||A smart running wheel system with adjustable exercise range and speed for precision rehabilitation of rats following ischemic stroke
||Department of BioMedical Engineering
Ischemic stroke rat
Dynamic exercise range running wheel
此滾輪平台的設計包含機構、硬體電路與軟體程式三個部分。機構設計包含滾輪主體與可調式運動區間側板，此側板下半部會切割一條寬為2公分的0~180度U形曲線，五組紅外線感測器可以依據老鼠個別的體能在U形曲線動態調整。硬體電路包含為微處理器、紅外線感測器與馬達的連接。軟體程式則利用紅外線感測訊號來偵測老鼠位置與控制滾輪速度。因此我們將此創新滾輪取名為動態訓練範圍滾輪(DEARW)。在此滾輪的各別化訓練過程分成三個階段：自由訓練(free training)、漸進式訓練(progressive training)與常規訓練(normal training)，首先為兩天的自由訓練讓老鼠自主的在滾輪上運動，並記錄平均速度與最大速度。接著三天的漸進式訓練從靜止逐漸上升到自由訓練的最大速度訓練，過程中會記錄老鼠運動範圍。接著利用自由訓練的平均速度與最大速度搭配漸進式訓練的運動範圍來進行兩週的常規訓練。跑步機組及傳統馬達式滾輪組在第一週以每分鐘10公尺、每次30分鐘、連續五天進行訓練，然後在第二及三週以每分鐘20公尺、每次30分鐘、連續五天進行訓練。在老鼠中大腦動脈暫時阻塞手術開刀前、開刀後第十四、第二十一及第二十八天，經尾部靜脈對老鼠抽血以測量皮質醇濃度(cortisol level)。在開刀後第七及第二十八天，對老鼠進行動作功能測試包括用來評估運動功能恢復的修改的神經學損傷嚴重程度評分表(Modified Neurological Severity Scores, mNSS)，前足位置錯誤測試(forelimb foot fault placing test)及滾筒運動測試(rotarod test)。在開刀後第二十八天，老鼠將於麻醉下將腦部取出進行用來評估損傷體積的尼氏染色(Nissl staining)及評估神經細胞損傷程度的蘇木精-伊紅染色(H&E-staining)。實驗老鼠隨機分配到偽對照組、控制組、動態訓練範圍滾輪組，跑步機組及傳統馬達式滾輪(MRW)等五組。中大腦動脈暫時阻塞手術開刀一週後在動態訓練範圍滾輪組，跑步機組及傳統馬達式滾輪(MRW)，進行連續三週的復健訓練後就皮質醇濃度、動作功能測試及尼氏染色和蘇木精-伊紅染色進行比較。
Background: An innovative animal running wheel of adopting an individualized design was implemented for the rehabilitation of rats following ischemic stroke by middle cerebral artery occlusion (MCAO) surgery.
Material and Methods: The design of the running wheel platform comprised three parts: the mechanism, the hardware circuit, and the software. The mechanism included the running wheel, and a side plate for exercise area adjustments. A U-curve with a width of 2 cm was drawn on the lower half of the side plate for the dynamic adjustments of five infrared (IR) sensors based on the physical fitness of the rats. The hardware circuit connected a microprocessor, the IR sensors, and a motor. The software was designed to detect the rat’s position and to control the speed of the running wheel based on the signals obtained from the IR sensors. This innovative running wheel was named the dynamic exercise range running wheel (DEARW). The individualized training process for this running wheel was divided into free training, progressive training and normal training, which consisted of 2 days of free training to record their average and maximum speeds, 3 days of progressive training for recording their exercise ranges, and 2 weeks of normal training based on their average speeds, maximum speeds, and exercise ranges. The treadmill and MRW groups trained at 10 m/min for 30 min daily for five consecutive days during the first week and 20 m/min for 30 min daily for five consecutive days during the second and third weeks. Blood samples were obtained from the tail veins of all rats before the operations and on Days 14, 21, and 28 post-surgery to measure cortisol levels. The motor function tests including Modified Neurological Severity Scores (mNSS), forelimb foot fault placing and rotarod tests were performed on Days 7 and 28 post-surgery. On Day 28 post-surgery, the rats were sacrificed under anesthesia, and their brains were removed for lesion volume determined using Nissl staining and levels of cell damage by using H&E staining. The rats were randomly assigned into five groups (i.e., sham, control, treadmill, motor running wheel (MRW), and DEARW) and subjected to MCAO surgery accordingly. After 1 week, the rats in the rehabilitation groups (i.e., the treadmill, MRW, and DEARW groups) began a 3-week rehabilitation training program for comparison of cortisol level, motor function tests, lesion volume determined using Nissl staining and levels of cell damage by using H&E staining.
Results: On Day 28 after surgery, the motor function evaluation (including Modified Neurological Severity Scores (mNSS), forelimb foot fault placing and rotarod tests), lesion volume determined using Nissl staining and levels of cell damage by using H&E staining of the DEARW and control groups differed significantly, indicating that this device is effective for ischemic stroke rats motor training. However, differences between the results of the control, treadmill and MRW groups were not significant, indicating the poor efficacy of treadmill and MRW rehabilitation. The level of cortisol in the treadmill group remained consistently high throughout the rehabilitation program, indicating that the electrically stimulated rats were under immense stress.
Conclusion: The outcomes of the rats in the proposed group, which were rehabilitated using the designed training system, were better than those of their control-group counterparts, indicating the advantages of the designed system and achieved concept of precision rehabilitation of rats following ischemic stroke with the use of DEARW technology for treating each rat’s rehabilitation process in an individualized manner. That may have new applications for clinical translation for unmet need of stroke rehabilitation in the future.
List of Tables IX
List of Figures X
Chapter 1. Background and Significance 1
1.1 Ischemic stroke and cardiovascular deconditioning 1
1.2 Exercise rehabilitation mechanisms 1
1.3 Animal stroke model 2
1.4 Animal training platforms 2
1.5 Running-wheel-based training mechanism 3
1.6 Dynamic Exercise Range Running Wheel(DEARW) 3
1.7 Motivation 3
1.8 Specific Aims 4
Chapter 2. Material and Methods 6
2.1 Mechanism design of the running wheel platform 7
2.2 Hardware circuit 9
2.3 Control software 9
2.4 Verification of system functions 12
2.5 Experimental model of stroke in rats 13
2.5.1 Middle Cerebral Artery Occlusion(MCAO) 13
2.5.2 Animals and grouping 13
2.5.3 Rehabilitation training 14
2.5.4 Assessment of ischemic stroke rehabilitation 15
18.104.22.168 Modified Neurological Severity Scores(mNSS) 15
22.214.171.124 Forelimb foot fault placing test 16
126.96.36.199 Rotarod test 16
188.8.131.52 Serum cortisol levels 17
184.108.40.206 Nissl staining 17
220.127.116.11 H&E staining 18
2.6 Statistical analysis 18
Chapter 3. Results 19
3.1 Motor performance of ischemic stroke rats in DEARW 19
3.2 Motor function test 20
3.3 Cortisol level 21
3.4 Tissue photographs and lesion volume 22
3.5 Tissue photos and brain damage score 23
Chapter 4. Discussion 26
4.1 Achievement in present study 26
4.2 Method of forced training 26
4.3 Level of stress 27
4.4 Comparison with Commercial Products 29
4.5 Limitation 30
4.6 Clinical translation 31
Chapter 5. Conclusion and Future Work 33
5.1 Conclusion 33
5.2 Future Work 34
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