期刊论文详细信息
BMC Veterinary Research
Evaluation of chitosan-GP hydrogel biocompatibility in osteochondral defects: an experimental approach
Luis CLC Silva2  Bruno Cogliati4  Raquel YA Baccarin1  Yara M Michelacci3  Edivaldo AN Martins2 
[1] Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, USP, São Paulo, SP, Brazil;Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, USP, São Paulo, SP, Brazil;Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, Rua Três de Maio, 100, São Paulo, 04044-020, SP, Brazil;Departamento de Patologia, Faculdade de Medicina Veterinária e Zootecnia, USP, São Paulo, SP, Brazil
关键词: Scaffold;    Biocompatibility;    Equine joint;    Type II collagen;    Proteoglycan;    Cartilage;    Chitosan-GP;   
Others  :  1119067
DOI  :  10.1186/s12917-014-0197-4
 received in 2013-08-08, accepted in 2014-08-15,  发布年份 2014
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【 摘 要 】

Background

Articular cartilage, because of its avascular nature, has little capacity for spontaneous healing, and tissue engineering approaches, employing different biomaterials and cells, are under development. Among the investigated biomaterials are the chitosan-based hydrogels. Although thoroughly studied in other mammalian species, studies are scarce in equines. So, the aim of the present study was to investigate the biocompatibility of chitosan-GP in horse joints submitted to high mechanical loads.

Results

An osteochondral defect was created by arthroscopy in the medial surface of lateral trochlea of talus of left or right leg, randomly selected, from six healthy geldings. The defect was filled up with chitosan-GP. The contralateral joint received an identical defect with no implant. The chondral fragment removed to produce the defect was collected, processed and used as the “Initial” sample (normal cartilage) for histology, immunohistochemistry, and metabolic labelling of PGs. After 180 days, the repair tissues were collected, and also analyzed. At the end of the experiment (180 days after lesion), the total number of cells per field in repair tissues was equal to control, and macrophages and polymorphonuclear cells were not detected, suggesting that no significant inflammation was present. These cells were able to synthesize type II collagen and proteoglycans (PGs). Nevertheless, the cell population in these tissues, both in presence of chitosan-GP and in untreated controls, were heterogeneous, with a lower proportion of type II collagen-positives cells and some with a fibroblastic aspect. Moreover, the PGs synthesized in repair tissues formed in presence or absence of chitosan-GP were similar to those of normal cartilage. However, the chitosan-GP treated tissue had an disorganized appearance, and blood vessels were present.

Conclusions

Implanted chitosan-GP did not evoke an important inflammatory reaction, and permitted cell growth. These cells were able to synthesize type II collagen and PGs similar to those synthesized in normal cartilage and in healing tissue without implant, indicating its chondrocyte nature.

【 授权许可】

   
2014 Martins et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Temeroff JS, Mikos AG: Review: tissue engeneering for regeneration of articular cartilage. Biomaterials 2000, 21:431-440.
  • [2]Lynn AK, Brooks RA, Bonfield W, Rushton N: Repair of defects in articular joints. J Bone Joint Surg Am 2004, 86:1093-1099.
  • [3]Beris AE, Lykissas MG, Papageorgiou CD, Georgoulis AD: Advances in articular cartilage repair. Injury 2005, 36:14-23.
  • [4]Grande DA, Southerland SS, Manji R, Pate DW, Schwartz RE, Lucas PA: Repair of articular cartilage defects using mesenchymal stem cells. Tissue Eng 1995, 1:345-353.
  • [5]Caplan AI, Correa D: The MSC: an injury drugstore. Cell Stem Cell 2011, 9:11-15.
  • [6]Garg T, Singh O, Arora S, Murthy R: Scaffold: a novel carrier for cell and drug delivery. Crit Rev Ther Drug Carrier Syst 2012, 29:1-63.
  • [7]Wenstrup RJ, Florer JB, Brunskill EW, Bell SM, Chervoneva I, Birk DE: Type V collagen controls the initiation of collagen fibril assembly. J Biol Chem 2004, 279:53331-53337.
  • [8]Martel-Pelletier J, Boileau C, Pelletier JP, Roughley PJ: Cartilage in normal and osteoarthritis conditions. Best Pract Res Clin Rheumatol 2008, 22:351-384.
  • [9]Roughley PJ, Lee ER: Cartilage proteoglycans: structure and potential functions. Microsc Res Tech 1994, 28:385-397.
  • [10]Michelacci YM, Mourão PAS, Laredo-Filho J, Dietrich CP: Chondroitin sulfates and proteoglycans from normal and arthrosic human cartilages. Connect Tissue Res 1979, 7:29-36.
  • [11]Mourão PAS, Michelacci YM, Toledo OMS: Glycosaminoglycans and proteoglycans of normal and tumoral cartilages of humans and rats. Cancer Res 1979, 39:2802-2806.
  • [12]Kate Y, Gospodarowicz D: Effect of exogenous extracellular matrices on proteoglycan synthesis by cultured rabbit costal chondrocytes. J Cell Biol 1985, I00:486-495.
  • [13]Pingguan-Murphy B, Lee DA, Bader DL, Knight MM: Activation of chondrocytes calcium signalling by dynamic compression is independent of number of cycles. Arch Biochem Biophys 2005, 444:45-51.
  • [14]Francis Suh J-K, Matthew HWT: Application of chitosan-based polysaccjaride biomaterials in cartilage tissue engineering: a review. Biomaterials 2000, 21:2589-2598.
  • [15]Hoemann CD, Sun J, Légaré A, McKee MD, Buschmann MD: Tissue engineering of cartilage using an injectable and adhesive chitosan-based cell-delivery vehicle. Osteoarthritis Cartilage 2004, 13:318-329.
  • [16]Choa JH, Kim S-H, Park KD, Jung MC, Yang WI, Han SW, Noh JY, Lee JW: Chondrogenic differentiation of human mesenchymal stem cells using a thermosensitive poly (N-isopropylacrylamide) and water-soluble chitosan copolymer. Biomaterials 2004, 25:5743-5751.
  • [17]Chen J-P, Cheng T-H: Thermo-responsive chitosan-graft-poly (N-isopropylacrylamide) injectable hydrogel for cultivation of chondrocytes and meniscus cells. Macromol Biosci 2006, 6:1026-1039.
  • [18]Chenite A, Chaput C, Wang D, Combes C, Buschmann MD, Hoemann CD, Leroux JC, Atkinson BL, Binette F, Selmani A: Novel injectable neutral solutions of chitosan form biodegradable gels in situ. Biomaterials 2000, 21:2155-2161.
  • [19]Hoemann CD, Sun J, McKee MD, Chevrier A, Rossomacha E, Rivard G-E, Hurtig M, Buschmann MD: Chitosan-glycerol phosphate/blood implants elicit hyaline cartilage repair integrated with porous subchondral bone in microdrilled rabbit defects. Osteoarthritis Cartilage 2007, 15:78-89.
  • [20]Ngoenkam J, Faikrua A, Yasothornsrikul S, Viyoch J: Potential of an injectable chitosan/starch/beta-glycerol phosphate hydrogel for sustaining normal chondrocyte function. Int J Pharm 2010, 391:115-124.
  • [21]Malette WG, Quigley HJ, Gaines RD, Johnson ND, Rainer WG: Chitosan: a new hemostatic. Ann Thorac Surg 1983, 36:55-58.
  • [22]Ueno H, Nakamura F, Murakami M, Okumura M, Kadosawa T, Fujinag T: Evaluation effects of chitosan for the extracellular matrix production by fibroblasts and the growth factors production by macrophages. Biomaterials 2001, 22:2125-2130.
  • [23]Sampaio LO, Bayliss MT, Hardingham TE, Muir H: Dermatan sulfate proteoglycan from human articular cartilage. Variation in its contents with age and structural comparison with a small chondroitin sulfate proteoglycan from pig laryngeal cartilage. Biochem J 1988, 254:757-764.
  • [24]Berto AGA, Sampaio LO, Franco CRC, Cesar RM Jr, Michelacci YM: A comparative analysis of structure and spatial distribution of decorin in human leiomyoma and normal myometrium. Biochim Biophys Acta 2003, 1619:98-112.
  • [25]Martins SAR, Campos MSQ, Berto AGA, Aguiar JAK, Michelacci YM: Proteoglycan synthesis by human corneal explants submitted to laser in situ keratomileusis (LASIK). Mol Vis 2007, 13:142-150.
  • [26]de Lima CR, dos Santos-Junior JA, Nazário ACP, Michelacci YM: Changes in glycosaminoglycans and proteoglycans of normal breast and fibroadenoma during the menstrual cycle. Biochim Biophys Acta 1820, 2012:1009-1019.
  • [27]Dietrich CP, Dietrich SMC: Eletrophoretic behaviour of acidic mucopolysaccharides in diamine buffers. Anal Biochem 1976, 70:645-647.
  • [28]Denuziere A, Ferrier D, Damour O, Domard A: Chitosan-chondroitin sulfate and chitosan-hyaluronate polyelectrolyte complexes: biological properties. Biomaterials 1998, 19:1275-1285.
  • [29]Hamilton V, Yuan Y, Rigney DA, Puckett AD, Ong JL, Yang Y, Elder SH, Bumgardner JD: Characterization if chitosan films and effects on fibroblast cell attachment and proliferation. J Mater Sci Mater Med 2006, 17:469-485.
  • [30]Abarrategi A, Lópiz-Morales Y, Ramos V, Civantos A, López-Durán L, Marco F, López-Lacomba JL: Chitosan scaffolds for osteochondral tissue regeneration. J Biomed Mater Res A 2010, 95:1132-1141.
  • [31]Lavertu M, Filion D, Buschmann MD: Heat-induced transfer of protons from chitosan to glycerol phosphate produces chitosan precipitation and gelation. Biomacromolecules 2008, 9:640-650.
  • [32]Hunziker EB: Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. Osteoarthritis Cartilage 2001, 10:432-463.
  • [33]Bertolo A, Mehr M, Aebli N, Baur M, Ferguson SJ, Stoyanov JV: Influence of different commercial scaffolds on the in vitro differentiation of human mesenchymal stem cells to nucleus pulposus-like cells. Eur Spine J 2012, 21(Suppl 6):826-838.
  • [34]Onishi H, Machida Y: Biodegradation and distribution of water-soluble chitosan in mice. Biomaterials 1999, 20:175-182.
  • [35]Peluso G, Petillo O, Ranieri M, Santin M, Ambrosio L, Calabro D, Avallone B, Balsamo G: Chitosan-mediated stimulation of macrophage function. Biomaterials 1994, 15:1215-1220.
  • [36]Usami Y, Okamoto Y, Minami S, Matsuhashi A, Kumazawa NH, Tanioka S, Shigemasa Y: Migration of canine neutrophils to chitin and chitosan. J Vet Med Sci 1994, 56:1215-1216.
  • [37]Usami Y, Okamoto Y, Takayama T, Shigemasa Y, Minami S: Chitin and chitosan stimulate canine polymorphonuclear cells to release leukotriene B4 and prostaglandin E2. J Biomed Mater Res 1998, 42:517-522.
  • [38]Chevrier A, Hoemann CD, Sun J, Buschmann MD: Chitosan-glycerol phosphate/blood implants increase cell recruitment, transient vascularization and subchondral bone remodeling in drilled cartilage defects. Osteoarthritis Cartilage 2006, 15:316-327.
  • [39]Barr ED, Pinchbeck GL, Clegg PD, Boyde A, Riggs CM: Post mortem evaluation of palmar osteochondral disease (traumatic osteochondrosis) of the metacarpo/metatarsophalangeal joint in thoroughbred racehorses. Equine Vet J 2009, 41:366-371.
  • [40]Van Grevenhof EM, Ducro BJ, Van Weeren PR, Van Tartwijk JM, Van den Belt AJ, Bijma P: Prevalence of various radiographic manifestations of osteochondrosis and their correlations between and within joints in dutch warmblood horses. Equine Vet J 2009, 41:11-16.
  • [41]Hunziker EB: Articular cartilage repair: are the intrinsic biological constraints undermining this process insuperable? Osteoarthritis Cartilage 1999, 7:15-28.
  • [42]Shortkroff S, Barone L, Hsu HP, Wrenn C, Gagne T, Chi T, Breinan H, Minas T, Sledge CB, Tubo R, Spector M: Healing of chondral and osteochondral defects in a canine model: the role of cultured chondrocytes in regeneration of articular cartilage. Biomaterials 1996, 17:147-154.
  • [43]Frisbie DD, Morisset S, Ho CP, Rodkey WG, Steadman JR, McIlwraith CW: Effects of calcified cartilage on healing of chondral defects treated with microfracture in horses. Am J Sports Med 2006, 34:1824-1831.
  • [44]Frisbie DD, Lu Y, Kawcak CE, DiCarlo EF, Binette F, McIlwraith CW: In vivo evaluation of autologous cartilage fragment-loaded scaffolds implanted into equine articular defects and compared with autologous chondrocyte implantation. Am J Sports Med 2009, 37(Suppl 1):71S-80S.
  • [45]Vacanti CA, Langer R, Schloo B, Vacanti JP: Synthetic polymers seeded with chondrocytes provide a template for new cartilage formation. Plast Reconstr Surg 1991, 88:753-759.
  • [46]Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L: Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 1994, 331:889-895.
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