Comparative Effects of Tissue Flossing and Proprioceptive Neuromuscular Facilitation on Hamstring Flexibility and Knee Range of Motion in Young Female Students
Phase: NA
Conditions: Hamstring Tightness, Muscle Tightness
Sponsor: Sakshi Negi

Official Title of Study: Comparative effects of tissue flossing and proprioceptive 
neuromuscular facilitation on hamstring flexibility and knee joint range of motion among 
young female students 
NCT Number: NCT07178132 
Document Type: 
• 
Study Protocol 
• 
Statistical Analysis Plan (SAP) 
Date: 19-09-2025 
Principal Investigator: 
• Sakshi Negi, MPT 
• Institution: Guru Nanak Dev University 
• Email- snegipt@gmail.com  
Ethics Approval: 
Approved by the Institutional Review Board of Guru Nanak Dev University, Amritsar  
Approval Number: 2250/HG 
Funding: None 
 
 
 
 
 
 
 
 
 

STUDY PROTOCOL 
AIMS AND OBJECTIVES OF THE STUDY 
• To analyze the effect of PNF in increasing hamstring muscle flexibility  
• To analyze the effect of kinetic tissue flossing in increasing hamstring muscle flexibility  
• To increase the joint range of motion 
• Reduce the risk of injury and enhance overall performance 
• To prevent muscle imbalance and promote optimal musculoskeletal health 
RESEARCH DESIGN 
• Study Design: A crossover study 
• Source of Data: Guru Nanak Dev University, Amritsar 
• Study population: Young female students with hamstring tightness 
• Sample Size: 60 subjects.  
• Selection of subjects: Random sampling technique 
SELECTION CRITERIA 
Inclusion criteria: 
1. Female subjects were included in the study  
2. Subjects with hamstring muscle tightness were included  
3. Subjects in the age group of 18-25 years were included  
Exclusion criteria: 
1. Male subjects 
2. Subject who doesn’t fulfill the age criteria 
3. Subject with hamstring tear or other knee ligament injury 
4. Any history of lower limb surgery dating back a year  
5. Subjects with Lower back pain or any neurological compromise in the lower limb 
6. Any acute injury like a sprain, strain, and open wound 
7. Subjects who had latex allergies 
8. Subjects with venous thrombotic disease, heart disease, respiratory illness, and 
dermatitis in their lower legs were excluded 
 

STUDY DESIGN: 
This study was designed as a randomized crossover trial. Participants were randomly assigned 
to one of two sequences: 
1. Tissue Flossing followed by PNF hold–relax stretching, or 
2. PNF hold–relax stretching followed by Tissue Flossing. 
Each participant therefore received both interventions, with a minimum washout period of one 
week between sessions to reduce the risk of residual effects. Following the washout, 
participants crossed over to the alternate intervention. This design allowed each participant to 
serve as their own control, thereby reducing inter-individual variability. 
INTERVENTION: 
KINETIC TISSUE FLOSSING 
➢ A floss band of 2.1 m in length, 5 cm in width, and 1.3 mm in thickness was used to 
conduct flossing. 
➢ The participant assumed a standing position and placed their dominant leg forward. 
➢ The floss band was wrapped around the thigh beginning at the superior pole of the 
patella a progressing to the upper thigh. 
➢ Compressive force on the hamstring muscle was produced through wrapping of the 
thigh with 50% overlap and stretch of about 50% of the band’s maximum stretch 
➢ Participants then performed one set of 20 active knee flexions, 10 standing knee hugs, 
and 10 lunges within 2-minutes 
➢ The band was then subsequently removed and the procedure was repeated once more 
after a 2-minute break  
PNF HOLD-RELAX 
➢ The subject lay supine with both legs extended and the non-dominant leg was stabilized 
➢ With no rotation at the hip, the therapist passively stretched the hamstring muscle until 
the subject felt a mild stretch sensation. 
➢ The subjects were instructed to perform isometric contraction of the hamstring for 8 
seconds followed by relaxation. 

➢ Immediately following the muscle relaxation, the therapist further stretched the 
hamstring until a mild yet tolerable stretch was reported. 
➢ The stretch was maintained for a further 30 seconds and the process was repeated three 
times.  
PRIMARY OUTCOMES: 
• Active Knee Extension Test (AKET, degrees) 
• Passive Knee Extension Test (PKET, degrees) 
• Straight-Leg Raise (SLR, degrees) 
• Sit-and-Reach Test (cm) 
STATISTICAL ANALYSIS PLAN (SAP) 
 
General Principles: 
• Software: analysed using SPSS (18) software 
• Statistical significance: two-sided p < 0.05 
• Continuous variables: mean ± SD 
Primary Outcomes: 
• Measures: AKET, PKET, SLR, Sit-and-Reach 
• 
Analysis:  
Within-intervention analysis: 
For each intervention (Tissue Flossing and PNF), paired-samples t-tests were used to compare 
pre- and post-intervention scores on the outcome measures (Active Knee Extension Test, Passive 
Knee Extension Test, Straight Leg Raise, and Sit-and-Reach). 
      Between-intervention comparison: 
To compare the relative effectiveness of Tissue Flossing versus PNF, paired-samples t-
tests were applied to the change scores from each intervention, since the same 
participants underwent both treatments.