Calcified Cartilage Research Articles

Osteopetrosis a Significant Bone Disease: A Case Study

Background: Osteopetrosis is a genetic condition that affects just a few people. illness characterised by osteosclerosis, Due to inadequate osteoclast resorption, there is destruction of the medullary canal, calcified cartilage, and hard bone. Patients with osteopetrosis are more likely to develop osteoarthritis. Only a few studies have documented a patient who underwent both a complete Arthroplasty of the hip and knee (TKA). In this article, a patient with osteopetrosis describes a case of left and right hip osteoarthritis.
 Clinical Findings: Walking problems, Calcium deficiency, the optic nerve in the brain is under pressure. Hearing loss, as well as distress, can lead to neurological issues.
 Diagnostic Evaluation: CBC Investigation, blood test, Hb-12.9%, Total RBC count-4.84million/cumm, RDW-13.2%, HCT-36.7%, Total WBC Count-26200 cu mm, Monocytes- 01%, Granulocytes- 30%, Lymphocytes- 65%, Total platelet count-3.5 Lacs/cu mm, ALT(SGPT)-17 U/L, AST (SGOT)-44 U/L.
 Peripheral Smear: RBCs-Normocytic Normochromic Platelets-Adequate on smear, No Hemiparasite seen, WBCs-Neutrophilic leucocytosis.
 Therapeutic Intervention: Calcium 1200mg, D3 8000IU, vitamin D supplements calcitriol 0.25mcg/ml, hormone replacement therapy, Bone marrow transplantation (BMT), Alendronate70mg, (Fosamax), Risedronate35mg OD (Actonel), Inj Boniva 3mg OD, Inj Zoledronic acid 4mg BD (Recast), various medications, and/or surgery.
 Outcome: After therapy, the child's behaviour improves. Osteoporosis is a condition in which excessive bone resorption causes decreased bone mass and skeletal fragility, with the most serious clinical consequence being increased fracture risk.
 Conclusion: The patient was hospitalised to AVBRH with a known case of Osteopetrosis illness and complained of difficulties awakening, discomfort, and neurological issues. After receiving appropriate therapy and surgery, his condition improved. According to this case study, in patients with osteopetrosis, total joint arthroplasty is an effective therapy for painful hip and knee osteoarthritis.

Enhanced carbonic anhydrase expression with calcification and fibrosis in bronchial cartilage during COPD

Pulmonary cartilage plays a crucial structural role determining the physiologic airway compressibility and distensibility, necessary for proper mechanical function. This functionality deteriorates with aging due to increased stiffness of both airway muscle and cartilage, as well as, decreased renewal capacity. Altered airway remodeling has been suggested as a pathogenic driver of chronic obstructive pulmonary disease (COPD) through mechanisms still incompletely understood. Using paraffin-embedded lung tissue sections from archived autopsy material from COPD with non-COPD age matched controls a histopathologic analysis focused on inflammation, fibrosis and calcification was performed with special stains (Masson’s trichrome and Von Kossa) and immunohistochemistry for carbonic anhydrase IV (CA IV) and Ki-67. COPD lung tissues showed increased peribronchial inflammation compared to the non-COPD. Coarse amphophilic crystalline deposits in bronchial cartilage were more frequently observed in COPD sections, which were compatible with early dystrophic calcification of the extracellular matrix and chondrocytes. Moreover, Von Kossa staining revealed a significant calcium deposition in the cartilages from COPD in comparison to the controls. Interestingly, Ki-67 immunostains demonstrated a higher overall proliferative rate, including epithelial cells, in COPD. Furthermore, Masson’s trichrome staining revealed relatively increased peribronchial collagen deposition associated with a fibrotic stromal response, which may be secondary to the inflammatory milieu in COPD. To further characterize the tissue microenvironment associated with dystrophic calcification, immunohistochemistry for CA IV was used, revealing significantly increased expression in chondrocytes and peribronchial tissue in COPD. Our findings demonstrate that dystrophic calcification of the extracellular matrix and chondrocytes can be linked to CA IV expression in COPD and suggest that pH changes in pulmonary tissue associated with inflammation and calcification may play an active role in COPD.

Role of Ciliary Protein Intraflagellar Transport Protein 88 in the Regulation of Cartilage Thickness and Osteoarthritis Development in Mice.

Mechanical and biologic cues drive cellular signaling in cartilage development, health, and disease. Primary cilia proteins, which are implicated in the transduction of biologic and physiochemical signals, control cartilage formation during skeletal development. This study was undertaken to assess the influence of the ciliary protein intraflagellar transport protein 88 (IFT88) on postnatal cartilage from mice with conditional knockout of the Ift88 gene (Ift88-KO). Ift88fl/fl and aggrecanCreERT2 mice were crossed to create a strain of cartilage-specific Ift88-KO mice (aggrecanCreERT2 ;Ift88fl/fl ). In these Ift88-KO mice and Ift88fl/fl control mice, tibial articular cartilage thickness was assessed by histomorphometry, and the integrity of the cartilage was assessed using Osteoarthritis Research Society International (OARSI) damage scores, from adolescence through adulthood. In situ mechanisms of cartilage damage were investigated in the microdissected cartilage sections using immunohistochemistry, RNAScope analysis, and quantitative polymerase chain reaction. Osteoarthritis (OA) was induced in aggrecanCreERT2 ;Ift88fl/fl mice and Ift88fl/fl control mice using surgical destabilization of the medial meniscus (DMM). Following tamoxifen injection and DMM surgery, the mice were given free access to exercise on a wheel. Deletion of Ift88 resulted in progressive reduction in the thickness of the medial tibial cartilage in adolescent mice, as well as marked atrophy of the cartilage in mice during adulthood. In aggrecanCreERT2 ;Ift88fl/fl mice at age 34 weeks, the median thickness of the medial tibial cartilage was 89.42 μm (95% confidence interval [95% CI] 84.00-93.49), whereas in Ift88fl/fl controls at the same age, the median cartilage thickness was 104.00 μm (95% CI 100.30-110.50; P < 0.0001). At all time points, the median thickness of the calcified cartilage was reduced. In some mice, atrophy of the medial tibial cartilage was associated with complete, spontaneous degradation of the cartilage. Following DMM, aggrecanCreERT2 ;Ift88fl/fl mice were found to have increased OARSI scores of cartilage damage. In articular cartilage from maturing mice, atrophy was not associated with obvious increases in aggrecanase-mediated destruction or chondrocyte hypertrophy. Of the 44 candidate genes analyzed, only Tcf7l2 expression levels correlated with Ift88 expression levels in the microdissected cartilage. However, RNAScope analysis revealed that increased hedgehog (Hh) signaling (as indicated by increased expression of Gli1) was associated with the reductions in Ift88 expression in the tibial cartilage from Ift88-deficient mice. Wheel exercise restored both the articular cartilage thickness and levels of Hh signaling in these mice. Our results in a mouse model of OA demonstrate that IFT88 performs a chondroprotective role in articular cartilage by controlling the calcification of cartilage via maintenance of a threshold of Hh signaling during physiologic loading.

Chondrocyte Adipogenic Differentiation in Softening Osteoarthritic Cartilage

A chondrocyte-to-osteoblast lineage continuum exists in the growth plate. Adipogenic differentiation of chondrocytes in vivo should be investigated. Here, unilateral anterior crossbite (UAC), which can induce osteoarthritic lesions in the temporomandibular joint (TMJ), was applied to 6-wk-old C57BL/6 mice. Matrix loss in TMJ cartilage was obvious, as demonstrated by safranin O staining, and the condylar cartilage elastic modulus values, detected by using atomic force microscopy (AFM), were reduced, indicating cartilage softening that might be linked with loss of the highly charged proteoglycan. By crossing the Rosa26/tdTomato (TdT) mice with Sox9;CreERT2 mice or with Col10;CreERT2 mice, we obtained the Sox9-TdT and Col10-TdT strains, respectively, in which the Sox9- or Col10-expressing cells, accordingly, were labeled by TdT. A few TdT-labeled cells in both strains expressed AdipoQ or DMP-1. The Sox9-TdT+AdipoQ+ cells were primarily located in the deep zone cartilage and then in the whole cartilage. Col10-TdT+AdipoQ+ cells, Sox9-TdT+DMP-1+ cells, and Col10-TdT+DMP-1+ cells were located in the deep zone region. UAC promoted AdipoQ and DMP-1 expression in cartilage. The percentages of Sox9-TdT+AdipoQ+ and Col10-TdT+AdipoQ+ cells to Sox9-TdT+ and Col10-TdT+ cells, respectively, were increased (both P < 0.05), implying that more chondrocytes were undergoing adipogenic differentiation in the UAC group, the cartilage of which was softened. The percentages of Sox9-TdT+DMP-1+ and Col10-TdT+DMP-1+ cells to Sox9-TdT+ cells and Col10-TdT+ cells, respectively, were increased (both P < 0.05), consistent with our report that UAC enhanced deep zone cartilage calcification, causing stiffening of the deep zone cartilage. Our present data demonstrated that TMJ chondrocyte descendants can become adipogenic in vivo in addition to becoming osteogenic. This potential was promoted in osteoarthritic cartilage, in which deep zone cartilage calcification-associated cartilage stiffening and proteoglycan loss-associated cartilage softening were both stimulated.

Association of Quantified Costal Cartilage Calcification and Long-Term Cumulative Blood Glucose Exposure: The Multi-Ethnic Study of Atherosclerosis.

AimsAnecdotal reports have suggested increased soft tissue calcification in individuals with long-term exposures to high blood glucose. The association of costal cartilage calcification (CCC), a reliably quantifiable marker obtainable from non-contrast cardiac computed tomography (CT) with cumulative fasting blood glucose (FBG) exposure, is unknown. In this study, we aimed to determine the association between quantified CCC and cumulative glucose exposure using non-contrast coronary artery calcium (CAC) scoring computed tomography (CT) images in the Multi-Ethnic Study of Atherosclerosis (MESA).MethodsThe volume of bilateral CCC was quantified in high-density pixels (threshold of Hounsfield Unit>180) using the CAC scoring CT images acquired in the 5th MESA exam. Prior long-term cumulative exposure to FBG was calculated by area under the FBG-time curve over ten years before the time of the CT exam.ResultsA total of 2,305 participants (mean age: 69, female/male: 1.3) were included in this study. The median CCC volume was lower in females than males (1158 mm3 [IQR: 1751] vs. 3054 mm3 [3851], p<0.001). In cross-sectional analysis, quantified CCC was associated with FBG (9% increase per SD) and HbA1c (7% increase per SD) at the CT exam only in female participants after adjustment for age, race, BMI, and glomerular filtration rate. Only in female participants, quantified CCC was also associated with prior cumulative FBG (3% increase per decile change). In the subgroup of females with zero CAC scores, the adjusted CCC was still associated with FBG (13% increase per SD) at the time of CT exam and with prior cumulative FBG exposure (4% increase per decile change) before the CT exam.ConclusionsThe CCC, a reliably quantified marker in non-contrast cardiac CT, is associated with 10-year cumulative FBG exposure only in female participants, even those with zero CAC.

Age related changes in thyroid and cricoid cartilages: An autopsy based radiological analysis

Forensic age estimation is an invaluable aspect of human identification. Out of these many means of age estimation, few regions with untapped potential for assistance in forensic age estimation are the age-related changes in the laryngeal cartilages. As the age advances, the thyroid and cricoid cartilages undergo gradual calcification in an individual. These age-related changes can be visualized in both the living and the dead using conventional radiography and can be objectively assessed. The objective of the present study was to evaluate the age-related changes in the laryngeal cartilages and the results may be utilized for age estimation in forensic examinations. The laryngeal cartilages were carefully dissected using standard procedures from 75 cadavers of age ranging from 17 to 65 years, during the post-mortem examination. The laryngeal cartilages were radiographed and replaced in the body cavity before culminating the post-mortem examination. The calcification of both cartilages was studied by using the standard grading method. Calcification scores of individual regions of both the laryngeal cartilages show a statistically significant positive correlation with chronological age (P < 0.05). Regression models derived from the degree of calcification of thyroid and cricoid cartilages showed standard error of estimates that ranged between 9.90 and 11.07 years. Considering the standard error of estimates of the regression analysis, the present study concludes that these regression models can be used in adjunct with other methods of age estimation such as the skeletal and dental age or when such methods are not viable as in cases of charred or mutilated remains.

A rabbit model to investigate temporomandibular joint osteochondral regeneration

The objective of this study was 2-fold: (1) to describe the rabbit temporomandibular joint (TMJ) anatomy and (2) to provide a detailed, step-by-step description of a minimally invasive approach to perform a standard osteochondral TMJ defect that can be used to investigate the regenerative potential of biomaterials. This study was performed in 2 steps. In the first, a total of 8 rabbit carcasses (n=16 joints) were used to study the normal TMJ anatomy and histology to develop a minimally invasive approach to access the articulating surface of the condyle to perform a standard osteochondral defect. In the second, the surgical procedure was performed in 10 live animals to evaluate the feasibility of the model and to evaluate the regenerative potential of a biodegradable light-cured hydrogel seeded with stem cells (results not shown). The cartilage of the mandibular condyle showed 4 layers: fibrous, proliferative, hypertrophic, and a zone of calcified cartilage. Positive safranin O staining was observed in the cartilage. The mean duration of the procedure (from incision to last stitch) was 35.5 (±9.21) minutes. All animals survived the procedures without any major complications. This animal model represents an easy and nonmorbid surgical approach to rabbit TMJ.

Evaluation of manubriosternal joint fusion and second costal cartilage calcification: Are they useful for estimating advanced age groups?

The objectives of this study were to evaluate the manubriosternal joint (MSJ) to estimate the minimum chronological age of complete fusion, and how changes in this joint influence age estimation. The study also focused on investigating the calcification patterns of the second costal cartilage (SCC), as they may be related to age. Between January and September 2020, individuals between 30 and 80years of age, who underwent chest tomography at our institution, were listed by sex and divided into five groups according to age with intervals of 10years. Twenty patients from each age group and sex were randomly selected, and 200 patients were evaluated. The relationship between age and changes in MSJ and SCC was evaluated. The incidence of complete fusion did not differ significantly between the 30 and 80 age groups. A significant difference was found between the age groups and MSJ degeneration (p < 0.001). The degree of MSJ degeneration and calcification in SCC significantly increased with age, but the range was wide. These values might be used to support traditionally accepted age-estimation methods but not as independent indicators of specific age ranges.

The interface region between articular cartilage and bone by μMRI and PLM at microscopic resolutions.

This dual-modality microscopic imaging study quantifies the interface region between the noncalcified cartilage and the subchondral bone plate, which includes the deep portion of the noncalcified articular cartilage and the zone of calcified cartilage (ZCC). This interface region is typically not visible in routine MRI but becomes visible in MRI with the application of an ultra-short echo time (UTE) sequence. A number of cartilage-bone blocks from a well-documented canine humeral head were harvested for imaging by microscopic MRI (μMRI) and PLM (polarized light microscopy). In μMRI, T2 anisotropic images were acquired by 2D gradient-echo, magnetization-prepared spin-echo and UTE sequences at the 0° and 55° (the magic angle) orientations at 11.7μm/pixel resolution. In PLM, quantitative optical retardation (nm) and collagen orientation (°) were mapped from the thin sections from the same μMRI specimens at 0.5-2μm pixel resolutions. The orientational and organizational architecture of the collagen matrix in this interface region was quantified and correlated between the complementary imaging. The magic angle effect as seen in the noncalcified cartilage was statistically confirmed in ZCC in μMRI, which was further supported by quantitative PLM. With an enhanced understanding of the tissue properties in this important interface region, it will potentially be possible to monitor the changes of this tissue region which is instrumental to the initiation and development of osteoarthritis and other joint diseases.

Sympathectomy aggravates subchondral bone changes during osteoarthritis progression in mice without affecting cartilage degeneration or synovial inflammation

Osteoarthritis (OA) pathogenesis involves the interaction of articular cartilage with surrounding tissues, which are innervated by tyrosine hydroxylase-positive (TH+) sympathetic nerve fibers suggesting a role of the sympathetic nervous system (SNS) during OA progression. We analyzed the effects of sympathectomy (Syx) in a murine OA model. Peripheral Syx was generated by 6-hydroxydopamine (6-OHDA) injections in male C57BL/6 mice. OA was induced in wild-type (WT) and Syx mice by destabilization of the medial meniscus (DMM). TH+fibers and splenic NE were analyzed to evaluate Syx efficiency. OA progression was examined by OARSI and synovitis scores and micro-CT. Expression of TH, α2A- and β2-adrenergic receptors (AR), and activity of osteoblasts (ALP) and osteoclasts (TRAP) was investigated by stainings. Syx resulted in synovial TH+fiber elimination and splenic NE decrease. Cartilage degradation and synovitis after DMM were comparably progressive in both WT and Syx mice. Calcified cartilage (CC) and subchondral bone plate (SCBP) thickness and bone volume fraction (BV/TV) increased in Syx mice due to increased ALP and decreased TRAP activities compared to WT 8 weeks after DMMWT and Syx mice developed osteophytes and meniscal ossicles without any differences between the groups. AR numbers decreased in cartilage but increased in synovium and osteophyte regions after DMM in both WT and Syx mice. Peripheral dampening of SNS activity aggravated OA-specific cartilage calcification and subchondral bone thickening but did not influence cartilage degradation and synovitis. Therefore, SNS might be an attractive target for the development of novel therapeutic strategies for pathologies of the subchondral bone.

Calcification and resorption of mouse Meckel's cartilage analyzed by von Kossa and tartrate-resistant acid phosphatase histochemistry and scanning electron microscopy/energy-dispersive X-ray spectrometry.

Meckel's cartilage is essential for the normal development of the mandible. The fate of the intermediate portion of Meckel's cartilage is unique as most of it disappears soon after birth except for the part that forms the sphenomandibular ligament. The mechanism of the disappearance of Meckel's cartilage is unknown; therefore, this study was designed to investigate the process of Meckel's cartilage degradation, focusing on cartilage matrix calcification and the appearance of chondroclasts. Developing mouse mandibles at embryonic days 15, 16, 17, and 18, and postnatal day 2 were processed for whole-mount staining with alcian blue and alizarin red. The mandibles on embryonic days 15, 16, 17, and 18 were fixed and embedded in paraffin. Adjacent sections were processed for von Kossa and tartrate-resistant acid phosphatase (TRAP) histochemistry and scanning electron microscopy/energy-dispersive X-ray spectrometry (SEM/EDS). Calcification and the element concentrations of calcium, phosphorus, and carbon were examined with von Kossa histochemistry and SEM/EDS. The involvement of chondroclasts was investigated using TRAP histochemistry. The results demonstrated that the intermediate portion of Meckel's cartilage is resorbed by chondroclasts after chondrocyte hypertrophy and cartilage matrix calcification and that the mineral concentration of calcified Meckel's cartilage is comparable to that of the surrounding bone. This study contributes to the understanding of the mechanism of Meckel's cartilage resorption and provides useful insights into the development of the mandible.

Оценка риска развития остеоартроза коленного сустава при протезировании голени (пилотное экспериментально-морфологическое исследование)

Objective was to obtain preliminary data on the knee osteoarthritis risk following canine tibial prosthetics using one-stage osseointegration, external fixation and compression loading. Material and methods The study was carried out on 5 intact (control group) and 3 experimental (experimental group) animals aged 1.8 ± 0.5 years with a body weight of 19 ± 1.2 kg. Osteotomy was performed at the boundary of the upper and middle third of tibia and a PressFit type construct implanted. A special device was used for bone fixation and compression loading of FN = 20 N. Paraffin sections of the articular cartilage and the underlying subchondral bone were used for histomorphometric examination. Results The zonal structure of the articular cartilage and cytoarchitectonics were shown to be maintained in all experimental animals with insignificant destructive changes in the form of impaired homogeneity of the intercellular substance in the upper third of the surface zone. There was a two-fold decrease in the thickness of the calcified cartilage and a 1.9‑fold decrease in the thickness of the subchondral bone. The volumetric density of trabeculae in the subchondral bone decreased to 22.31 ± 5.41 % in experimental animals and to 46.94 ± 1.94 % in controls. Complete absence of calcified cartilage and the subchondral bone were observed in one case with vessels and bone marrow pannus invading the noncalcified cartilage. Conclusion Structural changes in the contact zone of the articular cartilage and the subchondral bone seen in the knee following experimental canine tibial prosthetics indicated the risk of developing knee osteoarthritis.

Computed tomography examination of the os cordis in a lamb (Ovis aries Linnaeus, 1758)

Objective. The present paper aims to investigate the role of computed tomography as an imaging technique of diagnosis in the identification of the os cordis in ovine, and also how this anatomical structure is morphotopographically characterized in macro and microscopic contexts, seeking to contribute for its functional understanding. Materials and method. The heart of a young male ovine had been donated to the Laboratory of Animal Anatomy of the Surgery Department of the FMVZ/USP, first being subject to a post-mortem examination by means of the cardiac “shedding” (transverse cross sectioning of the heart). A tomographic examination of the anatomic specimen was carried out, as well as the dissection and histological analysis of the collected sample. Results. The results indicate the presence of an osseous structure of 6.39 mm in length, located in the aortic valve of the heart, next to a ring comprised by three valve flaps, tendinous cords, and papillary muscles. The histological findings consist of fibrous connective tissue, cancellous bone tissue, and calcified hyaline cartilage wherein the cardiomyocytes are attached. Conclusions. It is concluded that computed tomography, even though seldom applied to animals of zootechnical interest, poses as an effective tool for the visualization of the os cordis in lambs.

3D Printed Multiphasic Scaffolds for Osteochondral Repair: Challenges and Opportunities.

Osteochondral (OC) defects are debilitating joint injuries characterized by the loss of full thickness articular cartilage along with the underlying calcified cartilage through to the subchondral bone. While current surgical treatments can provide some relief from pain, none can fully repair all the components of the OC unit and restore its native function. Engineering OC tissue is challenging due to the presence of the three distinct tissue regions. Recent advances in additive manufacturing provide unprecedented control over the internal microstructure of bioscaffolds, the patterning of growth factors and the encapsulation of potentially regenerative cells. These developments are ushering in a new paradigm of ‘multiphasic’ scaffold designs in which the optimal micro-environment for each tissue region is individually crafted. Although the adoption of these techniques provides new opportunities in OC research, it also introduces challenges, such as creating tissue interfaces, integrating multiple fabrication techniques and co-culturing different cells within the same construct. This review captures the considerations and capabilities in developing 3D printed OC scaffolds, including materials, fabrication techniques, mechanical function, biological components and design.

Biomechanical Aspects of Osteochondral Regeneration: Implications and Strategies for Three-Dimensional Bioprinting.

Osteoarthritis is among the most prevalent of musculoskeletal disorders in the world that causes joint pain, deformity, and limited range of movement. The resulting osteochondral defect can significantly decrease the patient's quality of life, but current treatment options have not demonstrated the capacity to fully regenerate the entire osteochondral microenvironment. Structurally, the osteochondral unit is a composite system composed of three layers-articular cartilage, calcified cartilage, and subchondral bone. Collectively these distinct layers contribute to the distinct biomechanical properties that maintain the health and aid in load transfer during joint articulation. The purpose of this review was to examine the role of the osteochondral interface in tissue engineering. Topics of discussion include the biomechanics of the osteochondral unit and an overview of various strategies for osteochondral interface tissue engineering, with a specific focus on three-dimensional bioprinting. The goal of this review was to elucidate the importance of the osteochondral interface and overview some strategies of developing an interface layer within tissue engineered scaffolds. Impact Statement This review provides an overview of interface tissue engineering for osteochondral regeneration. It offers a detailed investigation into the biomechanics of the osteochondral unit as it relates to tissue engineering, and highlights the strategies that have been utilized to develop the osteochondral interface within tissue engineering scaffolds.

Noninvasive Detection of Arytenoid Cartilage Calcification Using Computed Tomography and Prediction of Prognosis in Laryngeal Contact Granuloma

Laryngeal contact granuloma (LCG) is a relatively uncommon disease with chronic inflammatory stimulation, and long-term reflux irritation is a vital factor for arytenoid cartilage calcification. Our investigation compared the severity of ipsilateral arytenoid cartilage calcification with the frequency of recurrence of LCG after surgical treatment. A retrospective chart review of prospectively gathered data over five years from 327 patients, including 153 subjects without laryngeal lesions, were age- and sex-matched normal controls, 103 patients with various other vocal cord lesions were in the laryngeal lesion group and 71 LCG patients met the diagnostic criteria pathologically. All subjects underwent laryngeal high-resolution computed tomography (HRCT) prior to therapeutic interventions. The computed tomography (CT) value and arytenoid cartilage calcification were obtained using image data before surgery, and their clinical significance was further analyzed. Seventy-one patients with LCG, including sixty-two males, were enrolled in the study. Among these cases, there were 67 patients with unilateral vocal cord lesions. Of the 103 eligible patients in the laryngeal lesion group, 87 had unilateral lesions, which including eighty-seven men. Of the 153 average subjects, 105 were male. The rate of arytenoid cartilage calcification in the LCG group was dramatically higher in the lesion side than in the laryngeal lesions and normal group (P < 0.01). Furthermore, the CT value (P < 0.01) and range of calcification (P < 0.01) were significantly higher in patients with LCG than in those with laryngeal lesions. Importantly, patients with high CT values and the calcification range of lesions in the arytenoid cartilage displayed a greater lesion size and recurrence rate than patients with low CT values and lesion areas (P < 0.01). Our results suggest that most patients with LCG present with calcification of the arytenoid cartilage. The more severe the calcification in the arytenoid cartilage, the greater the risk of granuloma size and recurrence in LCG after surgical treatment. CT and bone density testing of the arytenoid cartilage may be an essential method to evaluate the prognosis of LCG.

Costal Cartilage Calcification: Prevalence, Amount, and Structural Pattern in the General Population and Its Association with Age: A Cadaveric Study.

Importance: Costal cartilage calcification (CCC) of the cartilage graft, commonly used in reconstruction of nasal/auricular deformities, can cause poor surgical outcome, but structural and quantitative analyses are lacking. Objective: To compare the prevalence, amount, and structural pattern of CCC from individuals by gender and age, as measured by digital contact radiography. Design, Setting, and Participants: This is a cross-sectional cadaveric study (n = 92) of the seventh rib cartilage. CCC prevalence/amount/structural pattern (central [c]/peripheral [p]/diffuse [d]) was analyzed within three age groups: I (<40 years), II (40-70 years), and III (>70 years). Main Outcomes and Measures: Qualitative and quantitative CCC analyses were set in relation to gender/structural pattern/age. Results: CCC prevalence was gender independent (96.7%) and occurred in c/p/d: 12.4%/22.4%/65.2%. Structural CCC pattern differed between age groups (I: 80%; c/p/d: 26.7%/46.6%/6.7%; II: 100%; c/p/d: 18.0%/30.8%/51.2%; III: 100%; p/d: 2.6%/97.4%). The mean CCC amount (9.1%) was gender independent and showed a significant correlation with age (p = 0.001). The mean amount showed a significant difference between the structural pattern [d/p: 3.3 times higher (p = 0.006), d/c: 7.7 times higher (p < 0.001)] and age groups (I/II/III: 6.4/8.5%/10.9%), whereby the amount was higher in groups II (factor: 7.4; p < 0.001) and III (factor: 16.5; p < 0.001) compared with group I. Conclusions: These data show an age-/gender-independent high CCC prevalence in the general population (96.7%). CCC already occurred in young donors (<40 years) with a not negligible amount (6.4%). CCC increased with age and structural analysis showed a gender-/age-specific pattern, whereby males were prone to peripheral/females to central CCC. Diffuse CCC was observed as an age-independent sign for high CCC levels.

Therapeutic Role of Platelet Rich Plasma in Formaldehyde-Induced Arthritis in Adult Male Albino Rats

Abstract Introduction Osteoarthritis (OA) is a common health problem. Platelet-rich plasma (PRP) has been recognized to enhance articular cartilage metabolism. Aim of the work the study was designed to investigate the influence of PRP on cartilage healing after induction of arthritis. Material and methods Forty two adult male albino rats were used in this study. The rats were randomly divided into three groups: Group I (n = 18): the control group (Ia, Ib &amp; Ic) Ib &amp; Ic were injected intra-articularly with saline and left for 3 and 6 weeks. Group II (n = 12): arthritic group, in which osteoarthritis was induced by injection of 0.02ml 5% formaldehyde once in the right knee joints, left without treatment, and were sacrificed after three weeks (IIa) or after six weeks (IIb). Group III(n = 12): arthritis was induced as group II, one week later, the rats were intra-articularly injected with single dose of 0.3ml PRP in the same joint then were sacrificed three weeks (IIIa) or six weeks (IIIb) after formaldehyde injection. At the end of the study the right knee joints were taken, decalcified then processed for paraffin sections to be examined by light microscope using H&amp;E, toluidine blue and Masson’s trichrome (MTC) stains. Immunohistochemistry for caspase-3 enzyme was done to demonstrate apoptotic chondrocytes. Morphometric study was conducted to measure the thickness of the non-calcified cartilage, count the chondrocytes and synovial membrane inflammatory cells and Mankin's score. Then statistical analysis was done. Results The arthritic groups revealed irregular surface of the articular cartilage, loss of the articular matrix and bone eburnation. Moreover, there was apparent hypocellularity and disorganization of the chondrocytes. Osteoclasts and osteoblasts were seen invading the osteochondral junction. MTC stained sections of the synovial membrane showed deposition of thick collagen bundles with heavy inflammatory cell infiltrate and numerous blood vessels. The affinity of the articular cartilage to toluidine blue stain was apparently decreased while caspase-3 immunoreactivity was apparent in many chondrocytes .Group IIIa demonstrated almost similar histological findings as the control group; regular articular cartilage surface with regularly arranged chondrocytes in the different cartilage zones. Synovial membrane illustrated minimal inflammatory cell infiltrate with thin collagen bundles and small blood vessels in MTC stained sections. There was high affinity of the articular cartilage to the toluidine blue stain and few chondrocytes showed positive caspase-3 immunoreactivity. Group IIIb revealed continuous surface of the articular cartilage, yet with minimal fibrillation in some areas. Osteoblasts and osteoclasts were seen invading the calcified cartilage. Synovial membrane showed deposition of dense collagen bundles with some inflammatory cell infiltrate. Toluidine blue sections revealed decreased articular cartilage affinity to the stain while caspase-3 immunoreactivity was evident in many chondrocytes. The morphometric results and statistical analysis confirmed the histological findings. Conclusion Intra-articular injection of PRP demonstrated advantageous role on articular cartilage healing, however, these effects appeared to be transient. So the need of multiple injections of PRP has to be considered in cases of OA.

Role of Laryngeal Ultrasonography in screening for laryngeal diseases in symptomatic patients

Abstract Background Laryngoscopy is widely considered the most appropriate tool for evaluating the larynx, however, it is invasive and not all patients can tolerate it. Although laryngeal ultrasound has been proposed as a promising noninvasive technique that can play an important role in laryngeal imaging, there is lack of consensus regarding its role. Objective The aim of this study is to assess laryngeal ultrasound sensitivity compared to indirect laryngoscopy in screening for laryngeal diseases in symptomatic patients. Patients and methods 30 undiagnosed symptomatic patients underwent laryngeal ultrasound and indirect laryngoscopy. Their results were compared. Results Technical success rate of laryngeal ultrasound was (83%). It showed overall sensitivity of (78.9%) in detecting organic laryngeal diseases with high sensitivity in detecting laryngeal masses, reinke’s edema, vocal cord cysts and polyps (100%), vocal cord palsy (83.3%), intermediate sensitivity for vocal cord nodules (66.7%), and poor sensitivity for laryngitis and non organic voice disorders (0%). Conclusion Laryngeal ultrasound is a useful non invasive imaging tool that can be used with acceptable sensitivity in detecting organic laryngeal diseases excluding laryngitis, hindered mainly by complete thyroid cartilage calcification in older males. Consequently we recommend its use as complementary to clinical examination and mirror laryngoscopy, or as an alternative to indirect laryngoscopy when required and in routine neck ultrasound exams when appropriate.

Growth factors in the initial stage of bone formation, analysis of their expression in chondrocytes from epiphyseal cartilage of rat costochondral junction.

In endochondral ossification septoclasts and osteoclasts (also called chondroclasts) release growth factors deposited in non-calcified and calcified zones of the growth plate. They stimulate, within the metaphysis, initial stages of the bone formation. We have recently reported quantitation of several growth factors in calcified cartilage from calf costochondral junction. Data from the analogous human cartilage could possibly help to choose efficient combinations of growth factors for clinical applications, but the amount of the calcified cartilage needed for analysis of numerous growth factors would be difficult to collect. The estimation of growth factors expression in endochondral chondrocytes may, indirectly, indicate which of them play a leading role in the stimulation of osteoprogenitor cells in metaphysis. To test this hypothesis, we used rat chondrocytes to evaluate mRNA levels of several growth factors. Chondrocytes were isolated from proliferative and hypertrophic zones of the epiphyseal cartilage forming costochondral junctions of inbred Lewis rats. The total RNA was isolated from chondrocytes and the level of mRNA for bone morphogenetic proteins 1-7 (BMP-1-7), vascular endothelial growth factor A (VEGF-A), basic fibroblast growth factor (bFGF), growth/differentiation factor 5 (GDF-5), NEL-like protein 1 (NELL-1), transforming growth factor beta 1 (TGF-b1), mesencephalic astrocyte-derived neurotrophic factor (MANF), connective tissue growth factor (CTGF), osteoclast-stimulating factor 1 (OSTF-1) and insulin-like growth factor 1 (IGF-1) was evaluated using real-time PCR method. All studied factors were expressed. The highest level of mRNA was detected for CTGF, MANF, VEGF-A and TGF-b1. Expression was also quite high for BMP-1, BMP-2, BMP-5, BMP-6, BMP-7, IGF-1, GDF-5 and OSTF-1. Very low level of mRNA was detected for BMP-3, BMP-4 and NELL-1. Chondrocytes from the proliferative and hypertrophic zones of the growth plate produce factors involved in the cartilage metabolism and bone formation. The determination of these growth factors in humans could help to choose their optimal composition necessary for stimulation of bone formation in clinical practice. In rat the best stimulation of bone formation would presumably be achieved with a mixture of BMP-2, BMP-5, BMP-6 and BMP-7.