AIM: To evaluate the recovery and function of isolated rat pancreatic islets during in vitro culture with small intestinal submucosa (SIS).METHODS: Pancreatic islets were isolated from Wistar rats by standard surgical...AIM: To evaluate the recovery and function of isolated rat pancreatic islets during in vitro culture with small intestinal submucosa (SIS).METHODS: Pancreatic islets were isolated from Wistar rats by standard surgical procurement followed by intraductal collagenase distension, mechanical dissociation and Euroficoll purification. Purified islets were cultured in plates coated with multilayer SIS (SIS-treated group) or without multilayer SIS (standard cultured group) for 7 and 14 d in standard islet culture media of RPMI 1640. After isolation and culture, islets from both experimental groups were stained with dithizone and counted. Recovery of islets was determined by the ratio of counts after the culture to the yield of islets immediately following islet isolation. Viability of islets after the culture was assessed by the glucose challenge test with low (2.7 mmol/L) and high glucose (16.7 mmol/L)solution supplemented with 50 mmol/L 3-isobutyl-1-methylxanthine (IBMX) solution. Apoptosis of islet cells after the culture was measured by relative quantification of histone-complexed DNA fragments using ELISA.RESULTS: After 7 or 14 d of in vitro tissue culture, the recovery of islets in SIS-treated group was significantly higher than that cultured in plates without SIS coating. The recovery of islets in SIS-treated group was about twice more than that of in the control group. In SIS-treated group, there was no significant difference in the recovery of islets between short- and long-term periods of culture (95.8±1.0% vs 90.8±1.5%, P>0.05). When incubated with high glucose (16.7 mmol/L) solution,insulin secretion in SIS-treated group showed a higher increase than that in control group after 14 d of culture (20.7±1.1 mU/L vs11.8±1.1 mU/L, P<0.05). When islets were placed in high glucose solution containing IBMX,stimulated insulin secretion was higher in SIS-treated group than in control group. Calculated stimulation index of SIS-treated group was about 23 times of control group. In addition, the stimulation index of SIS-treated group remained constant regardless of short- and long-term periods of culture (9.5±0.2 vs 10.2±1.2, P>0.05).Much less apoptosis of islet cells occurred in SIS-treated group than in control group after the culture.CONCLUSION: Co-culture of isolated rat islets with native sheet-like SIS might build an extracellular matrix for islets and provide possible biotrophic and growth factors that promote the recovery and subsequent function of islets.展开更多
BACKGROUND: The ability to maintain isolated human islet preparation in tissue culture has recently been adopted by most islet transplant centers to improve the safety and practicality of islet transplantation. Howeve...BACKGROUND: The ability to maintain isolated human islet preparation in tissue culture has recently been adopted by most islet transplant centers to improve the safety and practicality of islet transplantation. However, maintaining islet viability and recovery remains a challenge in clinical setting. Extracellular matrix (ECM) is one of the most important components of islet microenvironment. The reconstruction of the cell-matrix relationship seems to be effective in improving the loss of differentiated islet structure and function. Small intestinal submucosa ( SIS ) , a naturally occurring ECM, has been investigated to be able to promote wound healing, tissue remodeling, and cell growth. The purpose of this study was to evaluate the recovery and function of isolated rat pancreatic islets after in vitro culture with SIS. METHODS: Pancreatic islets were isolated from Wistar rats by using standard surgical procurement followed by intra-ductal collagenase distension, mechanical dissociation, and EuroFicoll purification. Groups of purified islets were cultured in plates which were coated with multilayer SIS (SIS-treated group) or without ( standard cultured group) for 7 days and 14 days in standard islet culture conditions of RP-MI 1640 tissue culture media in humidified atmosphere containing 95% air and 5% CO2 at 37 ℃. The mean recovery of islets after the culture period was determined by sizing duplicate counts of a known volume and their viability was assessed by static incubation with low glucose (2.7 mmol) , high glucose (16.7 mmol) and high glucose solution supplemented with 50 μm 3-isobutyl-1-methylxanthine (IB-MX) solution. RESULTS: After 7 days and 14 days of in vitro tissue culture, the SIS-treated group showed a significantly higher recovery compared with those cultured under standard conditions. The recovery in the SIS-treated group was about two times of the control group cultured in standard conditions after 14 days culture. In the SIS-treated group, there was no statistically difference between the short and long periods of culture ( 95. 8 ± 1.0% vs. 90. 8±1. 5% , P 】 0.05). During incubation in high glucose (16.7 mmol) solution, there was a 2-3 fold increase in insulin secretion from both groups, but the SIS-treated group showed a higher increase than the standard cultured group after 14-day culture (20.7 ±1.1 mU/L vs. 11. 8 ±1.1 mU/L, P 【 0. 05). When islets were placed in the high glucose solution supplemented with IBMX, the stimulated insulin response in the SIS-treated group was higher than that in the standard cultured group in spite of the duration of the culture. The stimulation index of the SIS-treated group was about 2-3 times of the standard cultured group. In addition , after a long period of culture, the stimulation index of the SIS-treated group was statistically equivalent with that of the short period of culture (9.5 ±0.2 vs. 10.2 ±1.2, P】0.05). CONCLUSIONS: The co-culture of isolated rat islets with native sheet-like SIS can provide an excellent extracellular matrix, possible biotrophic and growth factors that promote the recovery and subsequent function of islets after in vitro tissue culture. In view of results of this study and rapid degradation of SIS in vitro, future studies will investigate the extended duration of culture and the effect of SIS on islets in vitro.展开更多
Objective To investigate the feasibility of using small intestinal submucosa ( SIS) graft for repair of anterior urethral strictures. Methods From June 2009 to August 2010,18 men ( mean age,38 yrs) with anterior ureth...Objective To investigate the feasibility of using small intestinal submucosa ( SIS) graft for repair of anterior urethral strictures. Methods From June 2009 to August 2010,18 men ( mean age,38 yrs) with anterior urethral strictures underwent urethroplasty using a four layer SIS as an onlay patch graft. SIS was used to展开更多
Anal fistula is one of the three greatest anorectal diseases with a high prevalence. The traditional treatments（e.g., surgery） for fistula have limitations due to damage to the internal anal sphincter of patients. W...Anal fistula is one of the three greatest anorectal diseases with a high prevalence. The traditional treatments（e.g., surgery） for fistula have limitations due to damage to the internal anal sphincter of patients. With recent advances in biomaterials, treatments based on biomaterial filling （e.g., scleraprotein injection, fistula plug） have emerged as novel therapies for fistula. The anal fistula plug （e.g., based on small intestinal submucosa （SIS）） has attracted increasing attention because of short term healing rate and biocompatibility. However, challenges remain for this method such as plug falling as observed in clinics. To address this, this paper analyzes the case of SIS falling under physiological condition from mechanical point of view using ANSYS simulation. It then proposes three new geometrical structures for fistula plug and compares their mechanical behavior （e.g., axial stress, reaction of constraint） with that of clinically used structure （cone shape）. Based on the simulation, it optimizes the geometric parameters of fistula plug. The approach developed here can help to improve the design of fistula plug for better clinical treatments.展开更多
Background：Pelvic organ prolapse （POP） is a major health concern that affects women.Surgeons have increasingly used prosthetic meshes to correct POP.However,the most common used is synthetic mesh,and absorbable mes...Background：Pelvic organ prolapse （POP） is a major health concern that affects women.Surgeons have increasingly used prosthetic meshes to correct POP.However,the most common used is synthetic mesh,and absorbable mesh is less reported.This research aimed to evaluate the clinical effectiveness of porcine small intestinal submucosa （SIS）.Methods：Consecutive forty POP patients who met the inclusion criteria underwent pelvic reconstruction surgery with SIS between March 2012 and December 2013.The patients＆#39; clinical characteristics were recorded preoperatively.Surgical outcomes,measured by objective and subjective success rates,were investigated.We evaluated the quality of life （QOL） using the Pelvic Floor Distress Inventory-20 （PFDI-20） and the Pelvic Floor Impact Questionnaire-7 （PFIQ-7）.Sexual QOL was assessed by the Pelvic Organ Prolapse/Urinary Incontinence Sexual Function Questionnaire-12 （P1SQ-12）.Results：At postoperative 12 months,the subjective recurrence rate （7.5%） was much lower than the objective recurrence rate （40.0%）.Postoperatively,no erosion was identified.One underwent a graft release procedure because of urinary retention,and one had anus sphincter reconstruction surgery due to defecation urgency.Another experienced posterior vaginal wall infection where the mesh was implanted,accompanied by severe vaginal pain.Estrogen cream relieved the pain.One patient with recurrence underwent a secondary surgery with Bard Mesh because of stage 3 anterior vaginal wall prolapse.Scoring system of PFDI-20 was from 59.150 ± 13.143 preoperatively to 8.400 ± 4.749 postoperatively and PFIQ-7 was from 73.350 ± 32.281 to 7.150 ± 3.110,while PISQ-12 was from 15.825 ± 4.050 to 12.725 ± 3.471.Conclusions：QOL and the degree of subjective satisfaction were significantly improved postoperatively.Anterior repair deserves more attention because of the higher recurrence rate.The long-term follow-up of the patient is warranted to draw firm conclusion.展开更多
Tympanic membrane perforationsoccur frequently in children,and can result in hearing loss,otorrhea,pain,and cholesteatoma.Due to the narrower ear canal in children,a postauricular incision is often needed to access th...Tympanic membrane perforationsoccur frequently in children,and can result in hearing loss,otorrhea,pain,and cholesteatoma.Due to the narrower ear canal in children,a postauricular incision is often needed to access the tympanic membrane for surgical repair.Endoscopic approaches are increasingly being used for tympanic membrane repair,reducing the need for postauricular incisions.As the need for a postauricular incision decreases,the demand for non-autologous grafting material has increased.Acellular porcine small intestinal submucosa (SIS) has been described in the literature as an alternative to commonly used autologous grafts,and is well suited for use with transcanal endoscopic ear surgery as a minimally invasive approach.This paper describes techniques for use of SIS in endoscopic tympanic membrane repair in children.展开更多
Acellular porcine small intestinal submucosa(SIS)has been used for esophagoplasty with success in a canine model.However,it did not lead to complete epithelialization.For better reconstruction,a cellular component is ...Acellular porcine small intestinal submucosa(SIS)has been used for esophagoplasty with success in a canine model.However,it did not lead to complete epithelialization.For better reconstruction,a cellular component is required.Moreover,promotion of angiogenesis with copper has been widely recognized by basic research as well as clinical studies.In this study,we have evaluated the feasibility and effectiveness of combined Cu and SIS(SIS-Cu patch)for the esophageal repair using a canine model.Eighteen male beagle dogs were subjected to surgical resection to produce cervical esophageal defects(5 cm in length,180°in range).SIS with Cu(5 or 25μmol L 1copper)or without Cu was patched on the esophageal defects.Barium esophagram and histology exam were carried out to evaluate the effectiveness of the therapy.As shown,the SIS-Cu graft promoted re-epithelialization,re-vascularization and muscular regeneration.SIS-Cu patch is more effective than SIS alone for esophageal repair,and the SIS+25μmol L 1Cu group demonstrated additional advantages over the SIS+5μmol L 1Cu.展开更多
Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound h...Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound healing still needs to be improved to meet clinical demands.To tackle this problem,platelet-rich plasma(PRP)is used due to its potency to promote proliferation,migration and adhesion of target cells.In this study,we applied PRP and SIS to skin wounds to explore their effects on wound healing by evaluating re-epithelialization,collagen production,angiogenesis and the inflammatory response.Methods:A1×1-cm full-thickness skin defectwas established in mice.Sixty mice were divided into four treatment groups:PRP+SIS,PRP,SIS and control.On days 3,5,7,10 and 14 post-surgery,tissue specimens were harvested.Haematoxylin and eosin,Masson’s trichrome,immunohistochemical and immunofluorescence double staining were used to visualize epidermal thickness,collagen and vascular regeneration and inflammation.Results:Wound contraction in the PRP and PRP+SIS groups was significantly greater,compared with the other groups,on days 3 and 5 post-surgery.A histological analysis showed higher collagen expression in the PRP and PRP+SIS groups on day 7,whichwas associated with a thicker epidermal layer on day 14.In addition,immunohistochemical staining showed that CD31-positive blood vessels and vascular endothelial growth factor expression in the PRP+SIS and PRP groups were significantly higher,compared with the control group.Furthermore,immunofluorescence double staining showed that the number of M1 and M2 macrophages in the PRP+SIS and PRP groups was higher,compared with the control and SIS groups alone,on day 3.However,on day 7,the number of M1 macrophages dramatically decreased in the PRP+SIS and PRP groups.The ratio of M2 to M1 macrophages in the PRP+SIS and PRP groups was 3.97 and 2.93 times that of the control group and 4.56 and 3.37 times that of the SIS group,respectively.Conclusion:Co-administration of SIS and PRP has a better effect on promoting angiogenesis,reepithelialization and collagen regeneration in managing acute wound healing than either agent alone.展开更多
There has been an increase in the incidence of hypopharyngeal and cervical esophageal cancer worldwide,and hence growing needs for hypopharyngeal and cervical esophageal tissue repair.This work produced a bi-layer com...There has been an increase in the incidence of hypopharyngeal and cervical esophageal cancer worldwide,and hence growing needs for hypopharyngeal and cervical esophageal tissue repair.This work produced a bi-layer composite scaffold with decellularized small intestine submucosa and polylactic-co-glycolic acid,which resembled the layered architectures of its intended tissues.The decellularized small intestine submucosa contained minimal residual DNA(52.5±61.2 ng/mg)and the composite scaffold exhibited satisfactory mechanical properties(a tensile modulus of 21.1±64.8 MPa,an ultimate tensile strength of 14.0±62.9MPa and a failure strain of 26.9±65.1%).The interactions between cells and the respective layers of the scaffold were characterized by CCK-8 assays,immunostaining and Western blotting.Desirable cell proliferation and phenotypic behaviors were observed.These results have provided an important basis for the next-step in vivo studies of the scaffold,and bode well for its future clinical applications.展开更多
目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱...目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱细胞基质胶和白藜芦醇(resveratrol,Rev)溶液配制成膜片状的负载白藜芦醇的SIS脱细胞基质(Rev/SIS)。取小型猪3只,在每只猪背建立4个4 cm×4 cm的全层皮肤缺损创面,按照不同治疗手段随机分为4组:生理盐水纱布组(对照组)、白藜芦醇组(Rev组)、SIS脱细胞基质组(SIS组)和负载白藜芦醇的SIS脱细胞基质组(Rev/SIS组)。分别记录术后第7、14、21、28天的创面愈合及术后第42天的瘢痕形成状况;采集创缘组织行组织学检查和免疫组化染色分析,评估愈合过程中的炎性浸润和再血管化水平,以及所形成瘢痕组织的皮肤厚度和胶原沉积情况;并分别在术后第3、7天检测各组创面中炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、M1型巨噬细胞标志物i NOS和M2型巨噬细胞标志物ARG-1的m RNA相对表达量。结果术后第7、14、21天,SIS组和Rev/SIS组创面残留率均显著低于对照组(P<0.05),术后第21天Rev/SIS组创面残留率低于SIS组(P<0.001),术后第28天除Rev/SIS组外其余3组创面均未完全愈合;术后第42天Rev/SIS组瘢痕面积最小,质地柔软,扁平且无明显色沉。Rev/SIS组TNF-a、IL-6 m RNA表达量均低于对照组(P<0.05),3种治疗手段均能降低创面i NOS m RNA表达(P<0.001),但以Rev/SIS组最为显著(P<0.05),且Rev/SIS组的ARG-1 m RNA表达高于其余3组(P<0.05),该组创面炎性浸润程度最低,再血管化水平最高。其余3组瘢痕的表皮、真皮厚度均低于对照组(P<0.01),也以Rev/SIS组改善最为显著(P<0.0001),且该组胶原沉积与正常皮肤相似,排列有序。结论尽管SIS脱细胞基质和白藜芦醇单独使用也可能产生抗炎效应,在促进创面愈合的同时,减少瘢痕形成面积,有效提高愈合质量;但二者联合应用的效果更为显著,Rev/SIS脱细胞基质为开发新型皮肤创面敷料提供了一种潜在选择。展开更多
目的:设计不同pH值下螯合锶(strontium,Sr)的脱细胞小肠黏膜下层(decellularized small intestinal submucosa,dSIS)海绵支架(Sr/dSIS),以该支架的理化性能和生物相容性为评价指标,为制备Sr/dSIS选择合适的pH值。方法:(1)Sr/dSIS制备及...目的:设计不同pH值下螯合锶(strontium,Sr)的脱细胞小肠黏膜下层(decellularized small intestinal submucosa,dSIS)海绵支架(Sr/dSIS),以该支架的理化性能和生物相容性为评价指标,为制备Sr/dSIS选择合适的pH值。方法:(1)Sr/dSIS制备及分组:将dSIS溶液与氯化锶溶液等体积混合,调节溶液pH分别为3、5、7、9,于37℃下充分反应后经冷冻干燥制得多孔支架,分别命名为Sr/dSIS-3、-5、-7、-9,以dSIS支架为对照;(2)理化性能评价:观察支架大体形貌,采用扫描电镜分析微观形貌并测定孔隙率和孔径,能谱分析表面元素,红外光谱分析官能团结构,原子吸收分光光度法测定螯合率,比重法检测吸水率,万能力学测试机评价压缩强度;(3)生物相容性评价:采用Calcein-AM/PI活细胞/死细胞染色法评价各组支架的毒性和促骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)增殖效果。结果:扫描电镜下各组支架具有三维多孔网络结构,孔径和孔隙率差异无统计学意义;能谱分析中Sr/dSIS-5、-7、-9组检测出锶元素的特征峰,且锶元素均匀分布于支架;官能团分析验证了Sr/dSIS-5、-7、-9组有螯合物形成;螯合率分析显示Sr/dSIS-7组锶离子螯合率最高,与其他组的差异有统计学意义(P<0.05);各组支架吸水性良好;Sr/dSIS-5、-7、-9组的压缩强度显著高于对照组(P<0.05);各组支架的生物相容性良好,Sr/dSIS-7组展现出最佳的促细胞增殖能力。结论:pH为7时,Sr/dSIS支架具有高锶离子螯合率以及更好的促BMSCs增殖效果,是制备Sr/dSIS支架的理想pH值。展开更多
With the development of tissue engineering,the required biomaterials need to have the ability to promote cell adhesion and proliferation in vitro and in vivo.Especially,surface modification of the scaffold material ha...With the development of tissue engineering,the required biomaterials need to have the ability to promote cell adhesion and proliferation in vitro and in vivo.Especially,surface modification of the scaffold material has a great influence on biocompatibility and functionality of materials.The small intestine submucosa(SIS)is an extracellular matrix isolated from the submucosal layer of porcine jejunum,which has good tissue mechanical properties and regenerative activity,and is suitable for cell adhesion,proliferation and differentiation.In recent years,SIS is widely used in different areas of tissue reconstruction,such as blood vessels,bone,cartilage,bladder and ureter,etc.This paper discusses the main methods for surface modification of SIS to improve and optimize the performance of SIS bioscaffolds,including functional group bonding,protein adsorption,mineral coating,topography and formatting modification and drug combination.In addition,the reasonable combination of these methods also offers great improvement on SIS surface modification.This article makes a shallow review of the surface modification of SIS and its application in tissue engineering.展开更多
Currently the standard surgical treatment for bladder defects is augmentation cystoplasty with autologous tissues,which has many side effects.Biomaterials such as small intestine submucosa(SIS)can provide an alternati...Currently the standard surgical treatment for bladder defects is augmentation cystoplasty with autologous tissues,which has many side effects.Biomaterials such as small intestine submucosa(SIS)can provide an alternative scaffold for the repair as bladder patches.Previous studies have shown that SIS could enhance the capacity and compliance of the bladder,but its application is hindered by issues like limited smooth muscle regeneration and stone formation since the fast degradation and poor mechanical properties of the SIS.Procyanidins(PC),a natural bio-crosslinking agent,has shown anti-calcification,anti-inflammatory and anti-oxidation properties.More importantly,PC and SIS can crosslink through hydrogen bonds,which may endow the material with enhanced mechanical property and stabilized functionalities.In this study,various concentrations of PC-crosslinked SIS(PC-SIS)were prepared to repair the full-thickness bladder defects,with an aim to reduce complications and enhance bladder functions.In vitro assays showed that the crosslinking has conferred the biomaterial with superior mechanical property and anti-calcification property,ability to promote smooth muscle cell adhesion and upregulate functional genes expression.Using a rabbit model with bladder defects,we demonstrated that the PC-SIS scaffold can rapidly promote in situ tissue regrowth and regeneration,in particular smooth muscle remodeling and improvement of urinary functions.The PC-SIS scaffold has therefore provided a promising material for the reconstruction of a functional bladder.展开更多
基金Supported by the Key Program of Science and Technique of Ministry of Education of the People's Republic of China, No.104169
文摘AIM: To evaluate the recovery and function of isolated rat pancreatic islets during in vitro culture with small intestinal submucosa (SIS).METHODS: Pancreatic islets were isolated from Wistar rats by standard surgical procurement followed by intraductal collagenase distension, mechanical dissociation and Euroficoll purification. Purified islets were cultured in plates coated with multilayer SIS (SIS-treated group) or without multilayer SIS (standard cultured group) for 7 and 14 d in standard islet culture media of RPMI 1640. After isolation and culture, islets from both experimental groups were stained with dithizone and counted. Recovery of islets was determined by the ratio of counts after the culture to the yield of islets immediately following islet isolation. Viability of islets after the culture was assessed by the glucose challenge test with low (2.7 mmol/L) and high glucose (16.7 mmol/L)solution supplemented with 50 mmol/L 3-isobutyl-1-methylxanthine (IBMX) solution. Apoptosis of islet cells after the culture was measured by relative quantification of histone-complexed DNA fragments using ELISA.RESULTS: After 7 or 14 d of in vitro tissue culture, the recovery of islets in SIS-treated group was significantly higher than that cultured in plates without SIS coating. The recovery of islets in SIS-treated group was about twice more than that of in the control group. In SIS-treated group, there was no significant difference in the recovery of islets between short- and long-term periods of culture (95.8±1.0% vs 90.8±1.5%, P>0.05). When incubated with high glucose (16.7 mmol/L) solution,insulin secretion in SIS-treated group showed a higher increase than that in control group after 14 d of culture (20.7±1.1 mU/L vs11.8±1.1 mU/L, P<0.05). When islets were placed in high glucose solution containing IBMX,stimulated insulin secretion was higher in SIS-treated group than in control group. Calculated stimulation index of SIS-treated group was about 23 times of control group. In addition, the stimulation index of SIS-treated group remained constant regardless of short- and long-term periods of culture (9.5±0.2 vs 10.2±1.2, P>0.05).Much less apoptosis of islet cells occurred in SIS-treated group than in control group after the culture.CONCLUSION: Co-culture of isolated rat islets with native sheet-like SIS might build an extracellular matrix for islets and provide possible biotrophic and growth factors that promote the recovery and subsequent function of islets.
文摘BACKGROUND: The ability to maintain isolated human islet preparation in tissue culture has recently been adopted by most islet transplant centers to improve the safety and practicality of islet transplantation. However, maintaining islet viability and recovery remains a challenge in clinical setting. Extracellular matrix (ECM) is one of the most important components of islet microenvironment. The reconstruction of the cell-matrix relationship seems to be effective in improving the loss of differentiated islet structure and function. Small intestinal submucosa ( SIS ) , a naturally occurring ECM, has been investigated to be able to promote wound healing, tissue remodeling, and cell growth. The purpose of this study was to evaluate the recovery and function of isolated rat pancreatic islets after in vitro culture with SIS. METHODS: Pancreatic islets were isolated from Wistar rats by using standard surgical procurement followed by intra-ductal collagenase distension, mechanical dissociation, and EuroFicoll purification. Groups of purified islets were cultured in plates which were coated with multilayer SIS (SIS-treated group) or without ( standard cultured group) for 7 days and 14 days in standard islet culture conditions of RP-MI 1640 tissue culture media in humidified atmosphere containing 95% air and 5% CO2 at 37 ℃. The mean recovery of islets after the culture period was determined by sizing duplicate counts of a known volume and their viability was assessed by static incubation with low glucose (2.7 mmol) , high glucose (16.7 mmol) and high glucose solution supplemented with 50 μm 3-isobutyl-1-methylxanthine (IB-MX) solution. RESULTS: After 7 days and 14 days of in vitro tissue culture, the SIS-treated group showed a significantly higher recovery compared with those cultured under standard conditions. The recovery in the SIS-treated group was about two times of the control group cultured in standard conditions after 14 days culture. In the SIS-treated group, there was no statistically difference between the short and long periods of culture ( 95. 8 ± 1.0% vs. 90. 8±1. 5% , P 】 0.05). During incubation in high glucose (16.7 mmol) solution, there was a 2-3 fold increase in insulin secretion from both groups, but the SIS-treated group showed a higher increase than the standard cultured group after 14-day culture (20.7 ±1.1 mU/L vs. 11. 8 ±1.1 mU/L, P 【 0. 05). When islets were placed in the high glucose solution supplemented with IBMX, the stimulated insulin response in the SIS-treated group was higher than that in the standard cultured group in spite of the duration of the culture. The stimulation index of the SIS-treated group was about 2-3 times of the standard cultured group. In addition , after a long period of culture, the stimulation index of the SIS-treated group was statistically equivalent with that of the short period of culture (9.5 ±0.2 vs. 10.2 ±1.2, P】0.05). CONCLUSIONS: The co-culture of isolated rat islets with native sheet-like SIS can provide an excellent extracellular matrix, possible biotrophic and growth factors that promote the recovery and subsequent function of islets after in vitro tissue culture. In view of results of this study and rapid degradation of SIS in vitro, future studies will investigate the extended duration of culture and the effect of SIS on islets in vitro.
文摘Objective To investigate the feasibility of using small intestinal submucosa ( SIS) graft for repair of anterior urethral strictures. Methods From June 2009 to August 2010,18 men ( mean age,38 yrs) with anterior urethral strictures underwent urethroplasty using a four layer SIS as an onlay patch graft. SIS was used to
基金supported by the Major International (Regional) Joint Research Program of China (11120101002)the National Natural Science Foundation of China(10825210 and 31050110125)the National 111 Project of China(B06024)
文摘Anal fistula is one of the three greatest anorectal diseases with a high prevalence. The traditional treatments（e.g., surgery） for fistula have limitations due to damage to the internal anal sphincter of patients. With recent advances in biomaterials, treatments based on biomaterial filling （e.g., scleraprotein injection, fistula plug） have emerged as novel therapies for fistula. The anal fistula plug （e.g., based on small intestinal submucosa （SIS）） has attracted increasing attention because of short term healing rate and biocompatibility. However, challenges remain for this method such as plug falling as observed in clinics. To address this, this paper analyzes the case of SIS falling under physiological condition from mechanical point of view using ANSYS simulation. It then proposes three new geometrical structures for fistula plug and compares their mechanical behavior （e.g., axial stress, reaction of constraint） with that of clinically used structure （cone shape）. Based on the simulation, it optimizes the geometric parameters of fistula plug. The approach developed here can help to improve the design of fistula plug for better clinical treatments.
文摘Background：Pelvic organ prolapse （POP） is a major health concern that affects women.Surgeons have increasingly used prosthetic meshes to correct POP.However,the most common used is synthetic mesh,and absorbable mesh is less reported.This research aimed to evaluate the clinical effectiveness of porcine small intestinal submucosa （SIS）.Methods：Consecutive forty POP patients who met the inclusion criteria underwent pelvic reconstruction surgery with SIS between March 2012 and December 2013.The patients＆#39; clinical characteristics were recorded preoperatively.Surgical outcomes,measured by objective and subjective success rates,were investigated.We evaluated the quality of life （QOL） using the Pelvic Floor Distress Inventory-20 （PFDI-20） and the Pelvic Floor Impact Questionnaire-7 （PFIQ-7）.Sexual QOL was assessed by the Pelvic Organ Prolapse/Urinary Incontinence Sexual Function Questionnaire-12 （P1SQ-12）.Results：At postoperative 12 months,the subjective recurrence rate （7.5%） was much lower than the objective recurrence rate （40.0%）.Postoperatively,no erosion was identified.One underwent a graft release procedure because of urinary retention,and one had anus sphincter reconstruction surgery due to defecation urgency.Another experienced posterior vaginal wall infection where the mesh was implanted,accompanied by severe vaginal pain.Estrogen cream relieved the pain.One patient with recurrence underwent a secondary surgery with Bard Mesh because of stage 3 anterior vaginal wall prolapse.Scoring system of PFDI-20 was from 59.150 ± 13.143 preoperatively to 8.400 ± 4.749 postoperatively and PFIQ-7 was from 73.350 ± 32.281 to 7.150 ± 3.110,while PISQ-12 was from 15.825 ± 4.050 to 12.725 ± 3.471.Conclusions：QOL and the degree of subjective satisfaction were significantly improved postoperatively.Anterior repair deserves more attention because of the higher recurrence rate.The long-term follow-up of the patient is warranted to draw firm conclusion.
文摘Tympanic membrane perforationsoccur frequently in children,and can result in hearing loss,otorrhea,pain,and cholesteatoma.Due to the narrower ear canal in children,a postauricular incision is often needed to access the tympanic membrane for surgical repair.Endoscopic approaches are increasingly being used for tympanic membrane repair,reducing the need for postauricular incisions.As the need for a postauricular incision decreases,the demand for non-autologous grafting material has increased.Acellular porcine small intestinal submucosa (SIS) has been described in the literature as an alternative to commonly used autologous grafts,and is well suited for use with transcanal endoscopic ear surgery as a minimally invasive approach.This paper describes techniques for use of SIS in endoscopic tympanic membrane repair in children.
基金supported by the National Natural Science Foundation of China(31271058 and 81100327)the National High Technology Research and Development Program of China(2012AA020503)
文摘Acellular porcine small intestinal submucosa(SIS)has been used for esophagoplasty with success in a canine model.However,it did not lead to complete epithelialization.For better reconstruction,a cellular component is required.Moreover,promotion of angiogenesis with copper has been widely recognized by basic research as well as clinical studies.In this study,we have evaluated the feasibility and effectiveness of combined Cu and SIS(SIS-Cu patch)for the esophageal repair using a canine model.Eighteen male beagle dogs were subjected to surgical resection to produce cervical esophageal defects(5 cm in length,180°in range).SIS with Cu(5 or 25μmol L 1copper)or without Cu was patched on the esophageal defects.Barium esophagram and histology exam were carried out to evaluate the effectiveness of the therapy.As shown,the SIS-Cu graft promoted re-epithelialization,re-vascularization and muscular regeneration.SIS-Cu patch is more effective than SIS alone for esophageal repair,and the SIS+25μmol L 1Cu group demonstrated additional advantages over the SIS+5μmol L 1Cu.
基金funded by National Natural Science Foundation of China,grant number 81671924,81272105National Key Research and Development Plan of China,grant number 2017YFC1103301+6 种基金Health and Medical Treatment Collaborative Innovation Major Special Projects of Guangzhou,grant number 201508020253Science and Technology Key Project of Guangdong Province,grant number 2014B020212010Science and Technology Planning Project of Guangdong Province of China,grant number 2015B020233012Military Medical Innovation Special Projects,grant number 18CXZ029Zhejiang Provincial Basic Public Welfare Research Project,grant number GJ19H140001and China’s National Key R&D Programs(NKPs)grant number 2018YFB0407204.
文摘Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound healing still needs to be improved to meet clinical demands.To tackle this problem,platelet-rich plasma(PRP)is used due to its potency to promote proliferation,migration and adhesion of target cells.In this study,we applied PRP and SIS to skin wounds to explore their effects on wound healing by evaluating re-epithelialization,collagen production,angiogenesis and the inflammatory response.Methods:A1×1-cm full-thickness skin defectwas established in mice.Sixty mice were divided into four treatment groups:PRP+SIS,PRP,SIS and control.On days 3,5,7,10 and 14 post-surgery,tissue specimens were harvested.Haematoxylin and eosin,Masson’s trichrome,immunohistochemical and immunofluorescence double staining were used to visualize epidermal thickness,collagen and vascular regeneration and inflammation.Results:Wound contraction in the PRP and PRP+SIS groups was significantly greater,compared with the other groups,on days 3 and 5 post-surgery.A histological analysis showed higher collagen expression in the PRP and PRP+SIS groups on day 7,whichwas associated with a thicker epidermal layer on day 14.In addition,immunohistochemical staining showed that CD31-positive blood vessels and vascular endothelial growth factor expression in the PRP+SIS and PRP groups were significantly higher,compared with the control group.Furthermore,immunofluorescence double staining showed that the number of M1 and M2 macrophages in the PRP+SIS and PRP groups was higher,compared with the control and SIS groups alone,on day 3.However,on day 7,the number of M1 macrophages dramatically decreased in the PRP+SIS and PRP groups.The ratio of M2 to M1 macrophages in the PRP+SIS and PRP groups was 3.97 and 2.93 times that of the control group and 4.56 and 3.37 times that of the SIS group,respectively.Conclusion:Co-administration of SIS and PRP has a better effect on promoting angiogenesis,reepithelialization and collagen regeneration in managing acute wound healing than either agent alone.
基金This study was funded by Ningbo Municipal Bureau of Science and Technology(Grant No.202002N3125).
文摘There has been an increase in the incidence of hypopharyngeal and cervical esophageal cancer worldwide,and hence growing needs for hypopharyngeal and cervical esophageal tissue repair.This work produced a bi-layer composite scaffold with decellularized small intestine submucosa and polylactic-co-glycolic acid,which resembled the layered architectures of its intended tissues.The decellularized small intestine submucosa contained minimal residual DNA(52.5±61.2 ng/mg)and the composite scaffold exhibited satisfactory mechanical properties(a tensile modulus of 21.1±64.8 MPa,an ultimate tensile strength of 14.0±62.9MPa and a failure strain of 26.9±65.1%).The interactions between cells and the respective layers of the scaffold were characterized by CCK-8 assays,immunostaining and Western blotting.Desirable cell proliferation and phenotypic behaviors were observed.These results have provided an important basis for the next-step in vivo studies of the scaffold,and bode well for its future clinical applications.
文摘目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱细胞基质胶和白藜芦醇(resveratrol,Rev)溶液配制成膜片状的负载白藜芦醇的SIS脱细胞基质(Rev/SIS)。取小型猪3只,在每只猪背建立4个4 cm×4 cm的全层皮肤缺损创面,按照不同治疗手段随机分为4组:生理盐水纱布组(对照组)、白藜芦醇组(Rev组)、SIS脱细胞基质组(SIS组)和负载白藜芦醇的SIS脱细胞基质组(Rev/SIS组)。分别记录术后第7、14、21、28天的创面愈合及术后第42天的瘢痕形成状况;采集创缘组织行组织学检查和免疫组化染色分析,评估愈合过程中的炎性浸润和再血管化水平,以及所形成瘢痕组织的皮肤厚度和胶原沉积情况;并分别在术后第3、7天检测各组创面中炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、M1型巨噬细胞标志物i NOS和M2型巨噬细胞标志物ARG-1的m RNA相对表达量。结果术后第7、14、21天,SIS组和Rev/SIS组创面残留率均显著低于对照组(P<0.05),术后第21天Rev/SIS组创面残留率低于SIS组(P<0.001),术后第28天除Rev/SIS组外其余3组创面均未完全愈合;术后第42天Rev/SIS组瘢痕面积最小,质地柔软,扁平且无明显色沉。Rev/SIS组TNF-a、IL-6 m RNA表达量均低于对照组(P<0.05),3种治疗手段均能降低创面i NOS m RNA表达(P<0.001),但以Rev/SIS组最为显著(P<0.05),且Rev/SIS组的ARG-1 m RNA表达高于其余3组(P<0.05),该组创面炎性浸润程度最低,再血管化水平最高。其余3组瘢痕的表皮、真皮厚度均低于对照组(P<0.01),也以Rev/SIS组改善最为显著(P<0.0001),且该组胶原沉积与正常皮肤相似,排列有序。结论尽管SIS脱细胞基质和白藜芦醇单独使用也可能产生抗炎效应,在促进创面愈合的同时,减少瘢痕形成面积,有效提高愈合质量;但二者联合应用的效果更为显著,Rev/SIS脱细胞基质为开发新型皮肤创面敷料提供了一种潜在选择。
基金supported by the National Natural Science Foundation of China(No.81571919)LiaoNing Revitalization Talents Program(No.XLYC1907124).
文摘With the development of tissue engineering,the required biomaterials need to have the ability to promote cell adhesion and proliferation in vitro and in vivo.Especially,surface modification of the scaffold material has a great influence on biocompatibility and functionality of materials.The small intestine submucosa(SIS)is an extracellular matrix isolated from the submucosal layer of porcine jejunum,which has good tissue mechanical properties and regenerative activity,and is suitable for cell adhesion,proliferation and differentiation.In recent years,SIS is widely used in different areas of tissue reconstruction,such as blood vessels,bone,cartilage,bladder and ureter,etc.This paper discusses the main methods for surface modification of SIS to improve and optimize the performance of SIS bioscaffolds,including functional group bonding,protein adsorption,mineral coating,topography and formatting modification and drug combination.In addition,the reasonable combination of these methods also offers great improvement on SIS surface modification.This article makes a shallow review of the surface modification of SIS and its application in tissue engineering.
基金This work was supported by the National Key R&D Program of China(No.2017YFC1104702)Sichuan Science and Technology Program(No.2019JDRC0020)1.3.5 project for disciplines of excellence,West China Hospital,Sichuan University(No.ZYJC18002).
文摘Currently the standard surgical treatment for bladder defects is augmentation cystoplasty with autologous tissues,which has many side effects.Biomaterials such as small intestine submucosa(SIS)can provide an alternative scaffold for the repair as bladder patches.Previous studies have shown that SIS could enhance the capacity and compliance of the bladder,but its application is hindered by issues like limited smooth muscle regeneration and stone formation since the fast degradation and poor mechanical properties of the SIS.Procyanidins(PC),a natural bio-crosslinking agent,has shown anti-calcification,anti-inflammatory and anti-oxidation properties.More importantly,PC and SIS can crosslink through hydrogen bonds,which may endow the material with enhanced mechanical property and stabilized functionalities.In this study,various concentrations of PC-crosslinked SIS(PC-SIS)were prepared to repair the full-thickness bladder defects,with an aim to reduce complications and enhance bladder functions.In vitro assays showed that the crosslinking has conferred the biomaterial with superior mechanical property and anti-calcification property,ability to promote smooth muscle cell adhesion and upregulate functional genes expression.Using a rabbit model with bladder defects,we demonstrated that the PC-SIS scaffold can rapidly promote in situ tissue regrowth and regeneration,in particular smooth muscle remodeling and improvement of urinary functions.The PC-SIS scaffold has therefore provided a promising material for the reconstruction of a functional bladder.
