INTRODUCTION:

A bone disorder called osteoporosis is characterized by aberrant bone apposition and resorption processes.^1,2 These disorders weaken bones and increase the risk of bone fracture by reducing bone mineral density (BMD), interfering with bone mineralization, and reducing bone strength.^3-6

Osteoporosis is a common degenerative disease that affects over 200 million people worldwide.⁷ According to new research from the International Osteoporosis Foundation (IOF), one in every four women in Indonesia between the ages of 50 and 80 is at risk of developing osteoporosis. Although oral bisphosphonates are the most commonly used osteoporosis treatment, they can increase the risk of osteonecrosis and digestive disorders.^8-11 As a result, it is necessary to innovate in osteoporosis therapy with minimal side effects.

Osthole bone marrow mesenchymal stem cells (OBMMSCs) are a combination of bone marrow mesenchymal stem cells (BMMSCs) and Cnidium monnieri coumarin derivatives with anti-inflammatory, anti-apoptotic, and anti-osteoporosis properties.¹² The development of OBMMSCs into osteoblasts is facilitated by the activation of cyclic adenosine monophosphate/protein kinase A/cAMP response element-binding protein or cAMP/PKA/CREB and Wnt/β-catenin-BMP signaling pathways.^13,14 In order to stimulate osteogenic signaling in mesenchymal stem cells and encourage osteogenesis in vitro, bone morphogenetic protein 9 (BMP-9) has considered as the most effective BMP.¹⁵ BMP-9 induces Smad1 and/or Smad phosphorylation, which is necessary for osteogenic proliferation and differentiation by binding with high affinity to activin receptor-like kinase 1 (ALK1) and endoglin in endothelial cells.¹⁶

The biomaterial nanohydroxyapatite (nHA), which is high in calcium and phosphate, is crucial for bone remodeling. Collagen I (ColI) is the primary organic component of bone and is essential for cell growth and differentiation. Multiwalled carbon nanotubes (MWCNTs) are carbon-based materials with high mechanical properties that are frequently used as scaffolding materials.¹⁴ The combination of nHA, ColI, and MWCNTs in the form of a scaffold can aid stem cell proliferation and osteogenic differentiation.

The potential to promote OBMMSC proliferation and differentiation into osteoblasts by the use of OBMMSCs in a BMP-9-loaded nHA/ColI/MWCNTs scaffold based on triad tissue engineering makes it a promising approach for osteoporosis therapy.

MATERIAL AND METHODS:

The writing of this scientific paper uses the literature review method, which aims to provide ideas and innovations related to osteoporosis therapy options. Literature search was conducted using several keywords, including “osthole bone marrow mesenchymal stem cells”, “bone marrow stem cells”, “bone morphogenetic protein-9”, “nanohydroxyapatite”, “collagen type I”, “multiwalled carbon nanotubes”, “osteoporosis” and “bone regeneration.” The databases used include ScienceDirect, ResearchGate, PubMed, and Google Scholar, with the literature criteria used including published in the last 10 years (2011–2021), considered as review and research journal, and published in English. The literature used includes reviews and research results to be able to describe theories and find the latest ideas and innovations. The literature was selected by title, abstract and full paper to test the eligibility of the literature. The writing of this scientific paper can be used as the basis for further scientific research to test the truth of existing theories. The literature review is written by looking for similarities in several pieces of literature and then summing them together. The current theories will later be correlated with each other to form a concept that is easy to understand. Reference writing in this scientific paper uses the Vancouver style by sorting citations in number.

Osteoporosis:

A bone disorder called osteoporosis results from an imbalance between bone resorption and bone apposition, which raises the risk of fracture due to decreased bone mineral density (BMD), impaired bone mineralization, and decreased bone strength.^1,3 Most people with osteoporosis are elderly and over 65 years, making them very vulnerable to the incidence of osteoporosis.¹⁷ The etiology of this disease includes the aging process, the consumption of certain drugs (including glucocorticoids and anti-epileptic drugs), smoking, genetics, and lack of physical activity. The presence of etiological factors initiates the pathogenesis of this disease, increasing in proinflammatory cytokine expressions such as tumor necrosis factor- α (TNF-α) and interleukin 1β/6 (IL-1β /6), which increases the expression of macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor κβ ligand (RANKL). This condition manifests in activating osteoclast precursors into mature osteoclast, resulting in bone resorption.¹

OBMMSCS (Osthole Bone Marrow Mesenchymal Stem Cells):

Various tissues and organs' stem cells are used in stem-cell therapy to treat or prevent diseases and conditions. The main source of stem cells is bone marrow. Stem cells can regenerate themselves and generate various cells to potentially replace diseased and damaged body parts while minimizing the risk of rejection and adverse reactions. The use of stem cells from the patient’s body is called mesenchymal stem cells (MSCs).^18,19 Osthole bone marrow mesenchymal stem cells (OBMMSCs) are a combination of bone marrow mesenchymal stem cells (BMMSCs) with coumarin compounds derived from the Cnidium monnieri plant. OBMMSCs have anti-inflammatory, antiapoptotic, antitumor, antibacterial, and antiosteoporosis properties.¹² OBMMSCs can stimulate osteogenesis by increasing autophagy activity, namely the ability of cells to regenerate themselves after cell destruction occurs. The autophagy process is aided by an increase in β-ecdysterone. By activating the Wnt/β-catenin-BMP pathway and the cAMP/PKA/CREB signaling pathways with the aid of extracellular signal-regulated kinase 1/2 (ERK1/2), OBMMSCs can prevent osteoporosis and enhance differentiation of OBMMSCs into osteoblasts.^13,14 OBMMSCs were able to inhibit osteoclastic activity in estrogen deficiency. OBMMSCs work in an osthole dose-dependent manner, i.e., the magnitude of the induction of differentiation of BMMSCs is influenced by the level of osthole used.²⁰

BMP-9:

BMP-9, a member of the TGF-superfamily, transmits signals to endothelial cells by binding with high affinity to the activin receptor-like kinase 1 (ALK1) enzyme. BMP-9 can stimulate bone formation in vivo and osteogenic signaling in mesenchymal stem cells in vitro. This process requires a high concentration of ALK1, ALK2, and BMP-9.¹⁵ Angiogenic, chondrogenic, and osteogenic processes are all directly impacted by BMP-9. According to recent research, BMP9 strongly binds to endoglin and ALK1 in endothelial cells and causes the phosphorylation of Smad1 and/or Smad5. Through ALK1, BMP-9 prevents aortic endothelial cell DNA synthesis and cutaneous microvascular endothelial cell migration and proliferation (HMVEC-ds). Furthermore, BMP-9 enters the blood circulation to maintain blood vessel maturation.¹⁶

nHA/ColI/MWCNTs:

One of the most important things to comprehend and create right away is bone tissue engineering. One way to advance bone tissue engineering is to learn more about scaffolds with good physicochemical properties and biocompatibility. Mechanical properties, optimal architecture for colonization and cell organization, biocompatibility, and good scaffold integration with bone tissue are all desirable characteristics of an ideal scaffold for bone tissue engineering. Hydroxyapatite (HA) is the primary inorganic constituent of bone and a biomaterial with high biocompatibility, and nano hydroxyapatite (nHA) is a biomaterial with inorganic components similar to those found in humans. Bone regeneration can be accomplished with the presence of components resembling a bone scaffold. The presence of nHA, which is a source of calcium and phosphate, makes it essential in the process of bone regeneration and remodeling. The main organic component of bone, collagen I (ColI), has been employed for development, culture, and differentiation. This organic component is an excellent starting point for the creation of biomaterial scaffolds. Single-walled carbon nanotubes (SWCNTs) and multi-walled carbon nanotubes are the two categories under which the substance known as carbon nanotubes (CNTs) is classified (MWCNTs). In bone tissue engineering, MWCNTs can be employed as a reinforcement material to enhance the physical and chemical characteristics of the scaffold due to their superior chemical, electrical, and mechanical qualities. Combining these biomaterial components is carried out to obtain beneficial properties effectively.¹⁴

DISCUSSION:

Mesenchymal stem cells have been widely used for tissue repair because of multipotent differentiation potential and easy access. Bone marrow mesenchymal stem cells (BMMSCs) derived from bone marrow stromal cells have attracted much attention because of their ability to be good therapy for skeletal diseases. BMMSCs that work by promoting osteoblast differentiation and inhibiting adipocyte differentiation are believed to reduce the development of osteoporosis. Osthole extracted from Cnidium monnieri can stimulate osteoblast differentiation and bone formation, which can be an effective alternative therapy for osteoporosis.²⁰

Several studies observed the potential of osthole in osteoporosis therapy through several mechanisms. Zhang et al. (2017) mentioned that this compound promotes osteoblasts differentiation through BMP signaling pathway that interacts with cAMP/CREB signaling to target the transcription factor osterix (Osx). Yu et al. (2021) stated that osthole could restore BMMSC function in individuals with osteoporosis by enhancing Fas and FasL binding leads to restoration of T cell function and regulation of cell death. Osthole also enhances BMMSC’s immunomodulatory function in healthy individuals. Zheng et al. (2019) proved that the addition of osthole to BMMSCs can increase autophagy, which protects osteoblast cells from tumour necroting factor-α (TNF-α) and oxidative stress-mediated apoptosis. Jin et al. (2020) observed the ability of osthole in inhibiting osteoclastogenesis and bone resorption through modulating Wnt3a/β-catenin-OPG signaling. Osteoclastogenesis is inhibited by osthole through increased OPG expression in BMMSCs. Osteoprotegerin or OPG expression of osteoblasts is upregulated by β-catenin that plays an important role in canonical Wnt pathway. The protein complexes Wnt3a, Frizzled8, and low density lipoprotein receptor related protein 5/6 (LRP5/6) inhibit glycogen synthase kinase-3β (GSK3β) that induces release of active β-catenin. As a consequence, β-catenin interacts with site2 and site4 of OPG promoter in order to induce OPG production and secretion as well as inhibiting receptor activator of RANKL and receptor activator of nuclear factor κβ (RANK) binding, thereby reducing osteoclast formation and activity.^13,20-22

Cytokine expression is crucial in bone remodeling. Inhibiting cytokine expression can have an impact on bone resorption.²³ BMP-9 is a cytokine that promotes bone formation. The mechanism by which BMP-9 influences osteogenesis is unknown, but BMP-9 is thought to stimulate osteoclast expression and induce bone formation.²⁴ BMP-9 stimulates osteogenesis by activating several signaling pathways, such as Smad-dependent and Smad-independent signaling, resulting in osteogenic factors expression such as RUNX2, alkaline phosphatase (ALP), and osteocalcin (OSC), which further induces osteoblastic differentiation.²⁵ The signaling pathway that mediates BMP-9 in the osteogenesis process is possible by activating certain osteogenic and key regulatory genes in osteoblast differentiation.²⁶ The influence of BMP-9 on osteogenesis suggests that BMP-9 may activate the Wnt/β-catenin pathway through suppressing β-catenin degradation. BMP-9 promotes osteogenic differentiation, which upregulates leucine-rich repeat-containing G-protein coupled receptor 6 (LGR6) and activation of canonical Wnt/β-catenin signaling. BMP-9 regulates LGR6 expression by preventing phosphorylation of LRP5/6, stabilizing LRP5/6 structure, and decreasing stability of β-catenin. Phosphorylation is inhibited by BMP-9 during osteoclastic differentiation induced by RANKL, whereas RANKL activates protein kinase B (AKT) signaling pathway, which promotes osteoclastic differentiation and survival. Macrophage colony stimulating factor (MCSF) along with RANKL increase osteoclast activity by promoting proliferation and differentiation.²⁷ BMP-9 significantly increases the occurrence of bone formation and microstructure, as well as bone strength. This condition can be achieved by regulating RUNX2 and COL-1 expression in bone, which results in the formation of endochondral and intramembranous bone.^26,27 In order to significantly enhancing bone formation in the process of osteogenesis, BMP-9 can also induce the process of angiogenesis. The formation of sinusoidal capillaries during the early stages of bone formation demonstrates the process of angiogenesis.²⁸

Several studies stated that the combination between nHA and ColI can be used in nanomedicine and tissue engineering area as a candidate. According to Cheng et al. (2013) facilitating the osteogenic differentiation and proliferation of MCET3-E1 osteoblast can be done by incorporating of CNTs into PLGA scaffold. In order to get better properties for regeneration applications and bone tissue engineering such as structural, mechanical, and chemical properties, the combination of nHA, Coll, and MWCNT scaffolds is expected to provide those properties. The use of MWCNT itself was to increase scaffold biocompatibility and its mechanical strength. Scaffolds for tissue engineering take a role to facilitate the growth, cellular attachment, and transport of the cellular metabolites because scaffolds have the sufficient porosity, appropriate pore size, and interconnected pores. The characterized of Coll-HA and MWCNT-Coll-HA by Jing et al. (2017) shows how the addition of MWCNT can increase new bone formation in bone defects.^14,29,30

Increased bone resorption occurs in conditions of osteoporosis. Osthole inhibits the expression of TNF-α as a proinflammatory protein that is generally overexpressed in osteoporosis. Impaired of activating the signaling pathway of nuclear factor κβ (NF-κβ) caused by the decrease of TNF-α can lead to the decreasing expression of nuclear factor of activated T-cell (NFATc1) and activator protein (AP-1).^31-33 The decrease in the expression of the two proteins was able to inhibit the activation process of osteoclast precursors to become mature osteoclasts so that bone resorption activity could be inhibited.^34,35 On the contrary, the role of osthole as an anti-inflammatory substance is also shown by its ability to induce apoptotic activity in the intrinsic and extrinsic pathways through the induction of FasL expression. Increased FasL expression is known to be able to induce an increase in caspase-8 expression resulting in chromosomal degradation.³⁶

On the other hand, caspase-8 can interact with Bid to induce Bcl-2 associated X protein (Bax) expression and lead to the release of cytochrome c into the cytosol. Cytochrome c bind to apoptotic protease-activating factor 1 (Apaf-1) and adenosine triphosphate (ATP) in order to form apoptosomes, thus activating caspase-3 as the major effector protein in the intrinsic apoptosis pathway, which will induce the formation of apoptotic bodies.^37-39 The ability of osthole to induce apoptosis is needed to regulate T cells so that the process of proinflammatory cytokine expression by T cells decreases and manifests in the inhibition of bone resorption processes. This condition is expected to inhibit the process of osteoporosis from the bones and help the therapy process to run optimally.

The synergy of BMP-9 as a growth factor and nHA/Col1/MWCNTs as a scaffold in the concept of triad tissue engineering with BMMSCs as cell culture is known to have the potential to produce good osteogenic proliferation and differentiation. The osteogenic differentiation process of BMMSCs is mediated by activating the cAMP and Wnt signaling pathways. Cyclic adenosine monophosphate, or cAMP, is an adenosine triphosphate derivative that acts as a second messenger in intracellular processes. Activation of the cAMP signaling pathway by G-coupled receptor proteins (GPCRs) can induce PKA activation, which is expressed by the increasing expression of Runx2, activating transcription factor-1 (ATF-1), and CREB, resulting in increased expression of ALP, OSX, and COL1A1.^40,41 This condition accompanied by Ca²⁺ ions obtained from nHA modulates ECM formation and bone mineralization.⁴²

However, the induction of osteogenic differentiation and proliferation is also caused by the activation of Wnt/β-catenin pathway. Wnt activation is the beginning of this signaling pathway, which later interacts with LRP5/6 and FZD and leading to inflated β-catenin level.^43,44 Increased expression of β-catenin usher the increased of various osteogenic marker protein expression such as Runx2, ATF4, and OPG.^45,46 The use of BMP-9 as a growth factor on the other hand, can help the process of cell differentiation through the activation of Smad1/5/8 so that Smad4 is activated. Activation of these proteins results in increased expression of Dlx5, which works together with Runx2, ATF4, and OPG to modulate BMMSCs differentiation.^47,48

On the contrary, by activating the Wnt signaling pathway can induce phosphoinositide 3-kinase (PI3K) protein expression. Increased PI3K protein expression resulted in increased Akt expression, which was manifested in mammalian target of rapamycin (mTOR) activation.⁴⁹ The activated mTOR protein was able to increase the expression of p60S7K so that protein synthesis activity increased.⁵⁰ Activation of mTOR also induces hypoxia-inducible factor-1α (HIF-1α) expression so that vascular endothelial growth factor (VEGF) expression increases, which manifests in an increase in the angiogenesis process that plays a role in increasing nutrient and oxygen supply to the injured area.^26,51 On the other hand, mTOR modulates cyclin A expression to activate the G2/M phase as well as upregulates the level of cyclin D1/E1 in order to activate the G1/S phase. Activating all these phases causes the cell cycle process to run and manifests in cell mitosis.^52,53

CONCLUSION:

According to this literature review, developing osteoporosis treatment options with minimal side effects are urgently needed. OBMMSCs and BMP-9-loaded nHA/COLI/MWCNTS scaffolds are potential candidate biomaterials that may induce osteogenesis by activating various signaling pathways in osteoporosis therapy. Further research is needed regarding the combination of OBMMSCs and BMP-9-loaded nHA/COLI/MWCNTS scaffolds as a biomaterial candidate in osteoporosis therapy.

CONFLICT OF INTEREST:

The authors declare there is no conflict of interest.

REFERENCES:

1. Föger-Samwald U, Dovjak P, Azizi-Semrad U, Kerschan-Schindl K, Pietschmann P. Osteoporosis: Pathophysiology and therapeutic options. EXCLI J. 2020;Jul 20;19:1017-1037. doi: 10.17179/excli2020-2591.

2. Bhagyashri T, Sakhare R, Suryvanshi U, Kore P, Mohite S, Magdum C. Osteoporosis: The Brittle Bone. Asian J. Pharm. Res. 2018;8(1):39-43. doi: 10.5958/2231-5691.2018.00008.4.

3. Kirsti Uusi-Rasi, Saija Karinkanta, Kari Tokola, Pekka Kannus, Harri Sievänen, Bone Mass and Strength and Fall-Related Fractures in Older Age. Journal of Osteoporosis. 2019;2019:1-6. doi: 10.1155/2019/5134690.

4. Aicha Z, Nour H, Samir D. Analysis of Osteoporosis risk factors in Menopausal women's of Algeria population. Asian J. Res. Pharm. Sci. 2020;10(2):79-84. doi: 10.5958/2231-5659.2020.00015.6.

5. Zohreh K, Monireh A, Ebrahim H. A Description of Osteoporosis Preventive Behaviors in Iranian Adolescent Girls. Asian J. Nur. Edu. and Research. 2016;6(1):1-4. doi: 10.5958/2349-2996.2016.00001.X.

6. Rijo G. Effectiveness of Self-Instructional module on prevention of Osteoporosis among middle aged women who are attending Orthopaedic outpatient department in selected hospital, Bangalore. Int. J. Nur. Edu. and Research. 2020;8(4):525-528. doi: 10.5958/2454-2660.2020.00116.7.

7. Sözen T, Özışık L, Calik B. An overview and management of osteoporosis. European Journal of Rheumatology. 2017; 4(1):46. doi: 10.5152/eurjrheum.2016.048.

8. Bhutani, G, Gupta M. Emerging therapies for the treatment of osteoporosis. J Midlife Health. 2013;4(3):147-152. doi: 10.4103/0976-7800.118991.

9. Banita R, VKSK Priyanka. Oral Contraceptive use and Fracture Risk in Women- A Systemic Review. Asian J. Nursing Education and Research. 2021;11(2):263-266. doi: 10.5958/2349-2996.2021.00063.X.

10. Jesni K, Jismi S, Megha C, Saranya S, Snehamol S, Tisha T, Lincy J, Sheba E. A study to assess the knowledge regarding risk factors and Preventive measures of Osteoporosis among Elderly patients in a selected Hospital Kidandoor. Int. J. of Advances in Nur. Management. 2021;9(2):211-213. doi: 10.5958/2454-2652.2021.00047.0.

11. Chen L, Ko N, Chen K. Medical Treatment for Osteoporosis: From Molecular to Clinical Opinions. International Journal of Molecular Sciences. 2019;20(9):2213. doi: 10.3390/ijms20092213.

12. Zhang Z, Leung W, Cheung H, Chan C. Osthole: A Review on Its Bioactivities, Pharmacological Properties, and Potential as Alternative Medicine. Evidence-Based Complementary and Alternative Medicine, 2015:1-10. doi: 10.1155/2015/919616.

13. Zhang Z, Leung W, Li G, Kong S, Lu X, Wong Y, Chan C. Osthole enhances osteogenesis in osteoblasts by elevating transcription factor osterix via cAMP/CREB signaling in vitro and in vivo. Nutrients, 2017;9(6). doi: 10.3390/nu9060588.

14. Zhang R, Li X, Liu Y, Gao X, Zhu T, Lu L. Acceleration of Bone Regeneration in Critical-Size Defect Using BMP-9-Loaded nHA/ColI/MWCNTs Scaffolds Seeded with Bone Marrow Mesenchymal Stem Cells. BioMed Res Int, 2019:1-10. doi: 10.1155/2019/7343957.

15. Wei Z, Salmon R, Upton P, Morrell N, Li W. Regulation of Bone Morphogenetic Protein 9 (BMP9) by Redox-dependent Proteolysis. Journal of Biological Chemistry. 2014;289(45):31150-31159. doi: 10.1074/jbc.M114.579771.

16. Suzuki Y, Ohga N, Morishita Y, Hida K, Miyazono K, Watabe T. BMP-9 induces proliferation of multiple types of endothelial cells in vitro and in vivo. Journal of Cell Science. 2010;123(10):1684-1692. doi: 10.1242/jcs.061556.

17. Ukon Y. et al., Molecular-Based Treatment Strategies for Osteoporosis: A Literature Review. International Journal of Molecular Sciences. 2019;20(2557):1. doi: 10.3390/ijms20102557.

18. Selvi S. Stem Cell Therapy. International Journal of Advances in Nursing Management. 2017;5(4):361-364. doi: 10.5958/2454-2652.2017.00077.4.

19. Nikolaeva L. The Ionic Balance of Bone Marrow. Research Journal of Pharmacy and Technology. 2020;13(2):877-881. doi: 10.5958/0974-360X.2020.00166.3.

20. Zheng X, Yu Y, Shao B, Gan N, Chen L, Yang D. Osthole improves therapy for osteoporosis through increasing autophagy of mesenchymal stem cells. Experimental Animals. 2019;68(4):453–463. doi : 10.1538/expanim.18-0178.

21. Yu Y, Chen M, Yang S, Shao B, Chen L, Dou L, … Yang D. Osthole enhances the immunosuppressive effects of bone marrow-derived mesenchymal stem cells by promoting the Fas/FasL system. Journal of Cellular and Molecular Medicine. 2021;25(10):4835–4845. doi: 10.1111/jcmm.16459.

22. Jin Z, Liao X, Da W, Zhao Y, Li X, Tang D. Osthole Inhibits Osteoclast Formation and Enhances Bone Mass of Bone Marrow Mesenchymal Stem cells by Activating β -catenin- OPG Signaling Pathway. 2020:1–19. doi: 10.21203/rs.3.rs-133397/v1.

23. Alyasiry A, Aljammali Z, Almosawy A, Alrubbaie S. Dental Health in Osteoporotic Women. Research J. Pharm. and Tech. 2015; 8(10):1383-1388. doi: 10.5958/0974-360X.2015.00248.6.

24. Fujioka-Kobayashi M, Marjanowski SD, Kono M, Hino S, Saulacic N, Schaller B. Osteoinductive potential of recombinant BMP-9 in bone defects of mice treated with antiresorptive agents. Int. J. Oral Macillofac Surg. 2019;25:S0901-5027(21)00289-7. doi: 10.1016/j.ijom.2021.08.014.

25. Khorsand B, Elangovan S, Hong L, Dewerth A, Kormann MS, Salem AK. A Comparative Study of the Bone Regenerative Effect of Chemically Modified RNA Encoding BMP-2 or BMP-9. AAPS J. 2017;19(2):438-446. doi: 10.1208/s12248-016-0034-8.

26. Wang X, Huang J, Huang F, Zong J, Tang X, Liu Y, Zhang Q, Wang Y, Chen L, Yin L, He B, Deng Z. Bone morphogenetic protein 9 stimulates callus formatting in osteoporotic rats during fracture healing. Mol Med Rep. 2017;15(5):2537-2545. doi: 10.3892/mmr.2017.6302.

27. Zhou Y, Yang Y, Jing Y, Yuan T, Sun L, Tao B, Liu J, Zhao H. BMP9 Reduces Bone Loss in Ovariectomized Mice by Dual Regulation of Bone Remodeling. JBMR. 2020;35(5):978-993. doi: 10.1002/jbmr.3957.

28. Xiao H, Wang X, Wang C, Dai G, Zhu Z, Gao S, He B, Liao J, Huang W. 2020. BMP9 exhibits dual and coupled roles in inducing osteogenic and angiogenic differentiation of mesenchymal stem cells. Biosci Rep. 2020;40(6):BSR20201262. doi: 10.1042/BSR20201262.

29. Cheng Q, Rutledge K, Jabbarzadeh E. Carbon nanotube-poly(lactide-co-glycolide) composite scaffolds for bone tissue engineering applications. Annals of Biomedical Engineering. 2013;41(5): 904–916. doi: 10.1007/s10439-012-0728-8.

30. Jing Z, Wu Y, Su W, et al. Carbon nanotube reinforced collagen/hydroxyapatite scaffolds improve bone tissue formation in vitro and in vivo. Annals of Biomedical Engineering. 2017;45(9):2075–2087. doi: 10.1007/s10439-017-1866-9.

31. Liu T, Zhang L, Joo D, Sun S. NF-κB signaling in inflammation. Signal Transduction and Targeted Therapy. 2017;2(1). doi: 10.1038/sigtrans.2017.23.

32. Lee J, Kim L, Choi J. Revisiting the Concept of Targeting NFAT to Control T Cell Immunity and Autoimmune Diseases. Frontiers in Immunology. 2018:9. doi: 10.3389/fimmu.2018.02747.

33. Mognol G, González-Avalos E, Ghosh S, Spreafico R, Gudlur A, Rao A, Damoiseaux R, Hogan P. Targeting the NFAT:AP-1 transcriptional complex on DNA with a small-molecule inhibitor. Proceedings of the National Academy of Sciences. 2019;116(20):9959-9968. doi: 10.1073/pnas.1820604116.

34. Pang M, Rodríguez‐Gonzalez M, Hernandez M, Recinos C, Seldeen K, Troen B. AP‐1 and Mitf interact with NFATc1 to stimulate cathepsin K promoter activity in osteoclast precursors. Journal of Cellular Biochemistry. 2019;120(8):12382-12392. doi: 10.1002/jcb.28504.

35. Kim J, Lin C, Stavre Z, Greenblatt M, Shim J. Osteoblast-Osteoclast Communication and Bone Homeostasis. Cells. 2020;9(9):2073. doi: 10.3390/cells9092073. doi: 10.3390/cells9092073.

36. Shen C, Pei J, Guo X, Zhou L, Li Q, Quan J. Structural basis for dimerization of the death effector domain of the F122A mutant of Caspase-8. Scientific Reports. 2018;8(1). doi: 10.1038/s41598-018-35153-5.

37. Dorstyn L, Akey C, Kumar S. New insights into apoptosome structure and function. Cell Death & Differentiation. 2018;25(7):1194-1208. doi: 10.1038/s41418-017-0025-z.

38. Xu X, Lai Y, Hua Z. Apoptosis and apoptotic body: disease message and therapeutic target potentials. Bioscience Reports. 2019;39(1). doi: 10.1042/BSR20180992.

39. Arif M, Syed A, Mahmood A, Khan S, Rizwan M, Munir A. Modeling of apoptosis through gene interaction network and analysis of gene expression pattern. Meta Gene. 2020;25:100730. doi: 10.1016/j.mgene.2020.100730.

40. Steven A, Friedrich M, Jank P, Heimer N, Budczies J, Denkert C, Seliger B. What turns CREB on? And off? And why does it matter?. Cellular and Molecular Life Sciences. 2020;77(20):4049-4067. doi: 10.1007/s00018-020-03525-8.

41. Ju T, Zhao Z, Ma L, Li W, Li S, Zhang J. Cyclic Adenosine Monophosphate-Enhanced Calvarial Regeneration by Bone Marrow-Derived Mesenchymal Stem Cells on a Hydroxyapatite/Gelatin Scaffold. ACS Omega. 2021;6(21):13684-13694. doi: 10.1021/acsomega.1c00881.

42. Sato C, Yamazaki D, Sato M, Takeshima H, Memtily N, Hatano Y, Tsukuba T, Sakai E. Calcium phosphate mineralization in bone tissues directly observed in aqueous liquid by atmospheric SEM (ASEM) without staining: microfluidics crystallization chamber and immuno-EM. Scientific Reports. 2019;9(1). doi: 10.1038/s41598-019-43608-6.

43. Houschyar K, Tapking C, Borrelli M, Popp D, Duscher D, Maan Z, Chelliah M, Li J, Harati K, Wallner C, Rein S, Pförringer D, Reumuth G, Grieb G, Mouraret S, Dadras M, Wagner J, Cha J, Siemers F, Lehnhardt M, Behr B. Wnt Pathway in Bone Repair and Regeneration – What Do We Know So Far. Frontiers in Cell and Developmental Biology. 2019;6. doi: 10.3389/fcell.2018.00170.

44. Maeda K, Kobayashi Y, Koide M, Uehara S, Okamoto M, Ishihara A, Kayama T, Saito M, Marumo K. The Regulation of Bone Metabolism and Disorders by Wnt Signaling. International Journal of Molecular Sciences. 2019;20(22):5525. doi: 10.3390/ijms20225525.

45. Choi H, Kim G, Yoo H, Song D, Chung K, Lee K, Koo Y, An J. Vitamin C Activates Osteoblastogenesis and Inhibits Osteoclastogenesis via Wnt/β-Catenin/ATF4 Signaling Pathways. Nutrients. 2019;11(3):506. doi: 10.3390/nu11030506.

46. Amarasekara D, Kim S, Rho J. Regulation of Osteoblast Differentiation by Cytokine Networks. International Journal of Molecular Sciences. 2021;22(6):2851. doi: 10.3390/ijms22062851.

47. Wang Y, Feng Q, Ji C, Liu X, Li L, Luo J. RUNX3 plays an important role in mediating the BMP9-induced osteogenic differentiation of mesenchymal stem cells. International Journal of Molecular Medicine. 2017;40;1991-1999. doi: 10.3892/ijmm.2017.3155.

48. Mostafa S, Pakvasa M, Coalson E, Zhu A, Alverdy A, Castillo H, Fan J, et al. The wonders of BMP9: From mesenchymal stem cell differentiation, angiogenesis, neurogenesis, tumorigenesis, and metabolism to regenerative medicine. Genes & Diseases. 2019;6(3):201-223. doi: 10.1016/j.gendis.2019.07.003.

49. Liu H, Li X, Lin J, Lin M. Morroniside promotes the osteogenesis by activating PI3K/Akt/mTOR signaling. Bioscience, Biotechnology, and Biochemistry. 2021;85(2):332-339.doi: 10.1093/bbb/zbaa010.

50. Tong X, Gu J, Song R, Wang D, Sun Z, Sui C, Zhang C, et al. Osteoprotegerin inhibit osteoclast differentiation and bone resorption by enhancing autophagy via AMPK/mTOR/p70S6K signaling pathway in vitro. Journal of Cellular Biochemistry. 2018:120(2):1630-1642. doi: 10.1002/jcb.27468.

51. Pal K, Madamsetty V, Dutta S, Wang E, Angom R, Mukhopadhyay D. Synchronous inhibition of mTOR and VEGF/NRP1 axis impedes tumor growth and metastasis in renal cancer. npj Precision Oncology. 2019;3(1). doi: 10.1038/s41698-019-0105-2.

52. Hsieh H, Zhang W, Lin S, Yang W, Wang J, Shen J, Zhang Y, et al. Systems biology approach reveals a link between mTORC1 and G2/M DNA damage checkpoint recovery. Nature Communications. 2018;9(1). doi: 10.1038/s41467-018-05639-x.

53. Lamm N, Rogers S, Cesare A. The mTOR pathway: Implications for DNA replication. Progress in Biophysics and Molecular Biology. 2019;147:17-25. doi: 10.1016/j.pbiomolbio.2019.04.002.

Received on 28.12.2021 Modified on 22.06.2022

Research J. Pharm. and Tech 2023; 16(1):459-464.

DOI: 10.52711/0974-360X.2023.00078