Tibial Growth Research Articles

Boron Supplementation Promotes Osteogenesis of Tibia by Regulating the Bone Morphogenetic Protein-2 Expression in African Ostrich Chicks.

The present study aimed to explore the effects of supplemental boron on osteogenesis of tibia and to investigate the possible relationship between additional boron and the expression of bone morphogenetic protein-2 (BMP-2) in tibia of ostrich chicks. Therefore, forty-eight African ostrich chicks (15days old) were supplemented with 0mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L, and 640mg/L of boron in drinking water for 75days. The paraffin sections of tibia used to measure histomorphometric parameters by hematoxylin and eosin (HE) staining, Masson's staining, and immunohistochemistry (IHC). Enzyme-linked immunosorbent assay was performed to assess the level of BMP-2, osteocalcin (BGP), glucocorticoids (GCs), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-B ligand (RANKL) in serum. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) technique was performed to detect the cell apoptosis. The results indicated that low dose of supplemental boron (40mg/L-160mg/L) in drinking water promotes bone development by increasing the mature ossein. The expression of BMP2 on 45days was higher than 90days. Serum level of BMP-2, BGP, and GCs changed significantly in groups with low dosage of boron, and OPG/RANKL ratio was upregulated from 0 to 160mg/L. Cell apoptosis was least in 40mg/L and 160mg/L groups. Taken together, low dose of boron supplemented in drinking water could promote osteogenesis and growth and development of tibia by regulating the expression and secretion of BMP-2 and providing a dynamically balanced environment for tibia growth, development, and reconstruction by regulating the concentrations of BGP, GCs, and OPG/RANKL ratio in serum.

Risk Factors for Arthrofibrosis in Tibial Spine Fractures: A National 10-Site Multicenter Study

Objectives:To identify risk factors for developing arthrofibrosis after TSF treatment.Methods:This was a retrospective, multi-center study across ten institutions of 448 patients <25 years old who presented to care with a tibial spine fracture between 1/2000 and 2/2019. Patient records were reviewed for a multitude of pre-operative, intra-operative, and post-operative characteristics. Patients were then separated into two cohorts based on if they suffered from post-treatment arthrofibrosis.Results:Chart review demonstrated that 43 (9.6%) of the patients suffered from post-treatment arthrofibrosis. There were no demographic differences observed between the two groups. However, based on MRIs at the time of injury, distal femoral and proximal tibial growth plates were more frequently closed in the arthrofibrosis group (17.6% vs 4.4%, p=0.023 for both comparisons). Additionally, there was no difference in Meyers & McKeever (MM) classification (p=0.597). All arthrofibrosis patients received operative treatment (p=0.003), though there was no difference in fixation technique (p=0.734). Intraoperatively, a higher number of screws were used in the arthrofibrosis group (p=0.002) with the placement of hardware more likely to be epiphyseal (p=0.007). Other operative parameters including number of sutures were not different. Post-operatively, arthrofibrosis patients were more likely to have been immobilized in a cast (p<0.001) with no difference observed for weight-bearing status. After multivariate regression, screw number (OR 8.9, CI 1.9-41.7, p=0.005) and immobilization in a cast (OR 7.8, CI 1.0-60.4, p=0.049) remained significant predictors of post-treatment arthrofibrosis.Conclusion:This serves as the largest study of tibial spine fractures to analyze risk factors for the development of post-treatment arthrofibrosis. Our study demonstrates that pre-operative factors were largely similar between groups, but that intra-operative decisions, including the number of screws used for fixation and placement of hardware in relation to the physis, were significant predictors of post-treatment arthrofibrosis. These findings may influence operative decision-making in tibial spine fracture patients. Additionally, post-operative immobilization in a cast should be avoided given the high risk of arthrofibrosis.Table 1.Characteristics of patients with post-operative arthrofibrosis Arthrofibrosis (N=43) No Arthrofibrosis (N=405) P-value Demographic Age (years)11.112.00.065Male: Female ration24:19282:1230.064BMI20.220.40.824 Injury Sports-related (%)13 (30.2)205 (50.4) 0.012 MM classification III/IV20 (64.5)134 (59.6)0.597Concomitant ACL injury (%)5 (22.7)46 (21.6)0.902Delay to surgery (days)24.224.30.989 Operative Received surgery (%)43 (100.0)325 (78.5) 0.003 Operative time (min)142.0123.90.209Treatment type (%) ARIF36 (83.7)269 (82.8)0.734ORIF7 (16.3)45 (13.8) Screws (%)22 (51.2)114 (35.5) 0.046 Number of Screws1.81.4 0.002 Screw/Suture Placement (%) 0.005 Transphyseal5 (17.9)94 (44.5) 0.007 Epiphyseal23 (82.1)117 (55.5) Post-operative 41 (97.6)305 (97.8) Immobilized21 (51.2)72 (23.7)0.955Cast immobilization16 (39.0)208 (68.2) <0.001 Non-weightbearing11 (34.4)72 (25.4)0.277

EFFECT OF ALOE VERA ON GROWTH OF RAT PUPS

ABSTRACT: &#x0D; Aim and Objective: Widely and commonly grown plant, Aloe vera has numerous health benefits. The breast fed neonates of mothers consuming the plant as dietary supplement are exposed to its effects. The objective of the research was to evaluate the effects of Aloe vera extract on growth of rat pups. Study design: Experimental study. Place of study: Anatomy department of Postgraduate Medical Institute, Lahore. Material and Method: A week old Wistar albino rats were fed with extracts of Aloe vera (low dose 50mg/kg b.w and high dose 500mg/kg b.w) daily for ten days through oral gavage. The weight of the pups was recorded and after euthanasia the lower limb region was dissected and tibia were removed. The length and weight of the bone was noted. Results: Pups fed with a high dose of Aloe vera extract showed a significant increase in their body mass (p&lt;0.05) and also demonstrated a significant increase in the length of tibia (p&lt;0.001). Conclusion: 500mg/kg b.w of Aloe vera extract resulted in a significant increase in the body weight and promoted the tibial bone growth.&#x0D;

Lactobacillus rhamnosus JYLR-005 Prevents Thiram-Induced Tibial Dyschondroplasia by Enhancing Bone-Related Growth Performance in Chickens.

Tibial dyschondroplasia (TD) is a leg disorder caused by the abnormal development of the tibia in fast-growing poultry. Lactobacillus rhamnosus (L. rhamnosus) strains have been reported to have effects on increasing bone growth and improving osteoporosis in animals. However, whether L. rhamnosus JYLR-005 can improve bone growth in TD chickens remains unclear. In this study, we noted that L. rhamnosus JYLR-005 could not reduce the suppression of the production performance of TD broilers (p > 0.05) but had a slight protective effect on the broiler survival rate (χ2 = 5.571, p = 0.062). However, for thiram-induced TD broiler chickens, L. rhamnosus JYLR-005 could promote tibia growth by increasing tibia-related parameters, including the tibia weight (day 11, p = 0.040), tibia length (day 15, p = 0.013), and tibia mean diameter (day 15, p = 0.035). Moreover, L. rhamnosus JYLR-005 supplementation improved the normal growth and development of the tibial growth plate by maintaining the morphological structure of the chondrocytes and restored the balance of calcium and phosphorus. Taken together, these findings provide a proof of principle that L. rhamnosus JYLR-005 may represent a therapeutic strategy to treat leg disease in chickens.

Human Fetal Growth and Age Estimation Based on Osteometry on the Diaphysis of Fetal Tibia

In dealing with fetal growth, earlier studies carried out on chemically preserved fetuses with very little sample size and measured only the length of fetal long bones. The present study is a pioneer one, taken on naturally macerated fetal tibia bones. There have been six new osteometric measurements introduced on human fetal tibia. Maximum possible osteometric measurements have been taken on a record number of 912 fetal tibia bones from 456 fetuses having age range between 11 weeks to 40 weeks of intrauterine life. Bilateral and bisexual differences have been analyzed. Growth of the fetal tibia has been analyzed, based on its osteometry. The fastest rate of growth is observed in the length of the shaft of fetal tibia in females during 11 weeks to 16 weeks of the intrauterine life of human fetus. For every 1 mm increase in the CRL, the Maximum Length of Tibia (t-ml) is increased by .246 mm in females. Regression values have been calculated to estimate human fetal age from the osteometric measurements on fetal tibia. The age estimation is a crucial factor in dealing with medico-legal cases.

A new standard radiographic reference for proximal fibular height in children

Background and purpose — To date there is a lack of studies defining the anatomical position of the proximal fibula. This is especially relevant when planning surgical interventions affecting the knee joint such as permanent or temporary epiphysiodesis to correct leg length discrepancies or angular deformities in growing patients. The goal of this study is to establish a standardized measurement technique and radiological reference values for the position of the proximal fibula in children.Patients and methods — 500 measurements were performed in calibrated long standing anteroposterior radiographs of 256 skeletally immature patients (8–16 years; 233 female, 267 male legs). As a radiographic reference in the frontal plane, the distance between the center of the proximal tibial growth plate and a line tangential to the tip of the fibular head and horizontal to the imaging plane was measured (dPTFH).Results — The average value of dPTFH in the studied population (median age 12 years) was –2.7 mm (SD 3, CI –3.0 to –2.5) and normally distributed (p = 0.1). There were no clinically significant sex or age-dependent differences. The inter-rater reliability analysis showed excellent ICC values (ICC = 0.88; CI 0.77–0.93).Interpretation — This study provides a new radiographic reference value to assess the position of the proximal fibula in relation to the proximal tibia in children and adolescents. This reference can aid preoperative decision-making as to whether additional fibular epiphysiodesis is necessary when performing tibial epiphysiodesis to correct moderate leg-length discrepancies.

ARTICULAR CARTILAGE THICKNESS IN PEDIATRIC AND ADOLESCENT KNEES DECREASES WITH AGE AND SKELETAL MATURITY: IMPLICATIONS FOR OSTEOCHONDRAL GRAFTING PROCEDURES

Background:Quantifying native cartilage thickness in pediatric and adolescent knees can help match donor and recipient sites for articular cartilage restoration procedures such as osteochondral autograft transplantation (OATS) and osteochondral allograft transplantation (OCA).Hypothesis/Purpose:The purpose of the current study was to quantify articular cartilage thickness in pediatric and adolescent knees using magnetic resonance imaging (MRI). We hypothesized that cartilage thickness is inversely correlated with skeletal maturity and age.Methods:One hundred and twenty MRI scans were evaluated in a cohort of patients 9 to 18 years old without osteochondral lesions, chondral wear or pathology, intraarticular fractures, history of knee surgery, or inflammatory arthropathy. Measurements of articular cartilage thickness at the medial femoral condyle, lateral femoral condyle, lateral trochlea, and patella were made on axial, coronal, and sagittal MRI scans (Figure 1). Skeletal maturity was categorized as ‘open’, ‘closing’, or ‘closed’ based on the status of the proximal tibial and distal femoral growth plates. Descriptive statistics was used to evaluate cartilage thickness by age and sex. Independent samples t-test, analysis of variance (ANOVA), and linear regression were performed to investigate for associations with sex, skeletal maturity, and age.Results:On the femur, cartilage was thickest at the lateral trochlea with mean articular thickness of 4.2 ± 1.4 mm in males and 3.6 ± 1.3 mm in females (p=0.015) (Table 1). Skeletally immature patients with open physes had significantly thicker cartilage at the medial femoral condyle, lateral femoral condyle, and lateral trochlea compared to patients with closing and closed physes (Figure 2). Linear regression analysis also revealed a significant association between femoral cartilage thickness and age (Figure 3). Age explained 63% of the variance at the medial femoral condyle (B=6.1, p<0.001), 64% of the variance at the lateral femoral condyle (B=4.9, p<0.001), and 68% of the variance at the lateral trochlea (B=8.2, p<0.001) (Table 2). In contrast, cartilage thickness at the patella did not significantly vary by age, sex, or skeletal maturity (Figures 2 and 4).Conclusion:There is a strong inverse association between increasing age and cartilage thickness of the femoral condyles and lateral trochlea. In particular, pediatric knees demonstrate relatively thick cartilage at the lateral trochlea that decreases with age. This information will help surgeons understand recipient site anatomy and identify appropriate donor site tissue for articular cartilage restoration procedures such as OATS and OCA in children and adolescents.Tables:Table 1.Mean articular cartilage thickness of the femur and patellaCombined (n=120)Male(n=40)Female(n=60) p valueMedial femoral condyle3.2 ± 1.03.5 ± 1.02.9 ± 0.80.001Lateral femoral condyle2.8 ± 0.72.9 ± 0.72.7 ± 0.70.056Lateral trochlea3.9 ± 1.44.2 ± 1.43.6 ± 1.30.015Median ridge of patella4.2 ± 0.94.4 ± 0.94.1 ± 0.90.080Medial facet of patella4.7 ± 1.04.8 ± 1.04.6 ± 0.90.320Lateral facet of patella4.6 ± 0.84.7 ± 0.84.5 ± 0.80.185Table 2.Linear regression of articular cartilage thickness of the femur and patella by ageR2 B p valueMedial femoral condyle0.636.1<0.001Lateral femoral condyle0.644.9<0.001Lateral trochlea0.688.2<0.001Median ridge of patella0.024.80.090Medial facet of patella0.045.50.032Lateral facet of patella0.0014.50.741Figures:Figure 1.Measurements of patellofemoral articular cartilage thickness Measurements of articular cartilage thickness were made on (A) coronal, (B, C) sagittal, and (D) axial magnetic resonance imaging (MRI) of the knee. For the medial femoral condyle, cartilage thickness was measured at five regions: midpoint of the medial condyle (MFC) and midpoints of Cahill zones 1 (MID1), 2 (MID2), B (MIDB) and C (MIDC). Similarly, measurements of cartilage thickness of the lateral femoral condyle were made at the midpoint of the lateral condyle (LFC) and midpoints of Cahill zones 4 (MID4), 5 (MID5), B (MIDB), and C (MIDC). Measurement of the lateral trochlea was made at midpoint of Cahill zone A (MIDA). Measurements on the patella were made at the midpoint of the medial facet (MF), midpoint of the lateral facet (LF), and the median ridge (MR) on axial and sagittal MRI scans. Figure 2.Mean articular cartilage thickness of the femur and patella by growth plate statusFigure 3.Mean articular cartilage thickness of the femur by ageFigure 4.Mean articular cartilage thickness of the patella by age

RISK FACTORS FOR ARTHROFIBROSIS IN TIBIAL SPINE FRACTURES: A NATIONAL 10-SITE MULTICENTER STUDY

Background:Tibial spine fractures are common pediatric injuries with similar mechanism of injury to anterior cruciate ligament tears. Post-operative arthrofibrosis remains the most common complication following treatment of this injury, and many patients require subsequent manipulation under anesthesia. Several prior studies have examined risk factors for the development of arthrofibrosis, but with small patient populations and varying reported predictors. Therefore, our objective was to identify risk factors for arthrofibrosis in the largest known cohort of pediatric tibial spine patients.Hypothesis/Purpose:Identify risk factors for developing arthrofibrosis after TSF treatment.Methods:This was a retrospective, multi-center study across ten institutions of 448 patients <25 years old who presented to care with a tibial spine fracture between 1/2000 and 2/2019. Patient records were reviewed for a multitude of pre-operative, intra-operative, and post-operative characteristics. Patients were then separated into two cohorts based on if they suffered from post-treatment arthrofibrosis.Results:Chart review demonstrated that 43 (9.6%) of the patients suffered from post-treatment arthrofibrosis. There were no demographic differences observed between the two groups. However, based on MRIs at the time of injury, distal femoral and proximal tibial growth plates were more frequently closed in the arthrofibrosis group (17.6% vs 4.4%, p=0.023 for both comparisons). Additionally, there was no difference in Meyers & McKeever (MM) classification (p=0.597). All arthrofibrosis patients received operative treatment (p=0.003), though there was no difference in fixation technique (p=0.734). Intraoperatively, a higher number of screws were used in the arthrofibrosis group (p=0.002) with the placement of hardware more likely to be epiphyseal (p=0.007). Other operative parameters including number of sutures were not different. Post-operatively, arthrofibrosis patients were more likely to have been immobilized in a cast (p<0.001) with no difference observed for weight-bearing status. After multivariate regression, screw number (OR 8.9, CI 1.9-41.7, p=0.005) and immobilization in a cast (OR 7.8, CI 1.0-60.4, p=0.049) remained significant predictors of post-treatment arthrofibrosis.Conclusion:This serves as the largest study of tibial spine fractures to analyze risk factors for the development of post-treatment arthrofibrosis. Our study demonstrates that pre-operative factors were largely similar between groups, but that intra-operative decisions, including the number of screws used for fixation and placement of hardware in relation to the physis, were significant predictors of post-treatment arthrofibrosis. These findings may influence operative decision-making in tibial spine fracture patients. Additionally, post-operative immobilization in a cast should be avoided given the high risk of arthrofibrosis.Table 1.Characteristics of patients with post-operative arthrofibrosis Arthrofibrosis (N=43)No Arthrofibrosis (N=405)P-valueDemographicAge (years)Male: Female ratioBMI11.124:1920.212.0282:12320.40.0650.0640.824InjurySports-related (%)MM classification III/IVConcomitant ACL injury (%)Delay to surgery (days)13 (30.2)20 (64.5)5 (22.7)24.2205 (50.4)134 (59.6)46 (21.6)24.3 0.0120.5970.9020.989OperativeReceived surgery (%)Operative time (min)Treatment type (%)ARIFORIFScrews (%)Number of ScrewsScrew/Suture Placement (%)TransphysealEpiphyseal43 (100.0)142.0 36 (83.7)7 (16.3)22 (51.2)1.8 5 (17.9)23 (82.1)325 (78.5)123.9 269 (82.8)45 (13.8)114 (35.5)1.4 94 (44.5)117 (55.5) 0.0030.209 0.734 0.0460.0020.0050.007Post-operativeImmobilizedCast immobilizationNon-weightbearing41 (97.6)21 (51.2)16 (39.0)11 (34.4)305 (97.8)72 (23.7)208 (68.2)72 (25.4) 0.955<0.0010.277

Intermittent hypoxia retards mandibular growth and alters RANKL expression in adolescent and juvenile rats.

Chronic intermittent hypoxia (IH), a common state experienced in obstructive sleep apnoea (OSA), retards mandibular growth in adolescent rats. The aim of this study was to elucidate the differential effects of IH on mandibular growth in different growth stages. Three-week-old (juvenile stage) and 7-week-old (adolescent stage) male Sprague-Dawley rats underwent IH for 3 weeks. Age-matched control rats were exposed to room air. Mandibular growth was evaluated by radiograph analysis, micro-computed tomography, real-time polymerase chain reaction and immunohistology. Tibial growth was evaluated as an index of systemic skeletal growth. IH had no significant impact on the general growth of either the juvenile or adolescent rats. However, it significantly decreased the total mandibular length and the posterior corpus length of the mandible in the adolescent rats and the anterior corpus length in the juvenile rats. IH also increased bone mineral density (BMD) of the condylar head in adolescent rats but did not affect the BMD of the tibia. Immunohistological analysis showed that the expression level of receptor activation of nuclear factor-κB ligand significantly decreased (in contrast to its messenger ribonucleicacid level) in the condylar head of adolescent rats with IH, while the number of osteoprotegerin-positive cells was comparable in the mandibles of adolescent IH rats and control rats. The animal model could not simulate the pathological conditions of OSA completely and there were differences in bone growth between humans and rodents. These results suggest that the susceptibility of mandibular growth retardation to IH depends on the growth stage of the rats.

Effect of fibrous diet and vitamin C inclusion on uniformity, carcass traits, skeletal strength, and behavior of broiler breeder pullets

This experiment studied the effect of broiler breeder nutritional strategies on uniformity, carcass traits, tibia parameters, and behavior during rearing and prebreeder periods (up to 22 wk of age). One-day-old pullets (n = 384) were randomly assigned to 4 treatments arranged as a 2 × 2 factorial, with 2 fiber levels (control vs. fibrous diet, 15% diluted in AMEn and nutrient content) and 2 vitamin C feed inclusions (0 vs. 200 mg/kg). At 6, 15, and 22 wk, blood sampling was carried out (4 birds/replicate) to determine serum alkaline phosphatase (ALP) levels, and behavior was observed by visual scan sampling. At 22 wk, carcass traits, tibia parameters, and intestinal morphology were assessed (2 birds/replicate), and tail- and wing-feather integrity of all birds were scored. Fibrous diet did not modify BW uniformity, mortality, or tibia growth when compared with control diet. Pullets fed the fibrous diet had lower tibia breaking strength, elastic modulus, and ash content values (P < 0.05). They also had lower ALP serum level at 6 and 22 wk (P < 0.05), their breast muscle was less developed (18.5 vs. 19.8%, P < 0.05), and their abdominal fat deposition was higher (1.14 vs. 0.87%, P < 0.05). At 15 and 22 wk, they performed, on average, 97% less grasping feather pecking and 45% less non–food object pecking behaviors, and their wing-feather score was lower (P < 0.05) at 22 wk. Tail- and wing-feather scores of the control treatments were reduced by vitamin C inclusion (tail: 0.30 vs. 1.15, P < 0.05; wing: 0.98 vs. 1.26, P < 0.05) at 22 wk. In conclusion, fibrous diet improves carcass traits (reduces breast muscle and increases abdominal fat deposition), deteriorates bone mineral deposition and thus skeletal strength, and reduces stereotypic behaviors, improving wing-feather integrity. Vitamin C inclusion improves tail- and wing-feather integrity of lower in feed allowance.

Discoidin domain receptor 1 regulates endochondral ossification through terminal differentiation of chondrocytes.

Chondrocytes in growth plates are responsible for longitudinal growth in long bones during endochondral ossification. Discoidin domain receptor 1 (Ddr1) is expressed in chondrocytes, but the molecular mechanisms by which DDR1 regulates chondrocyte behaviors during the endochondral ossification process remain undefined. To elucidate Ddr1-mediate chondrocyte functions, we generated chondrocyte-specific Ddr1 knockout (CKOΔDdr1) mice in this study. The CKOΔDdr1 mice showed delayed development of the secondary ossification center and increased growth plate length in the hind limbs. In the tibial growth plate in CKOΔDdr1 mice, chondrocyte proliferation was reduced in the proliferation zone, and remarkable downregulation of Ihh, MMP13, and Col-X expression in chondrocytes resulted in decreased terminal differentiation in the hypertrophic zone. Furthermore, apoptotic chondrocytes were reduced in the growth plates of CKOΔDdr1 mice. We concluded that chondrocytes with Ddr1 knockout exhibit decreased proliferation, terminal differentiation, and apoptosis in growth plates, which delays endochondral ossification and results in short stature. We also demonstrated that Ddr1 regulates the Ihh/Gli1/Gli2/Col-X pathway to regulate chondrocyte terminal differentiation. These results indicate that Ddr1 is required for chondrocytes to regulate endochondral ossification in skeletal development.

Analysis of Association between Morphometric Parameters of Growth Plate and Bone Growth of Tibia in Mice and Humans.

The purposes of this study are to evaluate which growth plate parameters are associated with bone growth in mice and to compare the mouse results with those in humans. The sagittal sections of the proximal growth plate of the mouse tibia from neonate to young adult stages were subjected to histomorphometric and functional analyses. The radiographic images of tibias of human patients until puberty were analyzed to obtain the tibia length and the proximal growth plate height. It was found that a linear correlation best modeled the relationship between the growth plate variables with the tibia growth rate and length. In mice, total height, resting zone height, combined height of the proliferation and prehypertrophic zones, proliferation activity, and the total width of tibia growth plate showed high linear correlation with tibia bone length and bone growth rate, but the hypertrophic zone height and the growth plate area did not. In both mice and humans, the total growth plate width of tibia was found to have the strongest correlation with tibia length and growth rate. The results validated that growth plate total height, the height of the resting zone and cell proliferation activity are appropriate parameters to evaluate the balance between growth plate activity and bone growth in mice, consistent with previous reports. The study also provided a new growth plate parameter candidate, growth plate width for growth plate activity evaluation in both mouse and human tibia bone.

The effects of experimentally irradiated pituitary gland on the growth of rat’s tibia, skull, maxilla and mandible

Background/Aim. The reaction of an organism to radiation depends on the level of irradiation and the sensitivity of the affected tissue cells. The biological effects on the cells and tissues are proportional to the absorbed radiation energy. The aim of our study was to examine the effects of hypofunction of the pituitary gland, previously irradiated with x-rays, on the growth of rat?s craniofacial and stomatognathic system as well as rat?s tibia. Methods. A total dose of x-rays of 27.92 Gy was applied in eight sessions in the period from 8 to 63 days of animal age on their heads and the effects of irradiation-induced hypofunction of the pituitary gland on the growth of rat?s craniofacial and stomatognathic system and tibia were examined. In order to differentiate the effects of the irradiated pituitary gland from the direct effects of x-rays on the head, the experiment was set as a double study. One group of animals had the pituitary gland protected with a lead plate set beneath the projection of the gland, while the second group was irradiated with the same dose, but without the protection. The control group of animals were non-irradiated. Results. Growth measurements of numerous parameters of the craniofacial and stomatognathic system as well as tibia in rats with the entire locally irradiated head showed a statistically significant delay, compared to both the non-irradiated control group and the group with the protected pituitary gland. Conclusion. The damage of the stomatognathic and craniofacial system was greater in animals with irradiated head without pituitary gland protection compared to those with the gland protection whereby the growth of the craniofacial system was most affected. Irraduition-induced pituitary gland hypofunction and irradiation itself showed a stronger effect on the lower jaw growth rate impairment compared to the upper jaw. Similarly, the delay in tibia growth was more pronounced in animals without pituitary gland protection during irradiation of their heads compared to non-protected ones.

Liver X Receptor activation regulates genes involved in lipid homeostasis in developing chondrocytes

Osteoarthritis (OA) is the most common type of arthritis and causes debilitating symptoms and decreased quality of life. A better understanding of the molecular mechanisms maintaining cartilage health is needed for developing novel therapeutic strategies. Liver X Receptors (LXRs) are nuclear receptors that have been previously shown to protect against OA. To better understand the regulatory mechanisms behind this effect, we systematically examined LXR's effects on growth plate chondrocyte gene expression. Primary chondrocytes isolated from the long bones of E15.5 mice were treated with the specific LXR agonist, GW3965, and RNA was isolated for Affymetrix microarrays. Bioinformatics analyses were performed using Gene Ontology (GO) and KEGG pathway analysis. Immunohistochemistry was conducted to examine protein localization of LXR and identified targets in GW3965-treated E15.5 tibiae compared to control. LXR activation in primary growth plate chondrocytes resulted in differential regulations of various genes involved in lipid metabolism. This pattern was compared to LXR activation in immature murine articular chondrocytes (IMACs), which revealed similar roles in lipid homeostasis. Immunohistochemical analysis of LXR and its identified targets Abca1 and Srebf1 revealed preferential protein localization to pre-hypertrophic and resting chondrocytes in GW3965-treated tibial growth plates compared to controls. Our findings show for the first time that LXR activation alters expression of lipid metabolism genes in growth plate chondrocytes, in part through activation of molecules responsible for cellular cholesterol efflux. This provides insight into potential mechanisms through which LXR regulates cellular metabolism to alter chondrocyte behavior and phenotype.

Transcriptome-based screening of intracellular pathways and angiogenesis related genes at different stages of thiram induced tibial lesions in broiler chickens

BackgroundThe Tibial dyschondroplasia (TD) in fast-growing chickens is mainly caused by improper blood circulation. The exact mechanism underlying angiogenesis and vascularization in tibial growth plate of broiler chickens remains unclear. Therefore, this research attempts to study genes involved in the regulation of angiogenesis in chicken red blood cells. Twenty-four broiler chickens were allotted into a control and thiram (Tetramethyl thiuram disulfide) group. Blood samples were collected on day 2, 6 (8- and 14-days old chickens) and 15 (23 days old chickens).ResultsHistopathology and hematoxylin and eosin (H&E) results showed that angiogenesis decreased on the 6th day of the experiment but started to recover on the 15th day of the experiment. Immunohistochemistry (IHC) results confirmed the expressions of integrin alpha-v precursor (ITGAV) and clusterin precursor (CLU). Transcriptome sequencing analysis evaluated 293 differentially expressed genes (DEGs), of which 103 up-regulated genes and 190 down-regulated genes were enriched in the pathways of neuroactive ligand receptor interaction, mitogen-activated protein kinase (MAPK), ribosome, regulation of actin cytoskeleton, focal adhesion, natural killer cell mediated cytotoxicity and the notch signalling pathways. DEGs (n = 20) related to angiogenesis of chicken erythrocytes in the enriched pathways were thromboxane A2 receptor (TBXA2R), interleukin-1 receptor type 1 precursor (IL1R1), ribosomal protein L17 (RPL17), integrin beta-3 precursor (ITGB3), ITGAV, integrin beta-2 precursor (ITGB2), ras-related C3 botulinum toxin substrate 2 (RAC2), integrin alpha-2 (ITGA2), IQ motif containing GTPase activating protein 2 (IQGAP2), ARF GTPase-activating protein (GIT1), proto-oncogene vav (VAV1), integrin alpha-IIb-like (ITGA5), ras-related protein Rap-1b precursor (RAP1B), tyrosine protein kinase Fyn-like (FYN), tyrosine-protein phosphatase non-receptor type 11 (PTPN11), protein patched homolog 1 (PTCH1), nuclear receptor corepressor 2 (NCOR2) and mastermind like protein 3 (MAML3) selected for further confirmation with qPCR. However, commonly DEGs were sarcoplasmic/endoplasmic reticulum calcium ATPase 3 (ATP2A3), ubiquitin-conjugating enzyme E2 R2 (UBE2R2), centriole cilia and spindle-associated protein (CCSAP), coagulation factor XIII A chain protein (F13A1), shroom 2 isoform X6 (SHROOM2), ras GTPase-activating protein 3 (RASA3) and CLU.ConclusionWe have found potential therapeutic genes concerned to erythrocytes and blood regulation, which regulated the angiogenesis in thiram induced TD chickens. This study also revealed the potential functions of erythrocytes.Graphical abstract1. Tibial dyschondroplasia (TD) in chickens were more on day 6, which started recovering on day 15. 2. The enriched pathway observed in TD chickens on day 6 was ribosome pathway, on day 15 were regulation of actin cytoskeleton and focal adhesion pathway. 3. The genes involved in the ribosome pathways was ribosomal protein L17 (RPL17). regulation of actin cytoskeleton pathway were Ras-related C3 botulinum toxin substrate 2 (RAC2), Ras-related protein Rap-1b precursor (RAP1B), ARF GTPase-activating protein (GIT1), IQ motif containing GTPase activating protein 2 (IQGAP2), Integrin alpha-v precursor (ITGAV), Integrin alpha-2 (ITGA2), Integrin beta-2 precursor (ITGB2), Integrin beta-3 precursor (ITGB3), Integrin alpha-IIb-like (ITGA5). Focal adhesion Proto-oncogene vav (Vav-like), Tyrosine-protein kinase Fyn-like (FYN).

Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in chronic renal insufficiency rats

Objective: To observe the effect of autophagy of tibial growth plate chondrocyte on apoptosis in chronic renal insufficiency (CRI) rats. Methods: Male 4-week-old SD rats were randomly divided into two groups: (1) Sham group: only the left ureter was exposed (n=10); (2) CRI group: the left ureter was ligated to cause CRI (n=10). The urine from all the rats was collected 6 weeks after the operation and the total protein content was measured. Then all the rats were sacrificed and the concentrations of creatinine and urea nitrogen in intracardiac blood were detected. The proximal tibia were fixed and decalcified to prepare histological sections, and the number of chondrocytes of column cells in the proliferative area of tibia growth plate was observed by saffron O staining. The expression rate of protein Light Chain-3, an autophagy marker of chondrocytes, was detected by immunofluorescence. The apoptosis rate of chondrocytes was detected by the method of TUNEL assay. The level of glycogenin-1, a glycogen formation marker of chondrocyte was detected by immunohistochemistry in chondrocytes. Results: The 24 h urine total protein was higher in CRI group [(163.5±11.3) mg vs (38.6±9.8) mg, t=25.620, P<0.001]. The levels of blood creatinine [(67.3±16.2) μmol/L vs (28.4±11.5) μmol/L, t=5.974, P<0.001] and urea nitrogen [(16.4±6.4) mmol/L vs (4.8±2.0) mmol/L, t=5.198, P<0.001] were higher in CRI group. The number of chondrocytes of column cells in the proliferating area of tibia growth plate was lower in CRI group (4.2±2.1 vs 9.1±3.8, t=3.109, P=0.006). The expression rate of LC-3 protein in chondrocytes of CRI group was lower [(27.2±12.6)% vs (51.4±18.2)%, t=3.457, P=0.003]. The level of glycogenin-1 of chondrocytes in CRI group increased significantly (6.1±2.5 vs 3.5±1.8, t=2.669, P=0.016). The apoptosis rate of chondrocytes in CRI group also increased [(17.2±4.8)% vs (5.1±3.4)%, t=6.505, P<0.001]. Conclusion: Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in CRI rats, which might be caused by the failure of glycogen degradation in chondrocytes.

Puerarin enhance vascular proliferation and halt apoptosis in thiram-induced avian tibial dyschondroplasia by regulating HIF-1α, TIMP-3 and BCL-2 expressions.

Tetramethyl thiuram disulfide (thiram) is a dithiocarbamate pesticide used for crop protection and storage. But, it's widespread utilization is associated with deleterious growth plate cartilage disorder in broilers termed as avian tibial dyschondroplasia (TD). TD results in non-mineralized and less vascularized proximal tibial growth plate cartilage causing lameness and poor growth performance. This study investigated the therapeutic potential of puerarin against thiram toxicity in TD affected chickens. One-day-old broiler chickens (n=240) were alienated into three equal groups i.e. control, TD and puerarin (n=80) and were offered standard feed. Additionally, TD and puerarin groups were offered thiram at 50mg/kg of feed from 4 to 7 days for TD induction followed by puerarin therapy at 120mg/kg to puerarin group only from 8 to 18 days for TD treatment. Thiram feeding to TD and puerarin group chickens caused lameness, mortality, and increased the aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA) levels and growth plate (GP) size and upregulated HIF-1α expression. Besides, the production parameters, alkaline phosphatase (ALP), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels and the expressions of TIMP-3 and BCL-2 were decreased (p<0.05). Puerarin alleviated lameness, enhanced angiogenesis and growth performance and serum and antioxidant enzymes, decreased apoptosis and recuperated GP width by significantly downregulating HIF-1α and upregulating the TIMP-3 and BCL-2 mRNA and protein expressions in puerarin group chickens (p<0.05). In conclusion, the toxic effects associated with thiram can be mitigated using puerarin.

Effect of In-Ovo Administration of L-Arginine on the Gross Anatomy of Tibia Bone, Alkaline Phospahtase and Growth Performance in Japanese Quail (Coturnix japonica)

Effect of In-Ovo Administration of L-Arginine on the Gross Anatomy of Tibia Bone, Alkaline Phospahtase and Growth Performance in Japanese Quail (Coturnix japonica)

Enhanced Bone Formation in Osteoporotic Mice by a Novel Transplant Combined with Adipose-derived Stem Cells and Platelet-rich Fibrin Releasates

Osteoporotic fracture is the main complication of osteoporosis (OP) and accounts for millions of injuries annually. Local intervention by intra-marrow injection has been a good option for preventing osteoporotic bone loss when the osteoporotic femoral fracture has been treated. In this study, tail vein transplantations were examined to evaluate the cell-based therapeutic approach for treating OP with adipose-derived stem cells (ADSCs) and platelet-rich fibrin releasates (PRFr) in an ovariectomized (OVX) mice model. Thirty-six 12-wk-old female ICR mice were randomly divided into six groups: untreated control; sham-operated; OVX-control; OVX-ADSCs; OVX-PRFr; and OVX-ADSCs+PRFr. Starting 8 wk after ovariectomy, the OVX mice received tail vein injections once each week for four consecutive weeks, then were evaluated radiographically and histopathologically 8 wk after the first injection. We also assessed changes to bone trabeculae in the proximal tibial growth plate. In OVX mice treated with ADSCs or PRFr alone, or with a combination of ADSCs and PRFr, the trabecular bone mineral density (BMD), bone volume ratios (BV/TV), and numbers (Tb.N) in the proximal tibia areas were significantly higher than that in the OVX-control group. Significant differences between OVX-treated mice and OVX controls were found for trabecular separation, but not for trabecular thickness. These results indicate that ADSCs or PRFr treatment enhances bone microarchitecture in OP. The treatment of bone loss of OVX mice with ADSCs+PRFr induced greater bone consolidation with bone tissue production (P < 0.01) when compared to the others. Thus, we conclude that the transplantation of ADSCs combined with PRFr might provide an alternative strategy for the treatment of various bone disorders in OP with an unlimited source of cells and releasates.

Identification and expression analysis of microRNAs in tibial growth plate of chicken through thiram toxicity.

Thiram is a widely known tibial dyschondroplasia (TD) inducer. TD, a common metabolic cartilage disease, presents in rapidly growing poultry birds. There are evidences that miRNAs are involved in diverse aspects of normal skeletal development, but very less is known about the role of miRNAs in TD. Therefore, this study aimed to determine which genes and pathways show differential expression between TD suffered chickens and normal chickens. We collected growth plates from ten-days-old TD chickens and control chickens and performed high-throughput RNA sequencing (RNA-Seq). Afterwards, target prediction, GO annotation and KEGG pathway analysis were carried out to understand the role of DEMs (differentially expressed microRNAs). We obtained 96,884,760 and 94,574,290 clean reads and identified 17 significant DEMs between the TD and control groups. Functional enrichment analysis of DEMs indicated that the putative targets of miRNAs were remarkably enriched in bone-related pathways, such as Notch, MAPK and Autophagy. Overall, this study provides detailed understanding about the pathogenesis of thiram induced TD and new insights towards the molecular mechanism of miRNAs.