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1.
In this study, the bioactivity of a novel BMP2-derived oligopeptide P24 was investigated by using the model of rabbit femoral defect after loaded in the biodegradable poly (lactic acid / glycolic acid / asparagic acid-co-polyethylene glycol) (PLGA-[ASP-PEG]). A 1.5-cm unilateral segmental bone defect was created in the left femoral diaphysis in each of the 30 new zealand white rabbits. The defects of 18 legs filled with BMP2-derived peptide P24 combined with PLGA-[ASP-PEG] scaffold serves as the experimental group, and the defects in the rest 12 rabbits filled with (PLGA-[ASP-PEG]) without P24 as control group. The bone-repairing capability in the target region of the two group was grossly, radiologically, histopathologically and biomechanically evaluated 4, 8 and 12 weeks after the operation. Our results showed that in each group, primary healing of incision was achieved in the two groups. Radiographically, in experimental group, defects were filled with induced callus within 8 weeks, and a cortical bone-like structure was observed in some animals at the 12th week. According to the standardized stage of bone defect repair, 9 (64.28%) achieved grade-4 healing. In contrast, little bone formation was seen in the defects even 12 weeks after the operation, and 5 (62.50%) had grade 0 healing in this group. Histologically, tissue engineering material was mostly absorbed and cartilage was found around implants in the experimental group at the 4th week; 8 weeks after operation, the engineering material was completely absorbed, and formation of woven bone was observed and typical trabecular bone structure could be seen. In control group, 8 weeks after operation, the defect was filled with fibrous tissues, and no bone-like structure was observed. Statistical analysis showed very significant difference in biomechanical indicators between the two groups (P〈0.05). It is concluded that new oligopeptide P24 can induce excellent bone regeneration and promote bone repair.  相似文献   

2.
Objective:To assess the effect of puerarin,a natural flavonoid found in Chinese Pueraria Lobata (Wild.) Ohwi,on promotion of new bone formation.Methods:Osteoblasts isolated from calvarial of newborn rats were cultured in vitro in the presence of puerarin at various concentrations.The viability of osteoblasts and alkaline phosphotase activity and mineral node formation were determined.In addition,osteoblasts seeded in theβ-tricaclium phosphate scalfolds as bone substitute were implanted in rat dorsal muscles.Half of the recipient rats received intramuscular injection of puerarin at 10 mg/(kg·d) for 7 days.Osteogenesis was analyzed by examining the histology after 4 weeks of implantation.Results:The viability of osteoblasts treated with puerarin at either 40 or 80μmol/L was significantly higher than that of the control(P<0.05 and P<0.01, respectively).Alkaline phosphatase and mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 mol/L when compared with that of the untreated cells.The puerarin-treated rats had a higher rate of bone formation in the osteoblast implants than the control rats(6.35%vs.1.32%,respectively, P<0.05).Conclusion:Puerarin was able to affect osteoblast proliferation and differentiation,and promote the new bone formation in osteoblast implants.  相似文献   

3.
Chen G  Hu YR  Wan H  Xia L  Li JH  Yang F  Qu X  Wang SG  Wang ZC 《中华医学杂志(英文版)》2010,123(17):2424-2431
Background The most important objective of transplant studies in the injured spinal cord has been to provide a favorable environment for axonal growth. Moreover, the continuing discovery of new grafts is providing new potentially interesting transplant candidates. Our purpose was to observe the morphological and functional repair effects of the co-transplantation of neural stem cell (NSC), Schwann ceils (SCs) and poly lactide-co-glycolide acid (PLGA) on the spinal cord injury of rats.Methods A scaffold of PLGA was fabricated. NSCs and SCs were cultured, with the NSCs labeled with 5-bromodeoxyuridine, and the complex of NSC/PLGA or NSC+SCs/PLGA were constructed. Thirty-six Wistar rats were randomly divided into three groups: group A (transplantation of PLGA), group B (transplantation of NSC/PLGA) and group C (transplantation of NSC+SCs/PLGA). The 3 mm length of the right hemicord was removed under the microscope in all rats. The PLGA or the complex of PLGA-celIs were implanted into the injury site. Basso-Beattie-Bresnahan (BBB)locomotion scores, motor and somatosensory evoked potential of lower limbs were examined to learn the rehabilitation of sensory and motor function at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. All the recovered spinal cord injury (SCI) tissues were observed with HE staining, immunohistochemistry, and transelectronmicroscopy to identify the survival, migration and differentiation of the transplanted cells and the regeneration of neural fibres at 4 weeks, 8 weeks,12 weeks and 24 weeks after injury.Results (1) From 4 weeks to 24 weeks after injury, the BBB locomotion scores of cell-transplanted groups were better than those of the non-cell-transplanted group, especially group C (P 〈0.05). The amplitudes of the somatosensory evoked potential (SEP) and motor-evoked potential (MEP) were improved after injury in groups B and C, but the amplitude of SEP and MEP at 4 weeks was lower than that at 12 weeks and 24 weeks after injury. Compared with group B, the amplitude of SEP and MEP in group C was improved. The amplitude of SEP and MEP was not improved after injury in group A. (2) HE staining revealed the volume of the scaffold decreased and the number of cells in the scaffold increased. Newly-grown capillaries also could be seen. Immunohistochemistry staining showed the transplanted NSCs could survive and migrate until 24 weeks and they could differentiate into neurons and oligodendrocytes. The regenerated axons were observed in the scaffold-cell complex with transelectronmicroscopy. The above manifestations were more extensive in group C.Conclusions The transplanted NSC can survive and migrate in the spinal cord of rats up to 24 weeks after injury, and they can differentiate into various neural cells. Co-transplantation of cells/PLGA can promote the functional recovery of the injured spinal cord. The effect of co-transplanting NSC+SCs/PLGA is better than transplanting NSC/PLGA alone.  相似文献   

4.
Objective To study the biocompatbility of PLGA with tissue cell after implantation into the injured spinal cord of rat. Methods Schwann eels and neural stem cells were harvested and cultured in vitro, Then we constructed PLGA-eell complex and implanted the complex into the injured site of T9 SCI models developed from Wistar rats. Then we observed the degradation of the porous scaffold and the growth of tissue eels in the scaffold through electron microscope at different time. Results The PLGA in the scaffold-cells complex gradually degenerated under scanning electron microscope.[第一段]  相似文献   

5.
《中华医学杂志(英文版)》2011,124(19):3048-3048

Background  The most important objective of transplant studies in the injured spinal cord has been to provide a favorable environment for axonal growth. Moreover, the continuing discovery of new grafts is providing new potentially interesting transplant candidates. Our purpose was to observe the morphological and functional repair effects of the co-transplantation of neural stem cell (NSC), Schwann cells (SCs) and poly lactide-co-glycolide acid (PLGA) on the spinal cord injury of rats.

Methods  A scaffold of PLGA was fabricated. NSCs and SCs were cultured, with the NSCs labeled with 5-bromodeoxyuridine, and the complex of NSC/PLGA or NSC+SCs/PLGA were constructed. Thirty-six Wistar rats were randomly divided into three groups: group A (transplantation of PLGA), group B (transplantation of NSC/PLGA) and group C (transplantation of NSC+SCs/PLGA). The 3 mm length of the right hemicord was removed under the microscope in all rats. The PLGA or the complex of PLGA-cells were implanted into the injury site. Basso-Beattie-Bresnahan (BBB) locomotion scores, motor and somatosensory evoked potential of lower limbs were examined to learn the rehabilitation of sensory and motor function at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. All the recovered spinal cord injury (SCI) tissues were observed with HE staining, immunohistochemistry, and transelectronmicroscopy to identify the survival, migration and differentiation of the transplanted cells and the regeneration of neural fibres at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury.

Results  (1) From 4 weeks to 24 weeks after injury, the BBB locomotion scores of cell-transplanted groups were better than those of the non-cell-transplanted group, especially group C (P <0.05). The amplitudes of the somatosensory evoked potential (SEP) and motor-evoked potential (MEP) were improved after injury in groups B and C, but the amplitude of SEP and MEP at 4 weeks was lower than that at 12 weeks and 24 weeks after injury. Compared with group B, the amplitude of SEP and MEP in group C was improved. The amplitude of SEP and MEP was not improved after injury in group A. (2) HE staining revealed the volume of the scaffold decreased and the number of cells in the scaffold increased. Newly-grown capillaries also could be seen. Immunohistochemistry staining showed the transplanted NSCs could survive and migrate until 24 weeks and they could differentiate into neurons and oligodendrocytes. The regenerated axons were observed in the scaffold-cell complex with transelectronmicroscopy. The above manifestations were more extensive in group C.

Conclusions  The transplanted NSC can survive and migrate in the spinal cord of rats up to 24 weeks after injury, and they can differentiate into various neural cells. Co-transplantation of cells/PLGA can promote the functional recovery of the injured spinal cord. The effect of co-transplanting NSC+SCs/PLGA is better than transplanting NSC/PLGA alone.

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6.
Objective: Research on enhancing early osseointegration of cementless implants to improve early fixation and reducing of risk of loosening. Methods : Thirty New Zealand rabbits were divided into two groups at random. BMP combined with DBM 30 mg was inserted around the prosthesis in 15 rabbits as experimental group, the remaining rabbits were served as control group. After 4, 8, and 12 weeks, five rabbits were sacrificed in each group. The humerus with the implants were retrieved. Bone ingrowth was analyzed by none-decalcification bone ground section and biomechanical test. Results: At the end of 4 and 8 weeks the osseointegration rates of BMP group were higher than those of control (P 〈 0.05). The ultimate shear strength between BMP treated implantation and the control was the same as the results in osseointegration rates at 4, 8 weeks (P 〈 0.05). However, there was no difference between the treated and untreated group in the osseointegration rate and ultimate shear strength at 12 weeks (P 〉 0.05). Conclusion: BMP combination can enhance bone growth into gaps around cementless implants, especially in the early postoperative period.  相似文献   

7.
Background Tissue engineering techniques combined with gene therapy have been recently used to improve osteogenesis. NEL-like molecule-1 (Nell-1), a novel growth factor, has been reported to have specificity for osteochondral lineage. The study assessed the osteogenic differentiation of rat bone marrow stromal cells (bMSCs) after Nell-1 gene modification and examined its ectopic bone formation ability in a nude mice model with tissue engineering technique. Methods bMSCs obtained from Fischer 344 rats were transduced with either AdNell-1 (Nell-1 group) or Ad-β-galactosidase (AdLacZ, LacZ group) or left untransduced (untransduced group). The expression of Nell-1 protein was determined by Western blotting and transfer efficiency was assessed, mRNA expressions of osteopontin (OP), bone sialoprotein (BSP) and osteocalcin (OC) were assessed by real-time PCR 0, 3, 7, 14, and 21 days after gene transfer. Alkaline phosphatase (ALP) activity was measured and von Kossa test was also conducted. Finally, with a tissue engineering technique, gene transduced bMSCs, combining with β-tricalcium phosphate (β-TCP) at a concentration of 2×10^7 cells/ml, were implanted at subcutaneous sites on the back of nude mice. Four weeks after surgery, the implants were evaluated with histological staining and computerized analysis of new bone formation. Results Under current transduction conditions, gene transfer efficiency reached (57.9±6.8)%. Nell-1 protein was detected in Nell-1 group but not in untransduced group and LacZ group. Induced by Nell-1, BSP and OP expression were increased at intermediate stage and OC expression was increased at later stage. ALP activity and the number of calcium nodules were highest in Nell-1 group. Four weeks after implanted into nude mice subcutaneously, the percentage of new bone area in Nell-1 group was (18.1±5.0)%, significantly higher than those of untransduced group (11.3±3.2)% and LacZ group (12.3±3.1)% (P〈0.05). Conclusions This study has demonstrated the ability of Nell-1 to induce osteogenic differentiation of rat bMSCs in vitro and to enhance bone formation with a tissue engineering technique. The results suggest that Nell-1 may be a potential osteogenic gene to be used in bone tissue engineering.  相似文献   

8.
Background A practical problem impeding clinical translation is the limited bone formation seen in artificial bone grafts.Low-pressure/vacuum seeding and dynamic culturing in bioreactors have led to a greater penetration into the scaffolds,enhanced production of bone marrow cells,and improved tissue-engineered bone formation.The goal of this study was to promote more extensive bone formation in the composites of porous ceramics and bone marrow stromal cells (BMSCs).Methods BMSCs/β-tricalcium phosphate (β-TCP) composites were subcultured for 2 weeks and then subcutaneously implanted into syngeneic rats that were split into a low-intensity pulsed ultrasound (LIPUS) treatment group and a control group.These implants were harvested at 5,10,25,and 50 days after implantation.The samples were then biomechanically tested and analyzed for alkaline phosphate (ALP) activity and osteocalcin (OCN) content and were also observed by light microscopy.Results The levels of ALP activity and OCN content in the composites were significantly higher in the LIPUS group than in the control group.Histomorphometric analysis revealed a greater degree of soft tissue repair,increased blood flow,better angiogenesis,and more extensive bone formation in the LIPUS groups than in the controls.No significant difference in the compressive strength was found between the two groups.Conclusion LIPUS treatment appears to enhance bone formation and angiogenesis in the BMSCs/β3-TCP composites.  相似文献   

9.
10.
Summary: To study the angiogenic potency of hypoxia-prestimulated bone marrow stromal cells(BMSCs) when transplanted into acute myocardial infarction models of rats. BMSCs were cultured under hypoxia condition for 24 h. Their expression of VEGF was investigated. The rat acute myocardial infarction models were made by coronary artery ligation and divided into 3 groups at random.In normoxia group, twice-passaged BMSCs were labeled with Bromodeoxyuridine (BrdU) and then implanted into the infarction regions and ischemic border of the recipients in 4 weeks. The rats in hypoxia group were implanted with hypoxia-prestimulated BMSCs. In control group, the model rats received only DMEM medium injection. Six-weeks after AMI, the infarction regions were examined to identify the angiogenesis and the expression of the VEGF. Our results showed that viable cells labeled with BrdU could be identified in the host hearts. The infarction regions in normoxia and hypoxia groups had a greater capillary density and increased VEGF expression than the regions in control group. The capillary density and VEGF expression in hypoxia group were higher than in normoxia group. It is concluded that the enhanced expression of VEGF in BMSCs could be induced by ex vivo hypoxia stimulation. BMSCs implantation promoted the angiogenesis in myocardial infarction tissue via supplying exogenic VEGF. Angiogenic potency of bone marrow stromal cells was improved by ex vivo hypoxia prestimulation though the enhanced VEGF expression.  相似文献   

11.
目的研究聚乳酸聚乙醇酸复合物( Po ( yclactic - co - glycolic acid ), PLGA )支架作为组织工程骨支架修复骨缺损的可行性。方法体外培养兔骨髓间充质干细胞(bone marrow stromal cells, BMSCs),在成骨诱导荆地塞米松等的诱导下,向成骨细胞转化,并使之与修饰后PLGA支架复合,植入兔桡骨缺损模型,作为实验组。对照组a为PLGA材料对照,对照组b为空白对照,通过影像学观察骨缺损的修复情况。结果地塞米松等诱导组细胞形态向类成骨细胞转化,碱性磷酸酶表达明显增高,并表达I型胶原。应用多聚赖氨酸修饰的PLGA与成骨诱导的BMSCs复合植入骨缺损后,与对照组相比影像学显示明显促进骨缺损的愈合。结论适当浓度成骨诱导剂可成功的将免骨髓间充质细胞向成骨细胞诱导,诱导后细胞与PLGA支架联合应用可较好修复兔桡骨缺损。  相似文献   

12.
负载BMP的新型组织工程骨的构建及骨缺损修复实验   总被引:2,自引:0,他引:2  
OBJECTIVE: To construct a new tissue-engineered bone with poly (D, L-lactide-co-glycolide) (PLGA), bone morphogenetic protein (BMP) and bone marrow-derived stem cells (BMSCs) and observe its effect in repairing segmental bone defects. METHODS: A 15-mm bone defect in the right radius was induced in New Zealand white rabbits, and the models were randomized into three groups to receive implantation of the tissue-engineered bone grafts constructed with PLGA carrying 5 mg BMP and about 1 x 10(6) BMSCs (experimental group), grafts of PLGA with about 1 x 10(6) BMSCs (control group), or grafts of exclusive PLGA (blank control group), respectively. The osteogenesis in the bone defect after the implantation on was evaluated X-ray films, and the histological changes of the tissues sampled from the bone defect 4, 8, and 12 weeks after operation were observed and new bone formation was measured by image analysis. RESULTS: The bone defect was completely repaired in the experimental group 12 weeks after the implantation, showing the best results among the 3 groups. The bone defects in the blank control group was filled with only fibrous and connective tissues at 12 weeks. CONCLUSION: This tissue-engineered bone constructed with PLGA, BMP and BMSCs possesses good ability in repairing segmental bone defect.  相似文献   

13.
目的 探讨以聚乳乙醇酸(poly-(D,L-lactide-co-glycolide),PLGA)、骨形态发生蛋白(BMP)、骨髓基质干细胞(BM-SCs)构建成新型组织工程骨并观察其在动物体内的成骨能力。方法 制作新西兰大白兔右桡骨中段15mm骨缺损实验模型,随机分为实验组、对照组和空白组,实验组植入同时负载5mgBMP及1×106个已向成骨细胞诱导的BMSCs的PLGA、对照组植入负载1×106个已向成骨细胞诱导的BMSCs的PLGA、空白组仅植入PLGA。术后对动物进行大体观察、摄X线片观察各组不同时相骨缺损修复情况、比较不同时相的骨缺损区X线阻射密度、并于术后第4、8、12周取出骨缺损区标本进行大体观察和组织学切片观察,图像分析骨小梁的生成数量。结果 实验组在12周内骨缺损完全修复,且同时期内新生骨的数量和质量显著优于对照组,空白材料组骨缺损主要由纤维结缔组织填充。结论 利用含BMP的PLGA支架与BMSCs复合构建的新型组织工程骨具有良好的骨缺损修复能力。  相似文献   

14.
目的:制备兔脱钙骨基质(DBM),研究复合骨髓间充质干细胞(BMSCs)的DBM的异位成骨能力。方法:制备DBM材料,将体外培养的ICR小鼠BMSCs与DBM复合培养后构建的组织工程骨植入ICR小鼠股部肌袋中,以单纯DBM植入组为对照,术后7d及28d行X线检测,并于术后7,14,28d取植入材料作组织学检测、钙含量测定。结果:DBM与BMSCs复合体植入后7d即有成骨细胞出现,随着时间增加成骨细胞增多,术后28d即有较成熟骨质形成,移植物钙含量随时间推移逐渐增高。单纯植入DBM术后14d出现少量成骨细胞,术后28d出现少量类骨质,钙含量持续处于较低水平。结论:DBM与BMSCs复合后具有很好的异位成骨能力,DBM是组织工程骨研究较理想的支架材料。  相似文献   

15.
骨形态发生蛋白-2基因治疗对假体-骨界面影响的实验研究   总被引:6,自引:0,他引:6  
Yan MN  Tang TT  Zhu ZA  Zhou XS  Jia QW  Yu CF  Lou JR  Dai KR 《中华医学杂志》2005,85(22):1521-1525
目的观察假体周围骨缺损重建中,骨形态发生蛋白(BMP)2基因治疗对假体骨界面骨整合的影响。方法14条成年Beagle犬,于双侧股骨外髁造成横向骨缺损,植入光滑面钛合金假体后保持假体周围3mm骨缺损。共28侧分成4组:空白对照组(2侧),缺损区未行处理;其余分3组为无细胞组(8侧)、细胞组(8侧)和基因组(10侧),采用压缩植骨技术重建股骨髁假体周围骨缺损,分别植入犬异体冻干骨、复合自体骨髓基质干细胞的冻干骨以及复合转BMP2基因自体骨髓基质干细胞的冻干骨。通过组织学、组织形态计量学及生物力学评估假体骨界面的愈合和整合。结果术后6周,基因组假体表面明显较多的新骨沉积,可见散在的假体与新骨间点状接触,假体骨接触率(BIC)达10%左右,而空白对照组、无细胞组和细胞组界面为厚薄不一的软组织,BIC为0;12周时,空白对照组界面仍是较厚的软组织,无细胞组和细胞组的界面主要为结缔组织纤维膜,少量点状骨接触,BIC均未超过10%,基因组的假体骨界面主要为骨组织,假体骨界面可见连续性骨接触,部分BIC达50%,远高于前两组(39.2±7.5比8.4±1.3、7.2±1.5,均P<0.01)。各组的界面推出强度随时间增加,基因组的强度在各个时间段均远高于前两组(1.40±0.22比0.09±0.04、0.08±0.04,均P<0.01)。结论BMP2基因治疗可明显提高假体骨界面的骨整合。  相似文献   

16.
目的:观察自制CPC/BMP复合人工骨对兔实验性股骨头缺血性坏死(ANFH)的修复作用,并探讨其临床应用的可行性。方法:24只健康成年兔分成CPC组12只和CPC/BMP组12只,在兔左侧股骨头建立ANFH骨缺损模型。将CPC作为BMP的载体制备成CPC/BMP复合材料后,植入骨缺损处,同时设立单纯CPC填充对照组。术后3及12周分批处死动物,每组每次处死6只。通过大体观察、X线摄片、HE染色、透射电镜等手段观察新骨形成和材料降解情况。综合评价CPC/BMP复合材料对ANFH骨缺损的修复能力。结果:CPC/BMP组3及12周时成骨情况明显优于单纯CPC组。CPC/BMP组3周时,CPC表面及内部可见较多新骨及幼稚骨细胞,CPC已被分解成多个区域(岛状分布),有板层骨形成;透射电镜观察可见成骨细胞侵入CPC内部,细胞浆内粗面内质网及线粒体增多扩张。CPC/BMP组12周时,光镜下见CPC降解,新生骨长入材料并与之相互分割包裹,骨小梁增粗; X线片显示新骨显影较清,材料密度减低,吸收降解现象明显;透射电镜观察可见大量成骨细胞散在于CPC颗粒之间,细胞内粗面内质网及线粒体增多扩张,较多毛细血管侵入材料内部。在材料降解速度方面,CPC/BMP组明显快于CPC组。结论:CPC是BMP的理想载体,CPC/BMP复合材料具有较强的传导成骨和诱导成骨活性,对实验性ANFH具有修复作用。在降解速度上CPC/BMP组明显快于单纯CPC组。  相似文献   

17.
骨不连与骨形成蛋白   总被引:1,自引:0,他引:1  
研究骨不连发生的原理及骨不连与骨诱导因子的关系是抗击有不连发生机理的重要课题。方法:将兔桡骨中段告成cm的缺损,A组术后即放入BMP,B组术后4周放入BMP,C组为在A组术后8周放入BMP。结果:发现BMP植入8周时形成骨小梁。B组和C组,骨缺损修复仍然取得满意的效果。  相似文献   

18.
目的:探讨多孔羟基磷灰石(hydroxyapatite,HA)支架作为人工骨替代材料的异位成骨效果。方法:将人胚胎间充质干细胞与多孔HA支架混合培养7 d,取出细胞支架复合体移植入比格犬背肌内为实验组,同时植入未与细胞共培养的多孔HA支架于比格犬背肌内为对照组,在植入第12周时取出体内异位成骨组织进行成骨效果检测。结果:植入第12周时的异位成骨组织内均有新生血管形成,新生骨组织实验组明显多于对照组。结论:多孔HA支架在体内有很好的异位成骨能力,人胚胎间充质干细胞能促进新骨的形成。  相似文献   

19.
PLGA膜包裹羟基磷灰石球眼眶植入的动物实验初步报告   总被引:4,自引:0,他引:4  
目的:观察PLGA膜包裹的羟基磷灰石球在兔眼眶血管化的情况,探讨Poly(lactide-co-glycolide acide75/25,简称PLGA)膜作为多孔羟基磷灰石球(CHA)包裹材料的可行性。方法:8只日本大耳兔分成4组,每组2只,分别对其右眼行眼球摘除术后,将以PLGA膜包裹的CHA植入眼眶肌锥腔内,分别于术后4,8,12,24周将2只球体取出作病理检查。结果:球结膜切口均甲级愈合,球体周围被结缔组织包裹紧密,且球体植入的时间越长,长入球体孔隙内的纤维血管组织越多;PGCA膜8周左右大部降解,其间未出现感染,球结膜裂开,球体脱出等并发症。结论:PLGA膜是一种可接受的CHA包裹材料,可减少对异体巩膜的需求,消除感染传染性疾病的潜在风险。  相似文献   

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