Accelerating effect of Shilajit on osteogenic property of adipose-derived mesenchymal stem cells (ASCs) – PubMed Black Hawk Supplements

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CONCLUSION: As Shilajit accelerates the differentiation of ASCs into the osteoblasts, without changing the physical properties of the Alg hydrogel, this combination may pave the way for more promising remedies considering bone defects.
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Accelerating effect of Shilajit on osteogenic property of adipose-derived mesenchymal stem cells (ASCs) - PubMed

Accelerating effect of Shilajit on osteogenic property of adipose-derived mesenchymal stem cells (ASCs)

Parisa Kangari et al. J Orthop Surg Res. .

Abstract

Background: Shilajit has been widely used remedy for treating a numerous of illness such as bone defects in Iran traditional folk medicine since hundreds of years ago. The aim of the present study was to explore the effect of Shilajit on the osteogenic differentiation of human adipose-derived mesenchymal stem cells (ASCs) in two- (2D) and three-dimensional (3D) cultures.

Materials and methods: ASCs were seeded in 3D 1% alginate (Alg) hydrogel with or without Shilajit (500 µg/mL) and compared with 2D cultures. Then, characterization was done using electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX), alkaline phosphatase (ALP) activity, alizarin red staining and Raman confocal microscopy.

Results: Adding Shilajit had no impact on the Alg scaffold degradability. In the 3D hydrogel and in the presence of osteogenic medium (OM), Shilajit acted as enhancer to increase ALP activity and also showed osteoinductive property in the absence of OM compared to the 2D matched groups at all time points (days 7 and 21 both P = 0.0006, for 14 days P = 0.0006 and P = 0.002, respectively). In addition, calcium deposition was significantly increased in the cultures exposed to Shilajit compared to 2D matched groups on days 14 (P < 0.0001) and 21 (P = 0.0003 and P = 0.003, respectively). In both 3D and 2D conditions, Shilajit induced osteogenic differentiation, but Shilajit/Alg combination starts osteogenic differentiation in a short period of time.

Conclusion: As Shilajit accelerates the differentiation of ASCs into the osteoblasts, without changing the physical properties of the Alg hydrogel, this combination may pave the way for more promising remedies considering bone defects.

Keywords: Adipose-derived mesenchymal stem cells; Alginate; Osteogenic differentiation; Regenerative medicine; Shilajit.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1

H-NMR spectrum of Shilajit. The H-NMR spectra shows the presence of functional groups related to organic compounds, especially polysaccharides in Shilajit

Fig. 2
Fig. 2

MSC characterization. A Flow cytometry indicated that ASCs were negative for CD45 and CD34 but positive for CD105 and CD73. B Isolated ASCs showed fibroblast-like shapes

Fig. 3
Fig. 3

Scanning electron micrographs. A Cells with mesenchymal stem cell phenotype expanded in Alg/ASCs. Osteoblast-like cells with short processes on B Shilajit/Alg/ASCs + OM, C Shilajit/Alg/ASCs and D Alg/ASCs + OM conditions

Fig. 4
Fig. 4

Energy-dispersive X-ray spectroscopy analysis. Calcium (Ca, A) and phosphorus (P, B) contents in 3D groups after 21 days. The highest level of Ca and P was observed on Shilajit/Alg/ASCs + OM. The strongest peaks of Ca and P were shown on Shilajit/Alg/ASCs + OM (**P < 0.01, ***P < 0.001, ****P < 0.0001)

Fig. 5
Fig. 5

The ALP activity assessment. The cells were cultured in 3D (A) and 2D (B) osteogenic conditions on 7, 14 and 21 days after differentiation and also, comparing 3D and 2D cultures was done on these time points (C). The highest level of ALP activity was indicated in Shilajit/Alg/ASCs + OM on 21 days after differentiation compared with other groups (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)

Fig. 6
Fig. 6

Detection calcium deposition. The calcium deposition assessment was done on 7, 14 and 21 days of differentiation in 3D (A) and 2D (B) groups and 3D conditions compared with matched 2D cultures in these time points (C). The highest level of calcium depositions was seen on Shilajit/Alg/ASCs + OM on 21 days after differentiation compared with other groups. D Micrographs of cells stained with alizarin red demonstrate the effects of shilajit on mineral deposition and nodule formation in 2D groups on 7, 14 and 21 days after differentiation. Qualitative observations showed that the number and size of the nodules increased in the presence of Shilajit (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)

Fig. 7
Fig. 7

Raman confocal microscopy. A Raman spectra from 3D groups 21 days after differentiation show the peaks assigned for mineral and organic compounds related osteogenesis. B Raman spectra of Shilajit at 785-nm excitation related to organic and inorganic components at 500–200 cm−1

Fig. 8
Fig. 8

The effect of Shilajit on ASCs proliferation/movement rate in Shilajit treated and untreated groups. A Images indicated cell proliferation enhancement at 8 and 12 h post-incubation in both treated and untreated groups compared to the beginning of the test (scale bar: 100 μm). B Healing/wounded area ratio in scratch healing analysis showed a significant increase at scratch healing present at treated compared to untreated group at both 8 h and 12 h after treatment. **P < 0.01. **P < 0.01

Fig. 9
Fig. 9

Degradation of scaffolds. Scaffolds prepared with or without Shilajit showed the same degradation rate

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References

    1. Habibovic P. Strategic directions in osteoinduction and biomimetics. Tissue Eng Part A. 2017;23(23–24):1295–1296. doi: 10.1089/ten.tea.2017.0430. – DOI – PubMed
    1. Chaparro O, Linero I. Regenerative medicine: a new paradigm in bone regeneration. Adv Tech Bone Regener. 2016;12:253.
    1. Kangari P, Talaei-Khozani T, Razeghian-Jahromi I, Razmkhah M. Mesenchymal stem cells: amazing remedies for bone and cartilage defects. Stem Cell Res Ther. 2020;11(1):1–21. doi: 10.1186/s13287-020-02001-1. – DOI – PMC – PubMed
    1. Usuelli FG, D’Ambrosi R, Maccario C, Indino C, Manzi L, Maffulli N. Adipose-derived stem cells in orthopaedic pathologies. Br Med Bull. 2017;124:1–24. doi: 10.1093/bmb/ldx030. – DOI – PubMed
    1. Furia JP, Lundeen MA, Hurd JL, Pearce DA, Alt C, Alt EU, Schmitz C, Maffulli N. Why and how to use the body’s own stem cells for regeneration in musculoskeletal disorders: a primer. J Orthop Surg Res. 2022;17(1):1–7. doi: 10.1186/s13018-022-02918-8. – DOI – PMC – PubMed

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Accelerating effect of Shilajit on osteogenic property of adipose-derived mesenchymal stem cells (ASCs) – PubMed