Cell Count and Cell Density Decrease as Age Increases in Cadaveric Pediatric Medial Menisci.

Purpose: To examine the histologic changes in terms of cellularity, cell density, and nuclear shape in medial meniscal cellularity during skeletal development using pediatric cadaver specimens. Methods: Medial menisci from 26 pediatric cadavers, 11 female and 15 male (total 36 menisci), were obtained from tissue bank. Mean age of female donors was 34 months (1-108 months) and of male donors was 52 months (1-132 months). Menisci were processed and embedded in paraffin blocks. Each tissue block containing 6 representative areas of meniscus (anterior root, anterior horn, body [n = 2], posterior horn, and posterior root) was sectioned at 4 microns and stained with hematoxylin and eosin for evaluation of chondrocyte nuclei. Each of the 6 representative areas was imaged at 10×; one image on peripheral one-third of section, the second image on central two-thirds of the section. FIJI imaging software was used to measure cell count, cell density, and nuclear morphology (1 = perfect circle). Data analysis included linear mixed models, Type II analysis of variance tests, and pairwise tests with the Tukey correction to assess statistical significance. Results: Peripheral meniscus was more cellular than central meniscus. The cell count was found to decrease by 14% per year of age. Peripheral cell count decreased at a rate similar to the cell count in the central meniscus. Meniscal cell density was 2× higher peripherally than centrally. Overall average cell density in all locations in the menisci decreased by an average of 14% per year of age. Conclusions: The results of this study reveal decreases in cell count, cell density, and circularity as age increases in cadaveric pediatric medial menisci. Clinical Relevance: To better understand the development of pediatric menisci at a cellular level and use this knowledge in the future on how to maintain the menisci in a younger, healthier state.

Stress-deprivation induces an up-regulation of versican and connexin-43 mRNA and protein synthesis and increased ADAMTS-1 production in tendon cells in situ.

Purpose: The proper function of the tenocyte network depends on cell-matrix as well as intercellular communication that is mechanosensitive. Building on the concept that the etiopathogenic stimulus for tendon degeneration is the catabolic response of tendon cells to mechanobiologic under-stimulation, we studied the pericellular matrix rich in versican and its predominant proteolytic enzyme ADAMTS-1, as well as Connexin-43 (Cx43), a major gap junction forming protein in tendons, in stress-deprived rat tail tendon fascicles (RTTfs).Materials and Methods: RTTfs were stress-deprived for up to 7 days under tissue culture conditions. RT-qPCR was used to measure mRNA expression of versican, ADAMTS-1, and Cx43. Protein synthesis was determined using Western blotting and immunohistochemistry.Results: Stress-deprivation (SD) caused a statistically significant up-regulation of versican, ADAMTS-1, and Cx43 mRNA expression that was persistent over the 7-day test period. Western blot analysis and immunohistochemical assessment of protein synthesis revealed a marked increase of the respective proteins with SD. Inhibition of proteolytic enzyme activity with ilomastat prevented the increased versican degradation and Cx43 synthesis in 3 days stress-deprived tendons when compared with non-treated, stress-deprived tendons.Conclusion: In the absence of mechanobiological signaling the immediate pericellular matrix is modulated as tendon cells up-regulate their production of ADAMTS-1, and versican with subsequent proteoglycan degradation potentially leading to cell signaling cues increasing Cx43 gap junctional protein. The results also provide further support for the hypothesis that the cellular changes associated with tendinopathy are a result of decreased mechanobiological signaling and a loss of homeostatic cytoskeletal tension.

Histologic Evaluation of Biopsy Specimens Obtained After Rotator Cuff Repair Augmented With a Highly Porous Collagen Implant.

PURPOSE: To histologically evaluate biopsy specimens from patients who previously underwent rotator cuff repair augmented with a highly porous collagen implant. METHODS: Biopsies of collagen implant/host-tissue constructs were obtained from 7 patients undergoing a second arthroscopic procedure at various time periods (5 weeks to 6 months) after arthroscopic rotator cuff repair augmented with a collagen implant overlay. The biopsy specimens were examined histologically for host-tissue ingrowth, host-tissue maturation, and host-implant biocompatibility. RESULTS: At the earliest time period (5 weeks), the biopsy revealed the presence of host cells (fibroblasts) within the interstices of the porous collagen implant. Cells were aligned along the linear orientation of the collagen implant structure, and there was evidence of early collagen formation. The 3-month biopsies showed increased collagen formation, maturation, and organization over the surface of the implant and evidence of the collagen implant. At 6 months, the newly generated tissue had the histologic appearance of a tendon, suggesting functional loading of the new generated host tissue. There was no evidence of any remnants of the collagen implant in the 6-month biopsy. There was no evidence of any inflammatory or foreign body reaction within any of the tissue samples. CONCLUSIONS: Biopsies of collagen implants retrieved from human rotator cuff repair subjects revealed cellular incorporation, tissue formation and maturation, implant resorption, and biocompatibility. CLINICAL RELEVANCE: The histologic observations from these clinical biopsies support the biocompatibility of this implant and its ability to promote new connective tissue with the histological appearance of tendon over the surface of the native cuff tendon.

Crimp length decreases in lax tendons due to cytoskeletal tension, but is restored with tensional homeostasis.

Collagen crimp morphology is thought to contribute to the material behavior of tendons and may reflect the local mechanobiological environment of tendon cells. Following loss of collagen tension in tendons, tenocytes initiate a contraction response that shortens tendon length which, in turn, may alter crimp patterns. We hypothesized that changes in the crimp pattern of tendons are the result of cell-based contractions which are governed by relative tautness/laxity of the collagen matrix. To determine the relationship between crimp pattern and tensional homeostasis, rat tail tendon fascicles (RTTfs) were either allowed to freely contract or placed in clamps with 10% laxity for 7 days. The freely contracting RTTfs showed a significant decrease in percent crimp length on both day 5 (3.66%) and day 7 (7.70%). This decrease in crimp length significantly correlated with the decrease in freely contracting RTTf length. Clamped RTTfs demonstrated a significant decrease in percent crimp length on day 5 (1.7%), but no significant difference in percent crimp length on day 7 (0.57%). The results demonstrate that the tendon crimp pattern appears to be under cellular control and is a reflection of the local mechanobiological environment of the extracellular matrix. The ability of tenocytes to actively alter the crimp pattern of collagen fibers also suggests that tenocytes can influence the viscoelastic properties of tendon. Understanding the interactions between tenocytes and their extracellular matrix may lead to further insight into the role tendon cells play in maintaining tendon heath and homeostasis. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:573-579, 2017.

Hypoxia inhibits primary cilia formation and reduces cell-mediated contraction in stress-deprived rat tail tendon fascicles.

BACKGROUND: Hypoxia, which is associated with chronic tendinopathy, has recently been shown to decrease the mechanosensitivity of some cells. Therefore, the purpose of this study was to determine the effect of hypoxia on the formation of elongated primary cilia (a mechanosensing organelle of tendon cells) in vitro and to determine the effect of hypoxia on cell-mediated contraction of stress-deprived rat tail tendon fascicles (RTTfs). METHODS: Tendon cells isolated from RTTfs were cultured under normoxic (21% O2) or hypoxic (1% O2) conditions for 24 hours. The cells were then stained for tubulin and the number of cells with elongated cilia counted. RTTfs from 1-month-old male Sprague-Dawley rats were also cultured under hypoxic and normoxic conditions for three days and tendon length measured daily. RESULTS: A significant (p=0.002) decrease in the percent of elongated cilia was found in cells maintained in hypoxic conditions (54.1%±12.2) when compared in normoxic conditions (71.7%±6.32). RTTfs in hypoxia showed a significant decrease in the amount of contraction compared to RTTfs in normoxia after two (p=0.007) and three (p=0.001) days. CONCLUSION: The decreased incidence of elongated primary cilia in a hypoxic environment, as well as the decreased mechanoresponsiveness of tendon cells under these conditions may relate to the inability of some cases of chronic tendinopathy to respond to strain-based rehabilitation modalities (i.e. eccentric loading).

High magnitude, in vitro, biaxial, cyclic tensile strain induces actin depolymerization in tendon cells.

BACKGROUND: the cytoskeleton is a dynamic arrangement of actin filaments that maintain cell shape and are vital in mediating the mechanobiological response of the cell. METHODS: to determine the cytoskeletal response to varying in vitro, biaxial stretch amplitudes, rat-tail tendon cells were paired into control and cyclically strained groups of 4.75, 9.5, or 12% strain at 1 Hz for 2 hours and the actin cytoskeleton stained. The cells were analyzed for actin staining intensity as a measure of relative depolymerization and for cell shape. Collagenase gene expression was measured in cells undergoing 12% cyclic strain at 1 Hz for 24 hours. RESULTS: there was no significant difference in the degree of actin staining intensity between the control group and cells strained at either 4.75 or 9.5%. However, cells strained at 12% demonstrated a significant decrease in actin staining intensity (depolymerization) compared to control cells, increased collagenase expression by 81%, and a clear shift towards a more rounded cell shape. CONCLUSION: the results of this study demonstrate that the previously reported induction of collagenase activity associated with the application of high magnitude, in vitro, tensile strains may actually be a result of cytoskeletal depolymerization, which causes loss of tensional homeostasis and alteration of cell shape.

Thermal energy enhances cell-mediated contraction of lax rat tail tendon fascicles following exercise.

BACKGROUND: the application of thermal energy (TE) has shown promise in the treatment of tendinopathy. However, the precise mechanism(s) of action of this therapy is unclear. The loss of tendon cell homeostatic tension, due to loading-induced laxity, produces catabolic changes associated with tendinopathy. This catabolic activity can be inhibited through the re-establishment of a normal tensile environment via a cellular contraction mechanism. We hypothesized that application of TE will enhance the contraction rate of lax rat tail tendon fascicles (RTTfs) in an in vitro model. METHODS: following loading, 10 lax RTTfs from each mature rat (n=5) were treated once daily for 7 days with TE by replacing the culture media at 37°C (control) with 42°C media. Using calibrated photographs, RTTf lengths were measured daily. Additional RTTfs were utilized to investigate any changes in material (n=12) and/or histological (n=12) properties with TE. RESULTS: TE significantly increased the contraction rate of RTTfs (p>0.001) without altering the material or histological properties. CONCLUSION: these results demonstrate that TE significantly enhances the contraction rate of previously exercised tendons. The ability to more quickly re-establish a normal mechanobiological environment, thus minimizing any catabolic changes, may explain the beneficial effects reported with applied TE in tendinopathy treatment.

Tendon mechanobiology: Current knowledge and future research opportunities.

Tendons mainly function as load-bearing tissues in the muscloskeletal system; transmitting loads from muscle to bone. Tendons are dynamic structures that respond to the magnitude, direction, frequency, and duration of physiologic as well as pathologic mechanical loads via complex interactions between cellular pathways and the highly specialized extracellular matrix. This paper reviews the evolution and current knowledge of mechanobiology in tendon development, homeostasis, disease, and repair. In addition, we review several novel mechanotransduction pathways that have been identified recently in other tissues and cell types, providing potential research opportunities in the field of tendon mechanobiology. We also highlight current methods, models, and technologies being used in a wide variety of mechanobiology research that could be investigated in the context of their potential applicability for answering some of the fundamental unanswered questions in this field. The article concludes with a review of the major questions and future goals discussed during the recent ORS/ISMMS New Frontiers in Tendon Research Conference held on September 10 and 11, 2014 in New York City.

Allografts in soft tissue reconstructive procedures: important considerations.

CONTEXT: Allografts offer several important advantages over autografts in musculoskeletal reconstructive procedures, such as anterior cruciate ligament reconstruction. Despite growing widespread use of allograft tissue, serious concerns regarding safety and functionality remain. We discuss the latest knowledge of the potential benefits and risks of allograft use and offer a critical review of allograft tissue regulation, management, and sterilization to enable the surgeon to better inform athletes considering reconstructive surgery options. EVIDENCE ACQUISITION: A review of sources published in the past 10 years is the primary basis of this research. STUDY DESIGN: Observational analysis (cohort study). LEVEL OF EVIDENCE: Level 3. RESULTS: Comparable outcome data for autografts and allografts do not support universal standards for anterior cruciate ligament reconstruction, and physician recommendation and bias appear to significantly influence patient preference and satisfaction. Sterilization by gamma and electron-beam irradiation diminishes the biomechanical integrity of allograft tissue, but radioprotective agents such as collagen cross-linking and free radical scavengers appear to have potential in mitigating the deleterious effects of irradiation and preserving tissue strength and stability. CONCLUSION: Allografts offer greater graft availability and reduced morbidity in orthopaedic reconstructive procedures, but greater expansion of their use by surgeons is challenged by the need to maintain tissue sterility and biomechanical functionality. Advances in the radioprotection of irradiated tissue may lessen concerns regarding allograft safety and structural stability.

Tendon Contraction After Cyclic Elongation Is an Age-Dependent Phenomenon: In Vitro and In Vivo Comparisons.

BACKGROUND: Tendons are viscoelastic tissues that deform (elongate) in response to cyclic loading. However, the ability of a tendon to recover this elongation is unknown. HYPOTHESIS: Tendon length significantly increases after in vivo or in vitro cyclic loading, and the ability to return to its original length through a cell-mediated contraction mechanism is an age-dependent phenomenon. STUDY DESIGN: Controlled laboratory study. METHODS: In vitro, rat tail tendon fascicles (RTTfs) from Sprague-Dawley rats of 3 age groups (1, 3, and 12 months) underwent 2% cyclic strain at 0.17 Hz for 2 hours, and the percentages of elongation were determined. After loading, the RTTfs were suspended for 3 days under tissue culture conditions and photographed daily to determine the amount of length contraction. In vivo, healthy male participants (n = 29; age, 19-49 years) had lateral, single-legged weightbearing radiographs taken of the knee at 60° of flexion immediately before, immediately after, and 24 hours after completing eccentric quadriceps loading exercises on the dominant leg to fatigue. Measurements of patellar tendon length were taken from the radiographs, and the percentages of tendon elongation and subsequent contraction were calculated. RESULTS: In vitro, cyclic loading increased the length of all RTTfs, with specimens from younger (1 and 3 months) rats demonstrating significantly greater elongation than those from older (12 months) rats (P = .009). The RTTfs contracted to their original length significantly faster (P < .001) and in an age-dependent fashion, with younger animals contracting faster. In vivo, repetitive eccentric loading exercises significantly increased patellar tendon length (P < .001). Patellar tendon length decreased 24 hours after exercises (P < .001) but did not recover completely (P < .001). There was a weak but significant (R (2) = 0.203, P = .014) linear correlation between the amount of tendon contraction and age, with younger participants (<30 years) demonstrating significantly more contraction (P = .014) at 24 hours than older participants (>30 years). CONCLUSION: Cyclic tendon loading results in a significant increase in tendon elongation under both in vitro and in vivo conditions. Tendons in both conditions demonstrated an incomplete return to their original length after 24 hours, and the extent of this return was age dependent. CLINICAL RELEVANCE: The age- and time-dependent contraction of tendons, elongated after repetitive loading, could result in transient alterations in the mechanobiological environment of tendon cells. This, in turn, could induce the onset of catabolic changes associated with the pathogenesis of tendinopathy. These results suggest the importance of allowing time for contraction between bouts of repetitive exercise and may explain why age is a predisposing factor in tendinopathy.

The basic science of platelet-rich plasma (PRP): what clinicians need to know.

Platelet-rich plasma (PRP) has been advocated for the biological augmentation of tissue healing and regeneration through the local introduction of increased levels (above baseline) of platelets and their associated bioactive molecules. In theory, the increased levels of autologous growth factors and secretory proteins provided by the concentrated platelets may enhance the wound healing process, especially in degenerative tissues or biologically compromised individuals. Although PRP has been increasingly utilized in the treatment of a variety of sports-related injuries, improvements in healing and clinical outcomes have not been universally reported. One reason for this may be the fact that all PRP preparations are not the same. Variations in the volume of whole blood taken, the platelet recovery efficacy, the final volume of plasma in which the platelets are suspended, and the presence or absence of white blood cells, and the addition of exogenous thrombin to activate the platelets or calcium chloride to induce fibrin formation, can all affect the character and potential efficacy of the final PRP product. This article will review the basic principles involved in creating PRP and examine the potential basic scientific significance of the individual blood components contained in the various forms of PRP currently used in sports medicine.

Microvasculature of the suspensory ligament of the forelimb of horses.

OBJECTIVE: To determine the microvascular anatomy of the suspensory ligament of the forelimb of horses. SAMPLE: 17 cadaveric forelimbs from 9 adult horses with no known history of forelimb lameness. PROCEDURES: The median artery of the forelimb was cannulated proximal to the antebrachiocarpal joint and injected with contrast medium for CT evaluation of the gross vasculature (n = 2) or India ink to evaluate the microvasculature (12). Routine histologic evaluation was performed on an additional 3 forelimbs to confirm the microvascular anatomy. RESULTS: The vascular supply of the suspensory ligament of the forelimb originated from branches of the medial and lateral palmar and palmar metacarpal vessels as well as the proximal and distal deep palmar arches. An abundant, longitudinally oriented microvascular supply was evident throughout the length of the suspensory ligament without distinct variation among the proximal, midbody, and distal regions. The intraligamentous blood supply originated from a periligamentous vascular plexus that surrounded the suspensory ligament throughout its length. Histologic findings indicated the presence of a periligamentous connective tissue plexus, which contained vessels that penetrated and anastomosed with an extensive network of intraligamentous vessels throughout the length of the suspensory ligament. CONCLUSIONS AND CLINICAL RELEVANCE: The suspensory ligament of the equine forelimb had an abundant intraligamentous microvascular supply throughout its entire length. The absence of an obvious hypovascular area suggested that regional variations in healing rates of the suspensory ligament are not associated with the microvascular anatomy.

Peak impact accelerations during track and treadmill running.

To determine whether peak vertical and horizontal impact accelerations were different while running on a track or on a treadmill, 12 healthy subjects (average age 32.8 ± 9.8 y), were fitted with a novel, wireless accelerometer capable of recording triaxial acceleration over time. The accelerometer was attached to a custom-made acrylic plate and secured at the level of the L5 vertebra via a tight fitting triathlon belt. Each subject ran 4 miles on a synthetic, indoor track at a self-selected pace and accelerations were recorded on three perpendicular axes. Seven days later, the subjects ran 4 miles on a treadmill set at the individual runner's average pace on the track and the peak vertical and horizontal impact magnitudes between the track and treadmill were compared. There was no difference (P = .52) in the average peak vertical impact accelerations between the track and treadmill over the 4 mile run. However, peak horizontal impact accelerations were greater (P = .0012) on the track when compared with the treadmill. This study demonstrated the feasibility for long-term impact accelerations monitoring using a novel wireless accelerometer.

Age-related changes in the cellular, mechanical, and contractile properties of rat tail tendons.

Tendon laxity following injury, cyclic creep, or repair has been shown to alter the normal homeostasis of tendon cells, which can lead to degenerative changes in the extracellular matrix. While tendon cells have been shown to have an inherent contractile mechanism that gives them some ability to retighten lax tendons and reestablish a homeostatic cellular environment, the effect of age on this process is unknown. To determine the effect of aging on cell number, cell shape, and tensile modulus on tendons as well as the rate of cell-mediated contraction of lax tendons, tail tendon fascicles from 1-, 3-, and 12-month-old rats were analyzed. Aging results in a decrease (p < 0.001) in cell number per mm(2): 1 m (981 ± 119), 3 m (570 ± 108), and 12 m (453 ± 23), a more flattened (p < 0.001) cell nuclei shape and a higher (p < 0.001) tensile modulus (MPa) of the tendons: 1 m (291 ± 2), 3 m (527 ± 38), and 12 m (640 ± 102). Both the extent and rate of contraction over 7 days decreased with age (p = 0.007). This decrease in contraction rate with age correlates to the observed changes seen in aging tendons [increased modulus (r(2) = 0.95), decreased cell number (r(2) = 0.89)]. The ability of tendons to regain normal tension following injury or exercise-induced laxity is a key factor in the recovery of tendon function. The decreased contraction rate as a function of age observed in the current study may limit the ability of tendon cells to retighten lax tendons in older individuals. This, in turn, may place these structures at further risk for injury or altered function.

Age-dependent effects of systemic administration of oxytetracycline on the viscoelastic properties of rat tail tendons as a mechanistic basis for pharmacological treatment of flexural limb deformities in foals.

OBJECTIVE: To describe the effect of systemically administered oxytetracycline on the viscoelastic properties of rat tail tendon fascicles (TTfs) to provide a mechanistic rationale for pharmacological treatment of flexural limb deformities in foals. SAMPLE: TTfs from ten 1-month-old and ten 6-month-old male Sprague-Dawley rats. PROCEDURES: 5 rats in each age group were administered oxytetracycline (50 mg/kg, IP, q 24 h) for 4 days. The remaining 5 rats in each age group served as untreated controls. Five days after initiation of oxytetracycline treatment, TTfs were collected and their viscoelastic properties were evaluated via a stress-relaxation protocol. Maximum modulus and equilibrium modulus were compared via a 2-way ANOVA. Collagen fibril size, density, and orientation in TTfs were compared between treated and control rats. RESULTS: Viscoelastic properties were significantly decreased in TTfs from 1-month-old oxytetracycline-treated rats, compared with those in TTfs from 1-month-old control rats. Oxytetracycline had no effect on the viscoelastic properties of TTfs from 6-month-old rats. Collagen fibril size, density, and orientation in TTfs from 1-month-old rats did not differ between oxytetracycline-treated and control rats. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed that systemically administered oxytetracycline decreased the viscoelastic properties of TTfs from 1-month-old rats but not those of TTfs from 6-month-old rats. The decrease in viscoelastic properties associated with oxytetracycline treatment does not appear to be caused by altered collagen fibril diameter or organization. The age-dependent effect of oxytetracycline on the viscoelastic properties of tendons may be related to its effect on the maturation of the extracellular matrix of developing tendons.

Re-establishment of cytoskeletal tensional homeostasis in lax tendons occurs through an actin-mediated cellular contraction of the extracellular matrix.

Cytoskeletal tensional homeostasis is known to be an important factor in controlling catabolic gene expression in tendon cells. Loss of cell tension in lax rat tail tendon fascicles (RTTfs) has been associated with an upregulation of MMP-13 gene expression and protein synthesis. To determine the role of the actin cytoskeleton in re-establishing tensional homeostasis in lax tendons, RTTfs were allowed to freely contract in vitro for 8 days. The cultured RTTfs contracted rapidly, reaching 50% of their initial length by 3 days. This contraction was associated with the presence of α-smooth muscle actin positive cells within the tendon. Disruption of the actin network by cytochalasian D caused an immediate and significant elongation of the contracted RTTfs. Subsequent removal of the cytochalasian D re-initiated the contraction process. When lax RTTfs were allowed to contract between fixed clamps in culture and become taut, they demonstrated a marked decrease in MMP-13 staining intensity when compared to freely contracting RTTfs. The ability of native tendon cells to contract lax tendons and re-establish their homeostatic "set point" with respect to collagenase production may be an important mechanism in the recovery of tendons elongated by injury, surgical positioning, or cyclic, viscoelastic creep secondary to repetitive exercise.

Comparison of the acute inflammatory response of two commercial platelet-rich plasma systems in healthy rabbit tendons.

BACKGROUND: Numerous studies have shown platelet-rich plasma (PRP) preparations differ with respect to the inclusion of certain blood components, which may affect the host's cellular response. HYPOTHESIS: This study evaluated the inflammatory effect of Biomet GPS III leukocyte-rich PRP (LR-PRP) versus MTF Cascade leukocyte-poor PRP (LP-PRP) after intratendinous injection in an animal model. The authors anticipated that LR-PRP would incite a greater acute inflammatory response than LP-PRP. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 17 skeletally mature New Zealand White rabbits were tested. In all cases, healthy patellar tendons were treated. In the control animals, one patellar tendon was injected with 2 mL autologous whole blood, and the other was injected with 2 mL sterile saline. Seven total tendons were injected with whole blood, and 7 tendons were injected with saline. In the experimental animals, one patellar tendon was injected with 2 mL LR-PRP, and the other was injected with 2 mL LP-PRP. Ten tendons were injected with LR-PRP, and 10 tendons were injected with LP-PRP. Animals were euthanized at 5 or 14 days after injection. Tendons were harvested and stained using hematoxylin and eosin and scored semi-quantitatively for total white blood cells (WBCs), mononuclear cells (macrophages and lymphocytes), polymorphonuclear cells (PMNs), vascularity, fiber structure, and fibrosis. RESULTS: At 5 days after injection, tendons treated with LR-PRP had significantly greater overall tendon scores (6.3 ± 1.79 vs 1.8 ± 1.64, P = .012), as well as mean scores for fiber structure (1.4 ± 0.22 vs 0.50 ± 0.50, P = .012), denoting disrupted composition, total WBCs (1.1 ± 0.89 vs 0.10 ± 0.22, P = .014), mononuclear cells (macrophages and lymphocytes) (0.80 ± 0.45 vs 0.10 ± 0.22, P = .014), vascularity (1.7 ± 0.27 vs 0.80 ± 0.16, P = .008), and fibrosis (1.0 ± 0.35 vs 0.3 ± 0.45, P = .037) compared with tendons treated with LP-PRP. Otherwise, there were no significant differences in mononuclear cells (P = .590), PMN cells (P = 1.00), total WBCs (P = .811), vascularity (P = .650), or total tendon score (P = .596) in any of the treatment groups at 14 days. CONCLUSION: Compared with leukocyte-poor Cascade PRP, leukocyte-rich GPS III PRP causes a significantly greater acute inflammatory response at 5 days after injection. There is no significant difference in the inflammatory response or cellularity regardless of the injection type at 14 days after intratendinous injection. CLINICAL RELEVANCE: Platelet-rich plasma injections are frequently prepared using commercial systems and are administered for clinical treatment of chronic tendinopathy. It is important to characterize the cellular responses elucidated by different injection preparations to further understand their effect on tissue healing and aid clinical decision making. Future investigations are necessary to apply these findings to the clinical setting.

Evaluation of the use of an autologous platelet-rich fibrin membrane to enhance tendon healing in dogs.

OBJECTIVE: To examine effects of an autologous platelet-rich fibrin (PRF) membrane for enhancing healing of a defect of the patellar tendon (PT) in dogs. ANIMALS: 8 adult dogs. PROCEDURES: Defects were created in the central third of the PT in both hind limbs of each dog. An autologous PRF membrane was implanted in 1 defect/dog, and the contralateral defect was left empty. Dogs (n = 4/time period) were euthanized at 4 and 8 weeks after surgery, and tendon healing was assessed grossly and histologically via a semiquantitative scoring system. Cross-sectional area of the PTs was also compared. RESULTS: Both treated and control defects were filled with repair tissue by 4 weeks. There was no significant difference in the histologic quality of the repair tissue between control and PRF membrane-treated defects at either time point. At both time points, the cross-sectional area of PRF membrane-treated tendons was significantly greater (at least 2.5-fold as great), compared with that of sham-treated tendons. At 4 weeks, the repair tissue consisted of disorganized proliferative fibrovascular tissue originating predominantly from the fat pad. By 8 weeks, the tissue was less cellular and slightly more organized in both groups. CONCLUSIONS AND CLINICAL RELEVANCE: A PRF membrane did not enhance the rate or quality of tendon healing in PT defects. However, it did increase the amount of repair tissue within and surrounding the defect. These results suggested that a PRF membrane may not be indicated for augmenting the repair of acutely injured tendons that are otherwise healthy.

In situ deflection of tendon cell-cilia in response to tensile loading: an in vitro study.

To determine if a correlation exists between tensile loading and the deflection of tendon cell-cilia in situ, rat-tail tendon fascicles were stained for tubulin and mounted in a loading device attached to the stage of a confocal microscope. Individual tendon cells (n = 13) were identified and sequential images taken at 0%, 2%, 4%, 6%, and 8% grip to grip strain. The change in ciliary deflection angle was then measured at each strain level. To determine the ability of cilia to return to their original orientation, additional fascicles were loaded to 6% strain and then unloaded to 0% and tendon cell-ciliary (n = 10) deflection angle measured. There was a weak (r(2) = 0.40) but significant (p < 0.0001) correlation between the change in deflection angle and applied strain. Tensile loading produced a change in deflection angle from 0% to 3% (p = 0.039) and from 3% to 6% (p = 0.001) strain. There was no change (p = 1.000) in deflection angle from 6% to 8% strain. Reducing the strain from 6% to 0% resulted in a change (p = 0.048) in angle towards the pre-load position. However, the angle did not return to the pre-strain position (p = 0.025). These results demonstrate that tensile loading produces in situ deflection of tendon cell-cilia and supports the concept that cilia are involved in the mechanotransduction response of tendon cells.