脾组织移植保留脾损伤患者脾功能的实验研究

自体脾组织移植已成为保留严重脾损伤患者脾功能的措施之一,但移植牌组织的功能如何,对机体究竟有多大影响,尚未定论。提高自体移植脾组织的功能已成为人们普遍关心的问题。因此,本文研究了动物自体移植脾组织的结构与功能;初步探索评价了促进自体移植脾组织生长的措施。     

去粘膜游离小肠段内自体脾组织移植实验研究和临床应用

本实验将犬自体脾组织移植于游离空肠段内,6月后处死取出移植脾组织进行大体形态学:光镜和电镜观察,结果显示:移植脾组织存活良好,再生明显,其重量为原移植重量的129.6%,组织形态学及超微结构与对照组无明显差异,经临床应用证明此方法是自体脾移植的理想方法。     

自体移植脾巨噬细胞C3b、Fc受体及蛋白质表达的实验研究

目的 探讨自体脾组织移植后的功能状况。方法 采用小鼠进行自体脾组织网膜内移植，术后6个月切取移植脾组织，检测巨噬细胞的Fc、C3b受体及蛋白表达。结果 自体移植脾巨噬细胞Fc受体的含量与原位脾相近，C3b受体的功能正常；蛋白质的表达与原位睥相同。结论 大网膜内自体移植脾组织的功能在细胞水平是正常的，移植脾组织具有原位脾的功能。     

脾脏移植的发展现状

随着对脾脏功能研究的开展和现代脾脏外科观念的深化．脾脏移植作为保留脾脏功能的重要手段已在不同层面得到广泛研究和实践。其中自体脾组织移植作为脾破裂后进行脾脏保留手术的主要措施已为大家所接受。在同种异体脾移植方面，我国学者做了许多研究工作，已使移植例数及术后生存期达到世界领先水平。     

脾损伤自体脾组织移植的临床应用

目的：探讨自体脾组织移植在临床中的应用。方法：总结32例脾外伤行全脾切除自体脾组织移植手术，其中采用大网膜囊内移植18例，去粘膜游离空肠段内移植12例，腹直肌鞘内移植2例。结果：术后随访均显示脾功能满意，尤以去粘膜游离空肠段内移植效果最好。结论：自体脾组织移植可作为严重脾外伤、全脾切除术后保留脾功能的一个重要有效手段，移植脾的功能恢复与血供有密切的关系。     

不同量自体脾组织移植抗肺炎球菌感染的研究

采用大鼠进行不同量脾组织网膜内移植，术后６月检测有关抗感染指标，结果显示气管内感染肺炎球菌后，４０％，６０％，８０％脾移植组和假手术组的存活率明显高于脾切除组，血中肺炎球菌廓清率在脾切除组中明显下降，而在各移植组则接近正常，结果表明，自体脾组织移植有一定上的抗感染能力，脾组织移植量以４０％～６０％为宜。     

自体移植脾巨噬细胞C_(3b)、Fc受体及蛋白质表达的实验研究

目的　探讨自体脾组织移植后的功能状况。方法　采用小鼠进行自体脾组织网膜内移植 ,术后 6个月切取移植脾组织 ,检测巨噬细胞的Fc、C3b受体及蛋白表达。结果　自体移植脾巨噬细胞Fc受体的含量与原位脾相近 ,C3b受体的功能正常 ;蛋白质的表达与原位脾相同。结论　大网膜内自体移植脾组织的功能在细胞水平是正常的 ,移植脾组织具有原位脾的功能。     

自体脾腹膜后移植在创伤性脾破裂中的临床应用

目的探讨自体脾组织移植在治疗创伤性脾破裂的应用.方法对本组于2000年1月至2005年4月22例脾破裂行全脾切除后,再行自体脾组织腹膜后移植术.通过检测外周血IgM、IgA、IgG水平和B超,CT、99mTc扫描来观察移植脾片成活和吞噬功能恢复情况.结果术后随访均显示移植脾存活良好,脾功能满意.结论自体脾组织移植可作为严重脾外伤全脾切除术后保留脾功能的一个重要有效手段.     

自体移植脾组织VEGF、KDR表达与血管再生的实验研究

目的 研究自体移植脾组织血管再生及VEGF、KDR表达规律，阐明VEGF、KDR对移植脾组织血管再生的调控作用，为脾脏外科临床及实验研究提供理论依据。方法 健康Wistar大鼠70只，体重100—120g，随机分为7组，每组10只中又设脾切除自体脾移植组5只，假手术组5只，分别于术后7，14，30，60，90，120，180d进行：(1)自体移植脾组织病理学检测；(2)大鼠行主动脉插管灌注墨汁，光镜观测再生血管并采用图像分析测定其密度；(3)免疫组化抗VEGF、KDR抗体染色，图像分析定量，阐明其表达规律及与血管再生的关系。结果 (1)自体脾组织移植术后7d即有血管从大网膜向脾组织内伸展，移植脾组织内血管密度逐渐增大，至术后180d血管再生接近正常；(2)自体脾组织移植术后7d、14d，VEGF、KDR阳性染色细胞密度迅速升高，术后60d达高峰，以后逐渐降低，至术后180d VEGF、KDR阳性染色细胞密度趋向正常。结论 自体脾组织大网膜内移植术是简便有效的脾移植方法；移植脾组织新生血管由大网膜再生而来；术后移植脾组织内VEGF、KDR表达量升高，促进血管形成，血管再生完成后恢复正常水平。     

脾移植的现状与展望

脾移植的现状与展望哈尔滨医科大学附属第一医院外科（１５０００１）姜洪池，吴业权同济医科大学器官移植研究所（４３００３０）夏穗生随着脾脏功能研究的深人，曾一度陷入低谷的脾移植有了新的发展，自体脾组织移植在国内外已被接受为脾破裂后进行脾保留手术的主要措施...     

Autotransplantation of splenic tissue

Autotransplantation of splenic fragments has already been carried out in humans. The optimal size of the particles and amount of tissue required for this procedure has yet to be found. In normal young pigs and miniature piglets, autologous splenic tissue was transplanted into the greater omentum. The regenerated splenic mass, splenic blood flow, and histology were studied six months later. Implanting small splenic particles produced comparable results to implanting thin slices of splenic tissue. The mass of regenerated splenic tissue was only 3.1 g after implanting the whole spleen and 4.5 g after transplanting half of the spleen, which means 5.3% and 7.8% respectively of the weight of control spleens. The blood flow per gram in the regenerated splenic tissue was much lower than in the normal spleen. The blood flow in the whole of the splenic tissue is important for the clearance function of the spleen. Six months after transplanting the whole or half of the spleen, the blood flow to the regenerated splenic tissue was only 1% of that in the control minipigs. When half of the spleen was left in situ, as a model for a partial splenectomy, and the other half transplanted, the regenerated mass was only 3.4% of all splenic tissue and the blood flow 1.5% of the total splenic blood flow. In this model the regenerated splenic mass was independent of the size of the implants and the mass of implanted tissue. The extremely low blood flow indicates an inadequate clearance function and thus the protective function would probably be negligible.     

Pancreatic transplantation. Absence of diabetes in dogs after total pancreatectomy and intrasplenic autotransplantation of pancreatic tissue.

Two simple, rapid, and reproducible techniques of pancreatic tissue preparation produced favorable results after just part of the gland was autotransplanted in totally pancreatectomized animals. It is probable that the short and less traumatic treatment in vitro was responsible for the sufficient yield of viable islets. Moreover, the results show convincingly that the complete separation of endocrine from exocrine pancreas is not mandatory to insure adequate endocrine secretion. The spleen, an organ of no vital importance having a rich blood supply and belonging to the portal circulation, was a very suitable experimental implantation site, which permitted thorough evaluation of the tissue preparation techniques. These favorable results were not conditioned by the splenic tissue itself, because similar results have been obtained in dogs when the tissues were implanted into the liver via the portal vein.     

Regeneration and function of autologous splenic grafts in pigs

The growth kinetics, perfusion and immune response of autotransplanted splenic tissue were studied in pigs. Splenic fragments were transplanted subfascially or in the greater omentum and regenerated to small splenules with a normal histologic structure 6 months later. The grafts showed a normal function of the white pulp after antigenic stimulation, and of the red pulp as demonstrated scintigraphically. The blood flow was reduced. The growth kinetic was less rapid than in rodents and resulted in a relatively lower mass of splenic tissue, but these results are closer to findings in human splenosis.     

Immunologic function against infection in splenic autotransplanted mice

The increasing recognition of the danger of overwhelming postsplenectomy infection (OPSI) has led surgeons to attempt to maintain splenic function after spleen injury. One technique they use when splenorrhapy or partial splenectomy are not feasible is the deliberate autotransplantation of splenic tissue. But the amount of splenic tissue necessary to prevent OPSI remains controversial, and opinions differ about the importance of the location and size of the splenic fragments implanted. The mice were divided into five groups, I. splenectomy, II. splenectomy +30% of the spleen implanted intraperitoneal site, III. splenectomy +50% implanted intraperitoneally, IV. splenectomy +50% implanted subcutaneously and V. Sham operation. This study assessed the blood flow of the splenic tissue, increasing weight of splenic mass, histology, the serum level of the immunoglobulins (IgG, IgA, and IgM), pneumococcal antibody titers after vaccination, and survival after intravenous pneumococcal challenge. This study demonstrated that intraperitoneal transplantation showed better regeneration and afforded better protection from OPSI than subcutaneous transplantation. And 30 to 50 percent of the whole splenic tissue mass protected against experimental pneumococcal sepsis. The splenic autotransplants developed in volume and blood supply after 8 weeks, and immunologic function against infection recovered at the same time.     

Experimental study on function of regenerated splenic tissue after splenic autotransplantation

To prevent postsplenectomy overwhelming sepsis, splenic autotransplantation has been clinically attempted. However, function of regenerated splenic tissue after splenic autotransplantation has not been completely understood. Changes in weigh of regenerated splenic tissue, splenic blood flow, splenic immune responses and phagocytic function were studied for one year after splenic autotransplantation using Sprague-Dawley rats. At one year after autotransplantation, the weight of regenerated splenic tissue was increased to 80% of the originally implanted spleen and the blood flow was increased to 80% of the control spleen. The counts of lymphocytes and macrophages in the regenerated splenic tissue were significantly low at eight weeks after transplantation, however lymphocytes was increased to 58.8% and macrophages was increased to 29.5% of the control spleen at 16 weeks after transplantation. The blast formation of splenic lymphocytes was lower at the early stage after transplantation, thereafter, it was increased at the later time after transplantation. Microangiography of the regenerated spleen showed new capillaries around the implanted tissue 2 weeks after transplantation. These results suggested that the transplanted splenic tissue was regenerated to the similar structure to normal spleen and immunological function was recovered close to the normal splenic tissue.     

Does survival depend on the amount of autotransplanted splenic tissue?

Susceptibility to Streptococcus pneumoniae infection was studied in 11 groups of rats allocated to sham operation, splenectomy, or splenic autotransplantation of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the removed spleen. Three months later, all rats were exposed intravenously to type 1 Streptococcus pneumoniae (median lethal dose, LD50, for control group). Survivors were killed 13 days after the bacterial challenge. Autopsy showed that more splenic tissue was recovered in rats that received less than 50% splenic tissue compared with those that received 50% or more. More survivors were found among sham-operated rats (47.5%; 95% confidence intervals, 32 to 68) and rats that had 40% splenic tissue implanted (35%; confidence interval, 20 to 54) or those that were found to have regenerated 40% splenic tissue. We conclude that 40% of the spleen should be autotransplanted to protect the rat optimally against infection after splenectomy.     

Cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted mice

BACKGROUND: Our department worked out a modified surgical form of spleen autotransplantation earlier, named "spleen apron method" introduced already into the clinical practice. Recently we tested the immunological changes in a group of patients autotransplanted with about 10-15% of their spleen, what was the at least always implantable amount after the severe splenic injuries. In the current work we aimed at measuring some cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted inbred mice two and eight months after the operations in order to get more unambiguous results than that we could obtain in our patients with this technique. MATERIALS AND METHODS: We divided 96 two months old Balb/c female mice into eight groups (n = 12/group). The group of controls, sham operated, splenectomized and autotransplanted animals with two and eight months of survival time after the operations. During the autotransplantation we inserted the same amount of spleen, five slices, "chips," about 10-15% of total mass of spleen, into the greater omentum similarly as it was used in the patients. The concentration of serum proteins were measured by laser nephelometry. The lymphocyte subsets were analyzed by flow cytometry. RESULTS: We found that two months after the operations the number of CD 19+ B-cells increased in the splenectomized but decreased in the autotransplanted animals. Eight months after the operations the number of both CD3+ T and CD19+ B lymphocytes decreased both in the splenectomized and autotransplanted animals compared to the controls and sham operated mice. However, the numbers of T and B cells were slightly but not significantly higher in the autotransplanted than in the splenectomized mice. The serum level of IgM was also decreased in the splenectomized and autotransplanted mice at both time points, however, eight months after the operations the concentration of IgM was significantly higher in the autotransplanted group than in the splenectomized animals. CONCLUSION: The effects of autotransplanted "chips" were different at the various ages of the animals. Additionally, they showed some immunological benefit being quantitatively in accordance to the amount of the transplanted spleen. The elevated level of serum IgM what we found in the autotransplanted mice even with this amount of transplanted spleen eight months after the operations, however, might have the potentially greatest importance compared to splenectomy. These experiments can prove that the attempts for autotransplantation may have real perspectives but their efficacy depends on the amount of the successfully transplanted (saved) mass of spleen.     

Splenic tissue autotransplantation in rabbits: no restoration of host defense

BACKGROUND: The loss of spleen may increase the incidence of overwhelming sepsis. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the effectiveness of regenerated splenic tissue in preventing infection. This study explored the effectiveness of splenic tissue autotransplantation in restoring host defense. MATERIALS AND METHODS: Rabbits were divided into three groups: splenic autotransplantation, sham operation, and total splenectomy. Histomorphology, T-lymphocyte count, serum lysozyme levels, hemolysin titers, and pneumococcal clearance were observed as read-out parameters over 24 weeks. RESULTS: Histological study showed that the white pulp was poorly developed and central arterioles were missing in the regenerated splenic tissue of the autotransplanted rabbits. The weight of regenerated spleens recovered 6 months later in the splenic autotransplantation group was 11% of that in the sham operation group and was significantly less than the weight at implantation. There was no significant difference in the number of T lymphocytes or level of serum lysozyme between the three groups. A poor antibody response by the rabbits in the splenic autotransplantation and total splenectomy groups was noted after the primary intravenous administration of sheep red blood cells compared to those of sham operation group. After the challenge with type 3 pneumococci intravenously, pneumococcal clearance from the bloodstream in the splenic autotransplantation group did not differ significantly from that in the total splenectomy group, but was markedly delayed compared with that in the sham operation group. CONCLUSIONS: The low quantity and poor quality of the regenerated splenic tissue contribute to the inferior immunoprotective ability of animals autotransplanted with one-third of the original spleen. This suggests that the regenerated spleen cannot compensate for the immunological function of the original one, especially host resistance to infection.     

Optimal site and amount of splenic tissue for autotransplantation.

Clinical and basic studies have documented a high susceptibility to pneumococcal infection in asplenic humans and animals. It has been suggested that autotransplantation of splenic tissue might be a method of providing host resistance when total splenectomy is necessary. However, the effect of splenic autograft has remained controversial. This study was performed to evaluate the most effective site and amount of splenic autograft using rats. Rats were divided into five groups for the purpose of determining the site of splenic autotransplantation: splenectomy, sham operation, implantation into the omental pouch, intraperitoneal implantation, and intramuscular implantation. For determining the amount for autotransplantation, the rats were divided into seven groups: splenectomy, sham operation, and implantations of 25, 50, 100, 200, or 300 mg of splenic tissue. All animals were challenged with Streptococcus pneumoniae type 6, 16 weeks after surgery. Howell-Jolly bodies appeared postsplenectomy, but disappeared in the implanted rats 16 weeks after the operation. Histologically, the implanted tissue was indistinguishable from that of a normal spleen. Pneumococcal clearance from the bloodstream and survival rate were significantly higher in rats implanted in the omental pouch as compared with splenectomized rats. Intraperitoneal and intramuscular implanted rats did not show a significant difference from the splenectomized rats. More than 50% of splenic tissue for autograft showed a significant increase in pneumococcal clearance and survival rate as compared with that of splenectomized rats. It was suggested that the most effective site of autotransplantation is the omental pouch and approximately 50% of the whole spleen would be necessary for prevention from sepsis.