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3D打印聚乳酸-羟基乙酸共聚物支架在软骨缺损修复方面的价值

Value of 3D Printed PLGA Scaffolds for Cartilage Defects in Terms of Repair.

作者信息

Fan Longkun, Teng Wei, He Jinqiu, Wang Dongni, Liu Chunhui, Zhao Yujia, Zhang Limin

机构信息

Cangzhou Central Hospital, No. 16, Xinhua West Road, Cangzhou City, Hebei Province, China.

Cangzhou Women and Children's Health Hospital, Fuyang North Avenue, Cangzhou City, Hebei Province, China.

出版信息

Evid Based Complement Alternat Med. 2022 Aug 3;2022:3561430. doi: 10.1155/2022/3561430. eCollection 2022.

DOI:10.1155/2022/3561430
PMID:35966730
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9365545/
Abstract

OBJECTIVE

To examine the poly (lactic-co-glycolic acid) and sodium alginate (SA) scaffolds produced by 3D printing technology, access the healing morphology of bones following PLGA/SA implantation within rat cartilage, and examine osteogenesis-related factors in rat serum to determine the efficacy of PLGA/SA scaffolds in healing animal cartilage injuries. To identify the potential of this material to repair a tissue engineering osteochondral injury.

METHODS

Polylactic acid-glycolic acid copolymer and sodium alginate were used as raw materials to create PLGA/SA scaffolds. We observed the scaffold's macrostructure and microstructure, and the scaffold's microstructure was observed through a scanning electron microscope (SEM). The mechanical toughness of a stent was assessed using a biomechanical device. Hematoxylin-eosin staining revealed immune rejection after embedding the scaffolds under the skin of SD rats. The CCK-8 cell proliferation test kit was used to measure cell proliferation. An experimental model of cartilage injury in the knee joint was created in rats. Rats were used to establish an experimental model of cartilage damage in the knee joint. 120 female rats aged 5 weeks were chosen at random from the pool and divided into the experimental and control groups. They were all completely anesthetized with an anesthetic before having the lateral skin of the knee articular cartilage incised. Implanted PLGA/SA scaffolds were not used in the control group and only in the experiment group. Both groups of rats had their muscles and skin sutured and covered in plaster bandages. On the third, seventh, fourteenth, twenty-first, twenty-eighth, and thirty-fifth days after the procedure, the two groups of rats were divided into groups. At various stages, bone tissue, blood samples, and cartilage were examined and evaluated. Immunohistochemistry was used to identify the local bone morphogenetic protein-2 (BMP2).

RESULTS

(1) PLGA/SA was successfully used to build an artificial cartilage scaffold. (2) Macroscopic and SEM observation results showed the material had increased density and numerous microvoids on the surface. (3) The result of the biomechanical test showed that the PLGA/SA scaffold had superior biomechanical characteristics. (4) The stent did not exhibit any noticeable immunological rejection, according to the results of the subcutaneous embedding experiment performed on rats. (5) The CCK-8 data demonstrated that as the cell development time rose, the number of cells gradually increased. However, there was not statistically significant difference between the growth of the cells in the scaffold extract and the control group ( > 0.05). (6) A successful rat model based on a cartilage defect of the medial knee joint has been built. (7) Observations of specimens revealed that the experimental group's bone tissue score was higher than that of the control group. (8) Using immunohistochemistry, it was found that the experimental group's BMP2 expression was higher on the 7th, 14th, and 28th days than it was in the control group ( < 0.05).

CONCLUSION

Strong mechanical and biological properties are present in stable, biodegradable PLGA/SA scaffolds that mimic cartilage. We demonstrated that the cartilage biomimetic PLGA/SA scaffold may repair cartilage and prevent negative reactions such as osteoarthritis in rat knee cartilage, making it suitable as a cartilage scaffolding material for tissue engineering. The PLGA/SA scaffold was also able to promote BMP2 expression in the bone healing zone when inserted into a knee cartilage lesion. Improved cartilage damage is the outcome of BMP2's promotion of bone formation and restriction of bone resorption in the bone healing zone.

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a150/9365545/cf59e5410803/ECAM2022-3561430.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a150/9365545/38180d0b6abb/ECAM2022-3561430.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a150/9365545/cf59e5410803/ECAM2022-3561430.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a150/9365545/38180d0b6abb/ECAM2022-3561430.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a150/9365545/cf59e5410803/ECAM2022-3561430.002.jpg
摘要

目的

研究通过3D打印技术制备的聚乳酸-乙醇酸共聚物(PLGA)与海藻酸钠(SA)支架,观察PLGA/SA植入大鼠软骨后骨的愈合形态,并检测大鼠血清中与成骨相关的因子,以确定PLGA/SA支架在修复动物软骨损伤方面的效果。评估该材料修复组织工程化骨软骨损伤的潜力。

方法

以聚乳酸-乙醇酸共聚物和海藻酸钠为原料制备PLGA/SA支架。观察支架的宏观和微观结构,通过扫描电子显微镜(SEM)观察支架的微观结构。使用生物力学装置评估支架的机械韧性。将支架埋植于SD大鼠皮下,苏木精-伊红染色观察免疫排斥反应。使用CCK-8细胞增殖检测试剂盒检测细胞增殖情况。建立大鼠膝关节软骨损伤实验模型。从大鼠群体中随机选取120只5周龄雌性大鼠,分为实验组和对照组。两组大鼠均在麻醉后切开膝关节外侧皮肤软骨。对照组不植入PLGA/SA支架,仅实验组植入。两组大鼠均缝合肌肉和皮肤,并用石膏绷带覆盖。术后第3、7、14、21、28和35天,将两组大鼠分组,在不同阶段对骨组织、血液样本和软骨进行检查和评估。采用免疫组织化学法鉴定局部骨形态发生蛋白-2(BMP2)。

结果

(1)成功利用PLGA/SA构建了人工软骨支架。(2)宏观和SEM观察结果显示,该材料密度增加,表面有许多微孔。(3)生物力学测试结果表明,PLGA/SA支架具有优异的生物力学特性。(4)大鼠皮下埋植实验结果显示,该支架未表现出明显的免疫排斥反应。(5)CCK-8数据表明,随着细胞培养时间延长,细胞数量逐渐增加。然而,支架提取物中细胞生长与对照组之间无统计学显著差异(>0.05)。(6)成功建立了基于内侧膝关节软骨缺损的大鼠模型。(7)标本观察显示,实验组骨组织评分高于对照组。(8)免疫组织化学检测发现,实验组在第7、14和28天的BMP2表达高于对照组(<0.05)。

结论

稳定、可生物降解的PLGA/SA支架具有强大的机械和生物学性能,可模拟软骨。我们证明,仿生软骨的PLGA/SA支架可修复大鼠膝关节软骨,并预防骨关节炎等不良反应,使其适合作为组织工程软骨支架材料。将PLGA/SA支架植入膝关节软骨损伤处时,还能够促进骨愈合区域BMP2的表达。骨愈合区域中BMP2促进骨形成并抑制骨吸收,从而改善软骨损伤。

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