Profiling and Bioinformatics Analysis of Differentially Expressed circRNAs in Spinal Ligament Tissues of Patients with Ankylosing Spondylitis.

Recent studies have reported that circular RNAs (circRNAs) play a crucial regulatory role in a variety of human diseases. However, the roles of circRNAs in ankylosing spondylitis (AS) remain unclear. In this study, we conducted circRNA expression profiling of the spinal ligament tissues of patients with AS by RNA sequencing (RNA-seq) and analyzed the potential functions of differentially expressed circRNA by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to investigate the potential mechanisms associated with AS. The results showed that a total of 1,172 circRNAs were detected in the spinal ligament tissue samples, of which 123 circRNAs were significantly differentially expressed by a fold change ≥ 1.5 and p value < 0.05. Among these, 57 circRNAs were upregulated, and 66 were downregulated. GO and KEGG analyses demonstrated that the differentially expressed circRNAs were mainly involved in the regulation of biological processes of peptidyl-serine phosphorylation and human immune system that may be related to AS. In addition, the circRNA/miRNA interaction networks were established to predict the potential roles of differentially expressed circRNAs by bioinformatics analysis. Taken together, these results revealed the expression profiles of circRNAs and the potential functions of the differentially expressed circRNAs in the spinal ligament tissue of patients with AS, which may provide new clues for understanding the mechanisms associated with AS, and proceed to identify novel potential molecular targets for the diagnoses and treatment of AS.

Involvement of Notch1/Hes signaling pathway in ankylosing spondylitis.

We aimed to investigate the role of Notch1/Hes signaling pathway in the pathogenesis of abnormal ossification of hip ligament in patients with ankylosing spondylitis (AS). 22 AS patients scheduled for artificial hip arthroplasty were randomly chosen as AS group. As controls, we used 4 patients diagnosed with transcervical fracture who underwent hip replacement surgery. Notch1 and Hes mRNA expressions were detected by real-time fluorescent quantitative polymerase chain reaction (RFQ-PCR). Immunohistochemistry (IHC) was used to detect Notch1 and Hes protein expression. Correlation analyses of Notch-l and Hes with AS-related clinical factors were conducted with spearman's correlation analysis and partial correlation analysis. RFQ-PCR results showed significant differences in Notch1 and Hes mRNA expressions between AS group and the control group (all P<0.05). IHC analysis further indicated positive nuclear signals of Notch1 and Hes protein, indicating functional activation of the Notch1 and Hes pathways. Semi-quantitative IHC showed a higher Notch1 and Hes expression levels in AS group compared to the control group (all P<0.05). Correlation analysis suggested that Hes protein expression was positively associated with the clinical course of the disease in AS patients. In conclusion, Notch1 and Hes overexpression was clearly detected in hip joint ligaments of AS patients, Hes protein expression was associated with the clinical course of AS. Taken together, we suggest that signaling pathways mediated by Notch1-Hes may contribute to ligament ossification of hip joints in AS patients.

Are myofibroblasts activated in idiopathic carpal tunnel syndrome? an immunohistochemical study.

OBJECTIVES: This study aims to investigate whether myofibroblasts participate in the fibrotic process of idiopathic carpal tunnel syndrome (CTS). PATIENTS AND METHODS: Forty patients (12 males, 28 females; median age 50.85 ± 11.2 years; range 30 to 71 years) who were operated in our clinic between March 2010 and August 2010 were included in the study. Twenty-five idiopathic CTS patients were assigned to the study group, and 15 trauma patients were assigned to the control group. Samples were taken from both transverse carpal ligament and subsynovial connective tissue (SSCT) of participants and they were analyzed by immunohistochemical method. Four immunohistochemical markers were used to analyze myofibroblast existence and vascular wall thickness (alpha smooth muscle actin [α-SMA]), collagen type IV antibodies, and T (CD3) and B (CD20) lymphocytes. RESULTS: The existence of myofibroblasts (α-SMA) in SSCT of patients who were in early phase of idiopathic CTS was shown through the positive reaction of their antibodies with fibroblasts. A significantly increased reaction of α-SMA and collagen antibodies in vascular structure of SSCT demonstrated increased vascular wall thickness and vascularity in the study group (p<0.01). No significant difference was detected between the two groups in terms of T and B lymphocyte antibody reaction (inflammation). CONCLUSION: The findings of this study indicate a potential for myofibroblasts to be activated during the early phase of the disease and contribute to the onset of disease. Further studies with larger sample sizes would be of great assistance in determining the role of myofibroblasts in idiopathic CTS.

Carpal tunnel syndrome is associated with high fibrinogen and fibrinogen deposits.

BACKGROUND: Idiopathic carpal tunnel syndrome (ICTS) is a common entrapment neuropathy. Some cases of ICTS are linked to mutations of the transthyretin gene, whereas others are associated with systemic amyloidosis. The majority of ICTS cases are of unknown etiology. OBJECTIVE: To study molecular mechanisms of ICTS development. METHODS: A total of 71 ICTS patients and 68 control subjects were included in the study. The fibrinogen level was determined before surgery and its deposition in the transversal carpal ligament (TCL) was detected by immunohistochemistry, Western blot, and mass spectrometry. Fibrinogen interaction with other proteins was studied by immunoprecipitation assay. RESULTS: Plasma levels of the proinflammatory and hemostatic protein fibrinogen are elevated in ICTS patients. Other measured systemic inflammatory markers were not affected, and local inflammatory responses in TCL were absent. ICTS patients have shorter bleeding times, probably because of the elevated plasma levels of fibrinogen. Polymorphisms of the fibrinogen B promoter region were previously associated with increased plasma fibrinogen, but this association was not observed among patients with ICTS. Interestingly, we detected fibrinogen deposits in the TCL, whereas transcriptional activity of the fibrinogen genes was low. Amyloidogenic proteins, including transthyretin and α-synuclein, were also found in the TCL, whereas their local transcriptional activity was rather high. Finally, we demonstrated that fibrinogen interacts with transthyretin and α-synuclein in TCL lysates. CONCLUSION: Our data indicate that fibrinogen and other aggregation-prone proteins have potentially important roles in the pathogenesis of ICTS.

The importance of pulsed lavage on interface temperature and ligament tension force in cemented unicompartmental knee arthroplasty.

BACKGROUND: Mechanical loosening is the most common cause of revision in unicompartmental knee arthroplasty. We determined the effect of bone lavage on tibial cement penetration and interface temperature with controlled ligament tension forces. We presumed pulsed lavage would allow increased cement penetration compared with syringe lavage. METHODS: Cemented unicompartmental knee arthroplasty was performed in 12 pairs of fresh-frozen knees. Lavage was performed using pulsed lavage on one side (A) and syringe lavage on the other (B). Cement penetration pressure, interface temperature, and ligament tension forces were continuously monitored during the operation. Screened radiographs were taken and cement penetration under the tibial plateau was measured. FINDINGS: The pulsed lavage group showed a mean cement penetration area of 187.24 (SD 36.37) mm², whereas 144.29 (SD 35.74) mm(2) was measured in the group with syringe lavage. Cement penetration pressure was 13.29 (SD 8.69) kPa in Group A and 20.21 (SD 7.78) kPa in Group B. Maximum interface temperatures of 46.99°C were observed in Group A and 45.02°C in Group B. INTERPRETATION: Our data showed pulsed lavage cleansing of the cancellous tibial bone substantially improved cement penetration compared with syringe lavage without reaching the temperature threshold for bone necrosis. We recommend the routine use of pulsed lavage to improve long-term fixation.

An immunohistochemical study of the extracellular matrix of entheses associated with the human pisiform bone.

The immunohistochemical labelling patterns of the extracellular matrix at the insertion of the flexor carpi ulnaris tendon and the entheses at both ends of the pisometacarpal and pisohamate ligaments were compared in order to relate the molecular composition of the attachment sites to their mechanical environment. Tissue was obtained from elderly dissecting room cadavers and labelled with a panel of monoclonal antibodies directed against collagens, glycosaminoglycans, proteoglycans and matrix proteins. All entheses were fibrocartilaginous and labelled positively for molecules typically associated with articular cartilage (type II collagen, chondroitin 6 sulphate, aggrecan and link protein). Labelling for type II collagen was most conspicuous at the attachment of the flexor carpi ulnaris tendon. In the ligaments, type II collagen labelling was always greater at the pisiform end. Matrilin 1 was universally present at all five entheses examined and fibromodulin labelling was most intense around the tidemark. Fibromodulin may thus be involved in anchorage and/or the control of mineralization at the hard-soft tissue interface of entheses. The greater prominence of fibrocartilage at the pisiform enthesis of the flexor carpi ulnaris tendon than at any ligament attachment may relate to the marked change in the tendon insertional angle that occurs with wrist movements. We also suggest that the more fibrocartilaginous character of the proximal compared with the distal ends of the ligaments relates to the fact that the pisiform is anchored in position and is thus at the centre of rotation of any movement of ligaments attached to it.

Analysis of estrogen binding sites of the posterior ligament of the human TMJ.

BACKGROUND: Conflicting results have been reported regarding the presence or absence of estrogen-binding sites (EBS) in the human temporomandibular joint (TMJ). The possible role of female sex hormones in the pathophysiology of internal derangement of the TMJ has been suggested to explain the prevalence of TMJ symptoms in female patients. PATIENTS AND METHODS: Posterior bilaminar tissue excised during TMJ articular disc repositioning and posterior ligament repair was taken from 28 patients (26 female, 2 male) for evaluation. Cryosections were stained using a monoclonal antibody (Mab) against EBS. To ensure efficacy of the antibody staining procedure, an internal positive control consisting of human breast tissue previously proven EBS-positive was used. No asymptomatic control TMJ tissue was available for our study. RESULTS: None (0%) of 28 TMJ tissue specimens showed nuclear-staining positive for the presence of EBS in the posterior bilaminar tissue of the TMJ. However, estrogen-binding sites associated with probable inflammatory cells were observed. Our results are consistent with the probability of positives as high as 0.1234 using a 95% confidence interval. CONCLUSIONS: The lack of EBS of the posterior ligament of the TMJ suggests that the role of estrogen contributing to internal derangement of the TMJ appears not to be significant.

Mapping lubricin in canine musculoskeletal tissues.

Lubricin, also known as superficial zone protein or PRG4, has many distinct biological functions, including lubrication, antiadhesion, and as a regulator of cell growth. This study investigated lubricin in canine musculoskeletal tissues using RT-PCR and immunohistochemistry. One or more variants were noted in canine flexor digitorum profundus (FDP) tendon, Achilles tendon, patellar tendon, A2 pulley, anterior cruciate ligament (ACL), knee lateral collateral ligament (LCL), articular cartilage, meniscus, muscle, and skin. We found 6 N-terminal lubricin splicing variants. The variants with larger sizes were identified in FDP tendon, ACL, LCL, A2 pulley, and cartilage. Lubricin was distributed both on the tissue surfaces and at the interface of fiber bundles within tissues, but this distribution varied by tissue type. We conclude that lubricin is present in many tissues; variations in splicing and physical distribution suggest that the variants of lubricin may play different roles in different locations.

The inferior glenohumeral ligament: a correlative investigation.

The inferior glenohumeral ligament (IGHL) was investigated by correlating the biomechanical properties, biochemical composition, and histologic morphology of its 3 anatomic regions (superior band, anterior axillary pouch, and posterior axillary pouch) in 8 human cadaveric shoulders. The overall biochemical composition of the IGHL appeared similar to other ligaments, with average water content of 80.9 +/- 2.5%, collagen content of 80.0 +/- 9.2%, and crosslinks of 0.715 +/- 0.13 mol/mol collagen. The proteoglycan content was highest in the superior band (2.73 +/- 0.7 mg/g dry weight) and may, in part, explain its viscoelastic behavior. Histologic analysis demonstrated longitudinally organized fiber bundles that were more uniform in the mid-substance but more interwoven and less uniformly oriented near the insertion sites. The superior band had the most pronounced fiber bundle interweaving, while crimping was more evident in the anterior axillary pouch. Elastin was identified in each of the regions. Tensile testing demonstrated a trend toward higher ultimate tensile stress (16.9 +/- 7.9 MPa) and tensile modulus (130.3 +/- 47.9 MPa) in the superior band compared to the axillary pouch. The mean ultimate tensile strain of the IGHL was 16.8 +/- 4.6%. These complex IGHL properties may help to explain its unique functions in stabilizing the shoulder in different arm positions and at different rates of loading, including the failure patterns seen clinically, as in Bankart lesions (insertion site) versus capsular stretching (ligament substance).

Molecular parameters indicating adaptation to mechanical stress in fibrous connective tissue.

The present study pursues the hypothesis that local compressive force and the occurrence of cartilage-specific transformation processes within the extracellular matrix of tendons and ligaments are directly correlated. We compare the pattern of certain marker molecules typical of (fibro)cartilage in select examples. Investigations are carried out of the extensor tendons of toes and fingers, the transverse ligament of the atlas, the transverse ligament of the acetabulum, and of the tendon of the superior oblique muscle and its trochlea. The marker molecules are detected with standardized immunohistochemical methods. The results show that certain molecules only occur under conditions of (relatively high) compressive stress, others being the result of tensile stress. The molecular spectrum of the molecules of the ECM allows qualifying conclusions as to the mechanical situation of a given part of the tissue. A quantifying statement about the intensity of compressive stress is not possible to make thus far, but the extension of the restructuring areas corresponds to the area of compressive stress. Depending on the intensity and duration of the local compressive strain, the molecules involved may be ordered chronologically according to their occurrence in the ECM. The glycosaminoglycans react at lower stress levels than the proteoglycans, which in turn react earlier than the collagens, especially with regard to the vanishing of type I collagen and the first occurrence of type II collagen. Of the glycosaminoglycans, dermatan sulfate and keratan sulfate occur first. These are detectable in virtually all cases. They are followed by chondroitin 4 sulfate. The last glycosaminoglycan, which occurs in already significantly fibrocartilaginous tissue, is chondroitin 6 sulfate. Under chronologically intensifying compressive stress in the increasingly fibrocartilaginous tissues, the proteoglycans versican and, to a lesser extent, tenascin--characteristic markers of fibrous tissue--can still be detected. However, their spatial expansion steadily decreases until they finally vanish. Contrastingly, aggrecan and link protein expression becomes increasingly prominent in such tissues. The spatial expansion of the adaptation zones in tendons and ligaments roughly corresponds with the zones subjected to compressive force; tensile stress alone does not result in a production of fibrocartilage. The questions asked at the beginning may thus be answered as follows: The molecular composition of the various fibrous connective tissues, such as tendons and ligaments, can be directly correlated with the respective tissue's mechanical function. As an expression of this regular interrelation, a ranking of certain ECM molecules may be set up that corresponds to the type, intensity, and duration of the mechanical stress. Grounded on this, it seems possible to prognosticate the occurrence of certain components in the ECM depending on the nature of the mechanical stress. The occurrence of certain molecules within the fibrocartilaginous tissue is of clinical importance in connection with various forms of rheumatoid arthritis and perhaps other diseases with an autoimmune-related etiology. Since a considerable part of the inflammatory destructions involved may at least indirectly result from autoimmune processes directed against the cartilage-type components of the ECM, every fibrocartilage constitutes a potential target to a certain degree. This applies particularly to those fibrocartilages whose ECM has a molecular composition closely resembling that of hyaline articular cartilage. Therefore, knowledge of the regional molecular composition allows a prediction of sites where clinical symptoms may occur in the course of rheumatoid arthritis.

Quantitative cytochemical evidence for local increases in bone turnover at the acromial enthesis of the human coracoacromial ligament.

OBJECTIVE: Enthesophytic bone outgrowths are found at many ligament attachment sites, and while their incidence is associated with many pathologies, the mechanism by which they form remains controversial. We hypothesized that changes in local cell behavior, provoked by mechanical alterations within the coracoacromial ligament (CAL), lead to acromial enthesophyte formation. We investigated whether cell behavior at acromial entheses is consistent with this. METHODS: We used quantitative enzyme cytochemistry to measure glucose 6-phosphate dehydrogenase (G6PD), alkaline phosphatase (ALP; osteoblastic activity), and tartrate-resistant acid phosphatase (TRAP; osteoclastic phenotype) activities in cells of the acromial attachment into the CAL in patients with rotator cuff tears. RESULTS: (1) Resident osteoblasts on the acromion's inferior aspect express elevated activity of G6PD and ALP, indicative of increases in osteogenic potential. (2) These activities are selectively raised at the "leading edge" of acromial bone CAL enthesis. (3) In contrast, distribution of TRAP-positive cells does not exhibit a spatial correlation with enthesis architecture. We also found that cells situated close to the CAL attachment into the acromion exhibited elevated levels of G6PD and ALP activity, but intriguingly, also showed higher TRAP activity than neighboring cells distant from entheses. CONCLUSION: These results suggest that the acromion in these patients undergoes bone accretion at the inferior attachment of the CAL, and that enthesial ligament cells close to the bone express characteristics consistent with enthesophyte formation at the leading edge of this bony spur's extension into the ligament.

The functional anatomy of the human anterior talofibular ligament in relation to ankle sprains.

The anterior talofibular ligament is the most commonly injured ligament in the ankle. Despite considerable interest in the clinical outcome of treatment protocols, we do not know whether the distinctive pattern of localization of the injuries relates to regional differences in the structure and molecular composition of the ligament. To address this issue, ligaments were examined by histology and immunohistochemistry. Differences in the structure of its two attachments (i.e. entheses) were evaluated with quantitative, morphometric techniques, and regional differences in the distribution of collagens, glycosaminoglycans and proteoglycans were determined qualitatively by immunolabelling. Morphometric analyses showed that bone density was less at the fibular attachment, but that enthesis fibrocartilage was more prominent. Immunohistochemistry revealed the presence of a fibrocartilage (containing type II collagen and aggrecan) at the site where the ligament wraps around the lateral talar articular cartilage in a plantarflexed and inverted foot: the fibrocartilage is regarded as an adaptation to resisting compression. We propose that avulsion fractures are less common at the talar end of the ligament because (1) bone density is greater here than at the fibular enthesis, and (2) stress is dissipated away from the talar enthesis by the 'wrap-around' fibrocartilaginous character of the ligament near the talar articular facet.

Do facet joint capsular ligaments contain estrogen receptors? Application to pathogenesis of degenerative spondylolisthesis.

Studies showing that degenerative spondylolisthesis is 4 to 5 times more common in females than in males have suggested that hormonal influences account for this gender difference. Estrogen has been shown to play a role in other instabilities, such as those of the anterior cruciate ligament and the shoulder capsular ligaments, and estrogen receptors have been identified in these tissues. We wanted to assess facet joint capsular ligaments for the presence of such receptors. Accordingly, we collected facet joint capsular ligaments from 14 consecutive patients undergoing lumbar spinal fusion. Tissue sample analysis was performed by immunohistology using prediluted estrogen monoclonal antibody (Vantana) and automated immunostaining on a Vantana instrument. None of the specimens analyzed contained estrogen receptors. We conclude that, though degenerative spondylolisthesis may have hormonal influences, estrogen seems not to play a direct role in its development.

Fibrocartilage in the transverse ligament of the human atlas.

STUDY DESIGN: Immunohistochemical investigation. OBJECTIVE: To determine whether molecules typical of articular cartilage are present in the transverse ligament and whether the ligament may be a target for an autoimmune response in rheumatoid arthritis. SUMMARY OF BACKGROUND DATA: In chronic rheumatoid arthritis there is often a marked instability of the atlantoaxial complex, and the transverse ligament can show degenerative changes that compromise its mechanical function. In some rheumatoid patients there can be an autoimmune response to cartilage link protein, aggrecan, and Type II collagen. METHODS: Transverse ligaments were removed from 13 cadavers and fixed in 90% methanol. Cryosections were immunolabeled with antibodies against proteoglycans (aggrecan, link protein, and versican), glycosaminoglycans (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, and keratan sulfate), and collagens (Types I, II, III, and VI). RESULTS: Labeling for aggrecan and link protein was characteristic of the fibrocartilages, but versican was only detected in the fibrous regions. Equally, Types I, III, and VI collagens and keratan, dermatan, and chondroitin-4-sulfates were found throughout the ligament, but labeling for Type II collagen and chondroitin-6-sulfate was restricted to the fibrocartilages. CONCLUSION: The presence of molecules typical of articular cartilage (aggrecan, link protein, and Type II collagen) in the transverse ligament explains why it can be a target for destruction in rheumatoid arthritis and also suggests that it is subject to constant compression against the dens rather than only at the extremes of movement.

Identification of sex hormone receptors in human and rabbit ligaments of the knee by reverse transcription-polymerase chain reaction: evidence that receptors are present in tissue from both male and female subjects.

Gender-related factors have been attributed to observed differences in the rate of injury to ligaments (e.g., anterior cruciate ligament) between male and female subjects. These differences may be a result of unique regulatory mechanisms within the tissue in response to the sex hormones estrogen and progesterone. These hormones, when bound to specific intracellular receptors (estrogen receptor and progesterone receptor, respectively), modulate gene expression within hormone-responsive tissue. The purpose of this study was to evaluate the expression of the estrogen and progesterone receptors in ligament tissue from male and female rabbits and humans by the sensitive molecular technique of reverse transcription-polymerase chain reaction. Total RNA was extracted from human anterior cruciate ligament tissue and from medial cruciate ligament, anterior cruciate ligament, patellar tendon, and synovium tissue of the New Zealand White rabbit by the newly developed TRIspin method. The total RNA was reverse transcribed and analyzed by polymerase chain reaction to assess the expression of estrogen and progesterone receptors. Our results demonstrate that estrogen and progesterone receptor transcripts are expressed in ligament tissue of male and female rabbits and humans and that alterations in receptor expression occur in ligaments during pregnancy. In the human samples, only a small percentage of the estrogen receptor appeared to be a nonfunctional mRNA splice variant, and the predominant form contained the estrogen-binding domain.

pO2 measurement in an experimental model of patellar tendon autograft pro-anterior cruciate ligament.

Thirty-four sheep were submitted to surgery substituting the native ACL with the central third of the patellar tendon, ten enter this study. The purpose was to find a possible relationship between tissue pO2 and healing processes considering also the biomechanical and histomorphological aspects of the grafts. Four of them were sacrificed under general anaesthesia after 6 months, and six after 1 year in order to perform tissue pO2 measurement and an analysis of microvessel density on specimens of the normal ACL and the graft. Our data showed higher pO2 values of the autografts after 6 months. After 1 year the data was comparable to those of native ACL. This was confirmed by a microvessel count of the histological specimens and the data was in relationship to biomechanical and histomorphological analysis. Tissue pO2 can be observed and recorded in "in vivo" ACL, and patellar tendon used as graft, with no injury to their integrity. The monitoring system might be considered as an experimental tool for indirect controls of the anterior cruciate substitutes.

Morphological and biochemical studies of the elastic fibres in the craniomandibular joint articular disc of the tight-skin mouse.

The tight-skin (TSK) mouse is characterized by the hyperplasia of loose connective tissues, and of excessive growth of cartilage and of bones including the mandible. Since the fibroelastic connective tissues of the craniomandibular joint (CMJ) are essential to the functions of this joint, the present histological study compared the presence and general distribution of elastic fibres in CMJ discal tissues of TSK and normal mice. The excised CMJs were processed for light microscopy. The tissues were fixed, demineralized, embedded in paraffin, sectioned and then stained with resorcin-fuchsin to demonstrate elastic fibres. There were no obvious histological differences in either the amount or the distribution of elastic fibres in the discs from the two groups. In both groups, elastic fibres were found in the disc and in many of the attachments of the disc to the mandible and squamosal bone. In addition to the morphological preparations, articular discs and samples of lung tissue were excised from other mice and subjected to a radioimmunoassay for desmosine in order to estimate the amounts of elastin in these tissues; the amount of elastin was significantly reduced in the TSK lung, but the amounts of elastin in the TSK and normal CMJ discal tissues were not significantly different statistically. These morphological and histochemical results suggest that the distribution and quantity of elastic fibres in the TSK mouse disc are not significantly different from those in the normal mouse articular disc. Moreover, these data may be interpreted to either suggest a differential effect on the elastic fibres in different TSK tissues, or to support the suggestion that abnormal degradation of elastic fibres may not be characteristic of the TSK mouse.

The intrinsic and extrinsic ligaments of the wrist. A correlation of collagen typing and histologic appearance.

20 fresh frozen human wrist specimens were dissected. Gross and histological examination and biochemical evaluation were performed on the intrinsic ligaments (scapho-lunate and luno-triquetral) and extrinsic ligaments (radio-scapho-capitate and radio-lunate). All ligaments were observed to have longitudinal collagen bundles. The intrinsic ligaments had large amounts of fibrocartilage near insertions and no elastin, while the extrinsic ligaments had little fibrocartilage and sparse amounts of elastin. The intrinsic ligaments were noted to have significantly more collagen Type 3 (41%) than the extrinsic ligaments (19%). These findings suggest that structural differences between the intrinsic and extrinsic ligaments of the wrist may in part account for the biomechanical observations that the intrinsic ligaments are stronger and elongate further prior to failure than the extrinsic ligaments.