Effect of Small Intestinal Submucosa Sponges on the Attachment and Proliferation Behavior of Schwann Cells

한갑수,강길선,고현아,장나금,송정은 한국고분자학회 2014 Macromolecular Research Vol.22 No.12

The aim of the study was to investigate the effects of small intestinal submucosa (SIS) sponges, a naturalbiodegradable polymer scaffold, on the adhesion behavior and the proliferation of Schwann cells (SCs). SIS spongeswere prepared by varying the contents of SIS powders (1%, 2%, and 3% SIS concentration) and SCs were seededon the sponges and incubated to investigate the adhesion and proliferation of SCs. A series of analytical process wasconducted to observe the water absorption rate of sponges and confirmed the proliferation and morphology of SCsby scanning electron microscopy (SEM), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)assay, reverse transcription polymerase chain reaction (RT-PCR), and histological evaluation. The best adhesionbehavior and the proliferation of SCs were observed on the sponge with 2% powder content. Also, analysis of messengerribonucleic acid (mRNA) expression using the neuron-specific enolase (NSE) and neurofilament protein (NF)nerve markers showed that SC gene expression was excellent in the 2% SIS sponge. In vivo hematoxylin and eosin(H&E) staining and immunofluorescence results showed that cells and tissue formed well in the pores of spongeswith 2% and 3% SIS powder contents, compared with formation in the 1% SIS sponge. The SIS sponges provided anunsuitable habitat for the adhesion and proliferation of cells in that the pore size was small. In this study, the adhesionbehavior and the proliferation could be changed depending on the pore size and SIS powder content in the sponges. Thus, an appropriate powder content of SIS should be used to prepare SIS sponge scaffolds.

Fabrication of Electrospun PLGA and Small Intestine Submucosa-blended Nanofibrous Membranes for Tissue Engineering and their Biocompatibility

Kihoon KIM,Jong Yoon LEE,Young Tai YOO,Hwi-Yool KIM 한국고분자학회 2016 한국고분자학회 학술대회 연구논문 초록집 Vol.41 No.2

Implantation of Bone Marrow Mesenchymal Stem Cells into Small Intestinal Submucosa Improves Bile Duct Injury in Rabbits

Li Ying,Wang Piao,Hu Xiao-dong,Zeng Jing-da,Fang Cheng,Gan Yu,Peng Fang-yi,Yang Xiao-li,Luo De,Li Bo,Su Song 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.5

BACKGROUND: Bile duct injury (BDI), which may occur during cholecystectomy procedures and living-donor liver transplantation, leads to life-altering complications and significantly increased mortality and morbidity. Tissue engineering, as an emerging method, has shown great potential to treat BDI. Here, we aimed to explore the application of small intestinal submucosa (SIS) matrix composites with bone marrow mesenchymal stem cells (BMSCs) to treat BDI in a rabbit model. METHODS: Rabbit-derived BMSCs were used as seed cells. Porcine SIS was used as the support material. Five centimetres of the common bile duct was dissected, and 1/3–1/2 of the anterior wall diameter was transversely incised to construct the rabbit BDI model. Then, SIS materials without/with BMSCs were inserted into the common bile duct of the BDI rabbits. After 1, 2, 4, and 8 weeks of implantation, the common bile duct was removed. Haematoxylin and eosin (HE) staining was used to assess pathological alterations in the common bile duct, while immunohistochemical staining and western blotting were used to detect expression of the epithelial cell markers CK19 and E-cadherin. Scanning electron microscopy was used to evaluate BMSC growth. RESULTS: Compared with BMSCs alone, SIS-attached BMSCs had increased growth. HE staining showed that the injured bile duct healed well and that the complex gradually degraded as the time from implantation increased. Immunohistochemical staining and western blotting showed that compared with the control group, the in vivo complex group had significantly elevated expression levels of CK19 and E-cadherin. CONCLUSION: BMSC implantation into SIS could improve BDI in rabbits, which might have clinical value for BDI treatment. BACKGROUND: Bile duct injury (BDI), which may occur during cholecystectomy procedures and living-donor liver transplantation, leads to life-altering complications and significantly increased mortality and morbidity. Tissue engineering, as an emerging method, has shown great potential to treat BDI. Here, we aimed to explore the application of small intestinal submucosa (SIS) matrix composites with bone marrow mesenchymal stem cells (BMSCs) to treat BDI in a rabbit model. METHODS: Rabbit-derived BMSCs were used as seed cells. Porcine SIS was used as the support material. Five centimetres of the common bile duct was dissected, and 1/3–1/2 of the anterior wall diameter was transversely incised to construct the rabbit BDI model. Then, SIS materials without/with BMSCs were inserted into the common bile duct of the BDI rabbits. After 1, 2, 4, and 8 weeks of implantation, the common bile duct was removed. Haematoxylin and eosin (HE) staining was used to assess pathological alterations in the common bile duct, while immunohistochemical staining and western blotting were used to detect expression of the epithelial cell markers CK19 and E-cadherin. Scanning electron microscopy was used to evaluate BMSC growth. RESULTS: Compared with BMSCs alone, SIS-attached BMSCs had increased growth. HE staining showed that the injured bile duct healed well and that the complex gradually degraded as the time from implantation increased. Immunohistochemical staining and western blotting showed that compared with the control group, the in vivo complex group had significantly elevated expression levels of CK19 and E-cadherin. CONCLUSION: BMSC implantation into SIS could improve BDI in rabbits, which might have clinical value for BDI treatment.

Rat Peripheral Nerve Regeneration Using Nerve Guidance Channel by Porcine Small Intestinal Submucosa

Yi, Jin-Seok,Lee, Hyung-Jin,Lee, Hong-Jae,Lee, Il-Woo,Yang, Ji-Ho The Korean Neurosurgical Society 2013 Journal of Korean neurosurgical society Vol.53 No.2

Objective : In order to develop a novel nerve guidance channel using porcine small intestinal submucosa (SIS) for nerve regeneration, we investigated the possibility of SIS, a tissue consisting of acellular collagen material without cellular immunogenicity, and containing many kinds of growth factors, as a natural material with a new bioactive functionality. Methods : Left sciatic nerves were cut 5 mm in length, in 14 Sprague-Dawley rats. Grafts between the cut nerve ends were performed with a silicone tube (Silicon group, n=7) and rolled porcine SIS (SIS group, n=7). All rats underwent a motor function test and an electromyography (EMG) study on 4 and 10 weeks after grafting. After last EMG studies, the grafts, including proximal and distal nerve segments, were retrieved for histological analysis. Results : Foot ulcers, due to hypesthesia, were fewer in SIS group than in Silicon group. The run time tests for motor function study were 2.67 seconds in Silicon group and 5.92 seconds in SIS group. Rats in SIS group showed a better EMG response for distal motor latency and amplitude than in Silicon group. Histologically, all grafts contained some axons and myelination. However, the number of axons and the degree of myelination were significantly higher in SIS group than Silicon group. Conclusion : These results show that the porcine SIS was an excellent option as a natural biomaterial for peripheral nerve regeneration since this material contains many kinds of nerve growth factors. Furthermore, it could be used as a biocompatible barrier covering neural tissue.

Evaluation of decellularization protocols for production of tubular small intestine submucosa scaffolds for use in oesophageal tissue engineering

Syed, O.,Walters, N.J.,Day, R.M.,Kim, H.W.,Knowles, J.C. Elsevier BV 2014 ACTA BIOMATERIALIA Vol.10 No.12

Small intestine submucosa (SIS) has emerged as one of a number of naturally derived extracellular matrix (ECM) biomaterials currently in clinical use. In addition to clinical applications, ECM materials form the basis for a variety of approaches within tissue engineering research. In our preliminary work it was found that SIS can be consistently and reliably made into tubular scaffolds which confer certain potential advantages. Given that decellularization protocols for SIS are applied to sheet-form SIS, it was hypothesized that a tubular-form SIS would behave differently to pre-existing protocols. In this work, tubular SIS was produced and decellularized by the conventional peracetic acid-agitation method, peracetic acid under perfusion along with two commonly used detergent-perfusion protocols. The aim of this was to produce a tubular SIS that was both adequately decellularized and possessing the mechanical properties which would make it a suitable scaffold for oesophageal tissue engineering, which was one of the goals of this work. Analysis was carried out via mechanical tensile testing, DNA quantification, scanning electron and light microscopy, and a metabolic assay, which was used to give an indication of the biocompatibility of each decellularization method. Both peracetic acid protocols were shown to be unsuitable methods with the agitation-protocol-produced SIS, which was poorly decellularized, and the perfusion protocol resulted in poor mechanical properties. Both detergent-based protocols produced well-decellularized SIS, with no adverse mechanical effects; however, one protocol emerged, SDS/Triton X-100, which proved superior in both respects. However, this SIS showed reduced metabolic activity, and this cytotoxic effect was attributed to residual reagents. Consequently, the use of SIS produced using the detergent SD as the decellularization agent was deemed to be the most suitable, although the elimination of the DNase enzyme would give further improvement.

혈관대치물 응용을 위한 소장점막하층의 체외 특성에 관한 기초연구

신재승,선경,지경수,박기동,조양현,민병주,황진욱,조종호 한국생체재료학회 2003 생체재료학회지 Vol.7 No.4

Various vascular diseases inspire us to develop the ideal vascular substitute. Small intestinal submucosa (SIS) is a feasible and wide available matrix as biomaterials for the vascular substitutes. An in-vitro preliminary study for the mechanical and biological properties of the heparin bonded SIS as a vascular substitute was preceded by the development of the anti-thrombogenecity technique using heparin bonding methods. The study groups were divided, e.g., SIS group for fresh SIS; SIS-GA group for SIS treated with glutaraldehyde (GA); and SIS-GA-Heparin group for SIS treated with glutaraldehyde and heparin. The quantity analysis for heparin of the heparin bonded SIS shows stable concentration, e.g., 0.339±0.084 g/cm2. GA treated heparinized tissue (SIS-GA-Heparin) reveals highest digestion resistance to collagenase among the study groups. The data of the study for maximal stress show highest in SIS-GA-Heparin group (0.55 ±0.19 kg/mm2), and lowest in SIS group (0.28±0.07 kg/mm2). The data of the study for maximal strain show highest in SIS group (78.69±8.59%), and lowest in SIS-GA-Heparin group (34.85±6.51%). As concerns degree of heparinization, digestion resistance to collagenase and mechanical properties, GA treated heparinized tissue (SIS-GA-Heparin) is superior to the other types of SIS as a vascular substitutes. According to this study, further studies for the antithrombogenecity and patency rate are needed.

창상드레싱을 위한 소장점막하조직을 이용한 시트의 제조와 이의 특성 결정

강길선,조선행,이해방,신혜원,김문식,김선화 한국생체재료학회 2004 생체재료학회지 Vol.8 No.3

Biological wound dressings are widely used as a temporary cover for an open wound to promote wound healing, control wound exudate, and decrease wound contamination as well as evaporative water loss. Small intestinal submucosa (SIS) is consisted with collagen and glycosaminoglycan and it contains some growth factors. Very recently, it has been recognized that SIS has been tested successfully for the application for biomaterials without xenograft immuno-rejection response. Growth factors consisted in SIS play an important part in full thickness wound healing. SIS sheet is made simply and is a type having proteins and growth factors in SIS safely. Crosslinked SIS sheet was characterized for the possibility of the wound dressing and tissue engineered scaffold. Separated SIS sheet crosslinked with 1-ethyl-(3,3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) solution (in distilled water : ethanol = 5 : 95) with 1, 5, 10, 50 and 100 mM concentration for 24 hours and lyophilized. SIS sheets were characterized by tensile strength, Fourier transform infrared (FT-IR), scanning electron microscopy (SEM), water uptake and in vitro test. From the result of SEM observation, we could confirm that the water uptake of SIS sheet was slightly decreased by using EDC crosslinking agent. It was observed that the concentration of EDC was 50 mM to get high tensile strength and in vitro characterization. In conclusion, this study suggests that SIS sheet could be useful for the applications to wound dressing.

Evaluation of the Therapeutic Potential In vitro and In vivo of the SIS/PLGA Scaffolds for Costal Cartilage Regeneration

차세롬,조선아,이선의,장나금,조성준,송정은,강길선 한국고분자학회 2016 Macromolecular Research Vol.24 No.5

In this study, porcine small intestinal submucosa (SIS) without immune response were used to make SIS/ PLGA scaffolds with different SIS content i.e. 0, 10, 20, 40, and 80 wt%. Further, the attachment and proliferation of costal cartilage cells (CCs) in SIS/PLGA scaffolds were characterized. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to verify the effects of SIS on extracellular matrix (ECM) formulation. The CCs specific gene expression was confirmed by polymerase chain reaction (PCR). In conclusion, the sGAG production was formed to be higher in the 20 wt% SIS/PLGA scaffolds, which showed enhanced CCs cell growth and proliferation compared with other scaffolds.

Preparation and Characterization of Swelled Small Intestinal Submucosa (SIS) sheet for Wound Healing

김선화,홍금덕,신혜원,김문석,조선행,강길선,이해방 한국고분자학회 2004 한국고분자학회 학술대회 연구논문 초록집 Vol.29 No.2

전층 피부 손상에의 생물학적 상처 치료용 소장점막하조직 드레싱

강조아,이일우,강길선,이해방,이상진 한국생체재료학회 2002 생체재료학회지 Vol.6 No.4

Biological wound dressings are widely used as a temporary coverage for an open wound to promote wound healing, control wound exudate, and decrease wound contamination as well as evaporative water loss. Very recently, it has been recognized that a porcine small intestinal submucosa (SIS) as a natural biomaterial mainly composed of collagen and glycosaminoglycan has been tested successfully for the application for biomaterials without xenograft immuno-rejection response. In this study, the possibility of SIS as a biological wound dressing in 15 mm diameter of full-thickness wounds made on the back of Sprague-Dawley (SD) rats had been investigated. Three types of wound dressing manufactured from SIS powder and fluid were applied with ointment, Carbopol gel, and Pluronic gel, respectively. The rate of wound contraction was measured daily for 10 days. The wound healing revealed better contraction rate for the SIS-treated wounds than those of control. Gel form showed better results than ointment form at same concentration of SIS powder. Especially, Carbopol gel condition with SIS fluid was better than SIS powder. In conclusion, powder and fluid shape of SIS may have positive effects on the wound contraction and the formation of dermis due to the stimulation of many kinds of bioactive substance as growth factor and cytokine in SIS for the wound healing.