高压氧联合超短波辅助治疗兔跟腱断裂的实验研究

doi:10.3877/cma.j.issn.1674-0785.2020.09.010

目的

探讨高压氧联合超短波辅助治疗跟腱断裂对跟腱愈合的影响。

方法

将80只新西兰白兔切断跟腱，制造模型，均行Kessler法缝合，并石膏超踝膝固定4周后拆除。随机分为高压氧组（A组）、超短波组（B组）、高压氧联合超短波组（C组）、对照组（D组），每组20只。A组、C组术后即刻行高压氧治疗，前3天每天2次，后4天每天1次，共治疗7 d。B组、C组术后每天给予超短波治疗，共6 d，D组不予治疗。术后分别于4周、8周各组分别处死10只白兔，取跟腱组织行生物力学检测（最大拉伸长度和最大抗拉力）、HE染色和免疫组织化学染色。采用方差分析比较术后4周和8周生物力学检测结果的组间差异，并采用LSD-t检验进行组间的两两比较。

结果

生物力学测试结果显示：术后4周A组、B组、C组跟腱平均最大抗拉力和平均最大拉伸长度与D组相比显著升高[最大抗拉力：（276.20±8.16）N vs（278.10±7.92）N vs（309.90±9.78）N vs（259.50±4.99）N；最大拉伸长度：（4.62±0.14）mm vs（4.67±0.16）mm vs（5.05±0.17）mm vs（4.18±0.32）mm]，差异均具有统计学意义（最大抗拉力：t=5.519、6.280、14.513，P均<0.001；最大拉伸长度：t=3.934、4.136、7.563，P=0.001、<0.001、<0.001），且C组更优于A组及B组，差异均具有统计学意义（最大抗拉力：t=8.366、7.989，P均<0.001；最大拉伸长度：t=6.164、5.190，P均<0.001）。术后8周A组、B组、C组跟腱平均最大抗拉力和平均最大拉伸长度与D组相比显著升高[最大抗拉力：（354.60±5.68）N vs（355.30±9.62）N vs（390.70±7.48）N vs（339.60±5.60）N；最大拉伸长度：（4.88±0.18）mm vs（5.10±0.16）mm vs（5.38±0.15）mm vs（4.46±0.23）mm]，差异均具有统计学意义（最大抗拉力：t=5.946、4.461、17.286，P均<0.001；最大拉伸长度：t=4.578、7.327、10.628，P均<0.001），且C组更优于A组及B组，差异均具有统计学意义（最大抗拉力：t=12.150、9.187，P均<0.001；最大拉伸长度：t=6.600、3.925，P<0.001、=0.001）。组织学观察结果：C组发现大量成纤维细胞在跟腱断端附近增生，并有少量炎性细胞。A组和B组中虽然可见大量成纤维细胞，但较C组少，且炎性细胞数量较多。D组镜下炎性细胞数量显著多于其他3组，成纤维细胞生长情况明显落后于其他3组，且排列杂乱，并见较多肉芽组织。免疫组织化学染色检查：术后可见Ⅰ型胶原纤维呈现阳性表达，且C组Ⅰ型胶原纤维表达数目最为丰富，B组略多于A组，D组最少。

结论

高压氧联合超短波可促进跟腱断裂后跟腱愈合生物学性能，促进Ⅰ型胶原纤维合成。

关键词: 跟腱断裂, 辅助治疗, 高压氧, 超短波, 生物力学

Objective

To investigate the effect of hyperbaric oxygen combined with ultrashort wave assisted treatment of Achilles tendon rupture on Achilles tendon healing.

Methods

Eighty New Zealand white rabbits were used for creating models by cutting off the Achilles tendon, and all ruptures were sutured by the Kessler method. After being fixed by plaster and ankle-knee fixation for 4 weeks, the rabbits were randomly divided into four groups: hyperbaric oxygen group (group A), ultrashort wave group (group B), hyperbaric oxygen combined with ultra-short wave group (group C), and control group (group D), with 20 rabbits in each group. Groups A and C were treated with hyperbaric oxygen immediately after surgery, twice a day for the first three days and once a day for the next four days, for a total of 7 days. Groups B and C were given ultrashort wave treatment every day for 6 days after operation. Group D was not treated. Ten white rabbits were sacrificed in each group at 4 and 8 weeks after operation. Achilles tendon tissue was taken for HE staining, immunohistochemical staining, and biomechanical detection (maximum tensile length and maximum tensile force). ANOVA was used to compare the results of biomechanical tests at 4 and 8 weeks after operation, and LSD-t test was used for comparisons between two groups.

Results

Biomechanical test results showed that the average maximum tensile force and length of the Achilles tendon in groups A, B, and C at 4 weeks postoperatively were significantly higher than those in group D [maximum tensile force: (276.20±8.16) N, (278.10±7.92) N, and (309.90±9.78) N vs (259.50±4.99) N, t=5.519, 6.280, and 14.513, P<0.001; maximum tensile length: (4.62±0.14) mm, (4.67±0.16) mm, and (5.05±0.17) mm vs (4.18±0.32) mm, t=3.934, 4.136, and 7.563, P=0.001, <0.001, and <0.001]; the two indicators in group C were significantly better than those in groups A and B (maximum tensile force: t=8.366 and 7.989, P<0.001; maximum tensile length: t=6.164 and 5.190, P<0.001). The average maximum tensile force and length of the Achilles tendon in groups A, B, and C at 8 weeks postoperatively were also significantly higher than those in group D [maximum tensile force: (354.60±5.68) N, (355.30±9.62) N, (390.70±7.48) N vs (339.60±5.60) N, t=5.946, 4.461, and 17.286, P<0.001; maximum tensile length: (4.88±0.18) mm, (5.10±0.16) mm, (5.38±0.15) mm vs (4.46±0.23) mm, t=4.578, 7.327, and 10.628, P<0.001]; the two indicators in group C were significantly better than those in groups A and B (maximum tensile force: t=12.150 and 9.187, P<0.001; maximum tensile length: t=6.600 and 3.925, P<0.001 and =0.001). Histologically, it was found that there were a large number of proliferating fibroblasts near the stump of the Achilles tendon, and there were a few inflammatory cells in group C. Although a large number of fibroblasts can be seen in groups A and B, they were fewer than those in group C, but the number of inflammatory cells was larger. In group D, the number of inflammatory cells was significantly more than those of the other three groups, and the growth of fibroblasts lagged significantly behind the other three groups. Immunohistochemical staining showed positive expression of type I collagen fibers, and the number of type I collagen fibers was most abundant in group C, followed sequentially by group B, group A, and group D.

Conclusion

Hyperbaric oxygen combined with ultrashort wave can promote the healing after Achilles tendon rupture, and promote the synthesis of typeⅠcollagen fibers.

Key words: Achilles tendon rupture, Adjuvant therapy, Hyperbaric oxygen, Ultrashort wave, Biomechanics

图2 缝合后的新西兰白兔后肢跟腱

图4 白兔后肢断裂后接受超短波治疗

图6 取出跟腱的生物力学检测

表1 各组兔子术后跟腱最大抗拉力和最大伸长度比较（±s）

组别	只数	最大抗拉力（N）		最大拉伸长度（mm）
组别	只数	4周	8周	4周	8周
D组	20	259.50±4.99^a	339.60±5.60^a	4.18±0.32^a	4.46±0.23^a
A组	20	276.20±8.16^b	354.60±5.68^b	4.62±0.14^b	4.88±0.18^b
B组	20	278.10±7.92^c	355.30±9.62^c	4.67±0.16^c	5.10±0.16^c
C组	20	309.90±9.78	390.70±7.48	5.05±0.17	5.38±0.15
F值		70.88	88.63	28.49	44.99
P值		<0.001	<0.001	0.001	<0.001

表1 各组兔子术后跟腱最大抗拉力和最大伸长度比较（±s）

组别	只数	最大抗拉力（N）		最大拉伸长度（mm）
组别	只数	4周	8周	4周	8周
D组	20	259.50±4.99^a	339.60±5.60^a	4.18±0.32^a	4.46±0.23^a
A组	20	276.20±8.16^b	354.60±5.68^b	4.62±0.14^b	4.88±0.18^b
B组	20	278.10±7.92^c	355.30±9.62^c	4.67±0.16^c	5.10±0.16^c
C组	20	309.90±9.78	390.70±7.48	5.05±0.17	5.38±0.15
F值		70.88	88.63	28.49	44.99
P值		<0.001	<0.001	0.001	<0.001

图7 术后4周跟腱断端镜下纤维结构图（HE染色，×40）　图a为A组染色图，图b为B组染色图，图c为C组染色图，图d为D组染色图（黑色←指示成纤维细胞；红色←指示炎性细胞）

图8 手术后4周免疫组织化学染色检测结果（HE×40）　图a为A组染色图，图b为B组染色图，图c为C组染色图，图d为D组染色图

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Hyperbaric oxygen combined with ultrashort wave for treatment of rabbit Achilles tendon rupture: an experimental study

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Hyperbaric oxygen combined with ultrashort wave for treatment of rabbit Achilles tendon rupture: an experimental study