Editorial Commentary: Elbow Injury Results When Pediatric and Adolescent Throwing Athletes Throw as Hard as Possible, and Weighted Baseball Training Should Be Banned for Youth Athletes.

We are in the middle of an epidemic involving pediatric and adolescent throwing athletes. Too many young athletes have elbow injuries, and it is unnerving how young surgical indications can present. Increased competition has led to increased demands before athletes have reached their full adult size. Evidence shows that higher ball velocity, higher shoulder external rotation angle, and higher arm speed are associated with increased medial elbow torque and elbow injury in this population. Pediatric and adolescent athletes should not try to throw as hard as possible, and weighted baseball training should be banned for youth athletes.

The safety and efficacy of 2 anterior-inferior portals for arthroscopic repair of anterior humeral avulsion of the glenohumeral ligament: cadaveric comparison.

BACKGROUND: Humeral avulsion of the glenohumeral ligament (HAGL) lesions are associated with shoulder instability. Arthroscopic repair of anterior HAGL lesions typically requires the placement of an anterior-inferior (5-o'clock) portal, with different variations of this portal described. The purpose of this study was to determine the efficacy of described anterior-inferior shoulder arthroscopy portals for arthroscopic anterior HAGL repair, as well as evaluate the safety of these portals with respect to the surrounding neurovascular structures. Additionally, we sought to evaluate the effect of arm adduction vs. standard abduction during anterior-inferior portal creation. METHODS: HAGL lesions were created and repaired using an all-arthroscopic technique in 12 cadaveric shoulders (matched pairs). Half of the repairs were performed using a standard 5-o'clock portal, whereas the other half of the matched pairs were repaired using a medialized 5-o'clock portal. Repairs were timed, and the number of anchor pullouts was recorded. The shoulders were subsequently dissected to measure the proximity of the portal to the cephalic vein, musculocutaneous nerve, axillary nerve, and lateral cord of the brachial plexus. RESULTS: The average time for HAGL repair was 18.0 ± 4.6 minutes. Repair times using the medial 5-o'clock portal (19.0 ± 3.3 minutes) vs. standard 5-o'clock portal (16.2 ± 5.8 minutes) were not significantly different (P = .37). From abduction to adduction, the cephalic vein distance from the standard 5-o'clock portal increased from 4.1 ± 4.7 mm to 5.2 ± 5.4 mm (P = .02); musculocutaneous nerve distance, from 14.4 ± 9.8 mm to 18.1 ± 10.8 mm (P = .005); axillary nerve distance, from 19.2 ± 9.6 mm to 19.8 ± 9.2 mm (P = .12); and distance of the lateral cord of the brachial plexus, 13.8 ± 6.6 mm to 16.7 ± 6.4 mm (P = .0006). CONCLUSIONS: The arm abduction angle significantly affects the distance of the cephalic vein, musculocutaneous nerve, and lateral cord of the brachial plexus from the anterior-inferior portal, regardless of which portal-standard or medial 5-o'clock portal-is chosen. This portal should be created with the arm in adduction. Arthroscopic HAGL repair can be performed safely, although accurate anchor placement remains a challenge. There was no advantage to use of the medial 5-o'clock portal. With a curved guide, the standard 5-o'clock portal allows for reproducible anchor placement and is recommended for anterior HAGL repairs.

Autologous thrombin preparations: Biocompatibility and growth factor release.

Platelet-rich plasma (PRP) has been investigated to promote wound healing in a variety of tissues. Thrombin, another essential component of wound healing, is sometimes combined with PRP to generate a fibrin clot in order to retain the sample at the delivery site and to stimulate growth factor release. Using a fully autologous approach, autologous serum (AS) with thrombin activity can be prepared using a one-step procedure by supplementing with ethanol (E+ AS) to prolong room temperature stability or prepared ethanol free (E- AS) by utilizing a two-step procedure to prolong stability. The objective of this study was to evaluate potential wound healing mechanisms of these two preparations using commercially available devices. A variety of tests were conducted to assess biocompatibility and growth factor release from PRP at various ratios. It was found that E- AS contained greater leukocyte viability in the product (97.1 ± 2.0% compared to 41.8 ± 11.5%), supported greater bone marrow mesenchymal stem cell proliferation (3.7× vs 0.8× at a 1:4 ratio and 3.6× vs 1.6× at a 1:10 ratio), and stimulated release of growth factors and cytokines from PRP to a greater extent than E+ AS. Of the 36 growth factors and cytokines evaluated, release of 27 of them were significantly reduced by the presence of ethanol in at least one of the tested configurations. It is concluded that the high concentrations of ethanol needed to stabilize point of care autologous thrombin preparations could be detrimental to normal wound healing processes.

Platelet-Rich Plasma: Fundamentals and Clinical Applications.

Platelet-rich plasma (PRP) is perhaps the most widely studied of the biologic therapies, with an ever-growing body of evidence supporting its safety and efficacy in decreasing inflammation and pain and promoting healing in the setting of both nonoperative and operative treatments. PRP is produced by the centrifugation of whole blood, isolating its constituent parts based on their unique densities. These density gradients can be selectively harvested so as to obtain different concentrations of various blood product components, such as platelets and leukocytes. A precise and consistent method for describing the essential characteristics of different PRP formulations is critical for both practical and research purposes. The concentration of platelets, method of activation, and the total number of red blood cells (RBCs), white blood cells (WBCs), and neutrophils relative to baseline values are all of particular importance in accurately describing a PRP formulation. The biologic activity of PRP is manifold: platelet α granules promote the release of various growth factors, including vascular endothelial growth factor and tissue growth factor ß, while inflammation is modulated through inhibition of the nuclear factor-κB pathway. PRP has been convincingly shown to be efficacious in the setting of patellar tendinopathies, knee osteoarthritis, and lateral epicondylitis. In fact, several recent randomized controlled trials have demonstrated the superiority of PRP over both corticosteroids and hyaluronic acid in treating knee OA-related symptoms. There is also substantial promise for the utility of PRP in treating partial hamstring tears and as an adjunct to rotator cuff (RC) repair, especially in the setting of small- to medium-sized tears, where it appears to exert substantial analgesic effects and promote enhanced rates of RC repair healing.

Platelet-Rich Plasma Devices Can Be Used to Isolate Stem Cells From Synovial Fluid at the Point of Care.

PURPOSE: To assess whether point-of-care devices designed for collecting cellular components from blood or bone marrow could be used to isolate viable stem cells from synovial fluid. METHODS: Male and female patients older than 18 years old with either an acute, anterior cruciate ligament (ACL) injury or knee osteoarthritis (OA) with a minimum estimated 20 mL of knee effusion volunteered. Ten patients with an ACL injury and 10 patients with OA were enrolled. Two milliliters of collected synovial effusion were analyzed and cultured for cellular content. The remaining fluid was combined with whole blood and processed using a buffy-coat based platelet-rich plasma (PRP) processing system. Specimens were analyzed for cell counts, colony-forming unit (CFU) assays, differentiation assays, and flow cytometry. RESULTS: ACL effusion fluid contained 42.1 ± 20.7 CFU/mL and OA effusion fluid contained 65.4 ± 42.1 CFU/mL. After PRP processing, the counts in ACL-PRP were 101.6 ± 66.1 CFU/mL and 114.8 ± 73.4 CFU/mL in the OA-PRP. Cells showed tri-lineage differentiation potential when cultured under appropriate parameters. When analyzed with flow cytometry, >95% of cells produced with culturing expressed cell surface markers typically expressed by known stem cell populations, specifically CD45-, CD73+, CD29+, CD44+, CD105+, and CD90+. CONCLUSIONS: Multipotent viable stem cells can be harvested from knee synovial fluid, associated with an ACL injury or OA, and concentrated with a buffy coat-based PRP-processing device. CLINICAL RELEVANCE: PRP devices can be used to harvest stem cells from effusion fluids. Methods to use effusion fluid associated with an ACL injury and OA should be investigated further.

Arthroscopic Subchondral Drilling Followed by Injection of Peripheral Blood Stem Cells and Hyaluronic Acid Showed Improved Outcome Compared to Hyaluronic Acid and Physiotherapy for Massive Knee Chondral Defects: A Randomized Controlled Trial.

PURPOSE: The purpose of this study was to evaluate the safety and efficacy of intra-articular injections of autologous peripheral blood stem cells (PBSCs) plus hyaluronic acid (HA) after arthroscopic subchondral drilling into massive chondral defects of the knee joint and to determine whether PBSC therapy can improve functional outcome and reduce pain of the knee joint better than HA plus physiotherapy. METHODS: This is a dual-center randomized controlled trial (RCT). Sixty-nine patients aged 18 to 55 years with International Cartilage Repair Society grade 3 and 4 chondral lesions (size ≥3 cm2) of the knee joint were randomized equally into (1) a control group receiving intra-articular injections of HA plus physiotherapy and (2) an intervention group receiving arthroscopic subchondral drilling into chondral defects and postoperative intra-articular injections of PBSCs plus HA. The coprimary efficacy endpoints were subjective International Knee Documentation Committee (IKDC) and Knee Injury and Osteoarthritis Outcome Score (KOOS)-pain subdomain measured at month 24. The secondary efficacy endpoints included all other KOOS subdomains, Numeric Rating Scale (NRS), and Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) scores. RESULTS: At 24 months, the mean IKDC scores for the control and intervention groups were 48.1 and 65.6, respectively (P < .0001). The mean for KOOS-pain subdomain scores were 59.0 (control) and 86.0 (intervention) with P < .0001. All other KOOS subdomain, NRS, and MOCART scores were statistically significant (P < .0001) at month 24. Moreover, for the intervention group, 70.8% of patients had IKDC and KOOS-pain subdomain scores exceeding the minimal clinically important difference values, indicating clinical significance. There were no notable adverse events that were unexpected and related to the study drug or procedures. CONCLUSIONS: Arthroscopic marrow stimulation with subchondral drilling into massive chondral defects of the knee joint followed by postoperative intra-articular injections of autologous PBSCs plus HA is safe and showed a significant improvement of clinical and radiologic scores compared with HA plus physiotherapy. LEVEL OF EVIDENCE: Level I, RCT.

Mobilized Peripheral Blood Stem Cells are Pluripotent and Can Be Safely Harvested and Stored for Cartilage Repair.

PURPOSE: The primary objective of this study was to reproduce and validate the harvest, processing and storage of peripheral blood stem cells for a subsequent cartilage repair trial, evaluating safety, reliability, and potential to produce viable, sterile stem cells. METHODS: Ten healthy subjects (aged 19-44 years) received 3 consecutive daily doses of filgrastim followed by an apheresis harvest of mononuclear cells on a fourth day. In a clean room, the apheresis product was prepared for cryopreservation and processed into 4 mL aliquots. Sterility and qualification testing were performed pre-processing and post-processing at multiple time points out to 2 years. Eight samples were shipped internationally to validate cell transport potential. One sample from all participants was cultured to test proliferative potential with colony forming unit (CFU) assay. Five samples, from 5 participants were tested for differentiation potential, including chondrogenic, adipogenic, osteogenic, endoderm, and ectoderm assays. RESULTS: Fresh aliquots contained an average of 532.9 ± 166. × 106 total viable cells/4 mL vial and 2.1 ± 1.0 × 106 CD34+ cells/4 mL vial. After processing for cryopreservation, the average cell count decreased to 331.3 ± 79. × 106 total viable cells /4 mL vial and 1.5 ± 0.7 × 106 CD34+ cells/4 mL vial CD34+ cells. Preprocessing viability averaged 99% and postprocessing 88%. Viability remained constant after cryopreservation at all subsequent time points. All sterility testing was negative. All samples showed proliferative potential, with average CFU count 301.4 ± 63.9. All samples were pluripotent. CONCLUSIONS: Peripheral blood stem cells are pluripotent and can be safely harvested/stored with filgrastim, apheresis, clean-room processing, and cryopreservation. These cells can be stored for 2 years and shipped without loss of viability. CLINICAL RELEVANCE: This method represents an accessible stem cell therapy in development to augment cartilage repair.

Biceps Tendon Changes and Pitching Mechanics in Youth Softball Pitchers.

With the lack of pitch count regulation, youth softball pitchers are experiencing unremitting high stresses on the anterior shoulder. The purpose of this study was to examine the association of acute changes in the long head of the biceps tendon with pitching kinematics and kinetics in youth softball pitchers following an acute bout of pitching. Twenty-three softball pitchers (12.17±1.50 yrs.; 160.32±9.41 cm; 60.40±15.97 kg) participated. To investigate the association between biceps tendon changes and kinematic and kinetic changes from pre- to post-simulated game, each biceps tendon measure was split into those whose biceps tendon thickness, width, and/or area increased pre- to post-simulated game, and those whose did not. There were significant differences in biceps tendon longitudinal thickness (Z=- 2.739, p=0.006) and pitch speed; as well as a difference between groups in biceps tendon transverse thickness and the amount of change in trunk rotation at the start of the pitching motion (p=0.017) and the amount of change in trunk flexion at ball release (p=0.030). This study illustrates the association of trunk and lower extremity kinematics and shoulder kinetics with morphologic changes in the biceps tendon with an acute bout of windmill softball pitching.

Lower Extremity Pain and Pitching Kinematics and Kinetics in Collegiate Softball Pitchers.

The primary aims of the study were (1) to examine kinematics and kinetics of those pitching with and without lower extremity pain in collegiate softball pitchers, and (2) to determine if there was an association between the lower extremity pain and lower extremity kinematics, trunk kinematics, and shoulder kinetics in collegiate softball pitchers. Thirty-seven NCAA Division I female collegiate softball pitchers (19.8±1.3 yrs,173.7±7.7 cm, 79.0±12.4 kg) participated. Participants were divided into two groups, those who were currently experiencing lower extremity pain and those who were not. Participants threw three rise ball pitches. Kinematic data were collected at 100 Hz using an electromagnetic tracking system. Mann-Whitney U tests revealed no significant kinematic or kinetic differences between pitchers with and without lower extremity pain. Additionally, there were no significant correlations between pain and recorded kinematic and kinetic variables. Considering there were no biomechanical differences observed between pitchers, coaches and athletic trainers should take caution with athlete assessment since athletes may not display altered biomechanics. Further examination into the duration and degree of pain is needed in an attempt to fully understand the implication of pain and pitching mechanics.

3-T MRI mapping is a valid in vivo method of quantitatively evaluating the anterior cruciate ligament: rater reliability and comparison across age.

OBJECTIVE: As biologic augmentation methods emerge, objective measures of soft tissues are necessary for developmental study. The purpose of this study was to develop a quantitative MRI mapping protocol for the ACL. The objectives were (1) to provide age-based T2 relaxation, T2* relaxation, and volume values in healthy individuals, (2) to establish the intra-rater and inter-rater reliability of ACL mapping, and (3) to determine whether 3-T or 7-T MRI is more appropriate for future clinical trials. MATERIALS AND METHODS: Thirty healthy participants, aged 18-62, asymptomatic for knee pathology and without history of knee injury underwent both a 3-T and 7-T MRI. Manual image mapping of the anterior cruciate ligament was performed by two observers and processed to obtain T2, T2*, and volume values. Analysis of variance and two-way random effects model were used to calculate statistical significance and intraclass correlation coefficients. RESULTS: Across all participants, 3-T and 7-T mean T2, T2* and volume values were 37.1 ± 7.9 and 39.7 ± 6.2 ms (p = 0.124), 10.9 ± 1.3 and 10.9 ± 0.9 ms (p = 0.981), and 2380 ± 602 and 2484 ± 736 mm3 (p = 0.551), respectively. The T2, T2*, and volume did not vary between age cohorts (p > 0.05). Excellent inter-rater and intra-rater reliability regarding T2 and T2* values was found. While ACL volume exhibited good inter-rater reliability and excellent intra-rater reliability. CONCLUSIONS: T2 relaxation values and ACL volume do not vary with age and therefore can be used as a quantifiable, non-invasive method to assess ACL graft maturation. 7-T MRI analysis was not superior to 3-T MRI analysis, suggesting that 3-T MRI is practical and capable for future comparative studies.

Editorial Commentary: Food and Drug Administration Regulation of Biologics in Orthopaedics: Am I the Only One Around Here Who Gives a Flip About the Rules?!?!

As orthobiologics have appeared and stayed a part of our clinical practices, at times it seems that we (orthopaedic surgeons) have not focused appropriate attention and/or interest on regulation. However, regulation has focused on us, as the Food and Drug Administration (FDA) has made it clear. Safety and efficacy are top priorities of the FDA and should be ours too. The FDA has transmitted their communications to industry and to clinicians; we are responsible for understanding their regulations and the FDA definitions of relevant terms, including "minimal manipulation" and "homologous" use. Finally, FDA "clearance" does not mean safe or efficacious, nor compliant with other federal regulations.

Quantification and Qualification of Stem Cells From Blood After Mobilization With Filgrastim, and Concentration Using a Platelet-Rich Plasma System.

PURPOSE: To determine the cellular composition of a product created with peripheral blood harvested after systemic mobilization with filgrastim and processed with one point-of-care blood concentrating system, i.e., a platelet-rich plasma (PRP) system. The second purpose was to compare mobilized platelet-rich plasma (M-PRP) with a concentrated bone marrow aspirate (cBMA) and a PRP created from the same subjects with the same PRP system. METHODS: Ten healthy volunteer subjects were recruited for collection and analysis of 3 tissue sources: non-treated peripheral blood, bone marrow aspirate, and filgrastim-mobilized peripheral blood, involving 4 doses of weight-based filgrastim. One point-of-care blood and bone marrow concentrating system was used to create 3 products: PRP, cBMA, and M-PRP. Automated hematologic analysis was performed on all products to quantify total red blood cells, white blood cells (WBCs), monocyte, platelet, and hematopoietic progenitor cell (HPC) concentrations. Flow cytometry was used to determine hematopoietic and mesenchymal progenitor cell populations. Lastly, concentrates were cultured and fibroblast colony-forming units (CFU-F) and morphology of adherent cells were evaluated. RESULTS: M-PRP contained a greater concentration of WBC (mean difference = 53.2 k/µL; P < .0001), monocytes (mean difference = 8.3 k/µL; P = .002), and a trend toward a greater concentration of HPC (mean difference = 200.5 /µL; P = .060) when compared with PRP. M-PRP contained a greater concentration of monocytes (mean difference = 5.5 k/µL; P = .017) and a trend toward a greater concentration of platelets (mean difference = 348 k/µL; P = .051) and HPC (mean difference = 193.4 /µL; P = .068) when compared with cBMA. M-PRP had a similar concentration of platelets to PRP (mean difference = 110 k/µL; P = .051) and PRP had a greater concentration than cBMA (mean difference = 458 k/µL; P = .003). cBMA remained the only product capable of producing CFU-Fs (446 ± 247 /mL) as neither the M-PRP nor PRP produced CFU-Fs. M-PRP produced colonies consistent with WBC. CONCLUSIONS: M-PRP, produced with filgrastim mobilized blood and a proprietary PRP system, contained more total WBCs, monocytes, platelets, and HPCs than cBMA and more WBCs, monocytes, and HPCs than PRP. CLINICAL RELEVANCE: Filgrastim mobilized PRP may be an alternative to cBMA for use as a point-of-care product for orthopaedic treatments.

Exercise-Mobilized Platelet-Rich Plasma: Short-Term Exercise Increases Stem Cell and Platelet Concentrations in Platelet-Rich Plasma.

PURPOSE: To evaluate the effects of vigorous short-term exercise on the platelet and other cellular components of 2 point-of-care blood-processing devices: a buffy coat-based platelet-rich plasma (PRP) product and a plasma-based PRP product. METHODS: Twenty healthy subjects (aged 21-45 years) participated in a 20-minute vigorous exercise regimen on an upright stationary bike at 70% to 85% of maximum target heart rate. Pre- and post-exercise blood was processed in either a plasma-based or automated buffy coat-based PRP system. Complete blood counts were used to compare the cellular components in whole blood and the PRP products. RESULTS: Exercise significantly increased the concentrations of platelets by over 20% in whole blood (P < .001) and in both PRP products (P = .002 and P = .018). Both devices performed consistently with pre- and post-exercise blood. Buffy coat-based PRP prepared after exercise was also significantly larger in volume and had a significantly higher concentration of mobilized hematopoietic stem cells (hematopoietic progenitor cells [HPCs], from 1.7/µL to 2.7/µL, P = .043). The concentrations of all white blood cell types were increased, which could be differentially collected in the devices studied. CONCLUSIONS: Exercise can be used to consistently alter the composition of PRP. Twenty minutes of vigorous exercise can increase platelet concentrations in plasma-based and buffy coat-based PRP products and can increase HPC concentrations and volume in buffy coat-based PRP. CLINICAL RELEVANCE: This study shows a nonpharmacologic method to increase platelet and HPC harvests from peripheral blood. This is important because it highlights a method for altering biological therapies with limited comorbidity.

The Influence of an Active Glove Arm in Softball Pitching: A Biomechanical Evaluation.

The purpose of this study was to determine whether glove arm kinematics during a windmill softball pitch impact pelvic and trunk kinematics as well as pitching arm shoulder kinetics. Thirty-Nine college softball pitchers (20.0±1.4 yrs.; 174.7±6.1 cm; 82.0±13.0 kg; 10.7±2.7 yrs. of experience) threw 3 pitches to a catcher while kinematic and kinetic data were collected. Pearson product moment correlations were run, and significant correlations found with glove arm kinematics, occurring before pelvis kinematics, trunk kinematics, and shoulder kinetics, were then put through a linear regression to identify whether there was any potential cause and effect. Results revealed that glove arm elbow flexion during phase 1 significantly predicted normalized shoulder rotation moment during phase 4 (t=2.60, p=0.013). Additionally, glove arm shoulder horizontal abduction during phase 1 significantly predicted normalized shoulder moment in phase 3 (t=- 2.40, p=0.021) and pelvic angular velocity during phase 3 (t=- 3.20, p=0.003). In conclusion, an active glove arm was predictive of a more efficient kinetic chain later in the windmill pitching motion and could possibly play a role in preventing injury by lessening pitching shoulder joint loads.

Biceps Tendon Changes in Youth Softball Pitchers Following an Acute Bout of Pitching.

Because pitch counts do not exist for softball, pitchers may throw between 1200 and 1500 pitches over 3-day tournaments. With this workload, a common symptom among softball pitchers is anterior shoulder pain. The purpose of this study was to examine long-head biceps tendon (LHBT) changes in youth softball pitchers following an acute bout of pitching. Nineteen softball pitchers (11.89±1.2 years; 158.23±9.71 cm; 61.59±14.76 kg) participated. Images of the LHBT were obtained prior to and immediately following a simulated game protocol. Repeated-measures MANOVA was used to determine if there was a difference in LHBT size before and after the simulated game protocol. There were significant increases in both transverse thickness (t 18=-2.76, p=0.013, 95% confidence interval=-0.050 to -0.007) and longitudinal thickness (t 18=-2.64, p=0.016, 95% confidence interval=-0.060 to -0.007) of the LHBT following an acute bout of pitching. Longitudinal and transverse thickness of the biceps tendon significantly increases following an acute bout of softball throwing. These changes may indicate an inflammation response of the biceps tendon and the biceps pulley.

Upper Extremity Pain and Pitching Mechanics in National Collegiate Athletic Association (NCAA) Division I Softball.

The purpose of this study was to evaluate pitching mechanics between female softball pitchers with upper extremity pain and those without upper extremity pain. Specifically, the trunk, shoulder and elbow kinematics and shoulder kinetics during the change-up softball pitch were examined. Fifty-five collegiate softball pitchers participated, divided into those with upper extremity pain (20.0±1.3 yrs.; 174.4±6.9 cm; 82.9±12.4 kg; 11.1±2.6 yrs. of experience; n=23) and those who were pain-free (19.9±1.4 yrs.; 173.8±6.9 cm; 81.4±12.5 kg; 10.0±2.5 yrs. of experience; n=32). Pitching mechanics were obtained via the trakSTAR™ electromagnetic tracking system (Ascension Technologies, Inc., Burlington, VT, USA). Mann-Whitney U tests revealed significant differences in shoulder horizontal abduction at foot contact (p=0.014, U=153, Z=2.450) and trunk lateral flexion at ball release (p=0.012, U=150, Z=-2.515); and between shoulder distraction force at ball release (p=0.034, U=168, Z=-2.124). The pain group illustrated greater shoulder horizontal abduction at foot contact, less trunk lateral flexion towards the throwing side at ball release, and greater shoulder distraction at ball release than the pain-free group. The differences in trunk and shoulder kinematics, and shoulder kinetics between groups allows for insight into further studies examining injury pervasiveness in softball pitching.

Viable Stem Cells Are in the Injury Effusion Fluid and Arthroscopic Byproducts From Knee Cruciate Ligament Surgery: An In Vivo Analysis.

PURPOSE: To examine the number of viable stem cells contained in the postinjury effusion fluid and the waste byproducts of arthroscopic cruciate ligament surgery. METHODS: This study included patients older than 18 years of age with acute (<5 weeks old) cruciate ligament injuries requiring arthroscopic surgery. The postinjury effusion fluid (effusion fluid), fat pad and cruciate ligament stump debridement tissue (byproduct tissue), and arthroscopic fluid collected during fat pad and/or stump debridement (byproduct fluid) were collected at the time of surgery from 30 individuals. Specimens were analyzed, investigating cell viability, nucleated cell counts, cell concentrations, colony-forming unit assays, and flow cytometry. Samples from the first 20 individuals were collected in small specimen containers, and samples from the last 10 individuals were collected in larger specimen containers. RESULTS: Cells of the injury effusion exhibited the greatest viability (86.4 ± 1.31%) when compared with the small volume harvest byproduct tissue (50.2 ± 2.5%, P = .0001), small volume harvest byproduct fluid (48.8 ± 1.88%, P = .0001), large volume harvest byproduct tissue (70.1 ± 5.6%, P = .0001), and large volume harvest byproduct fluid (60.3 ± 3.41%, P = .0001). The culture analysis of fibroblast colony-forming units found on average 1916 ± 281 progenitor cells in the effusion fluid, 2488 ± 778 progenitor cells in the byproduct tissue, and 2357 ± 339 progenitor cells in the byproduct fluid. Flow cytometry confirmed the presence of immature cells and the presence of cells with markers typically expressed by known stem cell populations. CONCLUSIONS: Viable stem cells are mobilized to the postinjury effusion at the time of cruciate ligament injury and can be found in the byproduct waste of cruciate ligament surgery. CLINICAL RELEVANCE: The methodology around effusion fluid and byproduct tissue capture during cruciate ligament surgery should be investigated further. Cell amounts available from these tissues with current technologies are not sufficient for immediate evidence-based clinical application.

Application of biologics in the treatment of the rotator cuff, meniscus, cartilage, and osteoarthritis.

Advances in our knowledge of cell signaling and biology have led to the development of products that may guide the healing/regenerative process. Therapies are emerging that involve growth factors, blood-derived products, marrow-derived products, and stem cells. Animal studies suggest that genetic modification of stem cells will be necessary; studies of cartilage and meniscus regeneration indicate that immature cells are effective and that scaffolds are not always necessary. Current preclinical animal and clinical human data and regulatory requirements are important to understand in light of public interest in these products.

Osteochondral Regeneration in the Knee Joint with Autologous Peripheral Blood Stem Cells plus Hyaluronic Acid after Arthroscopic Subchondral Drilling: Report of Five Cases.

BACKGROUND: Treatment of osteochondral defects (OCDs) of the knee joint remains challenging. The purpose of this study was to evaluate the clinical and radiological results of osteochondral regeneration following intra-articular injections of autologous peripheral blood stem cells (PBSC) plus hyaluronic acid (HA) after arthroscopic subchondral drilling into OCDs of the knee joint. CASE PRESENTATION: Five patients with OCDs of the knee joint are presented. The etiology includes osteochondritis dissecans, traumatic knee injuries, previously failed cartilage repair procedures involving microfractures and OATS (osteochondral allograft transfer systems). PBSC were harvested 1 week after surgery. Patients received intra-articular injections at week 1, 2, 3, 4, and 5 after surgery. Then at 6 months after surgery, intra-articular injections were administered at a weekly interval for 3 consecutive weeks. These 3 weekly injections were repeated at 12, 18 and 24 months after surgery. Each patient received a total of 17 injections. Subjective International Knee Documentation Committee (IKDC) scores and MRI scans were obtained preoperatively and postoperatively at serial visits. At follow-ups of >5 years, the mean preoperative and postoperative IKDC scores were 47.2 and 80.7 respectively (p = 0.005). IKDC scores for all patients exceeded the minimal clinically important difference values of 8.3, indicating clinical significance. Serial MRI scans charted the repair and regeneration of the OCDs with evidence of bone growth filling-in the base of the defects, followed by reformation of the subchondral bone plate and regeneration of the overlying articular cartilage. CONCLUSION: These case studies showed that this treatment is able to repair and regenerate both the osseous and articular cartilage components of knee OCDs.

Cells Remain Viable When Collected With an In-Line-Suction Tissue Collector From Byproducts of Anterior Cruciate Ligament Reconstruction Surgery.

Purpose: To investigate the viability of cells collected with an in-line-suction autologous tissue collector from the tissue byproducts of arthroscopic anterior cruciate ligament (ACL) reconstruction, to characterize cells from different tissue types, and to identify mesenchymal stem cells. Methods: Patients aged 14 to 50 years with ACL injuries requiring arthroscopic reconstruction surgery were offered enrollment and screened for participation. In total, 12 patients were enrolled in the descriptive laboratory study. Arthroscopic byproduct tissue was collected with an in-line-suction autologous tissue collector from 4 intraoperative collection sites for each patient: ACL stump, ACL fat pad, notchplasty debris, and tunnel drilling debris. All tissue samples were digested using collagenase, and the derived cellular populations were analyzed in vitro, characterizing cellular viability, proliferative potential, qualitative multipotent differentiation capacity, and cell-surface marker presence. Results: An equivalent mass of arthroscopic byproduct tissue was taken from each of the 4 intraoperative collection sites (1.12-1.61 g, P = .433), which all showed an average viability of at least 99.95% and high average total nucleated cells (≥1.37 × 107 cells/mL). No significant differences in collected mass (P = .433), cellular viability (P = .880), or total nucleated cells (P = .692) were observed between the 4 byproduct tissues. The byproduct tissues did exhibit significant differences in monocyte (P = .037) and red blood cell (P = .038) concentrations, specifically with greater values present in the ACL stump tissue. Cells from all byproduct tissues adhered to plastic cell culture flasks. Significant differences were found between colony-forming unit fibroblast counts of the 4 byproduct tissues when plated at 106 (P = .003) and 103 (P = .016) cells as the initial seeding density. There was a significant relationship found between both the starting concentration (χ2 = 32.7, P < .001) and the byproduct tissue type (χ2 = 30.4, P < .001) to the presence of ≥80% confluency status at 10 days. Cells obtained from all 4 byproduct tissues qualitatively showed positive tri-lineage (adipocyte, osteoblast, chondroblast) differentiation potential compared with negative controls under standardized in vitro differentiation conditions. Cells derived from all 4 byproduct tissues expressed cell-surface antigens CD105+, CD73+, CD90+, CD45-, CD14-, and CD19- (>75%), and did not express CD45 (<10%). There were no statistically significant differences in cell-surface antigens between the four byproduct tissues. Conclusions: This descriptive laboratory study demonstrated that cells derived from arthroscopic byproduct tissues of ACL reconstruction remain viable when collected with an in-line-suction autologous tissue collector and these cells meet the ISCT criteria to qualify as mesenchymal stem cells. Clinical Relevance: It is known that viable mesenchymal stem cells reside in byproduct tissue of anterior cruciate ligament reconstruction surgery (ACLR). Practical methods to harvest these cells at the point of care require further development. This study validates the use of an in-line-suction autologous tissue collector for the harvest of viable mesenchymal stem cells after ACLR.