Skip to main content
SpringerLink
Log in

Identification and characterization of neurotrophic factors in porcine small intestinal submucosa

  • Original Article
  • Biomaterials
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

“Small intestinal submucosa” (SIS) is a natural degradable biomaterial derived from the small intestine of vertebrates. Porcine SIS has been widely used in repairing various tissues and organs, especially in nervous tissues because intrinsic autonomic nerve plexuses are included in submucosal layer. However, little is known about active neurotrophic factors in this material. In this study, we proposed a method to prepare an extract from decellularized SIS and to identify and characterize the major neurotrophic factors in it. Decellularized SIS extract was obtained by sequential procedures: mechanical disassociation, degreasing, enzyme digestion, freeze-drying, freezer-milling, extracting, concentrating, and filtering. Contamination with cellular components in the SIS extract was evaluated by hematoxylin and eosin (H&E) staining and the content of genomic DNA. Major neurotrophic factors in SIS extract was assessed quantitatively by ELISA. The effects of SIS extracts on cells were evaluated by observation of neurite outgrowth from PC12 cells. Decellularized SIS extract was obtained successfully by the proposed method and no cellular component was found within it. Brain-derived neurotrophic factor (BDNF), erythropoietin (EPO), glial cellderived neurotrophic factor (GDNF), nerve growth factor (NGF), and platelet-derived growth factor (PDGF) were identified and characterized form SIS extract. The bioactivity of SIS extract was assessed in terms of neurite outgrowth from PC12 cells. In summary, we prepared a decellularized extract from porcine SIS and identified major neurotrophic factors present in it. SIS extract also had bioactivity with PC12 cells. From these results, we conclude that SIS may be useful to repair or improve nervous system function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. CD Prevel, BL Eppley, DJ Summerlin, JR Jackson, M McCarty, SF Badylak, Small intestinal submucosa: utilization for repair of rodent abdominal wall defects, Ann Plast Surg, 35, 374 (1995).

    Article  PubMed  CAS  Google Scholar 

  2. GC Lantz, SF Badylak, MC Hiles, AC Coey, LA Geddes, K Kokini, GE Sandusky, RJ Mor, Small intestinal submucosa as a vascular graft: a review, J Invest Surg, 6, 297 (1993).

    Article  PubMed  CAS  Google Scholar 

  3. CE Schmidt, JB Leach, Neural tissue engineering:strategies for repair and regeneration, Annu Rev Biomed Eng, 5, 293 (2003).

    Article  PubMed  CAS  Google Scholar 

  4. DH Oh, IY Lee, M Choi, SH Kim, H Son, Comparison of neurite outgrowth induced by erythropoietin (EPO) and carbamylated erythropoietin (CEPO) in hippocampal neural progenitor cells, Korean J Physiol Pharmacol, 16, 281 (2012).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  5. M Choi, SY Ko, SE Wang, SH Lee, DH Oh, YS Kim, H Son, Carbamylated erythropoietin promotes neurite outgrowth and neuronal spine formation in association with CBP/p300, Biochem Biophy Res Comm, 446, 79 (2014).

    Article  CAS  Google Scholar 

  6. A Jezierski, K Rennie, B Zurakowski, M Ribecco-Lutkiewicz, J Haukenfrers, A Ajji, A Gruslin, M Sikorska, M Bani- Yaghoub, Neuroprotective Effects of GDNF-expressing Human Amniotic Fluid Cells, Stem Cell Rev, 10, 251 (2014).

    Article  PubMed  CAS  Google Scholar 

  7. Z Gale, PR Cooper, BA Scheven, Glial cell line-derived neurotrophic factor influences proliferation of osteoblastic cells, Cytokine, 57, 276 (2012).

    Article  PubMed  CAS  Google Scholar 

  8. CH Ho, PW Liao, SS Yang, FS Jaw, YC Tsai, The use of porcine small intestine submucosa implants might be associated with a high recurrence rate following laparoscopic herniorrhaphy, J Formos Med Assoc, 13, 1 (2013).

    Article  Google Scholar 

  9. M Okada, TR Payne, H Oshima, N Momoi, K Tobita, J Huard, Differential efficacy of gels derived from small intestinal submucosa as an injectable biomaterial for myocardial infarct repair, Biomaterials, 31, 7678 (2010).

    Article  PubMed  CAS  Google Scholar 

  10. S Hong, G Kim, Electrospun micro/nanofibrous conduits composed of poly(-caprolactone) and small intestine submucosa powder for nerve tissue regeneration, J Biomed Mater Res B Appl Biomater, 94, 421 (2010).

    PubMed  Google Scholar 

  11. K Fujita, P Lazarovici, G Guroff, Regulation of the differentiation of PC 12 pheochromocytoma cells, Environ Health Perspect, 80, 127, (1989).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. S Negrini, R D’Alessandro, J Meldolesi, NGF signaling in PC12 cells: the cooperation of p75(NTR) with TrkA is needed for the activation of both mTORC2 and the PI3K signalling cascade, Biol Open, 2, 855 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  13. LA Greene, AS Tischler, Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor, Proc Natl Acad Sci USA, 73, 2424 (1976).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  14. LA Greene, Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium, J Cell Biol, 78, 747 (1978).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  15. MA Dichter, AS Tischler, LA Greene, Nerve growth factorinduced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line, Nature, 268, 501 (1977).

    Article  PubMed  CAS  Google Scholar 

  16. JC Luo, W Chen, XH Chen, TW Qin, YC Huang, HQ Xie, XQ Li, ZY Qian, ZM Yang, A multi-step method for preparation of porcine small intestinal submucosa (SIS), Biomaterials, 32, 706 (2011).

    Article  PubMed  CAS  Google Scholar 

  17. W Chen, C Li, S Wu, H Xie, J Luo, Effect of acellular process on small intestinal submucosa cell residue and growth factor content, Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi, 24, 94 (2010).

    PubMed  Google Scholar 

  18. SY Chun, GJ Lim, TG Kwon, EK Kwak, BW Kim, A Atala, JJ Yoo, Identification and characterization of bioactive factors in bladder submucosa matrix, Biomaterials, 28, 4251 (2007).

    Article  PubMed  CAS  Google Scholar 

  19. R Schicho, G Skofitsch, J Donnerer, Regenerative effect of human recombinant NGF on capsaicin-lesioned sensory neurons in the adult rat, Brain Res, 815, 60 (1999).

    Article  PubMed  CAS  Google Scholar 

  20. CJ Serpe, SC Byram, VM Sanders, KJ Jones, Brain-derived neurotrophic factor supports facial motoneuron survival after facial nerve transection in immunodeficient mice, Brain Behav Immun, 19, 173 (2005).

    Article  PubMed  CAS  Google Scholar 

  21. Y Su, BF Zeng, CQ Zhang, KG Zhang, XT Xie, Study of biocompatibility of small intestinal submucosa (SIS) with Schwann cells in vitro, Brain Res, 1145, 41 (2007).

    Article  PubMed  CAS  Google Scholar 

  22. S Morcuende, R Muñoz-Hernández, B Benítez-Temiño, AM Pastor, RR de la Cruz, Neuroprotective effects of NGF, BDNF, NT-3 and GDNF on axotomized extraocular motoneurons in neonatal rats, Neuroscience, 250, 31 (2013).

    Article  PubMed  CAS  Google Scholar 

  23. Sarina, Y Yagi, O Nakano, T Hashimoto, K Kimura, Y Asakawa, M Zhong, S Narimatsu, E Gohda, Induction of neurite outgrowth in PC12 cells by artemisinin through activation of ERK and p38 MAPK signaling pathways, Brain Res, 15, 61 (2013).

    Article  Google Scholar 

  24. JW Fawcett, RJ Keynes, Peripheral nerve regeneration, Annu Rev Neurosci, 13, 43 (1990).

    Article  PubMed  CAS  Google Scholar 

  25. DM Thompson, HM Buettner, Oriented Schwann cell monolayers for directed neurite outgrowth, Ann Biomed Eng, 32, 1120 (2004).

    Article  PubMed  Google Scholar 

  26. KN Kang, JY Lee, YD Kim, BN Lee, HH Ahn, B Lee, G Khang, SR Park, BH Min, JH Kim, HB Lee, MS Kim, Regeneration of completely transected spinal cord using scaffold of poly (D,L-lactide-co-glycolide)/small intestinal submucosa seeded with rat bone marrow stem cells, Tissue Eng Part A, 17, 2143 (2011).

    Article  PubMed  CAS  Google Scholar 

  27. HS Phillips, JM Hains, M Armanini, GR Laramee, SA Johnson, JW Winslow, BDNF mRNA is decreased in the hippocampus of individuals with Alzheimer’s disease, Neuron, 7, 695 (1991).

    Article  PubMed  CAS  Google Scholar 

  28. C Hock, K Heese, C Hulette, C Rosenberg, U Otten, Regionspecific neurotrophin imbalances in Alzheimer disease: decreased levels of brain-derived neurotrophic factor and increased levels of nerve growth factor in hippocampus and cortical areas, Arch Neurol, 57, 846 (2000).

    Article  PubMed  CAS  Google Scholar 

  29. S Peng, J Wuu, EJ Mufson, M Fahnestock, Precursor form of brain-derived neurotrophic factor and mature brain-derived neurotrophic factor are decrease, in the pre-clinical stages of Alzheimer’s disease, J Neurochem, 93, 1412 (2005).

    Article  PubMed  CAS  Google Scholar 

  30. DW Howells, MJ Porritt, JY Wong, PE Batchelor, R Kalnins, AJ Hughes, Reduced BDNF mRNA expression in the Parkinson’s disease substantia nigra, Exp Neurol, 166, 127 (2000).

    Article  PubMed  CAS  Google Scholar 

  31. C Zuccato, M Marullo, P Conforti, ME MacDonald, M Tartari, E Cattaneo, Systematic assessment of BDNF and its receptor levels in human cortices affected by Huntington’s disease, Brain Pathol, 18, 225 (2008).

    Article  PubMed  CAS  Google Scholar 

  32. T Nishio, N Sunohara, S Furukawa, Neutrophin switching in spinal motoneurons of amyotrophic lateral sclerosis, Neuroreport, 9, 1661 (1998).

    Article  PubMed  CAS  Google Scholar 

  33. YE Sun, H Wu, The ups and downs of BDNF in Rett syndrome, Neuron, 49, 321 (2006).

    Article  PubMed  CAS  Google Scholar 

  34. G Favalli, J Li, P Belmonte-de-Abreu, AH Wong, ZJ Daskalakis, The role of BDNF in the pathophysiology and treatment of schizophrenia, J Psychiatr Res, 46, 1 (2012).

    Article  PubMed  Google Scholar 

  35. E Castren, V Voikar, T Rantamaki, Role of neurotrophic factors in depression, Curr Opin Pharmacol, 7, 18 (2007).

    Article  PubMed  CAS  Google Scholar 

  36. DW Metzger, TF Moyad, T McPherson, SF Badylak, Cytokine and antibody responses to xenogeneic SIS trans-plants, Paper presented at the first SIS symposium, Orlando, Florida, 11-12 December (1996).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Clinical Research Institute, Daejeon St Mary’s Hospital, College of Medicine, Catholic University of Korea, 520-2 Daeheung-dong, Jung-gu Daejeon, 301-723, Korea

    Keum-Jin Yang, Ki Cheol Park & Hyunsu Choi

  2. Korea Research Institute of Standards and Science, 267 Gajeong-ro, Yuseong-gu, Daejeon, 305-340, Korea

    Jun-Hyuk Choi & Sang-Ryoul Park

  3. Department of Neurosurgery, Daejeon St Mary’s Hospital, College of Medicine, Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 137-701, Korea

    Il-Woo Lee & Hyung-Jin Lee

Authors
  1. Keum-Jin Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ki Cheol Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hyunsu Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jun-Hyuk Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sang-Ryoul Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Il-Woo Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hyung-Jin Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hyung-Jin Lee.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, KJ., Park, K.C., Choi, H. et al. Identification and characterization of neurotrophic factors in porcine small intestinal submucosa. Tissue Eng Regen Med 11, 372–378 (2014). https://doi.org/10.1007/s13770-014-0043-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13770-014-0043-6

Key words

Search

Navigation