Pathological changes of the skeletal muscle after local loosening therapies

doi:10.3969/j.issn.2095-4344.0086

Abstract

Abstract:

BACKGROUND: Local soft tissue lysis has been widely used in clinical rehabilitation treatment of chronic soft tissue injuries, and has achieved certain clinical outcomes. However, it is an invasive treatment, and there is little information about the pathological changes and prognosis of the local tissues post-treatment.
OBJECTIVE: To observe the repair process and pathologic changes of the muscles after local soft tissue lysis with several needles.
METHOD: Songjin-needle, Yinzhi-needle, Changyuan-needle, Xiaozhen-needle, Ren-needle and Pi-needle were inserted into the belly of rat gastrocnemius gastrocnemius with a depth of 0.5-1.0 cm, twice or thrice. Then the rats were killed at 3, 7, 14, 21 and 30 days after acupuncture. The corresponding parts of tissues were removed and fixed with paraformaldehyde, and stained with hematoxylin and eosin and immunohistochemistry to observe the histomorphological features and pathological changes under light microscope. The CellSens-Entry 1.15 software was used for image acquisition, and the number of fibroblast cells and microvessel density were calculated. The integral absorbance values of collagen type I were calculated by Image Pro plus 6 software.
RESULTS AND CONCLUSION: On day 3 after intervention, muscle fiber dissolution, necrosis and inflammatory cell infiltration were visible. On day 7, there was a reduction in inflammatory cells, fibroblast activation and proliferation, and granulation tissue hyperplasia. On day 14, there was local accumulation of collagen with substantial granulation tissues. On day 21, collagen fibers decreased and granulation tissues organized. On day 30, there was local scar formation. Immunohistochemical staining results showed that there was collagen type I in the surgical area after intervention. The microvessel density, fibroblast count and integral absorbance value increased firstly and then decreased, which showed significant differences among groups (P < 0.01). These findings imply that the new types of needles for local soft tissue lysis do certain damage to the gastrocnemius muscle, and the inflammatory changes of the local tissue and granulation tissue organized will result in local scar formation after loosening therapies.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Muscle, Skeletal, Cicatrix

CLC Number:

R318

Jing Ya-jun 1, Zhang Lei1, Zhang Shao-qun1, Liao Li-qing1, Yuan Shi-guo2, Li Yi-kai1. Pathological changes of the skeletal muscle after local loosening therapies[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(4): 535-541.

Figures/Tables 2

2.1 实验动物数量分析纳入大鼠108只，均进入结果分析，无死亡和感染等。 2.2 干预后局部组织苏木精-伊红染色观察松解干预后第3天，各组松解局部均可见不同程度的肌纤维排列紊乱、溶解、坏死，大量炎性细胞聚集；干预后第7天，炎性反应减弱，成纤维细胞活化、向创区迁移并大量增殖，不同程度的新生毛细血管形成，肉芽组织形成并开始增生；干预后第14天，成纤维细胞向纤维细胞转化，局部大量胶原纤维堆积、无序排列并伴有肉芽组织大量形成，干预后第21天，胶原纤维及新生毛细血管逐渐减少，肉芽组织开始机化；干预后第30天，局部胶原纤维束状排列，间质中存在大量胶原沉积，新生毛细血管减少，局部组织瘢痕但尚未稳定(图1)。 2.3 不同时间、不同直径及类型针具干预部位组织微血管密度比较所选6种针具松解干预后，不同时间各组微血管密度比较见表1，2：同一针具不同时间点间微血管密度比较差异有显著性意义(P < 0.01)，两类针具干预后，微血管密度变化呈现先增加后逐渐下降趋势，干预后第7天，微血管密度达到峰值(均值：带刃型为13.17，圆钝型为17.42)，其后逐渐下降。相同时间，同类型针具不同直径间微血管密度比较差异有显著性意义(P < 0.01)，同类型针具其直径越大则微血管密度值越大；3种带刃针具组间微血管密度两两比较差异有显著性意义(P < 0.01)，相同时间小针刀组微血管密度值最大，铍针组最小；3 种圆钝型针具组间微血管密度两两比较，松筋针组与银质针组、长圆针组间微血管密度比较差异有显著性意义(P < 0.05)，松筋针组微血管密度值最大，长圆针组最小。 2.4 不同时间、不同直径及类型针具干预部位组织成纤维细胞计数比较松解干预后，不同时间各组成纤维细胞计数比较见表3， 4，同一针具不同时间点间成纤维细胞计数比较差异有显著性意义(P < 0.01)，两类针具干预后，成纤维细胞计数变化呈现先增加后逐渐下降趋势，干预后第7天，成纤维细胞计数达到峰值(均值：带刃型为59.80，圆钝型为75.07)，其后逐渐下降。相同时间，同类型针具不同直径间成纤维细胞计数比较差异有显著性意义(P < 0.01)，同类型针具直径越大则其成纤维细胞数目越多。3 种带刃针具组间两两比较差异有显著性意义(P < 0.01)，其中小针刀组成纤维细胞数目最多，铍针组最少；3种圆钝型针具组间两两比较差异有显著性意义(P < 0.01)，其中松筋针组最多，长圆针组最少。成纤维细胞计数变化趋势与MVD变化趋势一致。 2.5 干预部位组织Ⅰ型胶原纤维免疫组织化学染色观察及积分吸光度值比较免疫组织化学染色图2显示，干预后伴随成纤维细胞活化，细胞外基质分泌及胶原蛋白合成，Ⅰ型胶原沉积区被染色为棕黄色。不同时间各组积分吸光度值比较见表5，6，同一针具不同时间点间积分吸光度值比较差异有显著性意义(P < 0.01)，两类针具干预后，Ⅰ型胶原纤维积分吸光度值变化呈现先增加后逐渐下降趋势，干预后第7天，积分吸光度值达到峰值(均值：带刃型为 6 507.13，圆钝型为7 140.57)，其后逐渐下降。相同时间，同类型针具不同直径间Ⅰ型胶原纤维积分吸光度值比较差异有显著性意义(P < 0.01)，同类型针具其直径越大则其Ⅰ型胶原纤维积分吸光度值越大，3 种带刃针具组间积分吸光度值两两比较差异有显著性意义(P < 0.01)，其中小针刀组Ⅰ型胶原纤维积分吸光度值最大，铍针组最小；3 种圆钝型针具组间积分吸光度值两两比较差异有显著性意义(P < 0.05)，其中松筋针组Ⅰ型胶原纤维积分吸光度值最大，长圆针组最小。积分吸光度值变化趋势与微血管密度变化、成纤维细胞计数变化一致。干预第30天，可见Ⅰ型胶原尚未完全降解吸收，不同程度的沉积于创区(P < 0.01)，是瘢痕形成的主要因素之一，实验的免疫组织化学染色与苏木精-伊红染色结果基本一致。"

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