Dose-Dependent Effects of Platelet-Rich Plasma Powder on Chondrocytes In Vitro.

BACKGROUND: Platelet-rich plasma (PRP) is widely used in sports medicine. However, neither preparation nor parameters for clinical application, such as concentration, timing, and number of applications, are standardized, making research and clinical utilization challenging. PURPOSE: To investigate the effect of varying doses of PRP powder in terms of different concentrations, timing, and number of applications on human chondrocytes in a reproducible cell culture model. STUDY DESIGN: Controlled laboratory study. METHODS: A standardized lyophilized platelet growth factor preparation (PRP powder) was used to stimulate human chondrocytes. Chondrocytes were cultivated for 2 weeks with different stimulation frequencies (2×, 3×, 6×) and different concentrations of PRP powders (0.5%, 1%, 5%). Cell proliferation and metabolic cell activity were analyzed on days 7 and 14. Phenotypic changes were visualized through live-dead staining. Chondrogenic differentiation was quantified with enzyme-linked immunosorbent assay to assess the synthesis of procollagen types 1 and 2. Furthermore, sulfated proteoglycans and glycosaminoglycans were analyzed. RESULTS: Human chondrocytes exhibited a significant dose- and time-dependent increase after 14 days in cell number (1% and 5% PRP powder vs unstimulated control: 7.95- and 15.45-fold increase, respectively; 2× vs 6× stimulation with 5% PRP powder: 4.00-fold increase) and metabolic cell activity (1% and 5% PRP powder vs unstimulated control: 3.27-fold and 3.58-fold change, respectively). Furthermore, cells revealed a significant increase in the amount of bone-specific procollagen type 1 (14 days, 1.94-fold) and sulfated glycosaminoglycans (14 days, 2.69-fold); however, no significant change was observed in the amount of cartilage-specific collagen type 2. CONCLUSION: We showed that chondrocytes exhibit a significant dose- and time-dependent increase in cell number and metabolic cell activity. The standardized use of growth factor concentrates in cell culture models can contribute to clinical knowledge in terms of dosage and timing of PRP applications. CLINICAL RELEVANCE: Problems with PRP, such as the absence of standardization, lack of consistency among studies, and unknown dosage, could be solved by using characterized PRP powder made by pooling and lyophilizing multiple platelet concentrates. The innovative PRP powder generates new possibilities for PRP research, as well as for the treatment of patients.

Medial Patellofemoral Ligament Reconstruction: Impact of Knee Flexion Angle During Graft Fixation on Dynamic Patellofemoral Contact Pressure-A Biomechanical Study.

PURPOSE: Objective evaluation of the optimal graft tension angle to fully restore patellofemoral contact pressure in reconstruction of the medial patellofemoral ligament (MPFL) in comparison to the native knee. METHODS: Twelve cadaveric knee specimens were fixed in a custom-made fixation device. A sensitive pressure film (Tekscan) was fixed in the patellofemoral joint, and patellofemoral contact pressure was assessed during a dynamic flexion movement from 0° to 90°. The MPFL was cut and measurements were repeated. Reconstruction of the MPFL was performed with the gracilis tendon subsequently fixed in the femur at 15°, 30°, 45°, 60°, 75°, and 90° of knee flexion under controlled tension (2 N). The sequence of the flexion angles was alternated. Pressure measurements were repeated after every fixation of the graft. RESULTS: No significant differences were seen in the overall patellofemoral contact pressure compared to the native knee (P > .05). However, medial patellofemoral pressure showed a significant increased patellofemoral contact pressure after MPFL reconstruction at a knee flexion angle during graft fixation of 15° (P = .027), 45° (P = .050, P = .044), and 75° (P = .039). Moreover, proximal/distal patellofemoral contact pressure revealed a significantly reduced contact pressure at 15° (P = .003), 30° (P = .009), 45° (P = .025), 75° (P = .021), and 90° (P = .022) of flexion distal after MPFL reconstruction compared with the intact knee. Lateral patellofemoral contact pressure was significantly reduced in all performed reconstructions (P < .05). CONCLUSIONS: The flexion angle during graft fixation for MPFL reconstruction did not have a significant impact on the overall patellofemoral contact pressure. However, selective medial, proximal, distal, and lateral patellofemoral contact pressure was significantly altered for all reconstructions. Fixation of the MPFL graft at 60° of flexion was able to most closely restore patellofemoral contact pressure compared with the intact knee. CLINICAL RELEVANCE: Based on the findings of the present study, fixation of the graft in anatomic reconstruction of the MPFL should be considered in 60° of flexion under low tension (2 N) to most closely restore patellofemoral contact pressure compared with the native knee.

Biomechanical evaluation of MPFL reconstructions: differences in dynamic contact pressure between gracilis and fascia lata graft.

PURPOSE: To evaluate the knee kinematics of the intact, MPFL-ruptured and MPFL-reconstructed knee and, moreover, to compare dynamic patellofemoral contact pressure of the gracilis tendon and the fascia lata as an alternative graft option for reconstruction of the MPFL. METHODS: Eight paired human cadaveric knees were fixed in a custom-made fixation device. Patellofemoral contact pressure was assessed during a dynamic flexion movement at 15°-30°-45°-60°-75° and 90° using a pressure-sensitive film (Tekscan). The medial patellofemoral ligament was cut, and measurements were repeated. Finally, reconstruction of the MPFL was performed using the gracilis tendon (group I) or a fascia lata graft (group II). Tunnel localization was performed under fluoroscopic control. Grafts were fixed at 30° of flexion, and pressure measurements were repeated. RESULTS: Incision of the medial patellofemoral ligament significantly reduced patellofemoral contact pressure at 15°, 30° and 45° of knee flexion compared to the intact knee (p < 0.05), whereas reconstruction of the MPFL using either gracilis tendon of the fascia lata was able to restore pressure distributions at 15° and 30° of knee flexion. However, in the hamstring group, reconstruction of the MPFL revealed a significantly reduced contact pressure at 45° of flexion (p = 0.038) compared to the intact knee. In the fascia lata group, a significant reduction in patellofemoral contact pressure was observed after MPFL reconstruction at 45°, 60°, 75° and 90° of knee flexion (p < 0.05). CONCLUSIONS: Anatomic reconstruction of the MPFL with either a gracilis or a fascia lata graft showed comparable patellofemoral pressure distributions which were closely restored compared to the native knee. Therefore, the fascia lata has shown to be a viable alternative to the gracilis tendon for reconstruction of the MPFL. However, anatomic reconstruction of the MPFL may lead to persistently altered patellofemoral contact pressure during knee flexion compared to the native knee independent of the tested graft.

Platelet-Rich Plasma Powder: A New Preparation Method for the Standardization of Growth Factor Concentrations.

BACKGROUND: Platelet-rich plasma (PRP) is widely used in sports medicine. Available PRP preparations differ in white blood cell, platelet, and growth factor concentrations, making standardized research and clinical application challenging. PURPOSE: To characterize a newly standardized procedure for pooled PRP that provides defined growth factor concentrations. STUDY DESIGN: Controlled laboratory study. METHODS: A standardized growth factor preparation (lyophilized PRP powder) was prepared using 12 pooled platelet concentrates (PCs) derived from different donors via apheresis. Blood samples and commercially available PRP (SmartPrep-2) served as controls (n = 5). Baseline blood counts were analyzed. Additionally, single PCs (n = 5) were produced by standard platelet apheresis. The concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor AB (PDGF-AB), transforming growth factor ß1 (TGF-ß1), insulin-like growth factor 1 (IGF-1), interleukin (IL)-1α, IL-1ß, and IL-1 receptor agonist (IL-1RA) were analyzed by enzyme-linked immunosorbent assay, and statistical analyses were performed using descriptive statistics, mean differences, 95% CIs, and P values (analysis of variance). RESULTS: All growth factor preparation methods showed elevated concentrations of the growth factors VEGF, bFGF, PDGF-AB, and TGF-ß1 compared with those of whole blood. Large interindividual differences were found in VEGF and bFGF concentrations. Respective values (mean ± SD in pg/mL) for whole blood, SmartPrep-2, PC, and PRP powder were as follows: VEGF (574 ± 147, 528 ± 233, 1087 ± 535, and 1722), bFGF (198 ± 164, 410 ± 259, 151 ± 99, and 542), PDGF-AB (2394 ± 451, 17,846 ± 3087, 18,461 ± 4455, and 23,023), and TGF-ß1 (14,356 ± 4527, 77,533 ± 13,918, 68,582 ± 7388, and 87,495). IGF-1 was found in SmartPrep-2 (1539 ± 348 pg/mL). For PC (2266 ± 485 pg/mL), IGF-1 was measured at the same levels of whole blood (2317 ± 711 pg/mL) but was not detectable in PRP powder. IL-1α was detectable in whole blood (111 ± 35 pg/mL) and SmartPrep-2 (119 ± 44 pg/mL). CONCLUSION: Problems with PRP such as absent standardization, lack of consistency among studies, and black box dosage could be solved by using characterized PRP powder made by pooling and lyophilizing multiple PCs. The new PRP powder opens up new possibilities for PRP research as well as for the treatment of patients. CLINICAL RELEVANCE: The preparation of pooled PRP by means of lyophilization may allow physicians to apply a defined amount of growth factors by using a defined amount of PRP powder. Moreover, PRP powder as a dry substance with no need for centrifugation could become ubiquitously available, thus saving time and staff resources in clinical practice. However, before transferring the results of this basic science study to clinical application, regulatory issues have to be cleared.

Reconstruction of 25 and 50 % subscapularis tears: a single anchor with a double-mattress suture is sufficient for the reconstruction.

PURPOSE: Evaluation of the biomechanical performance of repairs of 25 % (Fox/Romeo II) and 50 % (Fox/Romeo III) full-thickness subscapularis tears using a single-suture anchor. METHODS: Six pairs of human cadaver specimens were used for the testing. Artificial subscapularis tears were created in order to simulate a 25 % (6) and a 50 % (6) full-thickness tear. The reconstructions were made with a double-loaded suture anchor (5.5-mm Bio-Corkscrew with two No. 2 Fiberwire) creating a double-mattress suture repair. Reconstructions were cyclically loaded from 10 to 60 N. The load was increased stepwise up to 100 and 180 N. Cyclic displacement (means + standard dev.) as well as load-to-failure was determined, and mode of failure was recorded. RESULTS: In the reconstructed shoulders at 60 N, a mean cyclic displacement of 3.2 ± 0.7 mm was found in the 25 % tear, 2.6 ± 0.6 mm in the 50 % tear. At 100 N, 5.1 ± 1.2 mm was seen in the 25 % tear and 4.3 ± 0.3 mm in the 50 % tear. At highest load of 180 N, 7.6 ± 2.2 mm was recorded in the 25 % tear, 6.5 ± 0.8 mm was found in the 50 % tear. Ultimate failure load was 486 ± 167 N in the 25 % tear and 455 ± 213 N in the 50 % tear. Statistically significant differences between the tested repairs were seen neither in cyclic displacement nor in ultimate failure loads (p > 0.05). Mode of failure revealed bone fractures and anchor pull-out as major cause in the 25 % group, whereas failure of the suture-tendon interface was the major cause of failure in the 50 % group. CONCLUSIONS: Subscapularis repair using a single double-loaded suture anchor revealed similar biomechanical performance in 25 % compared to 50 % full-thickness subscapularis tears. With increased tear size, however, an optimized suture-tendon interface seems to become more relevant in order to decrease failure rate of the repair. CLINICAL RELEVANCE: A single double-loaded suture anchor provides sufficient biomechanical strength even in Fox/Romeo grade III tears of the subscapularis tendon. However, a modified suture configuration is recommended, especially in grade III tears as the suture-tendon interface is the weakest point of the construct.

The influence of the medial meniscus in different conditions on anterior tibial translation in the anterior cruciate deficient knee.

PURPOSE: The purpose of this study was the evaluation of knee laxity in the ACL-deficient knee with combined meniscal tear, meniscal suture and partial medial meniscectomy. METHODS: Kinematics of the intact knee were determined in 18 human cadaver specimens in response to a 134-N anterior tibial load (aTT) as well as a combined rotatory load of 10 Nm valgus and 4 Nm internal tibial rotation using a robotic/universal force moment sensor testing system. The anterior cruciate ligament was resected. Subsequently, a vertical bucket-handle medial meniscal tear was created followed by a standard meniscus repair using horizontal inside-out stitches or a partial medial meniscectomy. Knee kinematics were calculated following every sub-step. RESULTS: A significant increase of anterior tibial translation was found in the ACL-deficient knee compared to the intact knee at 30° and 90° of flexion (p = 0.001; p ≤ 0.001). Additional tear of the medial meniscus significantly increased anterior tibial translation (p = 0.01). In response to a simulated pivot shift, anterior tibial translation of the intact knee did not increase significantly after ACL resection (p = 0.067). However, ACL deficiency with an additional medial meniscus tear led to a significant increase compared to the intact knee at 0° of flexion (p = 0.009). CONCLUSIONS: Additional injury of the medial meniscus increased aTT as well as aTT under a combined rotatory load in the ACL-deficient knee whereas repair of the meniscus significantly decreased aTT. Therefore, the meniscus status does have a significant impact on knee kinematics in the ACL-deficient knee. The present biomechanical study further highlights the importance of preserving the meniscus especially in patients with additional ACL injuries.

Biomechanical evaluation of knee kinematics after anatomic single- and anatomic double-bundle ACL reconstructions with medial meniscal repair.

PURPOSE: To evaluate knee laxity after anatomic ACL reconstruction with additional suture repair of a medial meniscus tear. METHODS: Kinematics of the intact knee were determined in 12 human cadaver specimens in response to a 134-N anterior tibial load (aTT) and a combined rotatory load of 10 Nm valgus and 4 Nm internal tibial rotation (aTTPS) using a robotic/universal force moment sensor testing system. Subsequently, the ACL was resected following the creation of a standardized tear of the medial meniscus, a standard meniscus repair and an ACL reconstruction using an anatomic single-bundle (6) or an anatomic double-bundle technique (6). Knee kinematics were determined following every sub-step. RESULTS: Significant increase of aTT in the ACL-deficient knee was found (p ≤ 0.001) with a further increase in the ACL-deficient knee with additional medial meniscal rupture (p ≤ 0.001). ACL reconstructions significantly decreased aTT compared with the ACL and meniscus-ruptured knee. No significant differences were seen between the intact knee and the ACL-reconstructed knee with additional meniscal repair (p < 0.05). In response to a simulated pivot shift, aTTPS in the intact knee significantly increased in the ACL-deficient knee and meniscus-ruptured knee (p = 0.005). No significant differences in knee kinematics were found between SB as well as DB ACL reconstruction with additional medial meniscal repair compared with the intact knee. Comparison of SB versus DB ACL reconstruction did not reveal any significant differences in a simulated Lachman test or simulated pivot shift test (n.s.). CONCLUSIONS: aTT as well as aTTPS significantly increased with ACL deficiency compared with the intact knee; additional medial meniscal rupture further increased aTT. Anatomic ACL reconstruction with medial meniscal repair did not reveal significant differences in knee kinematics compared with the intact knee. Comparison of anatomic SB versus DB ACL reconstruction with additional repair of the medial meniscus did not show significant differences neither in a simulated Lachman nor in a simulated pivot shift test.

Three-dimensional evaluation of cyclic displacement in single-row and double-row rotator cuff reconstructions under static external rotation.

BACKGROUND: The double-row suture bridge repair was recently introduced and has demonstrated superior biomechanical results and higher yield load compared with the traditional double-row technique. It therefore seemed reasonable to compare this second generation of double-row constructs to the modified single-row double mattress reconstruction. HYPOTHESIS: The repair technique, initial tear size, and tendon subregion will have a significant effect on 3-dimensional (3D) cyclic displacement under additional static external rotation of a modified single-row compared with a double-row rotator cuff repair. STUDY DESIGN: Controlled laboratory study. METHODS: Rotator cuff tears (small to medium: 25 mm; medium to large: 35 mm) were created in 24 human cadaveric shoulders. Rotator cuff repairs were performed as modified single-row or double-row repairs, and cyclic loading (10-60 N, 10-100 N) was applied under 20° of external rotation. Radiostereometric analysis was used to calculate cyclic displacement in the anteroposterior (x), craniocaudal (y), and mediolateral (z) planes with a focus on the repair constructs and the initial tear size. Moreover, differences in cyclic displacement of the anterior compared with the posterior tendon subregions were calculated. RESULTS: Significantly lower cyclic displacement was seen in small to medium tears for the single-row compared with double-row repair at 60 and 100 N in the x plane (P = .001) and y plane (P = .001). The results were similar in medium to large tears at 100 N in the x plane (P = .004). Comparison of 25-mm versus 35-mm tears did not show any statistically significant differences for the single-row repairs. In the double-row repairs, lower gap formation was found for the 35-mm tears (P ≤ .05). Comparison of the anterior versus posterior tendon subregions revealed a trend toward higher anterior gap formation, although this was statistically not significant. CONCLUSION: The tested single-row reconstruction achieved superior results in 3D cyclic displacement to the tested double-row repair. Extension of the initial rupture size did not have a negative effect on the biomechanical results of the tested constructs. CLINICAL RELEVANCE: Single-row repairs with modified suture configurations provide comparable biomechanical strength to double-row repairs. Furthermore, as increased gap formation in the early postoperative period might lead to failure of the construct, a strong anterior fixation and restricted external rotation protocol might be considered in rotator cuff repairs to avoid this problem.

Objective measurement devices to assess static rotational knee laxity: focus on the Rotameter.

PURPOSE: The present article summarizes the development of a simple, objective, and non-invasive measurement device for tibiofemoral rotation to assess static rotational knee laxity. METHODS AND RESULTS: The device is based on the dial test with the patient lying prone and the knee flexed to 30°. From measurements of 30 healthy participants, the device achieved high inter- and intra-observer reliability and showed a high correlation of the measured results with the contralateral knees of the participants. Measurements of the device were also performed in a human cadaver study and revealed highly correlated results when compared to the simultaneous measurements of a knee navigation system, which was used as an invasive standard method to assess tibial rotation. In human cadaver specimens, it was shown that a simulated tear of the posterolateral bundle as well as a complete ACL tear led to a significant increase in isolated tibiofemoral rotation compared to the intact ACL. A retrospective case series investigated the clinical results as well as knee laxity measurements after ACL surgery in vivo. Rotational, as well as anteroposterior (AP), knee laxity was objectively assessed in 52 patients at a mean postoperative follow-up of 27 months by comparing the measured results with the results of the contralateral unaffected knee in each patient. The clinical results were comparable to the results reported in the literature. Moreover, rotational laxity was successfully restored after ACL reconstruction, whereas AP laxity showed significant differences compared to the contralateral knees although they were defined as clinically successful according to the IKDC classification. CONCLUSIONS: A non-invasive and objective knee rotational measurement device has been developed, which offers good potential for objective quality control in knee ligament injuries and their treatment. LEVEL OF EVIDENCE: Review article, Level IV.

Comparable biomechanical results for a modified single-row rotator cuff reconstruction using triple-loaded suture anchors versus a suture-bridging double-row repair.

PURPOSE: To compare the biomechanical properties and footprint coverage of a single-row (SR) repair using a modified suture configuration versus a double-row (DR) suture-bridge repair in small to medium and medium to large rotator cuff tears. METHODS: We created 25- and 35-mm artificial defects in the rotator cuff of 24 human cadaveric shoulders. The reconstructions were performed as either an SR repair with triple-loaded suture anchors (2 to 3 anchors) and a modified suture configuration or a modified suture-bridge DR repair (4 to 6 anchors). Reconstructions were cyclically loaded from 10 to 60 N. The load was increased stepwise up to 100, 180, and 250 N. Cyclic displacement and load to failure were determined. Furthermore, footprint widths were quantified. RESULTS: In the 25-mm rupture, ultimate load to failure was 533 ± 107 N for the SR repair and 681 ± 250 N for the DR technique (P ≥ .21). In the 35-mm tear, ultimate load to failure was 792 ± 122 N for the SR reconstruction and 891 ± 174 N for the DR reconstruction (P ≥ .28). There were no statistically significant differences for both tested rupture sizes. Cyclic displacement showed no significant differences between the tested configurations at 60 N (P = .563), 100 N (P = .171), 180 N (P = .211), and 250 N (P = .478) for the 25-mm tear. For the 35-mm tear, cyclic displacement showed significantly lower gap formation for the SR reconstruction at 180 N (P = .037) and 250 N (P = .020). No significant differences were found at 60 N (P = .296) and 100 N (P = .077). A significantly greater footprint width (P = .028) was seen for the DR repair (16.2 mm) compared with the SR repair (13.8 mm). However, both reconstructions were able to achieve complete footprint coverage compared with the initial footprint. CONCLUSIONS: The tested SR repair using a modified suture configuration was similar in load to failure and cyclic displacement to the DR suture-bridge technique independent of the tested initial sizes of the rupture. The tested DR repair consistently restored a larger footprint than the SR method. However, both constructs achieved complete footprint coverage. CLINICAL RELEVANCE: SR repairs with modified suture configurations might combine the biomechanical advantages and increased footprint coverage that are described for DR repairs without increasing the overall costs of the reconstruction.

Good results after fluoroscopic-guided intra-articular injections in the treatment of adhesive capsulitis of the shoulder.

The purpose of the present study was the prospective evaluation of the results of fluoroscopic-guided intra-articular cortisone injection series in the treatment of adhesive capsulitis of the shoulder. Twenty-five patients (9 m, 16 w) with a mean age of 49 ± 8 years and stage II frozen shoulder syndrome according to the Reeves classification were treated with an intra-articular cortisone injection series (3 injections at 0, 4, 12, weeks). Clinical examination, ASES score and SF 36 score were performed at 0, 4, 8, 12 weeks, 6 and 12 months. In the results, significant improvements were seen in flexion (99 ± 30°-119 ± 31°, P < .0001), abduction (72 ± 24-99 ± 34°, P < .0001), external rotation (14 ± 16°-28 ± 21°, P < .0001) and internal rotation already at first follow-up after 4 weeks of treatment. The results were confirmed at any other follow-up. ASES score improved from 28 ± 13 to 45 ± 18 after 4 weeks (P < .0001), 59 ± 21 after 8 weeks (P < .0001), 63 ± 25 at 3 months (P < .0001), 64 ± 28 (P < .0001) at 6 months and 73 ± 27 (P < .0001) points at final follow-up after 1 year. Evaluation of the SF-36 Score showed significant improvements in almost all categories (physical and mental) after 4 weeks of treatment (P < .05). In conclusion, a fluoroscopic-guided intra-articular injection series of cortisone is an effective treatment option in frozen shoulder syndrome leading to a fast pain reduction and increased range motion.