer growth and to explore its possible mechanism. Methods The animal model of the MCF-7 tumor was established firstly through injection of the cells into nude mice. The animals were divided into two groups before the implantation of the iodine-125 granule into the tumor mass: control group (n=40, to implant no-load seeds, non-iodine-125 radioelement) and experimental group 〔n=40, to implant iodine-125 seeds (1.48×107 Bq) when the length of tumor was 8-10 mm〕. The width and length of tumor, in order to calculate the volume, were measured every three days to observe tumor growth curve and to calculate the rate of the tumor inhibition. When the length of tumor was 15-20 mm in the control group, 30 nude mice were killed in every group to detect the weight of tumor and histopathological changes. Other ten nude mice of each group were remained to be observed the national life span and survival rate for 90 days. Results Within 90 days, the average survival time in control group and experimental group were significantly different (56.2 d vs. 74.8 d, P＜0.05). In control group the growth curve was continuously elevated, while experimental group showed a low flat curve. With iodine-125 treatment, the tumor growth decreased in experimental group with tumor inhibition rate 55.21%. The average tumor weight in control group and experimental group was (3.26±0.39) g and (1.46±0.17) g (t′=22.8962, P＜0.05). As compared with control group, under light microscope, the number of cancer cells was less, nuclear debris increased, and cancer structure was not obvious in experimental group. Conclusion This study suggests that iodine-125 seed may inhibit the growth of breast cancer, which may be involved in direct radiation breakdown of tumor cells or induction of apoptosis and inhabitation of tumor angiogenesis.
Objective To investigate the biocompatibility of acellular urinary bladder submucosa (AUBS). Methods The acellular collagen matrix of human urinary bladder submucosa was developed using freeze-thawed enzymatic treatment and freeze-drying technique. Human oral keratinocytes were cultured and seeded on AUBS at a density of 2×106/ml in vitro.The proliferation of the cells were observed. Pockets were created in the abdominal muscle wall of 18 SD rats. AUBS in size 1 cm×1 cm was implanted into the pocket. The grafts were observed by light microscope 3, 6, 10, 14, 21 and 28 days after operation. Results AUBSmainly consisted of collagen fibers with a three-dimensional network structure. After the oral keratinocytes were seeded, continous oral epithelium layer was formed on the surface of AUBS after 10 days in vitro. Histological observation of the grafted AUBS showed progressive cell infiltration at 6 days. New capillaries formed at 14 days. The collagen fibers arranged regularly at 28 days after implantation. Conclusion Freeze-dried AUBS may be used as a suitable scaffold for tissue regeneration, which can induce cell proliferation both in vivo and in vitro and has good biocompatibilty.