Remnant preservation provides good clinical outcomes after anterior cruciate ligament reconstruction.

PURPOSE: To evaluate the association of remnant preservation (RP) and non-RP (NRP) with patient-reported outcome measures and subsequent graft rupture at a minimum 2-year follow-up after anterior cruciate ligament (ACL) reconstruction. METHODS: Patients in this retrospective study underwent primary isolated ACL reconstruction by the RP or NRP technique with a four- to five-strand hamstring tendon graft. Multivariate linear or logistic regression and Cox regression analyses were performed to compare the physical and psychological outcomes by the International Knee Documentation Committee subjective knee form (IKDC-SKF) and the Japanese Anterior Cruciate Ligament questionnaire 25 (JACL-25), respectively; satisfaction rate; and prognosticators of graft rupture. RESULTS: In total, 120 patients (mean age, 30.6 ± 12.7 years; 54 RP, 66 NRP) with a mean follow-up of 3.2 ± 1.6 years were enrolled in this study. At the latest postoperative follow-up, the RP group showed a mean IKDC-SKF score of 92.3 ± 8.5 and mean JACL-25 score of 13.2 ± 11.2, while these scores in the NRP group were 86.4 ± 12.2 and 24.4 ± 19.5, respectively (P = 0.016 and 0.007, respectively). No significant differences were found in the return-to-sports rate (RP vs. NRP, 79.5% vs. 67.5%) or satisfaction rate (RP vs. NRP, 89.2% vs. 74.4%) (n.s.); however, a significant difference was found in the rate of return to the preinjury sports level (RP vs. NRP, 64.1% vs. 37.5%; P = 0.014). The graft rupture rate was significantly higher in the NRP than RP group (9/66 vs. 1/54; hazard ratio 9.29; 95% confidence interval 1.04-82.81). Younger age (≤ 18 years) was the other important risk factor for graft rupture (hazard ratio 8.67; 95% confidence interval 2.02-37.13). CONCLUSION: Patients who underwent ACL reconstruction with the RP technique obtained somewhat better physical and psychological results than those who underwent ACL reconstruction with the NRP technique. With respect to clinical relevance, patients treated with the RP technique may obtain better outcomes in terms of graft rupture and return to the preinjury sports level than those treated with the NRP technique, but with no differences in overall return to sports or satisfaction. LEVEL OF EVIDENCE: IV.

Validation and defining the minimal clinically important difference of the Japanese version of the IKDC Subjective Knee Form.

BACKGROUND: This study aimed to validate and determine the minimal clinically important difference (MCID) in the Japanese version of the International Knee Documentation Committee Subjective Knee Form (Japanese IKDC-SKF) for patients with anterior cruciate ligament (ACL) injuries. METHODS: This prospective study was performed using the COnsensus-based Standards for the selection of health status Measurement INstruments (COSMIN) guideline to assess the measurement properties of reliability, responsiveness, validity, and interpretability of the Japanese IKDC-SKF. RESULTS: A total of 152 patients were enrolled in the study. The Japanese IKDC-SKF had excellent internal consistency [Cronbach's alpha (95% confidence interval, CI) was 0.90 (0.88-0.92), and the test-retest reliability [intraclass correlation coefficient, ICC2,1 (95% CI)] was 0.97 (0.94-0.98). The content validity (r), which was interpretable between the Japanese IKDC-SKF and the Japanese Anterior Cruciate Ligament questionnaire 25 was -0.87 (P < 0.001). The construct validity for the IKDC-SKF was 86% (six of seven hypotheses were consistent with the results), also indicating high validity. Responsiveness of the Japanese IKDC-SKF was confirmed, with an acceptable area under the curve of 0.81 and a large effect size (Cohen's d of 0.8). There were no floor or ceiling effects. The MCID for the Japanese IKDC-SKF for ACL injury was 10.7. CONCLUSIONS: Our results indicate that the Japanese version of the IKDC-SKF is a reliable, valid, and responsive measurement instrument with which to evaluate the physical function of patients with ACL injuries.

Reliability assessment of the functional movement screen for predicting injury risk in Japanese college soccer players.

[Purpose] This study aimed to assess the reliability of the Functional Movement Screen and explore whether this evaluation tool can predict the risks of personal injuries in Japanese soccer players. [Participants and Methods] Seventy-five Japanese college soccer players who participated in our 1 year prospective cohort study underwent a Functional Movement Screen assessment. Demographic data, athletic characteristics, and types and frequency of injuries sustained, were analyzed with the assessment results. [Results] There was no significant difference in the mean Functional Movement Screen composite scores between genders. Although the Functional Movement Screen showed excellent inter-rater reliability (0.92), low overall internal consistency (0.35) was observed. A maximum score of 3 in straight leg raise occurred in 94% of the females and was considered a ceiling effect. None of the cut-off point scores of the Functional Movement Screen were associated with the number of overall injuries, lower limb injuries, and traumatic injuries, or time to return to play. The Functional Movement Screen composite score of ≤15 represented the maximum sensitivity of 76.92% and specificity of 34.78% with 0.56 in the area under the curve. [Conclusion] Functional Movement Screen composite scores do not have sufficient sensitivity and specificity for predicting injuries in Japanese college soccer players.

Reproducibility, responsiveness and validation of the Tampa Scale for Kinesiophobia in patients with ACL injuries.

BACKGROUND AND PURPOSE: Psychological factors including fear of pain, re-injury during movement (kinesiophbia) affect return-to-sport rates after anterior cruciate ligament (ACL) reconstructive surgery. Clinicians often encounter in the daily practice that athletes explain lack of self-confidence or psychological readiness during the sports activity. The Tampa Scale for Kinesiophobia (TSK) has been used to evaluate psychological outcomes in patients with ACL injuries in many countries and translated into Japanese version in 2013. However, no researchers validated its reliability, validity, and responsiveness of TSK for patients with ACL injury up to now. The purpose of this study was to evaluate the measurement properties of the Japanese version of the TSK (TSK-J) in patients with ACL injuries. STUDY DESIGN: Cohort study (Diagnostic); Level of evidence, 2. METHODS: This prospective study was performed in the department of orthopaedic surgery at the university hospital of Juntendo from Sep 2016 and Apr 2017. Patients who diagnosed with ACL injury with or without reconstruction surgery completed several patient-reported outcome measures (PROMs) were included in this study. The COnsensus-based Standards for the selection of health status Measurement INstruments (COSMIN) guidelines were used to evaluate reliability, validity, responsiveness, and interpretability of the TSK-J. RESULTS: 222 patients were included in this study. The TSK-J for ACL injured patients showed good internal consistency (Cronbach's alpha = 0.79) and excellent test-retest reliability (intra-class correlation coefficient, ICC2,1 = 0.90, 95% CI = 0.81 to 0.95). In addtion, the TSK-J was significantly but moderately correlated with the IKDC-SKF (r = - 0.49, P <0.001), VAS-Sports (r = - 0.48, P <0.001), and JACL-25 (r = 0.48, P <0.001). The effect size (ES) was small with the Cohen's d = - 0.2. The minimal important difference (MID) was - 1.3 points. No significant TSK-J score change was observed over 1-year after ACL reconstruction (r = - 0.12, P <0.001). There were no floor or ceiling effects. CONCLUSIONS: Our study demonstrated that the Japanese version of TSK has good reliability. However, its low validity and responsiveness indicate that it may not the best way to assess psychological factors for patients with ACL injury.

Leukocyte concentration and composition in platelet-rich plasma (PRP) influences the growth factor and protease concentrations.

BACKGROUND: Platelet-rich plasma (PRP) therapy has become an increasingly popular treatment for orthopaedics and sports-related injuries, and various clinically available PRP preparation methods exist. However, the differences in PRP quality among numerous preparation methods remain unclear. Specifically, the benefit of including leukocytes in the PRP product remains controversial, and few studies have been conducted to evaluate the effects of the interaction between platelets and leukocytes on the growth factor concentrations. The aim of the present study was to compare the biological characteristics of PRPs focusing on the leukocyte concentration and composition. METHODS: Leucocyte rich (LR)-PRP, leucocyte poor (LP)-PRP, and pure-PRP were prepared from the peripheral blood of 6 healthy male volunteers (mean age: 31.3 years). The concentrations of platelets, leukocytes, erythrocytes, growth factors (transforming growth factor-beta 1: TGF-ß1; fibroblast growth factor-basic: FGF-b; platelet-derived growth factor-BB: PDGF-BB; vascular endothelial growth factor: VEGF) and matrix metalloproteinase-9 (MMP-9) from each of the PRP samples were measured. Considering the interaction between platelets and leukocytes, correlations between platelets/leukocytes and growth factors/MMP-9 were analyzed using partial correlation coefficients. RESULTS: The platelet concentration did not differ among the three PRP preparation methods. Conversely, the leukocyte concentration was dramatically different: 14.9 ± 4.5 (10(3)/µl) in LR-PRP, 2.4 ± 1.3 (10(3)/µl) in LP-PRP, 0.2 ± 0.2 (10(3)/µl) in pure-PRP. The platelet concentration positively correlated with all growth factors. On the other hand, the leukocyte concentration positively correlated with PDGF-BB and the VEGF concentration, while it negatively correlated with FGF-b. Regarding catabolic factors, the MMP-9 concentration strongly correlated with the leukocyte concentration, while there was no correlation between the platelet and MMP-9 concentrations. CONCLUSIONS: These findings demonstrate that leukocytes strongly influence the quality of PRPs. Therefore, modifying the PRP preparation method according to the pathology is essential to achieve better clinical results with PRP therapy.

Cross-cultural adaptation, validity, reliability and responsiveness of the Japanese version of the Victorian Institute of sports assessment for patellar tendinopathy (VISA-P-J).

BACKGROUND: This study aimed to translate, adapt, and test the psychometric properties of the Japanese version of the Victorian Institute of sports assessment for patellar tendinopathy (VISA-P-J). METHODS: This prospective cohort study registered 43 participants ≥ 18 years old with a history of painful symptoms in the inferior pole of the patella to the proximal patellar tendon lasting ≥ 1 month for patellar tendinopathy. Pain in daily life and during sports activities, symptom classification, and patient global impression of change were assessed at the baseline, 1 week, and 12 weeks. The psychometric properties, test-retest reliability, standard error of measurement, internal consistency criterion validity, construct validity, responsiveness, and interpretability, of the VISA-P-J were calculated according to the COSMIN. RESULTS: The two-way random-effects, absolute agreement intraclass correlation coefficient for test-retest reliability of VISA-P-J was 0.87 (95% confidence interval: 0.78, 0.93), and the standard error of measurement of VISA-P-J was 0.89. The Cronbach's alpha for internal consistency of VISA-P-J was 0.81. A correlation between VISA-P-J and Roel's classification, Visual Analog Scale for pain (VAS)-Active Daily Living, and VAS-Sports (r = - 0.52, r = - 0.66, r = - 0.86, p < 0.01, respectively) was observed for criterion validity. All hypotheses of the hypothesis-testing method to evaluate construct validity and responsiveness of VISA-P-J were substantiated. The minimal clinically important difference of VISA-P-J was 7 points. CONCLUSION: We demonstrated that the VISA-P-J was a reliable, valid, and responsive assessment method for individuals with chronic pain in the patellar tendon.

Effect of Isokinetic Training with Blood Flow Restriction During Rest Interval Versus Exercise on Muscle Strength, Hypertrophy, and Perception: A Pilot Study.

Objectives: This study aimed to determine the effects of high-intensity isokinetic training with blood flow restriction during rest interval between set (rBFR) versus during exercise (eBFR) on muscle hypertrophy and increasing muscle strength and determine whether BFR-induced exercise pain is suppressed by rBFR. Materials and Methods: Fourteen arms (7 participants) were recruited for the study. We conducted the following interventions for each arm: eBFR (n=4), rBFR (n=5), and exercise only (CON, n=5). The participants performed elbow flexion training with a BIODEX device twice weekly for 8 weeks. This study training consisted of total four sets; each was performed until <50% peak torque was achieved twice consecutively. BFR pressure was set at 120 mmHg. Elbow flexor peak torque during concentric contraction (CC), isometric contraction (IM), and muscle cross-sectional area (CSA) were measured before and after the intervention. Numerical rating scale scores used to assess pain during exercise were determined during training. Results: Peak torque at the CC increased in the rBFR (p<0.05) and IM increased in the rBFR and CON (p<0.05), while CSA increased in the rBFR and CON (p<0.001). The pain during exercise was severe in the eBFR and moderate in the rBFR and CON. Conclusions: This study's showed that high-intensity isokinetic training with rBFR did not have a synergistic effect on increasing muscle strength and muscle size. Additionally, high-intensity isokinetic training with BFR when it may be best not to perform it during exercise, because it was induces severe pain and may inhibit increases in muscle strength.

The Accuracy of Ankle Eccentric Torque Control Explains Dynamic Postural Control During the Y-Balance Test.

Background: The Y-Balance Test (YBT), especially the posteromedial (PM) reach direction (PM-YBT), is able to identify dynamic postural control deficits in those who have ankle instability. However, there still exists a need to understand how sensorimotor function at the ankle explains the performance during the PM-YBT. Hypothesis/Purpose: The purpose of this study was to determine whether the ability to accurately control eccentric ankle torque explained PM-YBT performance. It was hypothesized that eccentric dorsiflexion/plantarflexion torque control would be positively related to the maximum reach distance (MRD) of PM-YBT. Study Design: Cross-sectional study. Methods: Twelve healthy subjects performed the PM-YBT, maximum voluntary isometric contractions (MVIC) for both dorsiflexion and plantarflexion muscle strength, and then the torque control testing of the ankle. The torque control testing provided a target torque level on a screen in front of the subject and passive rotations of the ankle joint in the sagittal plane at 10 deg/sec between plantarflexion to dorsiflexion. Subjects were then instructed to eccentrically contract the dorsiflexors and plantar flexors to generate torque while the ankle joint rotated. The accuracy of torque control during eccentric dorsiflexion and plantarflexion by calculating absolute errors, the area between the target torque and the produced torque were evaluated. Tibialis anterior and soleus muscle activities were simultaneously recorded during testing. A step-wise linear regression model was used to determine the best model predicted the MRD of the PM-YBT (PM-MRD). Results: A step-wise linear regression developed a model explaining only eccentric dorsiflexion torque control predicted higher PM-MRD score (R2 = 44%, F1,10 = 7.94, ß = -0.67, p = 0.02). Conclusion: The accuracy of torque control during eccentric dorsiflexion predicts better performance in the PM-YBT. Level of Evidence: 3b.

Effect of low-intensity exercise with blood flow restriction during rest intervals on muscle function and perception.

PURPOSE: We assessed the effects of low-intensity exercise with blood flow restriction (BFR) during rest intervals on recovery of muscle function and pain during exercise and rest intervals. METHODS: Participants were 10 males and study arms of the participants were randomly assigned into three conditions; low-intensity exercise with BFR during rest intervals (rBFR), low-intensity exercise with BFR during exercise (eBFR) and low-intensity exercise only (EO). The exercise task was elbow flexion until repetition failure at 30% of 1 RM and cuff pressure was 120 mmHg. The maximum voluntary isometric contraction (MVIC) and the muscle endurance (ME) were measured pre, post, 1, 24 and 48 h after the exercise. Pain during exercise and rest intervals were evaluated using the Numerical Rating Scale. RESULTS: MVIC and ME significantly decreased after exercise in all conditions. Pain during exercise was lower in rBFR (4.2 ± 2.9) (p = 0.007) and EO (4.4 ± 2.7) (p = 0.014) conditions compared to eBFR condition (6.7 ± 1.7), but the pain during rest intervals was more intense in rBFR condition (5.2 ± 1.6) compared to eBFR (1.5 ± 1.4) and EO (1.7 ± 1.2) conditions (p < 0.001). CONCLUSION: We discovered that recovery of muscle function was the same as BFR during rest intervals and BFR during exercise. Also, our results suggested that BFR itself may cause the perception of pain. Future studies are thus required to investigate the optimal dosage focusing on the pressure volume and intensity used in BFR during intervals.

Age-related changes in mechanical properties of semitendinosus tendon used for anterior cruciate ligament reconstruction.

BACKGROUND: Hamstring tendons are a popular choice for autografts in anterior cruciate ligament (ACL) reconstruction. However, there is increasing evidence that hamstring tendon autografts carry a high risk of revision and residual instability in young patients. To elucidate the reasons for the inferior outcome of the reconstructed ACL with hamstring tendon autografts in young patients, we investigated the Young's modulus and the extent of cyclic loading-induced slackening of the semitendinosus tendon used for ACL reconstruction across a broad range of ages. METHODS: Twenty-six male patients (aged 17-53 years), who were scheduled for ACL reconstruction surgery using the semitendinosus tendon autograft, participated in this study. The distal portion of the harvested semitendinosus tendon, which was not used to construct the autograft, was used for cyclic tensile testing to calculate the Young's modulus and the extent of slackening (i.e., increase in slack length). RESULTS: Spearman correlation analysis revealed that the Young's modulus of the semitendinosus tendon was positively correlated with the patient's age (ρ = 0.559, P = 0.003). In contrast, the extent of tendon slackening did not correlate with the patient's age. CONCLUSIONS: We demonstrated that the Young's modulus of the semitendinosus tendon increases with age, indicating that the semitendinosus tendon used for ACL reconstruction is compliant in young patients.

Study protocol for a randomised placebo controlled trial of platelet-rich plasma injection to prevent post-traumatic knee osteoarthritis after anterior cruciate ligament reconstruction.

INTRODUCTION: The elevated cytokine levels in patients suffering from anterior cruciate ligament (ACL) rupture may lead to acute post-traumatic arthritis (APTA) and post-traumatic osteoarthritis (PTOA). Due to its chondrogenerative and anti-inflammatory effect, platelet-rich plasma (PRP) therapy is expected to show a positive outcome in APTA and PTOA. The proposed trial aims to quantitatively measure the efficacy of PRP injection in arresting post-traumatic cartilage degeneration among patients after ACL reconstruction. METHODS AND ANALYSIS: This will be a single-blind, randomised, prospective, controlled clinical trial designed following the Consolidated Standards of Reporting Trials guidelines. After ACL reconstruction, 80 patients will be randomised to receive either leucocyte-poor PRP injection after joint aspiration or a placebo control group receiving only joint aspiration. Participants (age 20-49 years) will be those who have undergone ACL reconstruction within the past 2 weeks with a body mass index<35 and Kellgren Lawrence osteoarthritis grade<2. The primary outcome will include MRI-T2 values of knee cartilage at 6 months. The secondary outcomes will include pain assessment by Visual Analogue Scale, Knee injury and Osteoarthritis Outcome Score, blood and urine test, physical findings, measurements for muscle strength and joint stability. ETHICS AND DISSEMINATION: The study was approved by The Independent Ethics Committee for Clinical Trials of the Japanese Association for the Promotion of State-of-the-Art Medicine. Results of the trial and each of the outcomes will be shared via conferences and publication in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: jRCTb030200391.

Intra-Articular Injections of the Adipose-Derived Mesenchymal Stem Cells Suppress Progression of a Mouse Traumatic Knee Osteoarthritis Model.

OBJECTIVE: This study aimed to determine whether the intra-articular injection of human adipose-derived mesenchymal stem cells (ADSCs) protects against the progression of murine post-traumatic osteoarthritis. DESIGN: ADSCs were isolated from human abdomen or buttock adipose tissues. In in vitro study, ADSCs conditioned medium was added to human chondrocytes pre-treated with interleukin-1ß (IL-1ß), and resultant gene expression of target inflammatory genes was measured by real-time quantitative polymerase chain reaction. A mouse model of knee osteoarthritis was generated by unilaterally transecting the medial meniscus in the right hind limb of 20 female C57BL/6 mice. Mice were randomly assigned to 2 treatment groups that received 6 µl intra-articular injections of either phosphate-buffered saline (control) or 2 × 104 cells/µl of ADSCs 14, 28, and 42 days post-surgery. Mice were euthanized 84 days post-surgery and histological and micro-computed tomography evaluation of knee joints were analyzed. Hind limb weight-bearing distribution was measured pre-surgery and 28 and 84 days post-surgery. RESULTS: Conditioned medium from cultured human adipose-derived mesenchymal stem cells suppressed the expression of target inflammatory genes in chondrocytes pre-treated with IL-1ß, suggesting anti-inflammatory properties (P < 0.01). Histological analyses indicated that the progression of destabilization of medial meniscus-induced knee osteoarthritis was suppressed by the administration of ADSCs compared with control group at medial femorotibial joint in vivo. This protective effect was related to a reduction in articular cartilage loss. CONCLUSION: The intra-articular injection of ADSCs suppressed articular cartilage loss in a mouse model of knee osteoarthritis, possible through anti-inflammatory mechanisms.

Biological effects of stored platelet-rich plasma eye-drops in corneal wound healing.

BACKGROUND/AIMS: This study aimed to assess the efficacy and sterility of stored platelet-rich plasma (PRP) eye-drops for corneal epithelial wound healing compared with those of autologous serum (AS) eye-drops. METHODS: At our single institution, PRP and AS eye-drops were prepared using peripheral blood obtained from six healthy volunteers and stored at 4°C. Platelet and leucocyte counts and transforming growth factor (TGF)-ß1, epidermal growth factor (EGF), and fibronectin levels were assessed during storage for up to 4 weeks. Sterility was assessed by culturing 4-week poststorage samples. PRP, AS, and phosphate-buffered saline (PBS) eye-drop efficacies were compared using corneal epithelial wound healing assays in vitro and in vivo and monitoring wound areas under a microscope every 3 hours. RESULTS: Higher platelet and lower leucocyte counts were seen in PRP than in whole blood on the day of preparation. After storage, TGF-ß1, EGF, and fibronectin levels were significantly higher in PRP than in AS eye-drops. In vitro and in vivo, PRP eye-drops used on the day of preparation significantly promoted corneal epithelial wound healing compared with PBS. Moreover, PRP eye-drops stored for 4 weeks significantly promoted corneal wound healing compared with PBS and AS eye-drops. CONCLUSION: PRP eye-drops stored at 4°C for 4 weeks promoted corneal epithelial wound healing with higher levels of growth factors than those observed in AS eye-drops, while maintaining sterility, suggesting that this preparation satisfies the unmet medical needs in the treatment of refractory keratoconjunctival epithelial disorders.

Predictors of Effectiveness of Platelet-Rich Plasma Therapy for Knee Osteoarthritis: A Retrospective Cohort Study.

There has recently been growing interest worldwide in biological therapies such as platelet-rich plasma injection for the treatment of knee osteoarthritis. However, predicting the effectiveness of platelet-rich plasma therapy remains uncertain. Therefore, this retrospective cohort study was performed to assess a range of predictors for the effectiveness of platelet-rich plasma therapy in treating knee osteoarthritis. The study included 517 consecutive patients who underwent three injections of leucocyte-poor platelet-rich plasma therapy from 2016 to 2019 at a single institution. The treatment outcomes, including patient-oriented outcomes (visual analogue scale score and Knee Injury and Osteoarthritis Outcome Score), were analyzed and compared according to the severity of knee osteoarthritis based on Kellgren-Lawrence (KL) grading using standing plain radiographs. Fisher's exact test, univariate regression, and multivariate regression were used for data analysis. Patient-oriented outcomes were significantly improved 6 and 12 months after platelet-rich plasma therapy. The overall responder rate in patients who met the Outcome Measures in Rheumatology (OMERACT)-Osteoarthritis Research Society International (OARSI) responder criteria was 62.1%. The responder rate was significantly lower in patients with severe knee osteoarthritis (KL4, 50.9%) than in those with mild (KL2, 75.2%) and moderate (KL3, 66.5%) knee osteoarthritis. The multivariate logistic regression analysis revealed that deterioration of the knee osteoarthritis grade (increased KL grade) was a significant predictor of a worse clinical outcome (odds ratio, 0.58; 95% confidence interval, 0.45-0.75; p < 0.001). The relative risk for non-responders in severe (KL4) KOA was 2.1 (95% CI, 1.5-3.0) at 6 months and 2.3 (1.6-3.2) at 12 months compared with mild-to-moderate (KL2-3) KOA. The efficacy of platelet-rich plasma therapy was not affected by age, sex, body weight, or platelet count. This study revealed that the effectiveness of platelet-rich plasma therapy for the treatment of knee osteoarthritis is approximately 60% and that the effectiveness depends on the severity of knee osteoarthritis. This observation is useful not only for physicians but also for patients with knee osteoarthritis.

The immature platelet fraction affects the efficacy of platelet rich plasma therapy for knee osteoarthritis.

INTRODUCTION: Platelet-rich plasma (PRP) therapy is used to treat pathological conditions such as degenerative inflammatory diseases including osteoarthritis (OA) by enhancing tissue repair and promoting anti-inflammatory effects. Although PRP therapy for patients with knee OA improved pain and functional scores, the association of clinical outcomes and quality of PRP including cell composition and concentration is unclear. METHODS: Therefore, this study analyzed blood cell counts, including the immature platelet fraction (IPF), in peripheral blood and PRP of 144 patients with knee OA who underwent PRP therapy. The mean leukocyte and platelet concentrations in whole blood and PRP were analyzed using an XN-1000 automated hematology analyzer. Visual analogue scale (VAS) scores and knee injury and osteoarthritis outcome scores (KOOS) before and 1 month after a single PRP injection were also determined. RESULTS: Higher platelet and lower leukocyte concentration rates were observed in PRP compared with whole blood. The platelet concentration in whole blood was negatively correlated with VAS improvement. The percentage of IPF (IPF%) in whole blood was positively correlated with VAS improvement and KOOS (pain) improvement, whereas the IPF% in PRP tended to correlate with VAS improvement. Furthermore, multivariate logistic regression demonstrated the high IPF% in whole blood was significantly associated with VAS improvement. The low percentage of neutrophil (neutrophil%) in PRP was significantly associated with the VAS improvement and KOOS (ADL) improvement. CONCLUSIONS: Therefore, PRP efficacy for OA might depend on the patient's biological status.

Platelet-rich plasma (PRP) accelerates murine patellar tendon healing through enhancement of angiogenesis and collagen synthesis.

PURPOSE: Although platelet-rich plasma (PRP) therapy has become an increasingly popular treatment for sports-related injuries, the molecular mechanisms of PRP on tissue healing process remain poorly understood. The aim of the present study was to develop an experimental method quantifying the efficacy of PRP with murine patellar tendon injury model, leading to future elucidation of the mechanisms of PRP on healing processes. METHODS: Full-thickness defects were created in the central third of the murine patellar tendon. The prepared allogenic PRP gel was applied on the defect of the patellar tendon (PRP group), while the remaining mice served as the untreated control group. Mice were sacrificed at 2, 4, 6, 8, and 10 weeks after the operation, with histological sections obtained in each time point (n = 4 / time point / group). Semi-quantitative histological evaluation was performed in accordance with the Bonar score. The variables included in this scoring system were cell morphology, ground substance, collagen arrangement, and vascularity, with higher grades indicating worse tendon structures. In addition, the ratio of the collagen fibers to the entire tendon tissue (FT ratio) was measured using KS400 software as a quantitative histological evaluation. RESULTS: The total Bonar score in the PRP group was significantly lower than in control group. With regard to the variables in the Bonar score, the vascularity score was significantly higher in the PRP group at 2 and 4 weeks, while the collagen arrangement score was significantly lower in the PRP group at 8 weeks. Based on a quantitative evaluation, the recovery speed of the patellar tendon determined by FT ratio was significantly faster in the PRP group than in the control group at 6 and 8 weeks. CONCLUSIONS: We have developed an experimental method for histological and quantitative evaluation of the effects of PRP on tissue healing using murine patellar tendon injury model. The results of this study suggest that the local application of PRP could enhance the tissue-healing process both directly through action on localized cells and indirectly through the recruitment of reparative cells through the blood flow. Further investigations will be needed to confirm the mechanisms of PRP in tissue-healing processes with the development of this experimental model.

Quality comparison between two different types of platelet-rich plasma for knee osteoarthritis.

INTRODUCTION: Knee osteoarthritis (KOA), the most common form of osteoarthritis (OA) is a considerable health concern worldwide. Platelet-rich plasma (PRP) is a common therapeutic option for KOA. Different types of PRPs have varying efficacies. However, a comparative analysis of the qualities of these PRPs is lacking. METHODS: Two types of PRPs, including autologous protein solution (APS), and leukocyte-poor PRP (LP-PRP) along with whole blood (WB) and platelet-poor plasma (PPP) were characterized for platelet content, leukocyte content, and composition in 10 healthy volunteers (HV) (the controlled laboratory study) and 16 KOA patients (a retrospective observational study). Additionally, the levels of the platelet-derived growth factor (PDGF)-BB, and different cytokines were estimated in HV. RESULTS: In HV, the concentrations of platelets and leukocytes, levels of different cytokines, including interleukin 1 receptor antagonist (IL-1Ra), soluble TNF receptor type II (sTNF-RII), and IL-1ß, and the ratio of IL-1Ra/IL-1ß were significantly higher in APS, whereas the PDGF-BB was higher in LP-PRP than APS. In KOA patients, a higher concentration of platelets was observed in LP-PRP, and a higher concentration of leukocytes was observed in APS than LP-PRP. Following the PAW classification system, LP-PRP was classified as P2-B type in HV (51.3 × 104/µl) and KOA (53.4 × 104/µl), whereas APS was classified as P3-A type in HV (110.1 × 104/µl) and P2-A type in KOA (29.0 × 104/µl). In a retrospective observational study, the KOA patients who underwent APS injection had a higher incidence of arthralgia, and this arthralgia lasted for a longer time than LP-PRP injection in the same individual. DISCUSSION: The quality of the two PRPs differed distinctively depending on their preparation methods, which might affect their clinical efficacies and adverse events. Therefore, the characterization of these parameters should be prioritized while choosing PRP.

Platelet-rich plasma promotes recruitment of macrophages in the process of tendon healing.

INTRODUCTION: Researchers have investigated the use of platelet-rich plasma (PRP) therapy. However, the mechanisms through which PRP affects tissue repair remain unclear. We hypothesize that PRP promotes tissue repair through not only via direct manner on the local cells but also via indirect manner that encourage the recruitment of reparative cells such as macrophages (MPs), and it depends on the quality of PRP including the concentration of leukocytes. The aim of this study is to elucidate the actions of the MPs in the mechanisms of PRP on tissue repair processes. METHODS: Leukocyte-rich (LR) PRP and leukocyte-poor (LP) PRP were prepared from 12-week-old C57BL6 mice. Full-thickness defects were created in central third of patellar tendons of 12-week-old C57BL/6 mice for histologic analysis (n = 36) and 12-week-old B6.129P-Cx3cr1tm1Litt/J mice for flow cytometry analysis (n = 108). B6.129P-Cx3cr1tm1Litt/J mouse is GFP-positive only in the MP-linage cells thus MPs recruited to the repair tissue can be distinguished whether it had originated from administrated PRP or recruited from host mouse. Mice were treated either with LR-PRP, LP-PRP, or without PRP (control group). Histological analyses were performed to evaluate the tendon healing using Bonar score as semi-quantitative histological scoring system. Flow cytometric analyses were performed to count the number of GFP-positive cells around repaired patellar tendon. In addition, the ratio of pro-inflammatory MPs (M1)/anti-inflammatory MPs (M2) were analyzed in those GFP-positive cells. The statistical analysis was performed using GraphPad Prism ver6. P values < 0.05 were considered statistically significant. RESULTS: In LR-PRP and LP-PRP groups, all variables in Bonar score such as cell morphology, cellularity, vascularity, and collagen arrangement were significantly improved in comparison with control group, indicating that both PRPs promote tendon hearing. LP-PRP promoted the tendon healing significantly faster than that of LR-PRP on postoperative day 28 (P < 0.001). LR-PRP enhanced angiogenesis (vascularity: P < 0.001), while LP-PRP improved the collagen arrangement on postoperative day 28 (collagen arrangement: P < 0.01). In other variables such as cell morphology and cellularity score, there were no significant differences between LR-PRP and LP-PRP groups in any time points. Flow cytometric findings showed that recruitment of GFP-positive MPs in the LR and LP-PRP groups were significantly increased from postoperative day 4 compared with control group without PRP treatment (P < 0.001). The majority of GFP-positive MPs were M1 at the initiation of tendon healing phase, and M2 were gradually increased from postoperative day 4. The number of M1 was significantly high both in the LP- and LR-PRP groups (day 4 and 7, p < 0.001), but the number of M2 was high only in the LP-PRP group (day 7 and 14, P < 0.05) when it compared with control group. The M1/M2 ratio on postoperative day 7 was significantly lower in the LP-PRP group than those in the control group (P < 0.05). CONCLUSIONS: This study demonstrated that PRP enhanced the tendon healing and promoted the recruitment of MPs to the injured tissue. The subtypes of MPs were different depends on the types of PRPs, suggesting that leukocytes in PRP influence the effect of PRP therapy.

Intra-articular Injection of Pure Platelet-Rich Plasma Is the Most Effective Treatment for Joint Pain by Modulating Synovial Inflammation and Calcitonin Gene-Related Peptide Expression in a Rat Arthritis Model.

BACKGROUND: Platelet-rich plasma (PRP) has emerged as a treatment for osteoarthritis (OA). However, the effect that leukocyte concentrations in PRP have on OA remains unclear. PURPOSE: To clarify the optimal PRP formulation for OA treatment by comparing pure PRP, leukocyte-poor PRP (LP-PRP), and leukocyte-rich PRP (LR-PRP) in a rat arthritis model. STUDY DESIGN: Controlled laboratory study. METHODS: Knee arthritis was induced bilaterally in male Wistar rats with intra-articular injections of monosodium iodoacetate (MIA) on day 0. Rats were randomly assigned to 1 of 3 treatment groups (pure PRP, LP-PRP, and LR-PRP). On day 1, allogenic PRP was injected into the right knee of rats and phosphate-buffered saline was injected into the left knee as a control. Weight distribution on the hindlimbs was measured for 14 days to assess pain behavior. Rats were euthanized at day 5 or 14 for histological assessment of synovial tissue and cartilage. Immunohistochemical staining of calcitonin gene-related peptide (CGRP) and α-smooth muscle actin was performed to determine the mechanism of pain relief induced by the PRP preparations. RESULTS: In all groups, PRP increased the load-sharing ratio on PRP-injected knees, with pure PRP eliciting the largest effect among the 3 kinds of PRP (P < .05). Structural changes in the synovial tissue were significantly inhibited in the pure-PRP group compared with the control group after both 5 and 14 days (P < .001 and P = .025, respectively), whereas no significant difference was found between the control, LP-PRP, and LR-PRP groups. An inhibitory effect on cartilage degeneration was observed only in the pure-PRP group on day 14. Pure PRP also significantly inhibited expression of CGRP-positive nerve fibers in the infrapatellar fat pad compared with the other groups (P < .05). CONCLUSION: In an MIA-induced arthritis model, pure PRP injection was the most effective treatment for reduction of pain-related behavior and inhibition of synovial inflammation and pain sensitization. CLINICAL RELEVANCE: PRP formulations should be optimized for each specific disease. This study shows the superiority of pure PRP for treatment of arthritis and joint pain.