CONSTRUCTION OF TISSUE ENGINEERED COMPOSITE WITH THERMOSENSITIVE COLLAGEN HYDROGEL IN DYNAMIC CULTURE SYSTEM_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

XUFeixiang ¹ , ZHAOJinsong ² , GUOBin ¹ , MAJianchao ¹ ,  HUANGLanfeng ¹

1. Department of Orthopaedics, the Second Hospital of Jilin University, Changchun Jilin, 130041, P.R.China;
2. Department of Ophthalmology, the Second Hospital of Jilin University, Changchun Jilin, 130041, P.R.China;

Corresponding author：

HUANGLanfeng, Email: hlf@jlu.edu.cn

Keywords：

Dynamic culture; Tissue engineering; Extracellular matrix; Seeding cell; Scaffold material

DOI：

10.7507/1002-1892.20160061

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To explore the morphological and functional features of tissue engineered composite constructed with bone mesenchymal stem cells (BMSCs) as seeding cells, thermosensitive collagen hydrogel (TCH) and poly-L-lactic acid (PLLA) as the extracellular matrix (ECM) scaffolds in the dynamic culture system. Methods BMSCs were separated from long bones of Fischer344 rat, and cultured; and BMSCs at the 3rd generation were seeded on the ECM scaffold constructed with braided PLLA fiber and TCH. The BMSCs-ECM scaffold composite was cultured in the dynamic culture system which was designed by using an oscillating device at a frequency of 0.5 Hz and at swing angle of 70° (experimental group), and in the static culture system (control group) for 7 days. The general observation and scanning electron microscopy (SEM) observation were performed; total DNA content was measured at 0, 1, 3, and 7 days. Results PLLA was surrounded by collagen to form translucent gelatiniform in 2 groups; and compact membrane developed on the surface of PLLA. SEM observation showed that BMSCs had high viability and were fusiform in shape with microvilli on the surface of cells, and arranged in line; collagen and cells filled in the pores of PLLA fiber in the experimental group. The cells displayed a flat shape on the surface; there were less cells filling in the pores of PLLA fiber in the control group. At 1, 3, and 7 days, total DNA content in the experimental group was significantly higher than that in control group (P < 0.05). The total DNA content were increased gradually with time in 2 groups, showing significant difference between at 0 day and at 7 days (P < 0.05). Conclusion The ECM constructed with TCH and PLLA has good biocompatibility. The dynamic cultivation system can promote the cell proliferation, distribution, and alignment on the surface of the composite, so it can be used for tissue engineered composite in vitro.

Citation： XUFeixiang, ZHAOJinsong, GUOBin, MAJianchao, HUANGLanfeng. CONSTRUCTION OF TISSUE ENGINEERED COMPOSITE WITH THERMOSENSITIVE COLLAGEN HYDROGEL IN DYNAMIC CULTURE SYSTEM. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(3): 309-313. doi: 10.7507/1002-1892.20160061 Copy

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18.	Hwang YS, Chiang PR, Hong WH, et al. Study in vivo intraocular biocompatibility of in situ gelation hydrogels: poly (2-ethyl oxazoline)-block-poly (epsilon-caprolactone)-block-poly (2-ethyl oxazoline) copolymer, matrigel and pluronic F127. PLoS One, 2013, 8(7): e67495.
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24.	Sun T, Norton D, Vickers N, et al. Development of a bioreactor for evaluating novel nerve conduits. Biotechnol Bioeng, 2008, 99(5): 1250-1260.
25.	Liu M, Liu N, Zang R, et al. Engineering stem cell niches in bioreactors. World J Stem Cells, 2013, 5(4): 124-135.
26.	Valmikinathan CM, Hoffman J, Yu X. Impact of scaffold micro and macro architecture on schwann cell proliferation under dynamic conditions in a rotating wall vessel bioreactor. Mater Sci Eng C Mater Biol Appl, 2011, 31(1): 22-29.
27.	Guénard V, Kletman N, Morrissey TK, et al. Syngeneic Schwann cells derived from adult nerves seeded in semipermeable guidance channels enhance peripheral nerve regeneration. J Neurosci, 1992, 12(9): 3310-3320.
28.	Brook GA, Plate D, Franzen R, et al. Spontaneous longitudinally orientated axonal regeneration is associated with the Schwann cell framework within the lesion site following spinal cord compression injury of the rat. J Neurosci Res, 1998, 53(1): 51-65.
29.	Lietz M, Dreesmann L, Hoss M, et al. Neuro tissue engineering of glial nerve guides and the impact of different cell types. Biomaterials, 2006, 27(8): 1425-1436.
30.	Laulicht B, Gidmark NJ, Trepathi A, et al. Localization of magnetic pills. Proc Natl Acad Sci U S A, 2011, 108(6): 2252-2257.
31.	Miller C, Jeftinija S, Mallapragada S. Micropatterned Schwann cell-seeded biodegradable polymer substrates significantly enhance neurite alignment and outgrowth. Tissue Eng, 2001, 7(6): 705-715.

1. Liu M, Liu N, Zang R, et al. Engineering stem cell niches in bioreactors. World J Stem Cells, 2013, 5(4): 124-135.
2. Cen L, Liu W, Cui L, et al. Collagen tissue engineering: development of novel biomaterials and applications. Pediatr Res, 2008, 63(5): 492-496.
3. Schumann D, Ekaputra AK, Lam CX, et al. Biomaterials/scaffolds. Design of bioactive, multiphasic PCL/collagen type I and type II-PCL-TCP/collagen composite scaffolds for functional tissue engineering of osteochondral repair tissue by using electrospinning and FDM techniques. Methods Mol Med, 2007, 140: 101-124.
4. Zhang CS, Lv G. Repair of sciatic nerve defects using tissue engineered nerves. Neural Regen Res, 2013, 8(21): 1985-1994.
5. Gambardella A, Nagaraju CK, O'Shea PJ, et al. Glycogen synthase kinase-3alpha/beta inhibition promotes in vivo amplification of endogenous mesenchymal progenitors with osteogenic and adipogenic potential and their differentiation to the osteogenic lineage. J Bone Miner Res, 2011, 26(4): 811-821.
6. James AW. Review of signaling pathways governing MSC osteogenic and adipogenic differentiation. Scientifica (Cairo), 2013, 2013: 684736.
7. Köllmer M, Buhrman JS, Zhang Y, et al. Markers are shared between adipogenic and osteogenic differentiated mesenchymal stem cells. J Dev Biol Tissue Eng, 2013, 5(2): 18-25.
8. El-Mahgoub ER, Ahmed E, Afifi RA, et al. Mesenchymal stem cells from pediatric patients with aplastic anemia: isolation, characterization, adipogenic, and osteogenic differentiation. Fetal Pediatr Pathol, 2014, 33(1): 9-15.
9. Scott JB, Afshari M, Kotek R, et al. The promotion of axon extension in vitro using polymer-templated fibrin scaffolds. Biomaterials, 2011, 32(21): 4830-4839.
10. Lundborg G. A 25-year perspective of peripheral nerve surgery: evolving neuroscientific concepts and clinical significance. J Hand Surg (Am), 2000, 25(3): 391-414.
11. Konofaos P, Ver Halen JP. Nerve repair by means of tubulization: past, present, future. J Reconstr Microsurg, 2013, 29(3): 149-164.
12. Marquard LM, Sakiyama-Elbert SE. Engineering peripheral nerve repair. Curr Opin Biotechnol, 2013, 24(5): 887-892.
13. Quigley AF, Bulluss KJ, Kyratzis IL, et al. Engineering a multimodal nerve conduit for repair of injured peripheral nerve. J Neural Eng, 2013, 10(1): 016008.
14. Salgado AJ, Oliveira JM, Martins A, et al. Tissue engineering and regenerative medicine: past, present, and future. Int Rev Neurobiol, 2013, 108: 1-33.
15. Saracino GA, Cigognini D, Silva D, et al. Nanomaterials design and tests for neural tissue engineering. Chem Soc Rev, 2013, 42(1): 225-262.
16. Feinberg AW, Parker KK. Surface-initiated assembly of protein nanofabrics. Nano Lett, 2010, 10(6): 2184-2191.
17. Sollerr EC, Tzeranis DS, Miu K, et al. Common features of optimal collagen scaffolds that disrupt wound contraction and enhance regeneration both in peripheral nerves and in skin. Biomaterials, 2012, 33(19): 4783-4791.
18. Hwang YS, Chiang PR, Hong WH, et al. Study in vivo intraocular biocompatibility of in situ gelation hydrogels: poly (2-ethyl oxazoline)-block-poly (epsilon-caprolactone)-block-poly (2-ethyl oxazoline) copolymer, matrigel and pluronic F127. PLoS One, 2013, 8(7): e67495.
19. Johnson EO, Zoubos AB, Soucacos PN. Regeneration and repair of peripheral nerves. Injury, 2005, 36 Suppl 4: S24-29.
20. Chung TW, Yang MC, Tseng CC, et al. Promoting regeneration of peripheral nerves in-vivo using new PCL-NGF/Tirofiban nerve conduits. Biomaterials, 2011, 32(3): 734-743.
21. Liu H, Wen W, Hu M, et al. Chitosan conduits combined with nerve growth factor microspheres repair facial nerve defects. Neural Regen Res, 2013, 8(33): 3139-3147.
22. Qi BW, Yu AX, Zhu SB, et al. Chitosan/poly (vinyl alcohol) hydrogel combined with Ad-hTGF-beta1 transfected mesenchymal stem cells to repair rabbit articular cartilage defects. Exp Biol Med (Maywood), 2013, 238(1): 23-30.
23. Ding K, Yang Z, Zhang YL, et al. Injectable thermosensitive chitosan/beta-glycerophosphate/collagen hydrogel maintains the plasticity of skeletal muscle satellite cells and supports their in vivo viability. Cell Biol Int, 2013, 37(9): 977-987.
24. Sun T, Norton D, Vickers N, et al. Development of a bioreactor for evaluating novel nerve conduits. Biotechnol Bioeng, 2008, 99(5): 1250-1260.
25. Liu M, Liu N, Zang R, et al. Engineering stem cell niches in bioreactors. World J Stem Cells, 2013, 5(4): 124-135.
26. Valmikinathan CM, Hoffman J, Yu X. Impact of scaffold micro and macro architecture on schwann cell proliferation under dynamic conditions in a rotating wall vessel bioreactor. Mater Sci Eng C Mater Biol Appl, 2011, 31(1): 22-29.
27. Guénard V, Kletman N, Morrissey TK, et al. Syngeneic Schwann cells derived from adult nerves seeded in semipermeable guidance channels enhance peripheral nerve regeneration. J Neurosci, 1992, 12(9): 3310-3320.
28. Brook GA, Plate D, Franzen R, et al. Spontaneous longitudinally orientated axonal regeneration is associated with the Schwann cell framework within the lesion site following spinal cord compression injury of the rat. J Neurosci Res, 1998, 53(1): 51-65.
29. Lietz M, Dreesmann L, Hoss M, et al. Neuro tissue engineering of glial nerve guides and the impact of different cell types. Biomaterials, 2006, 27(8): 1425-1436.
30. Laulicht B, Gidmark NJ, Trepathi A, et al. Localization of magnetic pills. Proc Natl Acad Sci U S A, 2011, 108(6): 2252-2257.
31. Miller C, Jeftinija S, Mallapragada S. Micropatterned Schwann cell-seeded biodegradable polymer substrates significantly enhance neurite alignment and outgrowth. Tissue Eng, 2001, 7(6): 705-715.

Chinese Journal of Reparative and Reconstructive Surgery

CONSTRUCTION OF TISSUE ENGINEERED COMPOSITE WITH THERMOSENSITIVE COLLAGEN HYDROGEL IN DYNAMIC CULTURE SYSTEM

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