交联几丁糖治疗兔膝骨关节炎的实验研究
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摘要
目的探讨关节腔注射交联几丁糖治疗兔膝骨关节炎的效果。方法将32只新西兰白兔随机分为A、B、C、D 4组(n=8),其中A、B、C组采用切断左后肢前交叉韧带方法制备膝骨关节炎模型;4周后,A组关节腔内注射交联几丁糖0.6 mL 1次;B、C组分别注射几丁糖、生理盐水,0.3 mL/次,每2周1次;D组为假手术组,关节腔内注射等量生理盐水。造模术后8周,行软骨大体观察、扫描电镜观察、组织学观测(HE染色、番红-固绿染色以及Mankin评分)、Ⅱ型胶原免疫组织化学染色观测。结果大体及扫描电镜观察示D组软骨结构接近正常,A、B组软骨表面结构磨损较D组严重,C组最严重;组织学染色观察示,A、B组软骨结构相似,且优于C组,但尚未达D组正常结构。B、C组软骨大体评分及Mankin评分明显高于D组(P<0.05),A、B组比较差异无统计学意义(P>0.05)。免疫组织化学染色观察示D组Ⅱ型胶原阳性软骨细胞百分比明显高于B、C组(P<0.05),A、B组比较差异无统计学意义(P>0.05)。结论交联几丁糖能明显改善兔膝骨关节炎,延缓骨关节炎病理改变,且注射次数减少。
Objective To study the effect of intraarticular injection of crosslinked-chitosan in the treatment of knee osteoarthritis in rabbits. Methods Thirty-two New Zealand white rabbits were randomly divided into 4 groups(groups A, B, C, and D; 8 rabbits in each group). The knee osteoarthritis models were prepared by anterior cruciate ligament transection in the left hind in groups A, B, and C. At 4 weeks after operation, the rabbits were received intraarticular injection of 0.6 mL crosslinked-chitosan in group A, 0.3 mL chitosan(once per 2 weeks, for twice) in group B, and0.3 mL saline(once per 2 weeks, for twice) in group C. The rabbits in group D were treated with sham operation in the left hind, and received intraarticular injection of 0.3 mL saline(once per 2 weeks, for twice). At 8 weeks, the macroscopic observation, histological examination(HE staining, Safranin-fast green double staining, and Mankin score), scanning electron microscopy(SEM) observation, and immunohistochemical staining of collagen type Ⅱ were performed.Results Macroscopic and SEM observations showed that the cartilage in group D was basically the same as normal and better than that in groups A and B, and the abrasion of cartilage in group C was the most serious. The histological observation results in groups A and B were slightly similar and better than those in group C, but not up to the structure of group D. The macroscopic score and Mankin score of groups B and C were significantly higher than those of group D(P<0.05), and there was no significant difference between group A and group B(P>0.05). Immunohistochemical staining results showed that the collagen type Ⅱ positive percentage of chondrocytes was significantly higher in group D than that in groups B and C, and no significant difference was found between group A and group B(P>0.05). Conclusion The crosslinked-chitosan can significantly improve the osteoarthritis of the rabbit knee, delay the pathological changes of o steoarthritis, and decrease the frequency of injection.
Objective To study the effect of intraarticular injection of crosslinked-chitosan in the treatment of knee osteoarthritis in rabbits. Methods Thirty-two New Zealand white rabbits were randomly divided into 4 groups(groups A, B, C, and D; 8 rabbits in each group). The knee osteoarthritis models were prepared by anterior cruciate ligament transection in the left hind in groups A, B, and C. At 4 weeks after operation, the rabbits were received intraarticular injection of 0.6 mL crosslinked-chitosan in group A, 0.3 mL chitosan(once per 2 weeks, for twice) in group B, and0.3 mL saline(once per 2 weeks, for twice) in group C. The rabbits in group D were treated with sham operation in the left hind, and received intraarticular injection of 0.3 mL saline(once per 2 weeks, for twice). At 8 weeks, the macroscopic observation, histological examination(HE staining, Safranin-fast green double staining, and Mankin score), scanning electron microscopy(SEM) observation, and immunohistochemical staining of collagen type Ⅱ were performed.Results Macroscopic and SEM observations showed that the cartilage in group D was basically the same as normal and better than that in groups A and B, and the abrasion of cartilage in group C was the most serious. The histological observation results in groups A and B were slightly similar and better than those in group C, but not up to the structure of group D. The macroscopic score and Mankin score of groups B and C were significantly higher than those of group D(P<0.05), and there was no significant difference between group A and group B(P>0.05). Immunohistochemical staining results showed that the collagen type Ⅱ positive percentage of chondrocytes was significantly higher in group D than that in groups B and C, and no significant difference was found between group A and group B(P>0.05). Conclusion The crosslinked-chitosan can significantly improve the osteoarthritis of the rabbit knee, delay the pathological changes of o steoarthritis, and decrease the frequency of injection.
引文
1 Tang X, Wang S, Zhan S, et al. The prevalence of symptomatic knee osteoarthritis in China:results from the China Health and Retirement longitudinal study. Arthritis Rheumatol, 2016, 68(3):648-653.
2 Chen Q, Shao X, Ling P, et al. Recent advances in polysaccharides for osteoarthritis therapy. Eur J Med Chem, 2017, 139:926-935.
3 Kaderli S, Viguier E, Watrelot-Virieux D, et al. Efficacy study of two novel hyaluronic acid-based formulations for viscosupplementation therapy in an early osteoarthrosic rabbit model.Eur J Pharm Biopharm, 2015, 96:388-395.
4 Karbasi S, Fekrat F, Semnani D, et al. Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering. Adv Biomed Res, 2016, 5:180.
5 Utomo L, Eijgenraam SM, Meuffels DE, et al. Meniscal extrusion and degeneration during the course of osteoarthritis in the murine collagenase-induced osteoarthritis model. J Orthop Res, 2018,36(9):2416-2420.
6 van der Sluijs JA, Geesink RG, van der Linden AJ, et al. The reliability of the Mankin score for osteoarthritis. J Orthop Res,1992, 10(1):58-61.
7 Pelletier JP, Lascau-Coman V, Jovanovic D, et al. Selective inhibition of inducible nitric oxide synthase in experimental osteoarthritis is associated with reduction in tissue levels of catabolic factors. J Rheumatol, 1999, 26(9):2002-2014.
8 Boyce MK, Trumble TN, Carlson CS, et al. Non-terminal animal model of post-traumatic osteoarthritis induced by acute joint injury. Osteoarthritis Cartilage, 2013, 21(5):746-755.
9 Liu Z, Hu X, Man Z, et al. A novel rabbit model of early osteoarthritis exhibits gradual cartilage degeneration after medial collateral ligament transection outside the joint capsule. Sci Rep,2016,6:34423.
10 Hermeto LC, Rossi R, Jardim PH, et al. Comparison between two different experimental models of osteoarthritis in rabbits. Intraarticular collagenase injection and anterior cruciate ligament transection. Acta Cir Bras, 2016, 31(9):602-607.
11 Teeple E, Jay GD, Elsaid KA, et al. Animal models of osteoarthritis:challenges of model selection and analysis. AAPS J, 2013, 15(2):438-446.
12翟吉良,翁习生,邱贵兴.骨关节炎动物模型的建立及选择.中国矫形外科杂志,2007,15(11):843-845.
13肖志锋,林定坤.骨关节炎软骨内骨化病理研究进展.中国修复重建外科杂志,2016, 30(12):1556-1561.
14 Vila S. Inflammation in osteoarthritis. P R Health Sci J, 2017, 36(3):123-129.
15 Levillain A, Magoariec H, Boulocher C, et al. Effects of a viscosupplementation therapy on rabbit menisci in an anterior cruciate ligament transection model of osteoarthritis. J Biomech,2017,58:147-154.
16 Nahas RM, Porto L, Ikemoto RY, et al. Viscosupplementation in the treatment of post-traumatic arthritis of the knee for 12 months.Revista Brasileira de Medicina do Esporte, 2016, 22(6):465-470.
17 Ondresik M, Azevedo Maia FR, da Silva Morais A, et al.Management of knee osteoarthritis. Current status and future trends. Biotechnol Bioeng, 2017,114(4):717-739.
18 Zhou Y, Liu S, Ming J, et al. Sustained release effects of berberineloaded chitosan microspheres on in vitro chondrocyte culture.Drug Development and Industrial Pharmacy, 2017, 43(10):1703-1714.
19 Zhang C, Liao Q, Ming J H, et al. The effects of chitosan oligosaccharides on OPG and RANKL expression in a rat osteoarthritis model. Acta Cir Bras, 2017, 32(6):418-428.
20 Cheung RC, Ng TB, Wong JH, et al. Chitosan:an update on potential biomedical and pharmaceutical applications. Mar Drugs,2015, 13(8):5156-5186.
21 Rieger R,Boulocher C,Kaderli S,et al. Chitosan in viscosupplementation:in vivo effect on rabbit subchondral bone.BMC Musculoskeletal Disorders, 2017, 18:350.
22 Rieger R, Boulocher C, Hoc T. How does hyaluronic acid-based viscosupplements modify the subchondral bone in a rabbit model of early OA. Osteoarthritis and Cartilage, 2016, 24(1):S365-S365.
2 Chen Q, Shao X, Ling P, et al. Recent advances in polysaccharides for osteoarthritis therapy. Eur J Med Chem, 2017, 139:926-935.
3 Kaderli S, Viguier E, Watrelot-Virieux D, et al. Efficacy study of two novel hyaluronic acid-based formulations for viscosupplementation therapy in an early osteoarthrosic rabbit model.Eur J Pharm Biopharm, 2015, 96:388-395.
4 Karbasi S, Fekrat F, Semnani D, et al. Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering. Adv Biomed Res, 2016, 5:180.
5 Utomo L, Eijgenraam SM, Meuffels DE, et al. Meniscal extrusion and degeneration during the course of osteoarthritis in the murine collagenase-induced osteoarthritis model. J Orthop Res, 2018,36(9):2416-2420.
6 van der Sluijs JA, Geesink RG, van der Linden AJ, et al. The reliability of the Mankin score for osteoarthritis. J Orthop Res,1992, 10(1):58-61.
7 Pelletier JP, Lascau-Coman V, Jovanovic D, et al. Selective inhibition of inducible nitric oxide synthase in experimental osteoarthritis is associated with reduction in tissue levels of catabolic factors. J Rheumatol, 1999, 26(9):2002-2014.
8 Boyce MK, Trumble TN, Carlson CS, et al. Non-terminal animal model of post-traumatic osteoarthritis induced by acute joint injury. Osteoarthritis Cartilage, 2013, 21(5):746-755.
9 Liu Z, Hu X, Man Z, et al. A novel rabbit model of early osteoarthritis exhibits gradual cartilage degeneration after medial collateral ligament transection outside the joint capsule. Sci Rep,2016,6:34423.
10 Hermeto LC, Rossi R, Jardim PH, et al. Comparison between two different experimental models of osteoarthritis in rabbits. Intraarticular collagenase injection and anterior cruciate ligament transection. Acta Cir Bras, 2016, 31(9):602-607.
11 Teeple E, Jay GD, Elsaid KA, et al. Animal models of osteoarthritis:challenges of model selection and analysis. AAPS J, 2013, 15(2):438-446.
12翟吉良,翁习生,邱贵兴.骨关节炎动物模型的建立及选择.中国矫形外科杂志,2007,15(11):843-845.
13肖志锋,林定坤.骨关节炎软骨内骨化病理研究进展.中国修复重建外科杂志,2016, 30(12):1556-1561.
14 Vila S. Inflammation in osteoarthritis. P R Health Sci J, 2017, 36(3):123-129.
15 Levillain A, Magoariec H, Boulocher C, et al. Effects of a viscosupplementation therapy on rabbit menisci in an anterior cruciate ligament transection model of osteoarthritis. J Biomech,2017,58:147-154.
16 Nahas RM, Porto L, Ikemoto RY, et al. Viscosupplementation in the treatment of post-traumatic arthritis of the knee for 12 months.Revista Brasileira de Medicina do Esporte, 2016, 22(6):465-470.
17 Ondresik M, Azevedo Maia FR, da Silva Morais A, et al.Management of knee osteoarthritis. Current status and future trends. Biotechnol Bioeng, 2017,114(4):717-739.
18 Zhou Y, Liu S, Ming J, et al. Sustained release effects of berberineloaded chitosan microspheres on in vitro chondrocyte culture.Drug Development and Industrial Pharmacy, 2017, 43(10):1703-1714.
19 Zhang C, Liao Q, Ming J H, et al. The effects of chitosan oligosaccharides on OPG and RANKL expression in a rat osteoarthritis model. Acta Cir Bras, 2017, 32(6):418-428.
20 Cheung RC, Ng TB, Wong JH, et al. Chitosan:an update on potential biomedical and pharmaceutical applications. Mar Drugs,2015, 13(8):5156-5186.
21 Rieger R,Boulocher C,Kaderli S,et al. Chitosan in viscosupplementation:in vivo effect on rabbit subchondral bone.BMC Musculoskeletal Disorders, 2017, 18:350.
22 Rieger R, Boulocher C, Hoc T. How does hyaluronic acid-based viscosupplements modify the subchondral bone in a rabbit model of early OA. Osteoarthritis and Cartilage, 2016, 24(1):S365-S365.