Reduced serum and local LncRNA MALAT1 expressions are linked with disease severity in patients with non-traumatic osteonecrosis of the femoral head

Abstract

OBJECTIVE:

This study was performed to illustrate the potential relationship between reduced serum and local LncRNA MALAT1 expressions with disease severity in patients with non-traumatic osteonecrosis of the femoral head (ONFH).

METHODS:

A total of 104 patients with non-traumatic ONFH and 100 healthy controls were consecutively recruited from our hospital. Serum and local LncRNA MALAT1 expressions were detected using real-time polymerase chain reaction (RT-PCR). Radiographic progression was defined by Ficat classification. Clinical severity was evaluated by Visual Analog Scale (VAS) and Harris Hip Score (HHS). Receiver operating characteristic (ROC) curve was carried out to determine the diagnostic value of MALAT1 in the radiographic progression.

RESULTS:

Serum LncRNA MALAT1 expressions were significantly lower in non-traumatic ONFH patients than in healthy controls. In addition, local MALAT1 expressions in non-traumatic ONFH tissue were significantly lower in the affected area than in the non-affected area. Ficat grade 4 has significantly lower serum and local LncRNA MALAT1 expressions in comparison with grade 3, and Ficat grade 3 showed markedly decreased serum and local LncRNA MALAT1 expressions compared with grade 2. Serum and local LncRNA MALAT1 expressions were significantly and negatively associated with VAS and positively related to the HHS. Further ROC curve analysis indicated that serum MALAT1 may act as a decent indicator in the diagnosis of non-traumatic ONFH.

CONCLUSIONS:

Decreased serum and local MALAT1 expressions may reflect disease severity in non-traumatic ONFH patients.

Keywords

LncRNA MALAT1 non-traumatic osteonecrosis of the femoral head disease severity

1. Introduction

Osteonecrosis of the femoral head (ONFH) is a bone-destructive disease which results from an imbalance of the coagulation and fibrinolysis system as well as insufficient blood supply [1]. ONFH usually causes progressive femoral head collapse and secondary arthritis necessitating total hip arthroplasty [2]. ONFH is a multifactorial and disabling disease that involves multiple genetic and environmental factors [3, 4]. The incidence of ONFH has increased over the past decade. It is estimated that 20,000 to 30,000 new patients are diagnosed with ONFH annually in the United States and 150,000 to 200,000 in China [5, 6].

ONFH can be divided into traumatic and non-traumatic conditions. As a subtype of ONFH, non-traumatic ONFH often occurs after the treatment of inflammatory diseases by steroid use [7]. It has been reported that the onset and development of non-traumatic ONFH is closely correlated with various factors including human immunodeficiency virus infection, autoimmune diseases, alcohol abuse, use of glucocorticoids and coagulopathies [8]. So far, the pathophysiological process of ONFH remains unclear. However, bone damage and insufficient blood supply are thought to be the two main final events in ONFH.

It has been reported that the development of non-traumatic ONFH is usually accompanied by changes in the expression of different types of non-coding RNA, such as microRNAs (miRNAs/miR) [9]. However, the involvement of long non-coding (lnc)RNAs, a subgroup of non-coding RNAs composed of $>$ 200 nucleotides with pivotal roles in both normal physiological and pathological processes, is largely unknown [10].

The long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) [11], or Lnc-MALAT1, is an abundant and highly conserved lncRNA [12] and was first identified as a valuable indicator for lung cancer metastasis [13]. Lnc-MALAT1 is upregulated in various human cancers [14] and interacts with serine/arginine-rich proteins to regulate the subcellular localization of splicing regulatory proteins [15].

In recent years, lncMALAT1 has been proved to protect bone against damage and promote angiogenesis. On the one hand, previous studies demonstrated that MALAT1 was significantly down-regulated in the decreased osteogenic differentiation of BMSCs in steroid induced ONFH patients [16]. Moreover, lnc-MALAT1 could protect human osteoblasts from dexamethasone-induced injury through activation of Ppm1e-AMPK signaling [17].On the other hand, MALAT1 has been proved to promote angiogenesis in various conditions [18, 19]. For example, MALAT1 could promote angiogenesis in brain microvascular endothelial cells by upregulating VEGF-A and ANGPT2 [20]. It could furthermore promote angiogenesis of mesenchymal stem cells by inducing VEGF [21].

The previous studies listed above indicate that APN may play a pivotal role in protecting non-traumatic ONFH patients against bone damage. However, to our knowledge, no studies explored the association of serum or local MALAT1 expressions and disease severity in ONFH patients. Therefore, the scope of our study was to examine whether serum and local MALAT1 expressions may be potential protective biomarkers of nontraumatic ONFH.

2. Patients and methods

2.1 Study participants

From October 2018 to December 2019, a total of 104 patients with non-traumatic ONFH mainly caused by steroid and alcohol use were enrolled in our study. All patients received total hip arthroplasty or hip preserving surgery including core decompression, vascularized bone grafting, femoral varus osteotomy, and femoral rotational osteotomy. The diagnosis, classification, and staging of ONFH were based on the 2001 Japanese Orthopaedic Association (JOA) classification, which refers to findings on anteroposterior and lateral plain radiographs or magnetic resonance imaging scans [22]. Meanwhile, 100 individuals attending the regular health examination served as age- and sex-matched healthy controls. The healthy controls were enrolled based on a physical examination, with no steroid-induced ONFH or other related diseases. All subjects were Han people living in or near Linyi City in China.

The inclusion criteria were: ONFH was diagnosed based on X-ray, MRI and CT scanning; Steroid-induced ONFH patients were diagnosed on the basis of taking a mean daily dose of 16.6 mg or an equivalent maximum daily dose of 80 mg of prednisolone within one year; Alcohol-induced ONFH patients must have a history of alcohol intake $>$ 400 ml/week (320 g/week, any type of alcoholic beverage) of pure ethanol for more than six months. The exclusion criteria were: patients who did not meet the diagnostic criteria of steroid-induced or alcohol-induced ONFH; Patients with traumatic ONFH, dislocation of the hip joint, or other hip diseases; History or diagnosis of Paget’s disease, renal osteodystrophy, hyperparathyroidism, hypoparathyrodism, or bone metastasis. This study was carried out according to the principles of the Declaration of Helsinki. The ethical committee of the hospital approved the study and all participants provided written informed consent.

2.2 Sample preparation and quantitative real-time polymerase chain reaction (qRT-PCR)

Venous blood samples were collected from patients prior to surgery or other treatments. All serum samples were processed within 4 h after blood drawing. In short, whole blood was drawn into blood vessels without adding anticoagulants and was separated to obtain serum by centrifugation at 1,500 g for 15 min. For tissues, 104 pairs of affected tissues and adjacent non-affected tissues were obtained from patients. Total RNA was reverse-transcribed to cDNA using PrimeScript RT reagent kit with gDNA Eraser (TaKaRa, Japan) according to the manufacturer’s instructions. qRT-PCR was performed using FastStart Universal SYBR Green Master (ROX) (Roche, Basel, Germany) in an Applied Biosystems 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) according to the manufacturer’s protocol. The data were normalized to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the relative expression levels of each gene were analyzed using the 2 ${}^{-}$ Ct method. All experiments were performed in triplicate. The primer sequences used in this study are: MALAT1: Forward: 5’-CAGTGGGGAACTCTGACTCG-3’; Reverse: 5’-GTGCCTGGTGCTCTCTTACC-3’ GAPDH: Forward: 5’-TGTGTTGGCGTACAGGTCTTTG-3’; Reverse: 5’-GGAAATCGTGCGTGACATTAAG-3’. MALAT1 expression was normalized to GAPDH.

2.3 Radiographic assessment

Radiographic progression was evaluated using the Ficat and Arlet classification system [23], which comprises four grades. Grade 1: Normal or, at most, minor changes; Grade 2: Diffuse or localized osteoporosis, sclerosis, or cysts of the femoral head; Grade 3: Sequestrum, break in articular cartilage, normal or increased joint space; Grade 4: Decreased joint space, collapse of the femoral head, acetabular osteoarthrotic changes. Patients with grade $\geqslant$ 2 were enrolled in the current study. If ONFH occurred at both sides, the most severe side was selected for analysis. The results were read by one experienced radiologist and one senior orthopedist in our hospital who were blinded to the enrolled patients. Kappa value was calculated for consistency.

2.4 Clinical severity assessment

The clinical severity of ONFH patients was evaluated by the Visual Analog Scale (VAS) and Harris Hip Score (HHS). The VAS was 10 mm long with three anchors dividing three zones (green, yellow, and red). For each symptom, the green zone (0–30 mm) was labeled “mild symptoms”, the yellow zone (30–60 mm) “moderate symptoms” and the red zone (61–100 mm) “bad symptoms”. Patients were asked to mark any point on the zone according to their perception of the symptoms. The distance between the 0 mm mark and the placement of the “X” was measured to provide a numeric interpretation of their symptom perception [24]. The HHS questionnaire includes questions about pain, lameness, ability to walk and to get up from/sit down on a chair, range of motion and limb length discrepancy. The scores range from 0 to 100. In this study, scores of less than 70 (poor), 70–79 (average), 80–89 (good) and 90–100 (excellent) were found [25].

2.5 Statistical analyses

All statistical analyses were performed using GraphPad 6.0 software (GraphPad Software Inc., San Diego, CA, USA). Analysis was expressed as mean $\pm$ standard error. The difference between groups were analyzed using an independent sample t-test. Differences among subgroups were examined using an ANOVA test followed by a Bonferroni post hoc test. The correlation of serum and local lncMALAT1 with ONFH severity indices was analyzed using Spearman’s correlation test. Receiver operating characteristic (ROC) curves and Area under the ROC Curve (AUC) values were also calculated, according to the software’s instruction. Two-sided $P$ values less than 0.05 were considered statistically significant.

3. Results

3.1 Demographic statistics

Table 1 describes the demographic data and etiology of the 104 enrolled ONFH patients and 100 healthy controls. The age of the patients (44 females and 60 males) was 51.7 $\pm$ 11.1 years (range from 22 to 75 years). The disease duration of ONFH ranged from three to 250 months, with a median of 80.2 months. There were no significant differences between ONFH patients and healthy controls with regards to the age, sex distribution and body mass index.

Table 1
Demographic data

	ONFH patients ( $n=$ 104)	Healthy controls ( $n=$ 100)	$P$ value
Age (Y)	51.7 $\pm$ 11.1	50.9 $\pm$ 11.8	0.567
Sex (F/M)	44/60	42/58	0.688
BMI (kg/m ${}^{2}$ )	23.5 $\pm$ 20	23.4 $\pm$ 1.6	0.323
Disease duration (M)	80.2 (3–250)
VAS scores	5.0 $\pm$ 1.9
HSS scores	67.8 $\pm$ 9.5
Ficat phases (2/3/4)	33/37/34
Serum MALAT1 expression (relative expression)	0.60 $\pm$ 0.11	1.00 $\pm$ 0.06	$<$ 0.001

All data are given as mean $\pm$ SD or median.

3.2 MALAT1 expression in ONFH patients

Serum relative MALAT1 expressions in patients with non-traumatic ONFH were significantly lower than in the healthy control group (0.60 $\pm$ 0.11 vs. 1.00 $\pm$ 0.06, $P<$ 0.001) (Fig. 1A). In ONFH patients, local MALAT1 expressions in affected tissue were significantly lower than in the non-affected area (0.42 $\pm$ 0.13 vs. 1.00 $\pm$ 0.05, $P<$ 0.001) (Fig. 1B). 53 patients had used steroids before and 51 patients had a history of alcohol consumption. No significant differences of MALAT1 were found between the steroid group and alcohol group (0.60 $\pm$ 0.10 vs. 0.60 $\pm$ 0.10, $P=$ 0.975) (Fig. 1C).

Figure 1.

A. Relative MALAT1 expression between ONFH patients and healthy controls. B. Relative MALAT1 expression between affected and non-affected tissue in ONFH patients. C. Relative MALAT1 expression between steroid-induced ONFH and alcohol-induced ONFH. Data were expressed as mean $\pm$ SD.

Figure 2.

Plain X-ray of ONFH from Ficat grades 1–4.

3.3 Association of serum and local MALAT1 expressions with Ficat grades

The serum and local MALAT1 expressions of 104 non-traumatic ONFH patients with different Ficat grades are depicted in Fig. 3. Accordingly, the ONFH patients were divided into three groups based on the Ficat grading system. The ONFH group included 33 patients with grade 2, 37 patients with grade 3, and 34 patients with grade 4. Representative figures of ONFH from Ficat 1–4 were demonstrated as Fig. 2. ONFH patients with Ficat grade 4 had significantly lower serum relative MALAT1 expressions than grade 3 (0.54 $\pm$ 0.10 vs. 0.60 $\pm$ 0.10, $P=$ 0.007) (Fig. 3A). ONFH patients with Ficat grade 3 demonstrated significantly attenuated expressions of serum MALAT1 expression compared with Ficat grade 2 (0.60 $\pm$ 0.10 vs. 0.67 $\pm$ 0.10, $P=$ 0.008) (Fig. 3A). The MALAT1 expression in serum was negatively associated with Ficat grading ( $r=$ $-$ 0.477, $P<$ 0.001) by Spearman’s correlation analysis (Fig. 3C).

Figure 3.

A. Comparison of serum MALAT1 expression among different Ficat grades. B. Comparison of local MALAT1 expression among different Ficat grades. C. Correlation of serum MALAT1 expression with Ficat grades. D. Correlation of local MALAT1 expression with Ficat grades.

In contrast, ONFH patients with modified Ficat grade 4 had markedly lower local MALAT1 expression than in Ficat grade 3 (0.34 $\pm$ 0.10 vs. 0.41 $\pm$ 0.12, $P=$ 0.02) (Fig. 3B). ONFH patients with Ficat grade 3 showed drastically reduced local expression of MALAT1 compared with those with Ficat grade 2 (0.41 $\pm$ 0.12 vs. 0.51 $\pm$ 0.13, $P=$ 0.001) (Fig. 3B). The local MALAT1 expression was negatively associated with Ficat grading ( $r=$ $-$ 0.494, $P<$ 0.001) (Fig. 3D).

3.4 ROC curve analysis

A ROC curve analysis was conducted to explore the diagnostic value of whether serum MALAT1 could be used as a diagnostic marker for radiographic progression of ONFH. As shown in Fig. 4, for Ficat grade 2 vs grade 3 and Ficat grade 3 vs grade 4, MALAT1 exhibited both significant AUCs (Ficat grade 2 vs grade 3: AUC $=$ 0.681, $P=$ 0.009; Ficat grade 3 vs grade 4: AUC $=$ 0.667, $P=$ 0.016) (Fig. 4A and B). This finding indicates that serum MALAT1 may act as a decent diagnostic marker to assess radiographic progression both in early-medium and late stage ONFH.

Figure 4.

Area under curve of serum MALAT1 for evaluating radiographic progression by ROC curve analysis A. Ficat grade 2 vs grade 3. B. Ficat grade 3 vs grade 4.

3.5 Association of serum and local MALAT1 expressions with clinical severity

The correlation of serum and local MALAT1 expressions with clinical severity determined by VAS and HSS scores were examined. We found that both serum MALAT1 and local expressions were negatively associated with VAS scores (serum $r=$ $-$ 0.381, $P<$ 0.001; local $r=$ $-$ 0.449, $P<$ 0.001) (Fig. 5A–C). In addition, serum MALAT1 and local MALAT1 expressions were significantly related and positively associated with HSS scores (serum $r=$ $-$ 0.349, $P<$ 0.001; local $r=$ $-$ 0.336, $P<$ 0.001) (Fig. 5B–D). These findings imply that both higher serum and local MALAT1 expressions indicate less pain and better functions in ONFH patients.

Figure 5.

A. Correlation of serum MALAT1 expression with VAS. B. Correlation of local MALAT1 expression with VAS. C. Correlation of serum MALAT1 expression with HSS. D. Correlation of local MALAT1 expression with HSS.

4. Discussion

The present study explored the potential involvement of serum and local MALAT1 expression in patients with non-traumatic ONFH. To the our best of knowledge, this is the first investigation of serum and local MALAT1 expression involved in the progression of non-traumatic ONFH. We found that serum MALAT1 expression was significantly lower in non-traumatic ONFH patients than in sex- and age-matched healthy controls. In addition, local MALAT1 expression in ONFH tissue was significantly lower in the affected area than in the non-affected area. Moreover, serum and local MALAT1 expressions were negatively associated with radiographic progression and clinical severity. Further ROC curve analysis indicated that serum MALAT1 may act as a decent indicator in the diagnosis of non-traumatic ONFH.

Up to date, the normal diagnosis methods of non-traumatic ONFH consist of complaints of symptoms, worse function and radiographic alternations including X-ray, CT and MRI [26]. The complaints of non-traumatic ONFH include hip pain and restricted hip joint activity. However, non-traumatic ONFH at an early phase may develop with no manifestations of symptoms. Besides, radiographic progressions including decreased joint space, collapse of the femoral head and acetabular osteoarthritic changes are mostly indicators of late phase ONFH [27]. Therefore, brief and more effective methods that use valuable biomarkers may make a great difference for the diagnosis, treatment and prognosis of non-traumatic ONFH.

Epigenetics, an effective method for studying the interplay between environmental signals and the genome, has received a great deal of attention recently [28]. It has become apparent that non-coding RNAs play an important epigenetic role in non-traumatic ONFH including miRNA, lncRNA and circRNA. For example, HOTAIR acts as regulator on osteogenic differentiation and proliferation through modulating miR-17-5p and its target gene SMAD7 in non-traumatic ONFH [29]. MiR-181d can inhibit the differentiation of hBMSCs into osteoblasts by regulating the expression of SMAD3 [30]. hsa_circ_0000219 and hsa_circ_0005936 may regulate the progression of ONFH by mediating the proliferation and differentiation of BMSCs by sponging corresponding miRNAs [31]. However, none of these studies explored the relationship between their expressions with disease severity and progression in non-traumatic ONFH.

Circulating lncRNAs in serum or plasma have a good potential to serve as diagnostic or prognostic markers in various diseases, because they are stable, accessible and closely related to various diseases [32]. In our study, we comprehensively evaluated the serum MALAT1 levels in non-traumatic ONFH patients. We demonstrated that serum MALAT1 expressions were significantly lower in ONFH patients compared to healthy individuals. We also found that in non-traumatic ONFH patients, MALAT1 expression was more downregulated in the affected area than in the non-affected area. In addition, both serum and local MALAT1 expressions were negatively related to radiographic progression in ONFH patients.

Decreased bone formation and aberrant osteoclastic activity have been implicated in non-traumatic ONFH . A previous study showed dexamethasone-inhibited osteogenic differentiation of BMSC in a dose-dependent manner, leading to decreased MALAT1 [33]. Another study demonstrated that BMSCs-derived exosomal MALAT1 enhances osteoblast activity in osteoporotic mice by mediating the miR-34c/SATB2 axis [34]. Yi et al. found MALAT1 promoted Runx2-mediated osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting miR-30 [35]. Bone repair involves bone resorption through osteoclastogenesis and the stimulation of neovascularization and osteogenesis by endothelial progenitor cells, suggesting that EPC-derived exosomes can promote bone repair by enhancing recruitment and differentiation of osteoclast precursors through LncRNA-MALAT1 [36].

Although some novel findings were revealed in the current study, there are still some limitations that should be taken into account when interpreting the results. First, our study did not include an intensive analysis of biological functions, which will be crucial for elucidating the role of MALAT1 in protecting non-traumatic ONFH. Second, this is a cross-sectional study with a relatively small sample size, which might influence the present findings. A larger case-control study is necessary to solve those problems, which could make our conclusions more powerful. Although we obtained evidence for the association between MALAT1 and ONFH disease progression, the pathogenesis of non-traumatic ONFH was still not completely understood. Further studies should be carried out to verify the results in other ethnicities.

5. Conclusion

In conclusion, this study is the first to report the aberrant expression of MALAT1 in relation to non-traumatic ONFH patients. We found that reduced serum and local LncRNA MALAT1 expressions are linked with disease severity in patients with non-traumatic ONFH. These findings offer a new insight into understanding the pathogenesis, facilitating early diagnosis and guiding the treatment of the disease.

Footnotes

Acknowledgments

This work was supported by the Natural Science Foundation of Heilongjiang Province of China (JJ2018ZZ0082) and the National Natural Science Foundation of China (31872980). The authors thank LetPub (www.letpub.com) for their linguistic assistance during the preparation of this manuscript.

Conflict of interest

None to report.

References

Choi

Steinberg

Cheng

. Osteonecrosis of the femoral head: Diagnosis and classification systems. Curr Rev Musculoskelet Med. 2015; 8(3): 210-220.

Wang

Azeddine

Mah

Harvey

Rosenblatt

Seguin

. Osteonecrosis of the femoral head: Genetic basis. Int Orthop. 2019; 43(3): 519-530.

Moya-Angeler

Gianakos

Villa

Lane

. Current concepts on osteonecrosis of the femoral head. World J Orthop. 2015; 6(8): 590-601.

Moya-Angeler

Gianakos

Villa

Lane

. Current concepts on osteonecrosis of the femoral head. World J Orthop. 2015; 6(8): 590-601.

Al-Jabri

Tan

Tong

Shenoy

Kayani

Parratt

Khan

. The role of electrical stimulation in the management of avascular necrosis of the femoral head in adults: A systematic review. BMC Musculoskelet Disord. 2017; 18(1): 319.

Zhang

Xie

Zhao

Wang

Yang

Meng

. Association of ABCB1 C3435T polymorphism with the susceptibility to osteonecrosis of the femoral head: A meta-analysis. Medicine (Baltimore). 2017; 96(20): e6049.

Okazaki

Nagoya

Matsumoto

Mizuo

Sasaki

Watanabe

Yamashita

Inoue

. Development of non-traumatic osteonecrosis of the femoral head requires toll-like receptor 7 and 9 stimulations and is boosted by repression on nuclear factor kappa B in rats. Lab Invest. 2015; 95(1): 92-99.

Assouline-Dayan

Chang

Greenspan

Shoenfeld

Gershwin

. Pathogenesis and natural history of osteonecrosis. Semin Arthritis Rheum. 2002; 32(2): 94-124.

Yang

Weng

Tse

Chan

. Emerging roles of MicroRNAs in osteonecrosis of the femoral head. Cell Prolif. 2018; 51(1).

10.

Mercer

Dinger

Mattick

. Long non-coding RNAs: Insights into functions. Nat Rev Genet. 2009; 10(3): 155-159.

11.

Guo

Yan

Han

Shui

. Down-Regulation of lncrna MALAT1 attenuates neuronal cell death through suppressing Beclin1-Dependent autophagy by regulating mir-30a in cerebral ischemic stroke. Cell Physiol Biochem. 2017; 43(1): 182-194.

12.

Amodio

Raimondi

Juli

Stamato

Caracciolo

Tagliaferri

Tassone

. MALAT1: A druggable long non-coding RNA for targeted anti-cancer approaches. J Hematol Oncol. 2018; 11(1): 63.

13.

Diederichs

Wang

Boing

Metzger

Schneider

Tidow

Brandt

Buerger

Bulk

Thomas

Berdel

Serve

Muller-Tidow

. MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer. Oncogene. 2003; 22(39): 8031-8041.

14.

Zhang

Hamblin

Yin

. The long noncoding RNA Malat1: Its physiological and pathophysiological functions. RNA Biol. 2017; 14(12): 1705-1714.

15.

Tripathi

Ellis

Shen

Song

Pan

Watt

Freier

Bennett

Sharma

Bubulya

Blencowe

Prasanth

. The nuclear-retained noncoding RNA MALAT1 regulates alternative splicing by modulating SR splicing factor phosphorylation. Mol Cell. 2010; 39(6): 925-938.

16.

Wang

Yang

Chen

Ren

Wang

Zhao

Zhang

Song

. LncRNA expression profiling of BMSCs in osteonecrosis of the femoral head associated with increased adipogenic and decreased osteogenic differentiation. Sci Rep. 2018; 8(1): 9127.

17.

Fan

Zhang

Liu

Zhu

Zhao

Cui

. Long Non-Coding RNA MALAT1 Protects Human Osteoblasts from Dexamethasone-Induced Injury via Activation of PPM1E-AMPK Signaling. Cell Physiol Biochem. 2018; 51(1): 31-45.

18.

Thum

Fiedler

. LINCing MALAT1 and angiogenesis. Circ Res. 2014; 114(9): 1366-1368.

19.

Wang

. Angio-LncRs: LncRNAs that regulate angiogenesis and vascular disease. Theranostics. 2018; 8(13): 3654-3675.

20.

Ren

Wei

. LncRNA MALAT1 up-regulates VEGF-A and ANGPT2 to promote angiogenesis in brain microvascular endothelial cells against oxygen-glucose deprivation via targetting miR-145. Biosci Rep. 2019; 39(3).

21.

Song

Liu

Miao

Ren

Ding

Wang

Hou

Zhao

. Long Non-Coding RNA MALAT1 promotes proliferation, angiogenesis, and immunosuppressive properties of mesenchymal stem cells by inducing VEGF and IDO. J Cell Biochem. 2017; 118(9): 2780-2791.

22.

Sugano

Atsumi

Ohzono

Kubo

Hotokebuchi

Takaoka

. The 2001 revised criteria for diagnosis, classification, and staging of idiopathic osteonecrosis of the femoral head. J Orthop Sci. 2002; 7(5): 601-605.

23.

Ficat

Arlet

. Necrosis of the femoral head. In: Hungerford

, editor. Ischemia and necrosis of bone. Baltimore: Williams and Wilkins, 1980, pp. 53-74.

24.

Reed

Van Nostran

. Assessing pain intensity with the visual analog scale: A plea for uniformity. J Clin Pharmacol. 2014; 54(3): 241-244.

25.

Shi

Mau

Chang

Wang

Chiu

. Responsiveness of the Harris Hip Score and the SF-36: Five years after total hip arthroplasty. Qual Life Res. 2009; 18(8): 1053-1060.

26.

Liu

Zhang

Sun

Gao

. Corticosteroid-induced osteonecrosis of the femoral head: Detection, diagnosis, and treatment in earlier stages. Chin Med J (Engl). 2017; 130(21): 2601-2607.

27.

Larson

Jones

Goodman

Koo

Cui

. Early-stage osteonecrosis of the femoral head: Where are we and where are we going in year 2018? Int Orthop. 2018; 42(7): 1723-1728.

28.

Thiagalingam

. Epigenetic memory in development and disease: Unraveling the mechanism. Biochim Biophys Acta Rev Cancer. 2020; 1873(2): 188349.

29.

Wei

Zhao

Guo

Liu

. Long non-coding RNA HOTAIR inhibits miR-17-5p to regulate osteogenic differentiation and proliferation in non-traumatic osteonecrosis of femoral head. PLoS One. 2017; 12(2): e169097.

30.

Xie

Shi

. MiR-181d promotes steroid-induced osteonecrosis of the femoral head by targeting SMAD3 to inhibit osteogenic differentiation of hBMSCs. Eur Rev Med Pharmacol Sci. 2018; 22(13): 4053-4062.

31.

Xiang

Weng

. Changed cellular functions and aberrantly expressed miRNAs and circRNAs in bone marrow stem cells in osteonecrosis of the femoral head. Int J Mol Med. 2020; 45(3): 805-815.

32.

Stepien

Costa

Kurc

Drozdz

Cortez-Dias

Enguita

. The circulating non-coding RNA landscape for biomarker research: Lessons and prospects from cardiovascular diseases. Acta Pharmacol Sin. 2018; 39(7): 1085-1099.

33.

Huang

Wang

Song

. LncRNA-MALAT1 promotes osteogenic differentiation through regulating ATF4 by sponging miR-214: Implication of steroid-induced avascular necrosis of the femoral head. Steroids. 2020; 154: 108533.

34.

Yang

Lei

Wen

. LncRNA MALAT1 shuttled by bone marrow-derived mesenchymal stem cells-secreted exosomes alleviates osteoporosis through mediating microRNA-34c/SATB2 axis. Aging (Albany NY). 2019; 11(20): 8777-8791.

35.

Liu

Xiao

. LncRNA MALAT1 sponges miR-30 to promote osteoblast differentiation of adipose-derived mesenchymal stem cells by promotion of Runx2 expression. Cell Tissue Res. 2019; 376(1): 113-121.

36.

Cui

Sun

Xing

Hou

. EPC-derived exosomes promote osteoclastogenesis through LncRNA-MALAT1. J Cell Mol Med. 2019; 23(6): 3843-3854.

Reduced serum and local LncRNA MALAT1 expressions are linked with disease severity in patients with non-traumatic osteonecrosis of the femoral head

Abstract

OBJECTIVE:

METHODS:

RESULTS:

CONCLUSIONS:

Keywords

1. Introduction

2. Patients and methods

2.1 Study participants

2.2 Sample preparation and quantitative real-time polymerase chain reaction (qRT-PCR)

2.3 Radiographic assessment

2.4 Clinical severity assessment

2.5 Statistical analyses

3. Results

3.1 Demographic statistics

Table 1 Demographic data

5. Conclusion

Footnotes

Acknowledgments

Conflict of interest

References

Table 1
Demographic data